Solar UV reduces Cryptosporidium parvum oocyst infectivity in environmental waters
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To determine the effect of solar radiation on Cryptosporidium parvum in tap and environmental waters.Outdoor tank experiments and a cell culture infectivity assay were used to measure solar inactivation of C. parvum oocysts in different waters. Experiments conducted on days with different levels of solar insolation identified rapid inactivation of oocysts in tap water (up to 90% inactivation within the first hour). Increased dissolved organic carbon content in environmental waters decreased solar inactivation. The role of solar ultraviolet (UV) in inactivation was confirmed by long-pass filter experiments, where UV-B was identified as the most germicidal wavelength. Reductions in oocyst infectivity following solar radiation were not related to a loss of excystation capacity.Solar UV can rapidly inactivate C. parvum in environmental waters.This is the first study to assess natural sunlight inactivation of C. parvum oocysts in surface waters and drinking water using an infectivity measure and determines the wavelengths of light responsible for the inactivation. The findings presented here provide valuable information for determining the relative risks associated with Cryptosporidium oocysts in aquatic environments and identify solar radiation as a critical process affecting the oocyst survival in the environment.Keywords:
Infectivity
Cryptosporidium parvum
Tap water
Ultraviolet
To determine the effect of solar radiation on Cryptosporidium parvum in tap and environmental waters.Outdoor tank experiments and a cell culture infectivity assay were used to measure solar inactivation of C. parvum oocysts in different waters. Experiments conducted on days with different levels of solar insolation identified rapid inactivation of oocysts in tap water (up to 90% inactivation within the first hour). Increased dissolved organic carbon content in environmental waters decreased solar inactivation. The role of solar ultraviolet (UV) in inactivation was confirmed by long-pass filter experiments, where UV-B was identified as the most germicidal wavelength. Reductions in oocyst infectivity following solar radiation were not related to a loss of excystation capacity.Solar UV can rapidly inactivate C. parvum in environmental waters.This is the first study to assess natural sunlight inactivation of C. parvum oocysts in surface waters and drinking water using an infectivity measure and determines the wavelengths of light responsible for the inactivation. The findings presented here provide valuable information for determining the relative risks associated with Cryptosporidium oocysts in aquatic environments and identify solar radiation as a critical process affecting the oocyst survival in the environment.
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Cryptosporidium parvum
Tap water
Ultraviolet
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Animal infectivity studies demonstrate the efficacy of pulsed and advanced UV in inactivating Cryptosporidium oocysts. Cryptosporidium parvum oocysts are highly resistant to conventional chlorine‐based disinfectants. The authors tested two innovative electrotechnologies that use ultraviolet (UV) light and found that both pulsed UV and advanced UV inactivated Cryptosporidium oocysts. The advanced UV system achieved >4‐log inactivation as determined in animal infectivity studies using the neonatal mouse model. With the pulsed UV system, oocyst inactivation was also noted in the process control (non‐UV‐exposed oocysts), suggesting that some oocyst inactivation may have occurred independently of UV exposure. Irrespective of this, both technologies appear to be effective and novel ways to treat drinking water and to provide an additional significant barrier that helps protect public health.
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Cryptosporidium parvum
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Cell culture assays in various formats have been used to study the infectivity of Cryptosporidium spp. as well as to determine the infectivity of naturally occurring oocysts in water. Currently, cell culture assays for infectious Cryptosporidium spp. in water have largely been limited to practice in research laboratories. One obstacle to the routine use of Cryptosporidium cell culture assays for the analysis of water samples is the coordination of water sample collection and processing with readiness of cell culture monolayers. For most Cryptosporidium cell culture assays, monolayers are allowed to develop for 24 to 48 h to reach 80 to 100% confluence prior to inoculation. In this study, we used immunofluorescent assay microscopy to evaluate freshly confluent (2-day-old) and aged (8- to 67-day-old) HCT-8 cell monolayers for their ability to support Cryptosporidium parvum infection. HCT-8 monolayers as old as 67 days were clearly shown to support infection. In two of three experiments, aged monolayers (8- to 11-day-old and 11- to 22-day-old, respectively) developed the same number of C. parvum clusters of infection as freshly confluent monolayers. Results suggest that it may be possible to use cell monolayers from freshly confluent to 3 weeks old on hand for infectivity assays without having to schedule sample processing to coincide with development of freshly confluent monolayers. This would make Cryptosporidium cell culture assays much more feasible for water quality and utility laboratories.
