Revisiting the Gage–Bidwell Law of Dilution in Relation to the Effectiveness of Swimming Pool Filtration and the Risk to Swimming Pool Users from Cryptosporidium

The transfer of water from a swimming pool to the treatment location is key in determining the effectiveness of water treatment by filtration in removing turbidity and managing the risk from particulate material, including microbial pathogens, such as Cryptosporidium spp. A key recommendation for pool operators when dealing with an accidental faecal release (the likely main source of high Cryptosporidium oocyst concentrations in pools) is that the pool water should be filtered for at least six turnover cycles prior to use. This paper briefly outlines the theoretical basis of what has become known as the Gage–Bidwell Law of Dilution, which provides a basis for this recommendation, and extends the idea to account for the impact of filter efficiency. The Gage–Bidwell Law reveals that for each pool turnover 63% of the water resident in the pool at the start of the turnover period will have been recirculated. Building on this, we demonstrate that both filter efficiency and water-turnover time are important in determining filtration effectiveness and can be combined through a single parameter we term ‘particle-turnover’. We consider the implications of the Gage–Bidwell Law (as referred to in the original 1926 paper) for the dynamics of the ‘dirt’ content of pool water, whether in terms of a specific particle size range (e.g., Cryptosporidium oocysts) or turbidity.

Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts

Cryptosporidium oocysts are known for being very robust, and their prolonged survival in the environment has resulted in outbreaks of cryptosporidiosis associated with the consumption of contaminated water or food. Although inactivation methods used for drinking water treatment, such as UV irradiation, can inactivate Cryptosporidium oocysts, they are not necessarily suitable for use with other environmental matrices, such as food. In order to identify alternative ways to inactivate Cryptosporidium oocysts, improved methods for viability assessment are needed. Here we describe a proof of concept for a novel approach for determining how effective inactivation treatments are at killing pathogens, such as the parasite Cryptosporidium. RNA sequencing was used to identify potential up-regulated target genes induced by oxidative stress, and a reverse transcription quantitative PCR (RT-qPCR) protocol was developed to assess their up-regulation following exposure to different induction treatments. Accordingly, RT-qPCR protocols targeting thioredoxin and Cryptosporidium oocyst wall protein 7 (COWP7) genes were evaluated on mixtures of viable and inactivated oocysts, and on oocysts subjected to various potential inactivation treatments such as freezing and chlorination. The results from the present proof-of-concept experiments indicate that this could be a useful tool in efforts towards assessing potential technologies for inactivating Cryptosporidium in different environmental matrices. Furthermore, this approach could also be used for similar investigations with other pathogens.

Multicenter Evaluation of an ELISA for the Detection of Cryptosporidium spp. Antigen in Clinical Human Stool Samples

Human cryptosporidiosis remains underdiagnosed, and rapid/accurate diagnosis is of clinical importance. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive, time-consuming, and requires skillful experience. Thus, we aimed to evaluate the usefulness of a coproantigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, we evaluated the performances of a commercial ELISA (CoproELISA Cryptosporidium kit, Savyon Diagnostics, Israel) for the detection of Cryptosporidium spp. in random clinical stool samples through a multicenter study. The sensitivity and specificity for coproantigen ELISA were 98.86% and 94.32%, respectively. The coproantigen ELISA results indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis, and may be useful for large-scale epidemiological studies of cryptosporidiosis.

The Surveillance of cryptosporidium in some dairy farm

Apicomplexan protozoan parasites of the genus Cryptosporidium is infect the gastrointestinal tract on wall epithelium cell of animals and humans. The farm is important object with a high risk significant outbreak of zoonotic infections. We conducted by random sampling prevalence of Cryptosporidium based on dairy farms in the Selenge, Tuwaimags and near of Ulaanbaatar city. In the study, a total of 170 fecal sample from 131 cattle and 4-10 months aged calves, were infected 37.6% (64/170). Detection of Cryptosporidium oocyst with commonly used for fuchsin staining. Further research into molecular biology is needed. Сүүний үхрийн зарим аж ахуйд криптоспоридийн тандан судалгаа хийсэн дүн Криптоспорид хүн, мал амьтдын ходоод, гэдэсний ханын хучуур эсэд шимэгчилдэг эгэл биетэн.Cүүний чиглэлийн үхрийн аж ахуйд ариун цэврийн дэглэм алдагдвал зоонозын халдвар дэгдэх өндөр эрсдэлтэй. Бид санамсаргүй түүвэрлэлтийн аргаар Сэлэнгэ, Төв аймаг, Улаанбаатар хот орчмын зарим сүүний үхрийн аж ахуйг түшиглэн криптоспоридийн тандан судалгааг явууллаа. Уг судалгаанд үнээ-131, 4-10 сарын бойжилттой тугал-39, нийт 170 баасны дээжид криптоспоридийн ооцистыг илрүүлэхэд 37.6% (64/170)халдварлалттай байв. Цаашид үүсгэгчийн биологи, молекул биологийн судалгааг нарийвчлан хийх шаардлагатай. Түлхүүр үг: Баасны дээж, ооцист, сахрын ханасан уусмал, тугал, фуксины будаг

