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Fact Sheet Removal and Inactivation of Cryptosporidium in the Environment

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Fact Sheet Removal and Inactivation of Cryptosporidium in the Environment Fact Sheet Removal and inactivation of Cryptosporidium in the environment Introduction Cryptosporidium oocysts commonly occur in surface and exposure to 0.06 M ammonia causing 5 log10 inactivation recreational waters due to faecal contamination from wild after 8.2 days at 24°C or 55 days at 4°C (Jenkins et al. 1998). animals or human activity. Oocysts have many character- Considering this sensitivity, it is likely that some level of istics that allow them to persist in aquatic environments inactivation by ammonia will occur in and withstand water treatment processes. Direct exposure animal faeces even at low temperatures, to contaminated recreational or drinking water can cause particularly if animal waste is stored prior outbreaks. However, as outlined below, the environment to disposal (Hutchison et al. 2005) can also be hostile to oocysts. This fact sheet provides an overview of the processes that can remove or inactivate As mentioned, oocysts are extremely Cryptosporidium oocysts in catchments and surface waters. sensitive to desiccation, which can occur More information about Cryptosporidium, including the within faeces because of processing by identification and detection of infectious oocysts, is available dung beetles (Ryan et al. 2011) or once in the Cryptosporidium toolbox factsheet. oocysts have been washed into soil. Inactivation by desiccation can be rapid, The terrestrial environment with oocysts air-dried onto glass slides inactivated by 2.5 log10 within 2 hours The two main ways in which oocysts can enter surface and completely inactivated after 4 hours waters are by direct deposition (eg, animal excretion or (Robertson et al. 1992). In addition to sewage discharge) or by rainfall runoff from land. The desiccation, the physical, chemical and terrestrial environment can expose oocysts to temperature biological properties of soil may reduce extremes and desiccation, which are particularly effective oocyst survival (Ferguson et al. 2003). at inactivating oocysts. Cow faeces exposed to sunlight can Soils with low water potential (osmolality), reach internal temperature peaks between 40°C and 70°C in combination with temperature and once the air temperature exceeds 25°C (Li et al. 2005). freeze-thawing, resulted in enhanced Oocysts suspended in water are highly sensitive to these oocyst degradation, with degradation more rapid in dry soil compared with calf faeces or in temperatures, with 6 log10 inactivation after exposure to 60°C for 15 seconds or 55°C for 30 seconds. These rates water at low temperatures (Walker et al. 2001). Oocysts in may be slower in faeces due to the presence of solids and saturated loamy soil had comparable inactivation to oocysts in distilled water, with 0.93 log inactivation after 10 days fats, but an inactivation rate of 3.3 log10 /day in cow faeces 10 has been reported for daily temperature cycles typical of at 30°C (Nasser et al. 2007). In comparison, oocysts in dry spring/autumn in a Mediterranean climate (Li et al. 2005). In loamy soil had 2.5 log10 inactivation after 10 days at 32°C (Nasser et al. 2007). winter the inactivation rates are much slower, with 0.2 log10/ day when the internal faecal matrix temperature was 30°C Aside from inactivation while on land, oocysts can also be and 0.03 log10 / day when the matrix was at 20°C (Li et al. 2010). Although sensitive to heat, oocysts can survive short removed prior to or during transport into receiving waters. periods of freezing (Fayer and Nerad 1996). However, cycles The transport of oocysts through soils is highly dependent of freeze/thawing rapidly inactivate oocysts, particularly in on soil type and conditions. Under some conditions oocysts soil where mechanical contact with particles causes abrasion do not appear to attach to soil particles in water (Brookeset and fragmentation of oocysts (Jenkins et al. 1999, Kato et al. 2006, Dai and Boll 2003, Kaucner et al. 2005). However, al. 2002). Chemicals within faeces also cause significant oocysts will attach to clay loam and sandy loam in the inactivation. Oocysts are highly sensitive to ammonia, with presence of manure (Kuczynska et al. 2005). Irrespective Water Research Australia Research solutions though collaboration April 2016 Page 1 of binding, soils can act as a filter to limit oocyst mobility of 10. In contrast, these values were 10.8 hours and 6.5 (Tufenkji et al. 2004), with the majority of oocysts retained in hours respectively for water with high (12.3 mg/L) DOC and the top 2 cm of soil (Mawdsley et al. 1996). Oocysts can pass high (77 HU) colour (King et al. 2008). The presence of DOC through saturated macroporous soils, with oocyst removals reduces penetration of UV in water, protecting oocysts that are at sufficient depth. However, solar inactivation may still of 1 – 3 log10 under such conditions (Darnault et al. 2003). Removal of oocysts through soil appears to be variable, occur from UV-A, which has a longer wavelength than UV-B possibly