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Even extremely low dosages of ultraviolet light can be highly effective for inactivating Cryptosporidium oocysts. Recent studies have shown that Cryptosporidium parvum oocysts demonstrate high susceptibility to low dosages of medium‐pressure ultraviolet (UV) light. These investigations have raised several questions, which include determination of minimum medium‐pressure UV dosages necessary to inactivate C. parvum oocysts, elucidation of differences (if any) between medium‐ and low‐pressure UV light for inactivating C. parvum oocysts, and evaluation of medium‐pressure UV effectiveness in inactivating oocysts suspended in poorer quality water. To compare low‐ and medium‐pressure UV, the authors exposed oocysts suspended in deionized water to UV delivered by either medium‐ or low‐pressure UV lamps at bench scale using a collimated beam apparatus. The applied UV dosages ranged from 3 to 33 mJ/cm 2 , and oocyst inactivation was assessed using the neonatal mouse infectivity assay. At 3 mJ/cm 2 , medium‐pressure UV showed a 3.4‐log inactivation of oocysts, and low‐pressure UV showed a 3.0‐log inactivation, demonstrating medium‐ and low‐pressure UV did not differ significantly in inactivating C. parvum oocysts.
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Cryptosporidium parvum
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An in vivo SCID mouse infectivity assay was used to determine its capacity to detect the infectivity of low concentrations of Cryptosporidium parvum oocysts in water. This biological test can be applied to demonstrate oocysts infectivity in water samples derived from drinking water supply and/or environmental sources. Ein biologisches Testverfahren zum Nachweis infektiöser Cryptosporidium parvum Oozysten in niedrigen Konzentrationen im Wasser Ein In-vivo-SCID-Maus-Übertragbarkeitstest wurde auf seine Eignung hin untersucht, infektiöse Cryptosporidium parvum Oozysten in niedrigen Konzentrationen in Wasser nachzuweisen. Es zeigte sich, dass dieses biologische Testverfahren sowohl für Wasserproben aus dem Wasserversorgungssystem als auch für andere Umweltproben einsetzbar ist.
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Small numbers of Cryptosporidium parvum oocysts can contaminate even treated drinking water, and ingestion of oocysts can cause diarrheal disease in normal as well as immunocompromised hosts. Since the number of organisms necessary to cause infection in humans is unknown, we performed a study to determine the infective dose of the parasite in healthy adults.After providing informed consent, 29 healthy volunteers without evidence of previous C. parvum infection, as determined by the absence of anti-cryptosporidium-specific antibodies, were given a single dose of 30 to 1 million C. parvum oocysts obtained from a calf. They were then monitored for oocyst excretion and clinical illness for eight weeks. Household contacts were monitored for secondary spread.Of the 16 subjects who received an intended dose of 300 or more oocysts, 14 (88 percent) became infected. After a dose of 30 oocysts, one of five subjects (20 percent) became infected, whereas at a dose of 1000 or more oocysts, seven of seven became infected. The median infective dose, calculated by linear regression, was 132 oocysts. Of the 18 subjects who excreted oocysts after the challenge dose, 11 had enteric symptoms and 7 (39 percent) had clinical cryptosporidiosis, consisting of diarrhea plus at least one other enteric symptom. All recovered, and there were no secondary cases of diarrhea among household contacts.In healthy adults with no serologic evidence of past infection with C. parvum, a low dose of C. parvum oocysts is sufficient to cause infection.
