Cryptosporidium and Giardia are important parasites due to their zoonotic potential and impact on human health, often causing waterborne outbreaks of disease. Detection of (oo)cysts in water matrices is challenging and few countries have legislated water monitoring for their presence. The aim of this study was to investigate the presence and origin of these parasites in different water sources in Northern Greece and identify interactions between biotic/abiotic factors in order to develop risk-assessment models. During a 2-year period, using a longitudinal, repeated sampling approach, 12 locations in 4 rivers, irrigation canals, and a water production company, were monitored for Cryptosporidium and Giardia, using standard methods. Furthermore, 254 faecal samples from animals were collected from 15 cattle and 12 sheep farms located near the water sampling points and screened for both parasites, in order to estimate their potential contribution to water contamination. River water samples were frequently contaminated with Cryptosporidium (47.1%) and Giardia (66.2%), with higher contamination rates during winter and spring. During a 5-month period, (oo)cysts were detected in drinking-water (<1/litre). Animals on all farms were infected by both parasites, with 16.7% of calves and 17.2% of lambs excreting Cryptosporidium oocysts and 41.3% of calves and 43.1% of lambs excreting Giardia cysts. The most prevalent species identified in both water and animal samples were C. parvum and G. duodenalis assemblage AII. The presence of G. duodenalis assemblage AII in drinking water and C. parvum IIaA15G2R1 in surface water highlights the potential risk of waterborne infection. No correlation was found between (oo)cyst counts and faecal-indicator bacteria. Machine-learning models that can predict contamination intensity with Cryptosporidium (75% accuracy) and Giardia (69% accuracy), combining biological, physicochemical and meteorological factors, were developed. Although these prediction accuracies may be insufficient for public health purposes, they could be useful for augmenting and informing risk-based sampling plans.

Land use/Land cover (LULC) associated with Cryptosporidium sp. and Giardia sp. quantification and distribution can provide identification of the environmental circulation patterns of these parasites. The aim of this research was to relate the occurrence and circulation of these parasites to the LULC watershed with poor sanitation infrastructure and livestock as important economic activity. The study involved 11 municipalities in the state of São Paulo, located in southeastern Brazil. Sampling was carried out at the catchment sites of each water supply on a monthly basis, starting in December 2014 and lasting until November 2015, totalizing 128 samples. Protozoans were quantified according to the 1623.1 US. EPA Method. For watershed delimitation, the hydrographic network was extracted from the hydrology tool of ArcGIS 10.1. The frequency of occurrence of these pathogens and the high concentrations were evidenced in the municipality with the largest urban area (16.2%) and intense livestock activity (39%) near the catchment site. The municipality that showed the lowest frequency of occurrence presented the smallest urban area (0.87%) and absence of livestock activity near the catchment site. The high concentration of pathogens suggests a correlation between the impact on water supply networks and river basin degradation caused by urban activity and livestock.

Biofilms containing pathogenic organisms from the water supply are a potential source of protozoan parasite outbreaks and a significant public health concern. The aim of the present study was to demonstrate the simultaneous and multi-spatial occurrence of waterborne protozoan pathogens (WBPP) in substrate-associated biofilms (SAB) and compare it to surface water (SW) and sediments with bottom water (BW) counterparts using manual filtration and elution from low-volume samples. For scenario purposes, simulated environmental biofilm contamination was created from in-situ grown one-month-old SAB (OM-SAB) that were spiked with Cryptosporidium parvum oocysts. Samples were collected from the largest freshwater reservoirs in Luzon, Philippines and a University Lake in Thailand. A total of 69 samples (23 SAB, 23 SW, and 23 BW) were evaluated using traditional staining techniques for Cryptosporidium, and Immunofluorescence staining for the simultaneous detection of Cryptosporidium and Giardia. WBPP were found in 43% SAB, 39% SW, and 39% BW of the samples tested in the present study with SAB results reflecting SW and BW results. Further highlights were demonstrated in the potential of using low-volume samples for the detection of parasites in source water. Scanning electron microscopy of OM-SAB samples revealed a naturally-associated testate amoeba shell, while Cryptosporidium oocysts spiked samples provided a visual profile of what can be expected from naturally contaminated biofilms. This study provides the first evidence for the simultaneous and multi-spatial occurrence of waterborne protozoan pathogens in low-volume aquatic matrices and further warrants SAB testing along with SW and BW matrices for improved water quality assessment strategies (iWQAS).

