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In June 2015, an outbreak of cryptosporidiosis with 35 cases (23 probable and 12 laboratory-confirmed) occurred among 191 attendees of a residential rehabilitation holiday for paediatric organ transplant patients (n= 49) and their families at a hotel in Somogy county, Hungary. The overall attack rate was 18%. Most of the cases were transplanted children who experienced severe acute disease and required adjustment to their tacrolimus immunosuppression. A retrospective case-control study suggested an association between recreational water exposures and illness: cases were seven times more likely than controls to have swum in the children's pool (odds ratio 7.17; 95% confidence interval 2.9–17.2;P< 0.0001) and five times more likely to have used the jetted whirlpool (odds ratio 5.25; 95% confidence interval 2.1–13.1;P< 0.0001). This was the first outbreak of cryptosporidiosis in Hungary and it is especially unfortunate that it affected vulnerable children who experienced severe symptoms.Cryptosporidiumpresents specific infection control difficulties in treated recreational water venues; the link to a whirlpool is unusual and highlights the importance of the age-appropriate use of these facilities and reminding users not to immerse their heads or swallow the water. Cryptosporidiosis is more commonly linked to children’ pools where improved bather hygiene and promoting exclusion of diarrhoea cases could help to avoid similar outbreaks.

The recent increase of reported cyclosporiasis outbreaks associated with fresh produce has highlighted the need for understanding environmental transmission of Cyclospora cayetanensis in agricultural settings and facilities. Conducting such environmental investigations necessitates robust sample collection and analytical methods to detect C. cayetanensis in water samples. This study evaluated three sample collection methods for recovery of C. cayetanensis oocysts from water samples during seeded recovery experiments. Two filtration-based methods, dead-end ultrafiltration (DEUF) and USEPA Method 1623.1, were evaluated for oocyst recovery from irrigation water. A non-filter-based method, continuous flow centrifugation (CFC), was evaluated separately for recovery from creek water and spent produce wash water. Median C. cayetanensis recovery efficiencies were 17% for DEUF and 16–22% for Method 1623.1. The DEUF method proved to be more robust than Method 1623.1, as the recovery efficiencies were less variable and the DEUF ultrafilters were capable of filtering larger volumes of high-turbidity water without clogging. Median C. cayetanensis recovery efficiencies for CFC were 28% for wash water and 63% for creek water, making it a viable option for processing water with high turbidity or organic matter. The data from this study demonstrate the capability of DEUF and CFC as filter-based and non-filter-based options, respectively, for the recovery of C. cayetanensis oocysts from environmental and agricultural waters.

The raccoon (Procyon lotor) carnivore native to North America is a fast spreading, invasive species in the Europe now. At the moment, the highest population occupies areas near the German-Polish border. The data on the occurrence of Cryptosporidium spp. and microsporidia in raccoons is limited to North America's territory and is totally lacking in the case of their introduction to Europe. Therefore, the objective of this study was to investigate the occurrence of microparasites, i.e., Cryptosporidium spp. and microsporidia in the introduced raccoons obtained from localities in Poland and Germany. A PCR-based approach that permitted genetic characterization via sequence analysis was applied to raccoon fecal samples (n = 49), collected during 2012-2014. All fecal samples were simultaneously tested with the use of genetic markers, and DNA of microsporidia and Cryptosporidium spp. was detected among the examined raccoons. The results of our research confirmed the presence of Cryptosporidium skunk genotype and Enterocytozoon bieneusi NCF2 genotype. The results suggest a possible role of raccoons in the contamination of the environment, including urban areas, with pathogens of zoonotic significance as well as their role in the transmission and introduction of new genotypes of microparasites in the areas where P. lotor has not been observed yet. To our knowledge, there has been no literature data on the above genotypes detected previously in humans or animals from the examined study sites so far.

Current methods to identify the etiology of diarrhea require laboratory facilities for storage of pathogens, which is often challenging in low-resource settings. This study evaluated the efficacy of a low-cost method for preserving stool specimens for the detection of parasitic enteropathogens using Whatman 903 protein saver cards (Sigma-Aldrich, St. Louis, MO). Stool samples known to be positive by multiplex real-time polymerase chain reaction for Giardia lamblia, Cryptosporidium spp., and Entamoeba histolytica parasites were preserved on 232 Whatman cards. DNA was then extracted from cards using Chelex and Qiagen extraction protocols, and tested for these parasites using multiplex real-time PCR. We included stool samples known to have a higher parasite load (cycle threshold [ct]-value < 30) and those with a lower parasite load (ct values 30-35). Sensitivities and specificities were determined using DNA extracted directly from whole stool samples using Qiagen kits (QIAGEN, Hilden, Germany). For whole stool samples with ct values < 30, preserved directly on Whatman 903 protein saver cards for Giardia analysis, the sensitivity was 100% for both Qiagen and Chelex DNA extraction. For E. histolytica, this was 100% for sensitivity for Qiagen and 80% for Chelex DNA extractions, and for Cryptosporidium, this was 80% for Qiagen and 50% for Chelex DNA extraction. The specificity was 100% for all parasites for all extraction procedures. Given the high sensitivity for stool samples with higher parasite loads, we recommend the use of the Whatman 903 protein saver card for preserving fecal specimens for the analysis of Giardia and E. histolytica using Qiagen DNA extractions in low-resource settings.

Cryptosporidium is a protozoan parasite that causes the diarrhoeal disease, cryptosporidiosis. Although many species have been identified, the majority of human disease worldwide is caused by two species; Cryptosporidium parvum and Cryptosporidium hominis. In Australia, data from the National Notifiable Diseases Surveillance System (NNDSS) show that cryptosporidiosis outbreaks occur every few years. To better understand the transmission, trends and nature of cryptosporidiosis outbreaks in Western Australia, epidemiological and genomic data from three cryptosporidiosis outbreaks in 2003, 2007 and 2011 were reviewed. The 2007 outbreak was the largest (n = 607) compared with the outbreaks in 2003 (n = 404) and 2011 (n = 355). All three outbreaks appeared to have occurred predominantly in the urban metropolitan area (Perth), which reported the highest number of case notifications; increases in case notifications were also observed in rural and remote areas. Children aged 0–4 years and non-Aboriginal people comprised the majority of notifications in all outbreaks. However, in the 2003 and 2007 outbreaks, a higher proportion of cases from Aboriginal people was observed in the remote areas. Molecular data were only available for the 2007 (n = 126) and 2011 (n = 42) outbreaks, with C. hominis the main species identified in both outbreaks. Subtyping at the glycoprotein 60 (gp60) locus identified subtype IbA10G2 in 46.3% and 89.5% of C. hominis isolates typed, respectively, in the 2007 and 2011 outbreaks, with the IdA15G1 subtype was identified in 33.3% of C. hominis isolates typed in the 2007 outbreak. The clustering of cases with the IdA15G1 subtype in the remote areas suggests the occurrence of a concurrent outbreak in remote areas during the 2007 outbreak, which primarily affected Aboriginal people. Both the C. hominis IbA10G2 and IdA15G1 subtypes have been implicated in cryptosporidiosis outbreaks worldwide; its occurrence indicates that the mode of transmission in both the 2007 and 2011 outbreaks was anthroponotic. To better understand the epidemiology, sources and transmission of cryptosporidiosis in Australia, genotyping data should routinely be incorporated into national surveillance programmes.