Micromanipulation System for Isolating a Single Cryptosporidium Oocyst

In this paper, an integrated system for contact micromanipulation of Cryptosporidium oocysts is presented. The system integrates five actuators and a partially automated control system and contacts the oocyst using a drawn glass end effector with tip dimensions of 1 μ m. The system is intended to allow single cell analysis (SCA) of Cryptosporidium—a very harmful parasite found in water supplies—by isolating the parasite oocyst of 5 μ m diameter in a new environment. By allowing this form of analysis, the source of Cryptosporidium can be found and potential harm to humans can be reduced. The system must overcome the challenges of locating the oocysts and end effector in 3D space and contact adhesion forces between them, which are prominent over inertial forces on this scale. An automated alignment method is presented, using the Prewitt operator to give feedback on the level of focus and this system is tested, demonstrating alignment accuracy of <2 μ m. Moreover, to overcome the challenge of adhesion forces, use of dry and liquid environments are investigated and a strategy is developed to capture the oocyst in the dry environment and release in the liquid environment. An experiment is conducted on the reliability of the system for isolating a Cryptosporidium oocyst from its culture, demonstrating a success rate of 98%.

Cryptosporidiosis among Hemodialysis Patients in Jordan: First Preliminary Screening Surveillance

Few studies have reported the incidence of cryptosporidiosis among hemodialysis patients worldwide. Currently many molecular and immunological assays have been developed for the sensitive diagnosis of cryptosporidiosis, but still, the microscopic detection of the parasitic infective stage (oocysts) in stool specimens using modified acid stain is regarded as a reliable sensitive technique which is widely used in many clinical labs. In the present study, a total of 133 stool samples were collected from hemodialysis patients and were screened for Cryptosporidium oocyst using formalin-ether concentration and modified acid-fast staining technique. Clinical and demographic data were also collected and analyzed. Cryptosporidium oocysts were recovered in 15/133 (11%) of the investigated hemodialysis patients. The age of patients ranged from 25 to 80 years (mean: 57.84 ± 12.22). Most of the Cryptosporidium-positive cases were recovered from males (73.7%) residing in rural villages in Irbid city (86.6%). The most repeatedly reported symptoms in the Cryptosporidium-positive patients were gastrointestinal symptoms, including diarrhea (15%), nausea (24%), abdominal pain (23%) and bloating (17%), in addition to general fatigue (32%) and weight loss (19%). No statistically significant associations for certain clinical symptoms or risk factors were found. The present study is the first preliminary study in Jordan that provided a brief screening for the incidence of cryptosporidiosis among hemodialysis patients.

Cryptosporidium Oocyst Contamination in Drinking Water: A Case Study in Italy

The aim of this study was to evaluate the occurrence of Cryptosporidium oocysts in a drinking water treatment plant (DWTP) located in a rural area of northern Italy. Influent and effluent samples were collected at the DWTP over three years (2013–2016). In parallel, tap water samples from a public drinking fountain were collected as well. All samples were analyzed for the presence of Cryptosporidium spp. oocysts by a common method based on an immunomagnetic separation (IMS)/immunofluorescence assay (IFA), complemented by 4,6-diamidino-2-phenylindole (DAPI) staining. A reverse transcriptase-PCR (RT-PCR) protocol was added to evaluate oocyst viability. The results highlighted a high variability of oocyst concentrations across all samples (mean 4.3 ± 5.8/100 L) and a high variability in the percentage of DAPI-positive specimens (mean 48.2% ± 40.3%). Conversely, RT-PCR did not reveal the presence of viable C. parvum and C. hominis oocysts. A nested PCR targeting Cryptosporidium 18S ribosomal DNA, carried out in two water samples, confirmed the presence of a Cryptosporidium genotype associated with wild animals in the river and in tap water. The results obtained underline the vulnerability of the investigated surface water to Cryptosporidium spp. contamination. Although the recovered Cryptosporidium genotype is not a human pathogen, its presence demonstrates the existence of a potential pathogen Cryptosporidium spp. contamination risk. Moreover, these results underline the importance of also considering unconventional (not bacterial) biological contaminations (protozoa) in water resources in rural areas, including those of developed countries.