Manila Bay is one of the major propagation sites of edible bivalves in the Philippines. Studies have shown that bivalves might be contaminated with human pathogens like the protozoan parasite Cryptosporidium, one of the major causes of gastroenteritis in the world. In this study, Cryptosporidium from four species of edible bivalves were isolated using a combination of sucrose flotation and immunomagnetic separation. Using direct fluorescent antibody test, Cryptosporidium oocysts were found in 67 out of 144 samples collected. DNA sequence analysis of the 18S rRNA gene of the isolates detected C. parvum and C. hominis (major causes of human cryptosporidiosis) and C. meleagridis (causes infection in avian species). Analysis of the 60kDa glycoprotein gene further confirmed the genotypes of the Cryptosporidium isolates. This study is the first to provide baseline information on Cryptosporidium contamination of Manila Bay where bivalves are commonly cultured.

The effects of precipitation on the hygienic quality of water and blue mussels collected from five different localities in the urban areas in the Inner Oslofjord were investigated, with samples analysed for Escherichia coli, Salmonella spp., pathogenic Vibrio spp., Norovirus, Sapovirus, Cryptosporidium spp. and Giardia duodenalis. The sampling sites were located at varying distances from the outlet of combined sewer overflows (CSO)-impacted rivers/streams. In general, 1–3 log10 increases in fecal indicator bacteria and human pathogens were observed after heavy rainfalls. Blue mussels appeared to be a useful indicator of the impact of sewage at these sites, and generally a good correlation was identified between concentrations of E. coli and other human pathogens in the mussels. Provision of general advice to the public of avoiding areas near the outlets of CSO-impacted rivers after heavy rainfall may reduce the risk of gastroenteritis by bathers and others that may swallow water during recreational activities.

In order to establish seawater contamination by emerging protozoan parasites, we used qPCR to molecularly characterize and evaluate the parasitic burden of Giardia duodenalis, Cryptosporidium spp., Toxoplasma gondii, and Cyclospora cayetanensis in 1255 wild bivalve mollusks collected along the Tunisian coasts. T. gondii, G. duodenalis and C. cayetanensis were detected in 6.9% (99% CI=1.6–12.2%) pools of Ruditapes decussatus. None of the samples were found positive to Cryptosporidium spp.; 6.6% pools of R. decussatus were positive for T. gondii Type I, 1.6% for G. duodenalis assemblage A, and 1.6% for the association T. gondii Type I/C. cayetanensis/G. duodenalis assemblage A. R. decussatus harbored up to 77500 oocysts/sample of T. gondii, up to 395 cysts/sample of G. duodenalis, and 526 oocysts/sample of C. cayetanensis. These results provide the first evidence that the Tunisian coasts are contaminated by zoonotic protozoan parasites that can constitute a direct or indirect risk for human health.

Sewage is a major contributor to pollution problems involving human pathogens in tropical coastal areas. This study investigated the occurrence of intestinal protozoan parasites (Giardia and Cryptosporidium) in tropical recreational marine waters contaminated with sewage. The potential risks of Cryptosporidium and Giardia infection from recreational water exposure were estimated from the levels of viable (oo) cysts (DIC+, DAPI+, PI−) found in near-shore swimming areas using an exponential dose response model. A Monte Carlo uncertainty analysis was performed in order to determine the probability distribution of risks. Microbial indicators of recreational water quality (enterococci, Clostridium perfringens) and genetic markers of sewage pollution (human-specific Bacteroidales marker [HF183] and Clostridium coccoides) were simultaneously evaluated in order to estimate the extent of water quality deterioration associated with human wastes. The study revealed the potential risk of parasite infections via primary contact with tropical marine waters contaminated with sewage; higher risk estimates for Giardia than for Cryptosporidium were found. Mean risks estimated by Monte Carlo were below the U.S. EPA upper bound on recreational risk of 0.036 for cryptosporidiosis and giardiasis for both children and adults. However, 95th percentile estimates for giardiasis for children exceeded the 0.036 level. Environmental surveillance of microbial pathogens is crucial in order to control and eradicate the effects that increasing anthropogenic impacts have on marine ecosystems and human health.