SUMMARYWaterborne illness related to the consumption of contaminated or inadequately treated water is a global public health concern. Although the magnitude of drinking water-related illnesses in developed countries is lower than that observed in developing regions of the world, drinking water is still responsible for a proportion of all cases of acute gastrointestinal illness (AGI) in Canada. The estimated burden of endemic AGI in Canada is 20·5 million cases annually – this estimate accounts for under-reporting and under-diagnosis. About 4 million of these cases are domestically acquired and foodborne, yet the proportion of waterborne cases is unknown. There is evidence that individuals served by private systems and small community systems may be more at risk of waterborne illness than those served by municipal drinking water systems in Canada. However, little is known regarding the contribution of these systems to the overall drinking water-related AGI burden in Canada. Private water supplies serve an estimated 12% of the Canadian population, or ~4·1 million people. An estimated 1·4 million (4·1%) people in Canada are served by small groundwater (2·6%) and surface water (1·5%) supplies. The objective of this research is to estimate the number of AGI cases attributable to water consumption from these supplies in Canada using a quantitative microbial risk assessment (QMRA) approach. This provides a framework for others to develop burden of waterborne illness estimates for small water supplies. A multi-pathogen QMRA ofGiardia, Cryptosporidium, Campylobacter, E. coliO157 and norovirus, chosen as index waterborne pathogens, for various source water and treatment combinations was performed. It is estimated that 103 230 AGI cases per year are due to the presence of these five pathogens in drinking water from private and small community water systems in Canada. In addition to providing a mechanism to assess the potential burden of AGI attributed to small systems and private well water in Canada, this research supports the use of QMRA as an effective source attribution tool when there is a lack of randomized controlled trial data to evaluate the public health risk of an exposure source. QMRA is also a powerful tool for identifying existing knowledge gaps on the national scale to inform future surveillance and research efforts.

Cryptosporidium species are the most common cause of gastrointestinal illness in treated recreational water venues. In order to protect public health during swimming, Cryptosporidium-sized microsphere removals by high-rate sand filtration with six coagulants were evaluated with a 5.5 m3 pilot-scale swimming pool. A sand filter without coagulation removed 20–63% of Cryptosporidium-sized microspheres. Cryptosporidium-sized microsphere removals exceeded 98% by sand filtration with five of the six tested coagulants. Continuously feeding coagulants A, B, and F (i.e., organic polymers) led to coagulant accumulation in the system and decreased removals over time (

Cryptosporidium is a protozoan parasite that infects humans and various animal species. The environmental stability and the low infectious dose of Cryptosporidium facilitate its transmission by water and food. Discharge of untreated wastewater may result in waterborne or foodborne Cryptosporidium outbreaks, therefore a suitable treatment may prevent its dissemination. Most studies on the prevalence of Cryptosporidium oocysts in wastewater have reported a concentration range between 10 and 200 oocysts/L and a prevalence of 6 to 100%. Activated sludge has been found to be ineffective for the removal of Cryptosporidium oocysts. Stabilization ponds and constructed wetlands are efficient for the reduction of Cryptosporidium from wastewater, especially when the retention time is longer than 20 days at suitable sunlight and temperature. High rate filtration and chlorine disinfection are inefficient for the reduction of Cryptosporidium from effluents, whereas ultrafiltration and UV irradiation were found to be very efficient for the reduction of Cryptosporidium oocysts. Adequate tertiary treatment may result in high quality effluent with low risk of Cryptosporidium for unrestricted irrigation and other non-potable applications.

SUMMARYIn March 2013, a veterinary student tested positive forCryptosporidium; four classmates reported similar gastrointestinal symptoms. We aimed to identify source(s) and risk factors forCryptosporidiuminfection in university persons symptomatic between 21 January and 14 April 2013. Sixty-four (79%) students from a cohort of 81 fourth-year veterinary students completed questionnaires, identifying 13 cases; four wereCryptosporidium parvumGP60 subtype IIaA16G1R1b, two were IIdA24G1, seven did not submit stool samples. Thirteen cases attended the university's field clinic before symptom onset (13/37 attendees, 35%); 11 visited at least one of four farms where students recalled seeing calves with diarrhoea.C. parvumsubtype IIaA16G1R1b was identified in calves at one of the farms. Entering pens of calves with diarrhoea [relative risk (RR) 7·6, 95% confidence interval (CI) 1·7–33·5] and eating in clinic cars (RR 9·1, 95% CI 1·3–65·8) were associated with being a case. Washing hands at least twice per farm visit (0 cases,P= 0·03) was protective. This outbreak investigation was notable for rapid and effective collaboration between public health, veterinary and environmental sectors, leading to swift identification of a microbiological and epidemiological link between cases, infected calves and their farms. We recommend frequent hand-washing using proper technique and dissuasion from eating in clinic cars to minimize possible exposure to contaminated surfaces.

Introduction Diarrhoea continues to cause significant morbidity in Egypt. Aim To determine the frequency and distribution of different enteropathogens in acute diarrhoeal episodes, utilising an expanded testing regimen, and to correlate clinical signs and symptoms associated with the detected pathogens. Material and methods The case-control study enrolled 356 patients < 5 years old with acute diarrhoea and 356 age and sex-matched healthy controls. Both cases and controls underwent a full history and physical examination, and provided two rectal swab specimens and a stool sample. Laboratory analysis included stool culture, microscopy, and indirect methods. Results Rotavirus was detected in 11% of patients. Enterotoxigenic Escherichia coli (ETEC), Campylobacter, Shigella, and Salmonella were detected in 7%, 3.7%, 1.1%, and 1.4% of patients, respectively; and in 11.1%, 3.1%, 0.6%, and 0.6% of controls, respectively, with no significant statistical difference. Cryptosporidium was detected in 3.9% of cases. Mixed infection was detected in 5.9% of cases and 0.9% of controls, with a significant difference (p < 0.001). No pathogen was detected in 66.3% of cases and in 83.5% of controls. Rotavirus infection was associated with recurrent vomiting, dehydration, and hospitalisation. Bacterial diarrhoea was associated with vomiting (52%) in ETEC infections, fever (80%) in Salmonella infections, mucus (100%) and blood (50%) in stools of Shigella infections, and convulsions (15%) in Campylobacter infections. Conclusions Rotavirus is a prominent cause of diarrhoea among Egyptian children. Despite utilising an expanded testing regimen, more work is still needed for identification of other enteropathogens that constitute other causative agents of diarrhoea.