due to variation in the health of oocysts within the and may be able to better penetrate surface waters, and from oocyst population, with the observed transport not consistent resuspension events such as warm water inflows. with colloid theory (Santamaria et al. 2011). Vegetation can also be effective at removing Cryptosporidium (Tate et al. Oocysts in water are not exposed to the same temperature 2004), with vegetation on sandy loam removing 1 – 2 log extremes as those on land. Inactivation still occurs, but the 10 rate is relatively slow, requiring 16 – 24 weeks at 15°C for 1 oocysts/meter compared with a reduction of 2 – 3 log10 oocysts/meter on silty clay or loam (Atwill et al. 2002). log10 oocyst inactivation (Keegan et al. 2008). Inactivation rates increase for higher water temperatures, with more than Surface waters 3 log10 inactivation after 12 weeks at 20°C, 8 weeks at 25°C and 3 weeks at 30°C (King et al. 2005). The rate of inactivation was In addition to providing a medium for oocyst transmission the same in sterile reservoir water and tap water. to a host, aquatic environments can support oocyst survival by providing a thermal buffer against temperature extremes. Oocysts surviving thermal or solar inactivation can be removed However, many factors can remove or inactivate oocysts in by biological interactions, especially at interfaces such as water, including particle interactions, temperature, sunlight sediments and biofilms. Despite the potential importance of exposure and predation by protozoans or zooplankton. biological interactions for the removal of oocysts from water, this field has been little studied. A wide range of organisms Oocyst transport in reservoirs is predominantly driven isolated from reservoir water can ingest oocysts, including by inflows, particularly from rainfall (Brookes et al. rotifers, ciliates, amoebae, gastrotrichs and platyhelminths 2004). Inflow temperature, velocity, insertion depth and (King et al. 2007). The kinetics of removal has not been entrainment rate all determine the distribution of oocysts determined using environmentally relevant oocyst numbers. in lakes and reservoirs (Brookes et al. 2004). The position of Predation may lead to removal of oocysts through degradation oocysts can influence survival, with oocysts at the surface of ingested oocysts or by causing clumping of excreted exposed to sunlight, whereas oocysts at depth can escape oocysts that are not digested (King et al. 2007). The infectivity sunlight inactivation but be removed by predation. The of oocysts excreted by freshwater predators has not been sedimentation of individual oocysts is relatively slow (0.27 – determined. However, predation of oocysts by zooplankton 0.35 µm/s) under laboratory conditions (Dai and Boll 2006, isolated from a waste stabilisation pond caused 0.7 to >2 Log10 Medema et al. 1998). Such a sedimentation rate makes it inactivation depending on the types of zooplankton present in unlikely that single oocysts will settle in large water bodies. the microcosm experiment (Salas Iglesias 2014). In addition to However, attachment to particles or organic aggregates can providing an environment for interaction between predators greatly increase sedimentation rates by 100-fold (Medema and oocysts, biofilms might also directly interact with oocysts. et al. 1998, Searcy et al. 2005). Oocyst sedimentation rates The attachment to and detachment of oocysts from biofilms measured in Lake Burragorang were 57.9 – 115.7 µm/s, likely can be variable and appears to be influenced by season due to oocyst aggregation with other oocysts or particles (Wolyniak et al. 2010). Of significance, oocysts embedded (Hawkins et al. 2000). Oocysts that enter sediment can at the bottom of a biofilm retain infectivity up to two times potentially be remobilised by re-suspension events such as better than oocysts at the top of a biofilm or oocysts with no turbulence from underflows (Michallet and Ivey 1999). biofilm following sunlight exposure (DiCesare et al. 2012). Another potential source of oocyst removal in freshwaters is Sunlight, in particular ultraviolet (UV) light, can be harmful by filter feeders such as bivalve molluscs, which concentrate to a wide variety of organisms and is a potent environmental oocysts from contaminated waters (Graczyk et al. 2001, Izumi stress for Cryptosporidium inactivation. Different et al. 2006). Only 1-3% of oocysts are retained following wavelengths of UV light cause different types of damage, ingestion, with the excreted oocysts retaining infectivity (Izumi with UV-B (280nm to 320nm) causing DNA damage and UV-A et al. 2006). Based on the high clearance rate, it is possible (320nm-400nm) causing damage to cellular components that detection of low numbers of oocysts in mollusc tissue such as lipids and proteins (Caldwell 1971, Friedberg et al. represents previous contamination events, while high numbers 1995, Malloy et al. 1997, Ravanat et al. 2001). Solar radiation of oocysts represents a recent event. can inactivate oocysts suspended in tap water and reservoir waters, with UV-B causing approximately two thirds of the observed inactivation (King et al.
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