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Free-Living Amebae (FLA) and Cryptosporidium oocysts occasionally share the same environment. From 2004 to 2016, Cryptosporidium was responsible for 60% of 905 worldwide waterborne outbreaks caused by protozoan parasites. The aim of this study was to evaluate interactions between C. parvum oocysts and two common FLAs (Acanthamoeba castellanii and Vermamoeba vermiformis) in a water environment. Encystment and survival of FLAs were evaluated by microscopy using trypan blue vital coloration. Oocysts were numerated on microscopy. Interactions were studied over time in conditions both unfavorable and favorable to phagocytosis. Potential phagocytosis was directly evaluated by several microscopic approaches and indirectly by numeration of microorganisms and oocyst infectivity evaluation. Occasional phagocytosis of C. parvum by FLAs was documented. However, oocyst concentrations did not decrease significantly, suggesting resistance of oocysts to phagocytosis. A temporary decrease of oocyst infectivity was observed in the presence of A. castellanii. The effect of these interactions on C. parvum infectivity is particularly interesting. The biofilm condition could favor the persistence or even the proliferation of oocysts over time. This study demonstrated interactions between C. parvum and FLAs. Further knowledge of the mechanisms involved in the decrease of oocyst infectivity in the presence of A. castellanii could facilitate the development of new therapeutic approaches.Interactions entre amibes libres et Cryptosporidium parvum : étude expérimentale.Les amibes libres et les oocystes de Cryptosporidium partagent parfois le même environnement. Entre 2004 et 2016, Cryptosporidium a été responsable de 60 % des 905 épidémies d’origine hydrique dans le monde causées par des parasites protozoaires. Le but de cette étude était d’évaluer les interactions entre les oocystes de C. parvum et deux espèces d’amibes libres communes (Acanthamoeba castellanii et Vermamoeba vermiformis) en environnement aquatique. L’enkystement et la survie des amibes libres ont été évalués par microscopie en utilisant une coloration vitale au bleu trypan. Les oocystes ont été comptés au microscope. Les interactions ont été étudiées au cours du temps dans des conditions à la fois défavorables et favorables à la phagocytose. La phagocytose potentielle a été évaluée directement par plusieurs approches microscopiques et indirectement par la numération des micro-organismes et l’évaluation de l’infectiosité des oocystes. Une phagocytose occasionnelle de C. parvum par amibes libre a été documentée. Cependant, les concentrations d’oocystes n’ont pas diminué de manière significative, ce qui suggère une résistance des oocystes à la phagocytose. Une diminution temporaire de l’infectivité des oocystes a été observée en présence d’A. castellanii. L’effet de ces interactions sur l’infectiosité de C. parvum est particulièrement intéressant. La condition biofilm pourrait favoriser la persistance ou même la prolifération des oocystes de C. parvum au fil du temps. Cette étude a démontré des interactions entre C. parvum et amibes libres. Une meilleure connaissance des mécanismes impliqués dans la diminution de l’infectiosité des oocystes en présence d’A. castellanii pourrait faciliter le développement de nouvelles approches thérapeutiques.
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As Cryptosporidium parvum continues to cause waterborne disease, despite extensive efforts by drinking water suppliers and regulators, it is important to have reliable and convenient methods for detection of this pathogen in wastewater discharges, environmental source waters and finished drinking water supplies. In order to better understand the health risks of this organism, it is necessary that detection methods be able to distinguish between infectious and non-infectious Cryptosporidium oocysts in these environmental samples. Cryptosporidium infectivity assay systems based on infections in mice and on in vitro infections in continuous mammalian cell lines are available. Currently, these methods are impractical for routine analysis of water samples because they are tedious, lengthy and costly. These methods rely on careful microscopic examination or further analysis by PCR and then characterisation of the amplified DNA. Practical and affordable non-microscopic methods to determine Cryptosporidium infectivity are much needed for environmental analysis. A cell culture infectivity detection system was developed for infectious Cryptosporidium oocysts that does not rely on microscopic examination of samples to score results, is applicable to a variety of samples and has the potential to be used for routine water monitoring and other environmental or biomedical analysis. Using a chemiluminescent immunoassay, the discrete foci of developmental stages of Cryptosporidium in cell cultures are clearly visible as discrete objects in an image of the entire cell culture layer in a dish or on a slide. These objects are directly countable with the unaided eye and their identity can be further confirmed or verified by microscopic examination.
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Water disinfection
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Abstract The authors compared the viability and infectivity of Cryptosporidium parvum oocysts in chlorinated tap water at various storage durations (i.e., 2 wk, 4 wk, 6 wk, or 8 wk) and at 2 cool temperatures (i.e., 10[ddot]C and 4[ddot]C), using in vitro (excystation) and in vivo (suckling mouse) methods. After 8 wk, mean oocyst excystation decreased to 33.4% and 26.7% at 10[ddot]C and 4[ddot]C, respectively. Suckling mice infectivity was higher after storage at 10[ddot]C than after storage at 4[ddot]C. These data suggest that Cryptosporidium parvum oocysts can survive and remain infectious for 8 wk in cool chlorinated tap water.
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