Cryptosporidium and Giardia protozoa are zoonotic parasites that cause human gastroenteritis and can be transmitted to human through the fecal-oral route and water or food. Several species belong to these genera and their resistant forms occur in water, but only some of them are infectious to human. Health risk depends on the occurrence of infectious Cryptosporidium and Giardia species and genotypes in water, and only molecular techniques allow detecting them, as well as enable to identify the contamination source. In this work, genotyping and phylogenetic analysis have been performed on the basis of 18S rDNA and s-giardin genes sequences of Cryptosporidium and Giardia, respectively, in order to provide the molecular characterization of these parasites detected earlier in five natural water bodies in Poland and to track possible sources of their (oo)cysts in water. Genotyping revealed a high similarity (over 99 up to 100 %) of analyzed sequences to cattle genotype of C. parvum isolated from cattle and human and to G. intestinalis assemblage B isolated from human. The sequences obtained by others originated from patients with clinical symptoms of cryptosporidiosis or giardiasis and/or with the infection confirmed by different methods. The contamination of three examined lakes is probably human-originated, while the sources of contamination of two remaining lakes are wild and domestic animals. Obtained phylogenetic trees support suggestions of other authors that the bovine genotype of C. parvum should be a separate species, as well as A and B assemblages of G. intestinalis.

Removing protozoa from a water supply using coagulation, flocculation, dissolved air flotation (DAF) and filtration on a bench scale was evaluated. Calcium carbonate flocculation with and without immunomagnetic separation (IMS) was chosen to detect Giardia spp. cysts and Cryptosporidium spp. oocysts in the studied samples. The results indicated that DAF removed between 1.31 log and 1.79 log of cysts and between 1.08 log and 1.42 log of oocysts. The performance was lower in filtration, with the removal of 1.07 log-1.44 log for cysts and 0.82 log-0.98 log for oocysts. The coagulation, flocculation, DAF and filtration steps removed more than 2.2 log of cysts and oocysts from the water studied. However, protozoa were detected in the filtered water, even with turbidity values of 0.2 NTU. The recovery of the detection method met the international criteria and was higher when there was no IMS. Including the third acid dissociation in the IMS was critical to improve the performance of the protocol tested. However, there was an increase in the technical and analytical complexity and costs. It was also observed that the efficiency of the treatment was linked to the performance of the selected method of detecting protozoa.

SUMMARYOpen-water swimming is increasingly popular, often in water not considered safe for bathing. Limited evidence exists on the associated health risks. We investigated gastrointestinal illness in 1100 swimmers in a River Thames event in London, UK, to describe the outbreak and identify risk factors. We conducted a retrospective cohort study. Our case definition was swimmers with any: diarrhoea, vomiting, abdominal cramps lasting ⩾48 h, nausea lasting ⩾48 h, with onset within 9 days after the event. We used an online survey to collect information on symptoms, demographics, pre- and post-swim behaviours and open-water experience. We tested associations using robust Poisson regression. We followed up case microbiological results. Survey response was 61%, and attack rate 53% (338 cases). Median incubation period was 34 h and median symptom duration 4 days. Five cases had confirmed microbiological diagnoses (four Giardia, one Cryptosporidium). Wearing a wetsuit [adjusted relative risk (aRR) 6·96, 95% confidence interval (CI) 1·04–46·72] and swallowing water (aRR 1·42, 95% CI 1·03–1·97) were risk factors. Recent river-swimming (aRR 0·78, 95% CI 0·67–0·92) and age >40 years (aRR 0·83, 95% CI 0·70–0·98) were protective. Action to reduce risk of illness in future events is recommended, including clarification of oversight arrangements for future swims to ensure appropriate risk assessment and advice is provided.

Cryptosporidium is a major cause of diarrheal disease worldwide. In developing countries, this infection is endemic and in children, associated with growth faltering and cognitive function deficits, with the most severe impact on those aged

This study compared the effectiveness and sustainability of five selected cost-effective home water treatment systems in removing Cryptosporidium and Giardia spp. from water sources. These systems included: silver impregnated porous pot (SIPP), biosand filter combined with zeolite (BSF-Z), biosand filter without zeolite (BSF-S), bucket filter (BF) and ceramic candle filter (CCF). The USEPA Methods 1623 were used for the isolation and the detection of the protozoan parasites. The flow rates of the devices ranged between 0.05 and 160.5 L/h. The average turbidity of the environmental intake water samples ranged between 1.47 and 42.93 NTU before filtration and between 0.05 and 14.49 NTU after filtration. The performance of the SIPP in removing the parasites (98–100% of both oocysts and cysts from synthetic water; 96–99.6% oocysts and 96.6–99.8% cysts from the environmental water sources) and in removing viral indicator (97.7–100%) was found to be significantly higher (p

SUMMARYFaecal samples from 804 cattle and buffaloes and 165 diarrhoeal children of Ismailia province were investigated by an immunochromatographic screening test and PCR to determine prevalences and distributions ofCryptosporidiumspp. Results were analysed statistically for clustering of animal and human cases.Cryptosporidiumherd prevalence was 73·3% and individual animal prevalence 32·3%.C. parvumwas the dominant species in animals (65·7%). Young calves watered with canal or underground water were at particular risk of infection. Detection rates were higher when calves showed diarrhoea, fever and dehydration. HumanCryptosporidiumprevalence was 49·1%.C. hominisdominated in humans (60·5%), followed byC. parvum(38·3%). Living in villages, drinking underground water and having contact with animals were risk factors. Cluster analysis revealed differences in the distribution of infections between animals and humans and suggests different transmission dynamics.