Cryptosporidium spp., as a genus of protozoan intestinal parasites, is recognized as responsible for cryptosporidiosis. The present study was conducted to provide an overview of the prevalence of Cryptosporidium based on water. In this regard, some databases such as Scopus, PubMed, Embase, and Web of Science were screened in order to retrieve the related citations from 1 January 1983 to 10 September 2019. The pooled prevalence of Cryptosporidium spp. was calculated by using a random effect model (REM) based on defined subgroups, including countries, water type, treatment conditions (treated and untreated), economic condition, World Health Organization (WHO) regions, and method of detection. In contrast, this index for treated and untreated water was 25.7% and 40.1%, respectively. Also, the overall prevalence of Cryptosporidium spp. among all water types was defined as 36 (95% CI: 31.4–40.7). The rank order of prevalence of Cryptosporidium spp. based on water type was wastewater (46.9%) > surface water (45.3%) > raw water (31.6%) > drinking water (25.5%) > reservoirs water (24.5%) > groundwater (18.8%) > swimming pool water (7.5%) > marine water (0.20%). Identifying the key contributing factors to Cryptosporidium spp. survival can help provide solutions at both local and global scales.

Constructed wetlands have been recognized as offering a removal treatment option for high concentrations removal of chemical and biological contaminants in domestic wastewater. The enteric protozoan parasite Cryptosporidium is considered one of the highly resistant to treatment and highly infectious organisms to humans and animals. Moreover, some species of Cryptosporidium are known to have a zoonotic nature. In this investigation a pilot scale for domestic wastewater treatment system was used, consisting of the following steps in series: (1) up-flow anaerobic sludge blanket (UASB) reactor, (2) free water surface (FWS) wetland unit, and (3) sub-surface flow (SSF) wetland unit. This treatment system was fed with domestic wastewater to assess its efficiency in removing Cryptosporidium oocysts. The obtained Cryptosporidium oocysts were detected and enumerated by two different staining techniques 'acid fast trichrome (AFT) and modified Ziehl Neelsen (MZN) stains'. Polymerase chain reaction (PCR) technique was also used to detect Cryptosporidium DNA in wastewater samples. Results revealed that anaerobic treatment (using UASB reactor) could remove about 53.1% of Cryptosporidium oocysts present in raw wastewater. The in-series connection between the two wetland units allowed complete elimination of Cryptosporidium oocysts as the first (FWS) wetland unit removed 95.9% of the oocysts present in anaerobically treated wastewater and the remaining portion of oocysts was completely removed by the second (SSF) wetland unit. Cryptosporidium oocysts were detected in 95.8% of raw wastewater samples with a mean count of 43.8 oocysts/l when AFT stain was used while they were detected in only 87.5% of raw wastewater samples with a mean count of 35.6 oocysts/l when MZN stain was used. Polymerase chain reaction (PCR) technique was able to detect Cryptosporidium DNA in only 45.8% of raw wastewater samples. Positive PCR results were only achieved in wastewater samples containing 52 oocysts or more per liter.

Knowledge gaps in the exposure parameters for recreational water activities make quantitative risk assessment related to water recreation difficult. Therefore, the annual exposure frequency and single exposure duration for the recreational water activities of residents from ten cities in the North and South of China were investigated. Questionnaire interviews were carried on recreational water activities comprising swimming (SW), boating (BA), playing in interactive fountains (PF), and watching fountains (WF). Quantitative microbial risk assessment for the exposure of urban residents to Cryptosporidium and Giardia was also performed. For the four recreational water activities, the participation rates of urban residents in SW and WF were higher than the others. For SW and BA, the mean annual exposure frequency and single exposure duration for males were significantly higher than those for females. PF and WF showed the opposite. The annual exposure frequency for above 35-year-old residents was higher than that for young residents (18–35 years). However, the single exposure duration for young residents was highest in SW, BA, and PF. The mean annual exposure frequency and single exposure duration for North China residents were higher than those for South China residents in all recreational water activities, except for SW. Overall, the annual exposure frequency and single exposure duration in recreational water activities for all urban residents followed a lognormal distribution. In the four recreational water activities, the total annual infection risk of male exposure to Cryptosporidium was 1.0 × 10⁻², with the confidence intervals between 95 and 5% of [4.3 × 10⁻⁴, 3.7 × 10⁻²], whereas that for females was 6.8 × 10⁻³ and [4.2 × 10⁻⁴, 2.4 × 10⁻²]. Also, the annual infection risk of males to Giardia was 8.8 × 10⁻³ and [5.1×10⁻⁴, 3.2×10⁻²], and that of females was 5.3 × 10⁻³ and [4.0 × 10⁻⁴, 1.8 × 10⁻²]. These results demonstrated that SW and PF made the highest contribution to the total annual infection risk. Sensitivity analysis highlighted that the characterization of exposure parameters plays a critical role in health risk assessment, which may provide a scientific basis for recreational water quality standards formulation.