Filter Backwash Water (FBW) from water treatment plants (WTP) is composed of raw water waste, chemicals and microorganisms. Inappropriate disposal of this residue impacts negatively in the environment and in the health of human populations. Aiming to characterize the FBW from one WTP of Blumenau-SC, physiochemical, microbiological and parasitological assessments and tests with different flocculants polymers were performed in order to propose strategies for treatment and reuse of this residue. Subsequently treated liquid is discharged into the Itajaí-Açu River (Class 2). Physicochemical and microbiological analyses showed results higher than those permitted by CONAMA Resolution n° 430/2011 and Giardia duodenalis (Assembly B) cysts and Cryptosporidium spp. oocyst positivity was observed, characterizing as polluted and contaminated residue that shouldn't be released in the hydric body. The anionic flocculant polymer showed satisfactory results in the turbidity sample reduction (99.49%), which may be a promising alternative in the treatment of this residue. Resumo A água de retrolavagem dos filtros (ARF) de estações de tratamento de água (ETA) é composta por resíduos da água bruta, produtos químicos e microrganismos. Seu descarte inadequado impacta negativamente o meio ambiente e a saúde da população humana. Objetivando caracterizar a ARF da ETA de Blumenau-SC, foram realizadas avaliações físico-químicas, microbiológicas e parasitológicas e testes com diferentes polímeros floculantes, afim de propor estratégias para tratamento e reuso deste resíduo para posteriormente, o líquido tratado, ser lançado no Rio Itajaí-Açu (Classe 2). Grande parte das análises físico-químicas e microbiológicas apresentaram resultados superiores aos determinados pela Resolução CONAMA n° 430/2011, além de positividade para cistos de Giardia duodenalis (Assembleia B) e oocistos de Cryptosporidium spp., caracterizando um resíduo poluído e contaminado que não deve ser lançado in natura no corpo hídrico. O polímero floculante aniônico apresentou resultados satisfatórios na redução da turbidez da amostra (99,49%), podendo ser uma alternativa promissora no tratamento deste resíduo.

The production of recombinant antigens in Escherichia coli and specific polyclonal antibodies for diagnosis and therapy is still a challenge for world-wide researchers. Several different strategies have been explored to improve both antigen and antibody production, all of them depending on a successful expression and immunogenicity of the antigen. Gene fusion technology attempted to address these challenges: fusion partners have been applied to optimise recombinant antigen production in E. coli, and to increase protein immunogenicity. Taking a 12-kDa surface adhesion antigen from Cryptosporidium parvum (CP 12) by example, the novel H fusion partner was presented in this work as an attractive option for the development of recombinant immunogens and its adjuvant-free immunisation. The H tag (of only 1 kDa) efficiently triggered a CP 12-specific immune response, and it also improved the immunisation procedure without requiring coadministration of adjuvants. Moreover, polyclonal antibodies raised against the HCP 12 fusion antigen detected native antigen structures displayed on the surface of C. parvum oocysts. The H tag proved to be an advanced strategy and promising technology for the diagnosis and therapy of C. parvum infections in animals and humans, allowing a rapid and simple recombinant production of the CP 12 antigen., The financial support of Fundacao para a Ciencia e Tecnologia (FCT), Portugal, is acknowledged: Project PTDC/CVT/103081/2008 (co-financed by COMPETE) and grant SFRH/BD/46482/2008 (POPH-QREN) to Costa SJ. We would like to thank Lurdes Delgado and Sonia Soares for the Cryptosporidium oocysts isolation from fecal samples, and also to Hitag (R) Biotechnology, Ltd for kindly providing the H and Fh8 tag sequences used in this work.

Cryptosporidium andersoni and Cryptosporidium muris infections have been found in the mice and/or cattle. The oocysts of C. andersoni and C. muris have been sporadically detected in human feces, but the infectious capacity and features have been unknown, because of the scarcity of reports involving human infections. To assess the infectivity and the clinical and pathological features of C. andersoni and C. muris in primates, an experimental infectious study was conducted using cynomolgus monkeys. The monkeys were orally inoculated with oocysts of two different C. andersoni Kawatabi types and C. muris RN-66 under normal and immunosuppressive conditions. The feces of the monkeys were monitored for about 40 days after the administration of oocysts using the flotation method, but no shedding oocysts were observed under either both normal or immunosuppressive conditions. Gross and histopathological examinations were performed on the immunosuppressive monkeys, but these revealed no evidence of Cryptosporidium infections, even though the monkeys were subjected to immunosuppressive conditions. It is hypothesized that C. andersoni and C. muris pose little danger of infection in primates even under immunosuppressive conditions.

A quantitative microbial risk assessment (QMRA) with Monte Carlo simulation was used to estimate risks of Cryptosporidium infection from drinking water. Four different approaches to assess Cryptosporidium oocyst removal by filtration were compared using three relationships between turbidity removal and oocyst removal proposed in the literature, and an empirical model based on oocyst log removal credits associated with filtered effluent turbidity. Using a large database of a full-scale water treatment plant in Brazil, two of the turbidity removal-based models returned 1–3 log unit removal of Cryptosporidium oocysts and median values of Cryptosporidium annual infection risk estimates of approximately 10−2pppy (per person per year). The other model based on turbidity removal and the empirical model resulted, respectively, in 2–3.5 and 0–4 log removal of Cryptosporidium oocysts and corresponding infection risk estimates around 10−2pppy, close therefore to the health-based targets of the WHO guidelines for drinking water. The results of this study indicate that the empirical model appears to be particularly suitable for QMRA studies and that similar models should be tested in other site-specific assessments of treatment performance.

A study was conducted to assess the impact of media aging on Cryptosporidium parvum transport and retention in granular filters. Column experiments were conducted with sand, anthracite, and granular activated carbon (GAC) filters. All columns possessed an identical filter design and grain size distribution. Oocyst removal was evaluated with both new and aged media. The variation in performance among new media was attributed to the shape of the tested granular media and to the concentration of extractable metals on the surface of collector grains. Media aging significantly increased the treatment performance of sand (26%) and GAC (31%) but had no influence on the perfor- mance of anthracite. This enhanced performance of aged media correlated well with the extent of biofilm development. The best removal performance was observed with aged GAC (71 � 2%), which supported the highest amount of biomass according to polysaccharide and protein measurements. The results of this investigation are of particular interest because they suggest that media aging increases the removal of protozoan parasites in full-scale granular filters. DOI: 10.1061/(ASCE)EE.1943-7870.0000672. © 2013 American Society of Civil Engineers. CE Database subject headings: Water treatment; Filtration; Filters; Biofilm; Abatement and removal. Author keywords: Water treatment; Granular filtration; Media aging; Cryptosporidium; Biofilm.

SUMMARYPrevious evidence has suggested an association between cryptosporidiosis and consumption of unfiltered drinking water from Loch Katrine in Scotland. Before September 2007, the water was only micro-strained and chlorinated; however, since that time, coagulation and rapid gravity filtration have been installed. In order to determine risk factors associated with cryptosporidiosis, including drinking water, we analysed data on microbiologically confirmed cases of cryptosporidiosis from 2004 to 2010. We identified an association between the incidence of cryptosporidiosis and unfiltered Loch Katrine drinking water supplied to the home (odds ratio 1·86, 95% confidence interval 1·11–3·11, P = 0·019). However, while filtration appears to be associated with initially reduced rates of cryptosporidiosis, evidence suggests it may paradoxically make those consumers more susceptible to other transmission routes in the long-term. These findings support implementation of similar treatment for other unfiltered drinking-water supplies, as a means of reducing cryptosporidiosis associated with drinking water.

Giardia and Cryptosporidium species are a serious problem if present in water supplies. The removal of these protozoans and the adaptation of existing protocols are essential for supplying drinking water to developing countries. Considering this, the aim of this study is to evaluate, on a bench level, the removal of Giardia spp. cysts and of Cryptosporidium spp. oocysts from water with high turbidity, using polyaluminium chloride as a coagulant. Filtration using mixed cellulose ester membranes, followed, or not, by purification through immunomagnetic separation (IMS) was used for detecting protozoans. By evaluating the adopted protocol, without using IMS, retrievals of 80% of cysts and 5% of oocysts were obtained, whereas by using IMS, recoveries of 31.5% of cysts and 5.75% of oocysts were reached. When analyzing the coagulant performance, a dosage of 65 mg L−1 showed contamination from protozoans in all the samples of filtered water. A dosage of 25 mg L−1 presented protozoans in 50% of the filtered water samples. The results showed an improved performance for the 25 mg L−1 dosage; therefore, the control of coagulation and adaptation of detection protocols must be evaluated according to the features of raw water and availability of local resources.

SUMMARYInsights into transmission dynamics of enteropathogens in children attending daycare are limited. Here we aimed at identifying daycare centre (DCC) characteristics associated with time-clustered occurrence of enteropathogens in DCC-attending children. For this purpose, we used the KIzSS network, which comprises 43 DCCs that participated in infectious disease surveillance in The Netherlands during February 2010–February 2013. Space–time scan statistics were used to identify clusters of rotavirus, norovirus, astrovirus, Giardia lamblia and Cryptosporidium spp. in a two-dimensional DCC characteristic space constructed using canonical correlation analysis. Logistic regression models were then used to further identify DCC characteristics associated with increased or decreased odds for clustering of enteropathogens. Factors associated with increased odds for enteropathogen clustering in DCCs were having indoor/outdoor paddling pools or sandpits, owning animals, high numbers of attending children, and reporting outbreaks to local health authorities. Factors associated with decreased odds for enteropathogen clustering in DCCs were cleaning child potties in designated waste disposal stations, cleaning vomit with chlorine-based products, daily cleaning of toys, extra cleaning of toys during a suspected outbreak, and excluding children with gastroenteritis. These factors provide targets for reducing the burden of gastrointestinal morbidity associated with time-clustered occurrence of major enteropathogens in DCC attendees.

Transport ofCryptosporidiumin runoff will contaminate the surrounding water body.Cryptosporidium-sized microspheres and inactiveCryptosporidiumparvum oocysts removal using an active carbon (AC) filter with/without coal gangue from the runoff was evaluated. Chemical composition of coal gangue was determined, and its performance as a water treatment material was tested. Results showed SiO2was the dominant chemical component of coal gangue. Removal was only 20% for microspheres and 24% for oocysts by AC filtration alone. The removal for both was increased to more than 98% by addition of coal gangue powder to the filter. Furthermore, gangue reuse turned commercially worthless material to a water treatment material as well helping prevent gangue from occupying agriculture/industry land.

SUMMARYA total of 207 wild rodents were caught on nine pig farms, five chicken farms and five non-farm locations in Sweden and surveyed for a selection of bacteria, parasites and viruses.Lawsonia intracellulariaand pathogenicYersinia enterocoliticawere only detected in rodents on pig farms (9% and 8% prevalence, respectively) which indicate that these agents are more likely to be transmitted to rodents from pigs or the environment on infected farms.Brachyspira hyodysenteriae(1%),Brachyspira intermedia(2%),Campylobacter jejuni(4%),Campylobacter upsaliensis(2%), leptospires (7%) and encephalomyocarditis virus (9%) were also detected from rodents not in contact with farm animals.GiardiaandCryptosporidiumspp. were common, although no zoonotic types were verified, andSalmonella entericawas isolated from 1/11 mice on one farm but not detected by PCR from any of the rodents.Trichinellaspp. andToxoplasma gondiiwere not detected.

Gastrointestinal side effects affect ∼20% of renal transplant recipients [1]. Among such side effects, diarrhoea is common [2] but under-recognized [3]. Diarrhoea may relate to immunosuppressive drugs or infectious complications. The latter may be caused by one of the usual microbial pathogens that also cause gastroenteritis in immunocompetent patients, such as campylobacter, salmonella or viruses. Cytomegalovirus (CMV) colitis, for example, is seen as a cause of diarrhoea, usually within the first one or two post-transplant years. Once these usual suspects have been excluded, the clinician is left with a long differential diagnosis of other causes of diarrhoea in the immunosuppressed host. We describe the interesting case of a 49-year-old renal transplant recipient who presented with an unusual cause of diarrhoea 2.5 years after transplantation.

Treatment of drinking water at the point of use (POU) has demonstrated health benefits for people who have access only to microbially contaminated drinking water. In this work, the ceramic siphon POU water filter was evaluated for its ability to reduce indicator microorganisms in test waters. During batch challenge tests, the filter reduced Escherichia coli in filtered water by 7 log10 (99.999987%) and bacteriophage MS2 by 0.12 log10 (24.0%). Next, a novel continuous flow dosing system allowing sewage-amended feed water to constantly pass through the filters allowed for determination of changes in microbial reductions over time and total volume of water filtered. E. coli B, MS2 and fluorescent microspheres (as a surrogate for Cryptosporidium oocysts) were seeded into test water and dosed to filters at 10, 25 and 50% of the filter's volume lifespan. Microbial removal efficacy decreased as the volume of water filtered increased and test filters did not achieve their volume lifespan before physically failing. The ceramic siphon household water filter is effective in reducing E. coli and surrogates for Cryptosporidium in water, but filter modifications may be needed to achieve acceptable levels of virus removal and to reach the target 7,000 L volume lifespan of the filter.

A case-control study was designed to investigate the role of different Cryptosporidium spp. in Swedish dairy herds with and without calf diarrhoeal problems. Faecal samples were collected from preweaned calves, young stock and cows. Cryptosporidium oocysts were detected by sodium chloride flotation and epifluorescence microscopy. Molecular diagnostics were used to identify Cryptosporidium species. Samples containing C. parvum were further analysed to determine subtypes. Calf faecal samples were also analysed for rotavirus, coronavirus and Escherichia coli F5+. Total protein was assessed in 1- to 8-day-old calves. A questionnaire was used to identify differences in management routines. Cryptosporidium infection was diagnosed in all herds, with equal prevalence in case and control herds in all three age groups. Cryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium ryanae and Cryptosporidium andersoni were all identified, as were rotavirus, coronavirus and E. coli F5+. C. ryanae and C. andersoni were only detected in non-diarrhoeal samples, whereas the other pathogens were detected in both diarrhoeal and non-diarrhoeal samples. Diarrhoea was more common in case herd calves. Disinfection of single pens was more common in case herds and several other management routines seemed to differ although results were not significant.

This study was undertaken to investigate the removal of pathogenic microorganisms and their indicators in a laboratory scale biological treatment system that simulated the secondary treatment process of a wastewater treatment plant (WWTP). Four groups of microorganisms including bacteria, viruses, protozoa and helminths as well as the selected indicators were employed in the investigation. The results demonstrated that approximately 2–3 log10 removal of the microbial indicators was achieved in the treatment process. The log removal of Clostridium perfringens spores was low due to their irreversible adsorption to sludge flocs. The laboratory treatment system demonstrated a similar removal capability for Escherichia coli and the bacterial indicators (total coliforms, enterococci and particles

SUMMARYAn outbreak of cryptosporidiosis associated with exposure to outdoor swimming-pool water affected an estimated 800–1000 individuals. PCR products were obtained from faecal specimens from 30 individuals who tested positive forCryptosporidiumoocysts. RFLP and sequencing analyses showed that all individuals were infected withCryptosporidium parvum. Among the infected individuals, five had just swum in an adjacent indoor pool during the same period, and had no identified contact with individuals linked to the outdoor pool. With the use of subgenotyping based on analysis of three mini- and microsatellite loci, MS1, TP14, and GP15, we could identify two sources of exposure. One subtype was associated with the outdoor pool and another with the indoor pool. These data demonstrate that the use of mini- and microsatellite loci as markers for molecular fingerprinting ofC. parvumisolates are valuable in the epidemiological investigation of outbreaks.

Currently, a major challenge on producing high-quality drinking water is to monitor pathogens, such as Giardia, Cryptosporidium and enteric viruses. Due to limitations of the analytical methods available to detect pathogens in water, the research of surrogate indicators is an up-to-date subject. In view of these aspects, a pilot scale study was performed to evaluate the association between microbiological and physical indicators and the presence of Giardia spp and Cryptosporidium sp in the effluent of upflow and downflow slow sand filters. The results showed that efficient bacterial removal could indicate suitable protozoa removal. Although coliforms and Escherichia coli do not present the appropriate physiological profile for an "ideal" indicator, they are still good references for drinking water microbiological quality, specifically for slow sand filtration. The results also point out to the need of deeper researches about the use of anaerobic spores as routine indicator. Regarding the control of Cryptosporidium outbreaks, the expectation that a single indicator will satisfy all purposes is unreal. It may be more useful to know the advantages and disadvantages of several indicators, and integrate them appropriately. Atualmente, um grande desafio para a produção de água de elevado padrão de qualidade é o monitoramento de patogênicos, como Giardia, Cryptosporidium e vírus entéricos. Devido a limitações nos métodos analíticos para a detecção de patogênicos na água, a pesquisa de indicadores substitutos é um tema atual. Em vista desses aspectos, foi realizado um estudo em escala piloto para a avaliação da associação entre indicadores microbiológicos e físicos e a presença de Giardia spp e Cryptosporidium sp no efluente de filtros lentos de areia de escoamentos ascendente e descendente. Os resultados mostraram que uma remoção bacteriana eficiente pode indicar adequada remoção de protozoários. Embora coliformes e Escherichia coli não apresentem o perfil fisiológico apropriado de um indicador "ideal", eles são ainda boas referências para a qualidade microbiológica da água, especificamente para a filtração lenta. Os resultados também chamam a atenção para o necessário aprofundamento de pesquisas sobre o uso de esporos anaeróbios como um indicador rotineiro. Quanto ao controle de surtos de Cryptosporidium, a expectativa de que um único indicador satisfará todos os objetivos não é realista. Seria mais útil conhecer as vantagens e desvantagens de diversos indicadores e utilizá-los apropriadamente de forma integrada.

SUMMARYSerological responses toCryptosporidium-specific antigens (15/17 and 27 kDa) were compared among populations in four areas of the Czech Republic that use drinking water from clearly defined sources: (1) wells in a fractured sandstone aquifer, (2) riverbank infiltration, or (3) two different filtered and chlorinated surface waters. Among persons surveyed in the area with riverbank-infiltration water, 33% had a strong serological response to the 15/17-kDa antigen group whereas, in the other three areas, over 72% of persons had a strong response. These response differences suggest thatCryptosporidiumexposures and infection were lower in the area with bank infiltration. The large percentage of the study population with a strong serological response to both antigens suggests high levels of previous infections that may have resulted in protective immunity for cryptosporidiosis. This may be one reason why no waterborne cryptosporidiosis outbreaks and few cases of cryptosporidiosis have been reported in the Czech Republic.

SUMMARYCryptosporidiuminfects millions of people worldwide causing acute gastroenteritis, but despite its remarkable epidemiological and economic impact, information on the epidemiological trends of human cryptosporidiosis is still scarce in most countries. Here we investigate a panel of 486 cases collected in Galicia (NW Iberian Peninsula) between 2000 and 2008, which sheds new light on the epidemiology in this region of the South Atlantic European façade. Incidence rates in Galicia are one order of magnitude higher than those reported in other regions of Spain, suggesting that this parasite remains largely underdiagnosed in this country, and are also larger than those typical of other European countries with available data. Two species dominate our dataset,Cryptosporidium hominis(65%) andC. parvum(34%). The sex ratio of patients infected by either species was 0·5, butC. hominiswas significantly more common in younger males.C. parvuminfections were more acute and required more specialized medical attention, which suggests a differential adaptation of each species to human hosts. The parasites display strong seasonal and geographical variation.C. parvumincidence peaked during summer and was mainly detected in rural areas whileC. hominisinfections were more frequent in autumn and exhibited a more even geographical distribution. Such differences probably reflect their distinct sources of infection – C. parvumis mainly zoonotic andC. hominisanthroponotic – and the effects of climatic variables, like temperature and rainfall.

In this study, we examined the prevalence and molecular characteristics of Cryptosporidium in buffalo, dairy cattle and sheep in different farms at Kafr El Sheikh Province, Egypt. Rectal fecal samples, including 466 samples from buffalo, 1697 from cattle and 120 from sheep, were collected from different ages and screened by modified Ziehl-Neelsen acid-fast microscopy for detection of Cryptosporidium oocysts. All studied farms were positives with an overall prevalence of 1.29% in buffalo (4.17% in claves versus 0.48% in adults), 7.07% in cattle (6.90% in calves versus 10.20% and 6.10% in heifers and adults, respectively) and 2.50% in sheep (4.40% in lambs versus 1.30% in adults). PCR-RFLP analyses of small-subunit rRNA genes from positive specimens revealed the occurrence of C. parvum and C. ryanae in buffalo; C. parvum, C. ryanae, C. bovis and C. andersoni in cattle and only C. xiaoi in sheep. Genotypes distribution showed that C. ryanae was the dominant species (60.0%) followed by C. parvum (40.0%) in buffalo calves. Meanwhile, in cattle calves, C. parvum was the commonest species (74.23%) followed by C. ryanae (16.10%) and C. bovis (9.70%). Subtyping of C. parvum based on sequence analysis of the polymorphic 60 kDa glycoprotein gene locus showed the presence of subtypes IIdA20G1 and IIaA15G1R1 in both buffalo and cattle calves, addressing the potential role of calves in zoonotic cryptosporidiosis in Egypt.

Wastewater samples collected from ten wastewater facilities across the US were analyzed to determine the occurrence of indigenous Cryptosporidium oocysts using methods based on modifications of USEPA Method 1622. Wastewater facilities participating in this study ranged in size from 0.6 to 193 mgd average daily flow. A total of 289 wastewater samples were analyzed over a 15-month period. ColorSeed is a commercial product containing gamma-irradiated Cryptosporidium oocysts that have been permanently stained with a Texas Red dye. ColorSeed was used as an internal control with each sample to assess method performance. In 500 ml sample volumes, mean ColorSeed recoveries in raw influents and primary effluents were 26.1±17.7% and 33.0±17.9%, respectively. In 10 liter volumes of secondary effluent, mean ColorSeed recovery was 25.0±16.6%. Volumes analyzed for tertiary effluent samples ranged from 14.8 l to 131.3 l resulting in a mean ColorSeed recovery of 48.8±14.5%. Indigenous oocysts were detected in 30% of raw influents, 46% of primary effluents, 58% of secondary effluents and 19% of tertiary effluents analyzed. Indigenous oocyst concentrations ranged from

Cryptosporidium spp. in diarrheic calves less than 30 days old from farms across Northern Ireland were examined over a year period by microscopic, genotyping, and subtyping techniques to characterize the transmission dynamics. Cryptosporidium oocysts were detected in 291 of 779 (37.4%) animals. The prevalence rates of rotavirus, coronavirus, and Escherichia coli K99+ were lower as seen in 242 of 806 (30.0%), 46/806 (5.7%), and 16/421 (3.8%) of animals, respectively. Of the 224 Cryptosporidium-positive specimens available for molecular analysis, Cryptosporidium parvum was identified in 213 (95.1%) specimens, Cryptosporidium bovis in eight (3.6%), and Cryptosporidium deer-like genotype in three (1.3%). Sequence analysis of the 60-kDa glycoprotein gene identified 16 IIa subtypes and a new subtype family, with 120 of the 216 (55.6%) positive specimens having the subtype IIaA18G3R1. Eight of the IIa subtypes were previously seen in humans in Northern Ireland. Several subtypes were temporally or geographically unique. The genetic diversity in calves in Northern Ireland was much greater than that reported from other areas. This work demonstrates the utility of genotyping and subtyping tools in characterizing the transmission of Cryptosporidium spp. in calves and humans.

Aims: The aims of this study were to validate a portable continuous flow centrifuge (PCFC) as an alternative concentration step of US-EPA Method 1623 and to demonstrate it's efficacy for recovery of low numbers of protozoa from large volumes of various water matrices. Methods and Results: Recoveries of Cryptosporidium parvum oocysts, Giardia intestinalis cysts and Encephalitozoon intestinalis spores spiked into 10–1000 l volumes of various water matrices were evaluated during in-house and collaborative trials. Spiked protozoa were either approved standards or diluted stock samples enumerated according to USEPA Method 1623. Cryptosporidium recoveries exceeded method 1623 criteria and substantially high recoveries were observed for Giardia and E. intestinalis. Conclusions: Portable continuous flow centrifuge methodology exceeded method 1623 acceptance criteria for Cryptosporidium and could be easily adopted for other protozoa. Significance and Impact of the Study: The PCFC could be adopted as an alternative user-friendly concentration method for Cryptosporidium and for monitoring of large volumes of source and tap water for accidental or deliberate contamination with protozoa and potentially with other enteric pathogens. It is anticipated that PCFC would also be equal or superior to filtration for protozoa monitoring in wastewater and effluents.

Laboratory dual media filtration experiments were conducted (a) in direct filtration mode using model raw water moderate in turbidity and low in DOC, and (b) in conventional filtration mode treating water moderate in turbidity and high in DOC. Model simulations of filter performance for the removal of particles provided hypotheses for the experimental studies of dual media filtration. An increase in alum dose in direct filtration mode, while improving filter performance, also showed some disadvantages, including rapid development of head loss. Suboptimal dose in direct filtration significantly impaired the filter performance. In conventional mode, the effect of alum dose on the filter performance, while obvious, was not as dramatic as in direct filtration. Ripening indicated by particle counts occurred earlier than by turbidity and breakthrough of particle counts started earlier than breakthrough of turbidity, suggesting that turbidity can be used as a more conservative monitor of filter performance during the ripening period to minimise the risk of passage of small particles, while particle counts can be considered a more sensitive indicator of deteriorating filter performance during the breakthrough period. The lower sand layer served as a multiple barrier for particles when the performance of the anthracite layer was not effective.

This article consolidates and updates available information on Cryptosporidium removal by filtration, with a focus on optimal and suboptimal filtration performance. Emphasis was placed on integrating various strategies for optimizing filtration processes with the most current information regarding pathogen removal by filtration. As this review underscores, in response to regulations such as the Interim Enhanced Surface Water Treatment Rule (IESWTR) and the Long Term 2 ESWTR, there is a need to develop methods to compare the relative effectiveness of treatment alternatives and to understand the most significant operational effects on filter performance. In many cases, filters operated at or near optimal conditions can achieve in the vicinity of 3-log (or better) removal of Cryptosporidium. Operational conditions such as coagulant upsets and end-of-run (early breakthrough) filtration may significantly decrease oocyst removal by drinking water filters, even when filter effluent turbidities are low (

Drinking water plays a major role in the spread of intestinal protozoa pathogenic for human, namely Giardia lamblia and Cryptosporidium parvum. Surface water is contaminated with Giardia or Cryptosporidium in two major ways: wastewater treatment plants may discharge effluent with cysts or by fecal pollution by various animals that live on the watersheds. So, they are of great concern to the water treatment industry because they are known to have caused a number of waterborne outbreaks of disease. Since, the identifying of Giardia cysts and Cryptosporidium oocysts require. special technique and lab personnel specially trained in microbiology. Particle counting is another physical technique for enumeration of Giardia cysts and Cryptosporidium oocysts by instrument depending on their size distribution. This study aims at evaluating the use of particle counter as a monitoring tool for particle removal in a conventional pilot water treatment plant. The results revealed that coagulation, sedimentation, and filtration removed the most percentage of the both parasites. The average removals of both parasites by coagulation and sedimentation were 76% and 71%, while filtration process only removed about ≥99.5%.On the other hand, the average removal of turbidity was 90.7% by coagulation and sedimentation and 95.2% by filtration. The average over-all removals of turbidity, Giardia, and Cryptosporidium were 99.53%, 99.95%, and 99.89%, respectively. While, the average log removal of parasites and turbidity were 3.3 log for Giardia, 3.02 log for Cryptosporidium, and 2.38 log for turbidity. A significant strong correlation was found between total particle removal and cyst and oocyst sizes removals, while a significant correlation between turbidity and parasites removal were observed. So, particle counter and turbidimeter can be used as surrogate indicator of Giardia and Cryptosporidium removal. The study recommended reducing the risk of raw water infection by Giardia and Cryptosporidium through the multiple barrier approach to treatment, including watershed protection, optimization of chemical pretreatment, filtration, disinfection, and a properly operated and maintained distribution system. It should be installed on-line instruments – particle counting and turbiditimeter monitoring – at various stages of the water treatment process to evaluate water purification plant performance for Giardia and Cryptosporidium removal and enable the operator to develop optimal operations conditions to achieve maximum particulates removal.

The goal of this project was to evaluate the removal of emerging pathogens and pathogen indicators by pilot-scale coagulation, flocculation, sedimentation, and granular media filtration. The emerging pathogens included Cryptosporidium parvum oocysts, Encephalitozoon intestinalis spores, Escherichia coli 0157:H7, and Aeromonas hydrophila. Bacteriophage MS2 and turbidity were used as pathogen indicators. This work revealed that some emerging pathogens were removed much more effectively than others. A. hydrophila was removed more effectively than C. parvum, and E. intestinalis spores and E. coli 0157:H7 were the least effectively removed. For the water tested in this study, the results suggest that a change in filter effluent turbidity requirements from 0.5 to 0.3 ntu would not achieve a significant improvement in the reliability of pathogen removal. However, by setting filter effluent turbidity goals below 0.2 ntu, significant improvements in microbiological quality could be obtained. In general, the pilot-plant data suggest that good removal of both turbidity and natural organic matter (NOM) decreases the risk of achieving poor emerging pathogen removal. In other words, optimizing the coagulation process for reduction of turbidity and NOM would improve removal of emerging pathogens.

Because much rier Cryptosporidium against attention of the this limitations has pathogen. oocysts been of directed and The many the ripening chemical fear toward of or waterborne filtration disinfectants maturation as disease the period for primary inactivating outbreaks, of the barfilCryptosporidium oocysts and the fear of waterborne disease outbreaks, much attention has been directed toward filtration as the primary barrier against this pathogen. The ripening or maturation period of the filter run may be the Achilles heel of this barrier. Several research groups have already examined Cryptosporidium removal during the filter-ripening period and found removals that were typically only slightly less than those of the same filters following the ripening period (Huck et al, 2000; Health Stream , 1999; Emelko et al, 1998; Patania et al, 1995). Many studies only seeded oocysts for a brief period (if at all) of the preceding filter run and allowed few oocysts to be present in the backwash remnant water that is responsible for a significant portion of the turbidity peak that follows backwashing. Amburgey and co-workers (2001a, 2001b) did seed the entire preceding filter runs of both conventional and biological filters and performed extensive sampling during the ripening period to more accurately assess the risk of Cryptosporidium passage. These researchers found that biological filters usually had effluent oocyst concentrations approximately equal to those found later in the filter run, but the conventional filter had oocyst concentrations from 3 to 18 times higher than the biological filters at the peak of the ripening period following an efficient backwash procedure. Hall et al (2000) found similar results following continuously seeded conventional filter runs. Overall, there appears to be only a brief period of increased Cryptosporidium passage during the ripening period of optimized pilot-scale filter runs, but this may not be the case for full-scale filters operating at suboptimal coagulation conditions and/or employing less efficient backwash techniques. Several enhancements of the normal filter-operating process have been explored in recent years in response to the risks encountered during the filter

This study evaluates the occurrence of Cryptosporidium oocysts and Giardia cysts in reclaimed effluents if method 1623 with the Envirochek capsule filters (standard and high-volume [HV] filters) and a modified version of the Information Collection Rule method (ICR) with the polypropylene yarn-wound cartridge filter are used. The recovery efficiency of the analytical methods was evaluated with samples of reagent, tap, and reclaimed water by using flow cytometer-sorted spike suspensions. (Oo)cyst recovery efficiency determined filter performance and method reproducibility in the water matrix tested. Method 1623 with the Envirochek HV capsule filter generated significantly higher recovery rates than did the standard Envirochek filter and the modified ICR method. Notwithstanding, large variations in recovery rates (>80%) occurred with samples of reclaimed water, and none of the water quality parameters analyzed in the reclaimed effluents could explain such variability. The highest concentrations of indigenous oocysts were detected by method 1623 with the HV filter, which provided a sufficient number of oocysts for further confirmation of infectious potential. Confirmation of species and potential infectivity for all positive protozoan samples was made by using a nested PCR restriction fragment polymorphism assay and the focus detection method most-probable-number assay, respectively. The methodology and results described in the present investigation provide useful information for the establishment of pathogen numeric standards for reclaimed effluents used for unrestricted irrigation.