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Disinfection Monroe L. Weber-Shirk School of Civil and Environmental Engineering Overview Why does getting this right matter? Disinfection Options Poisson Distribution of Pathogens Disinfection-by-products Tastes and odors Real pathogens Getting the right dose Probability of ingesting k pathogens Implications for dose dependency WaterBorne Disease Outbreaks Chlorine sources Metals Water Ammonia Organics The case for Chlorine Gas Bleach Onsite production Chemistry Chicks law CT Problems with Disinfection Disinfection mechanisms Chlorine Ozone Irradiation with Ultraviolet light Iodine Silver Chlorine It kills stuff Residual Recontamination Regrowth Hypochlorinators Chlorine free Some European cities Chlorine saves lives… If you accept the “Chlorine eliminated Typhoid Line” Then you will likely recommend chlorination as the first line of defense in the Global South But in small systems (in the Global South) Chlorine dose is generally not controlled based on a target residual dose Surface water may currently be untreated and hence have high turbidity that correlates with high chlorine demand that contains pathogens embedded in organic particles WHO on alternative disinfectants: Iodine, silver, copper, quaternary ammonium compounds none of them are considered suitable for long-term use to disinfect drinking water Iodine is difficult to deliver to water and can cause adverse health effects However, iodine, either dissolved in water or in the form of an iodinated exchange resin, has been used for short-term water treatment Silver and copper are difficult to deliver to water and are only bacteriostatic. Quaternary ammonium compounds are limited in availability, costly and not effective against viruses and parasites. Silver as a Disinfectant… Silver is used as a bacteriostatic agent for point-of-use or household water treatment by storing water in vessels composed of silver or passing water through porous or granular filter media impregnated with silver Many microbes including viruses, protozoan cysts, oocysts, and bacterial spores, are not inactivated at silver concentrations employed for point-of-use drinking water treatment Bacteria may develop silver resistance Therefore, silver is not recommended for routine disinfection of household water Chlorine Disinfection Mechanisms* Oxidation of membrane-bound enzymes for transport and oxidative phosphorylation Oxidation of cytoplasmic enzymes Oxidation of cytoplasmic amino acids to nitrites and aldehydes Oxidation of nucleotide bases Chlorine substitution onto amino acids (more likely) DNA mutations DNA lesions *It is possible that none of these mechanisms have been documented Chick’s Law The death of microorganisms is first order with respect to time Thus, the remaining number of viable microorganisms, N, decreases with time, t, according to: dN kN dt where k is an empirical constant descriptive of the microorganism, pH and disinfectant used. Integrating with respect to time, and replacing limits (N = No at t = 0) yields: 1 N kt pC* kt ln kt N N0e ln 10 N0 EPA Pathogen Inactivation Requirements Safe Drinking Water Act SDWA requires 99.9% inactivation for Giardia and 99.99% inactivation of viruses Giardia is more difficult to kill with chlorine than viruses and thus Giardia inactivation determines the CT Concentration x Time EPA Credits for Giardia Inactivation Treatment type Credit Conventional Filtration 99.7% Direct Filtration* 99% Disinfection f(time, conc., pH, Temp.) * No sedimentation tanks EPA Disinfection CT Credits To get credit for 99.9% inactivation of Giardia: Contact time (min) chlorine pH 6.5 pH 7.5 (mg/L) 2°C 10°C 2°C 10°C 0.5 300 178 430 254 1 159 94 228 134 Inactivation is a function of _______, time ____________ concentration pH temperature ______, and ___________. Where did these numbers (to 3 significant digits) come from? CT equation for Giardia CCl tcontact 0.2828 pH 2.69 CCl0.15 0.933 tcontact 0.2828 pH 2.69 CCl0.85 0.933 pC* tcontact CCl0.85 0.2828 pH 2.69 0.933 Chicks Law! T -5 T -5 T -5 pC * pC * CCl = Free Cl2 Residual [mg/L] tcontact = Time required [min] pH = pH of water T = Temperature, degrees C pC* = -[Log(fraction remaining)] Note: These equations are NOT dimensionally correct! Disinfectant Limitations Disinfection by products Tastes and Odors Real pathogens Getting the right dose Disinfection Byproducts Contaminant MCLG1 MCL (mg/L)2 (mg/L)2 Potential Health Effects from Ingestion of Water Bromate zero Increased risk of cancer 0.010 Sources of Contaminant in Drinking Water Byproduct of drinking water disinfection (plants that use ozone) Chlorite 0.8 1.0 Anemia; infants & young children: nervous system effects Byproduct of drinking water disinfection (plants that use chlorine dioxide) Haloacetic acids (HAA5) n/a6 0.060 Increased risk of cancer Byproduct of drinking water disinfection Total Trihalomethanes (TTHMs) none7 ---------n/a6 0.10 ---------0.080 Liver, kidney or central nervous system problems; increased risk of cancer Byproduct of drinking water disinfection Disinfectants MRDLG Contaminant 1 MRDL1 (mg/L) Potential Health Effects from Ingestion of Water Sources of Contaminant in Drinking Water (mg/L)2 2 Chloramines (as Cl2) MRDLG =41 MRDL =4.01 Eye/nose irritation; stomach discomfort, anemia Water additive used to control microbes Chlorine (as Cl2) MRDLG =41 MRDL =4.01 Eye/nose irritation; stomach discomfort Water additive used to control microbes Chlorine dioxide (as ClO2) MRDLG =0.81 MRDL =0.81 Anemia; infants & young children: nervous system effects Water additive used to control microbes Maximum Residual Disinfectant Level (MRDL) - The highest level of a disinfectant allowed in drinking water. There is convincing evidence that addition of a disinfectant is necessary for control of microbial contaminants. Maximum Residual Disinfectant Level Goal (MRDLG) - The level of a drinking water disinfectant below which there is no known or expected risk to health. MRDLGs do not reflect the benefits of the use of disinfectants to control microbial contaminants. Tastes and Odors: Taste Thresholds C O l Cl N H 2 N Cl H C ch l 2 lo de rofo ch rm di loro ch p h ca loro eno rb be l io on t nze do et ne fo rac rm h lo r O H Taste threshold (mg/L) id e Complaints of the chlorine taste should not be discounted Chlorine taste may prevent some consumers from using treated water Need to convince consumers that the chemical taste is healthy 1 0.1 0.01 0.001 0.0001 Chlorine Taste Acceptance The introduction of chlorine into a community that has always drunk water without the addition of chemicals can be difficult. Reducing the amount of chlorine added might increase the social acceptance of chlorination. With less chlorine, there is less breakthrough of chlorinated compounds into the finished water, and therefore less of a chemical taste. If the water doesn’t taste strongly like chemicals, more people are likely to subscribe to the use of chlorine for disinfection. Inactivation of Shielded Pathogens Many of the studies measuring inactivation of pathogens by disinfectants were conducted using dispersed pathogens What happens if the pathogens are embedded in an organic particle? Faecal contamination potentially contains pathogens embedded in protective organic matter. 0.36 mg/L average free Cl2 at pH 6 What do you conclude? pC* Cell Associate virus was inside fetal rhesus kidney derived cells 4 3.5 3 2.5 2 1.5 1 0.5 0 dispersed cell associated model 2nd model 0 50 Time (min) 86 100 CCl t to get pC* of 4 is (86 min)*(0.36 mg/L)=31 (min mg/L) Conclusions from Virus in Kidney Cells The rate of virus deactivation dropped significantly when the virus particles were inside kidney cells The deactivation of embedded virus particles can not be described by a single first order reaction (________________) Chicks Law is violated What is controlling the rate of virus deactivation? Scales of the Embedded Virus Location Dispersed Inside cell with 1000 nm disrupted cell wall 1 mm Inside intact cell Deactivation rate Very fast Slow Very slow Virus particles are about 20 nm HOCl are about 0.2 nm Mass Transport and Chlorine Protection Chlorine must diffuse through cell contents to reach virus Organic material inside the cell reacts with chlorine before it gets to the virus Scale this up to a Faecal Aggregate Turbid water could easily have organic particles that are 10 or even 100 mm in diameter The amount of organic matter in a small particle and the slow diffusion would provide long term protection for embedded pathogens 10 mm Getting the Right Dose: WHO on Chlorination exposed Chlorine compounds usually destroy pathogens after 30 minutes of contact time, and free residual chlorine (0.2–0.5 mg per liter of treated water) can be maintained in the water supply to provide ongoing disinfection. Several chlorine compounds, such as sodium hypochlorite and calcium hypochlorite, can be used domestically, but the active chlorine concentrations of such sources can be different and this should be taken into account when calculating the amount of chlorine to add to the water. The amount of chlorine that will be needed to kill the pathogens will be affected by the quality of the untreated water and by the strength of the chlorine compound used. If the water is excessively turbid, it should be filtered or allowed to settle before chlorinating it (___________________________) Remove particles first! Pathogen Poisson Process Probability Suppose we have an average pathogen concentration of C in our drinking water Suppose we drink volume V What is the probability that we will ingest k pathogens? Suppose V=1L, C=2/L, what is probability of ingesting exactly 2 pathogens? k 2 CV CV 2 2 P e P e 0.27 k k! k 2! CV=15 Probability of ingesting exactly k pathogens 0.12 1 0.1 0.8 0.08 0.6 0.06 0.4 0.04 0.2 0.02 0 Cumulative probability Probability of k Pathogens 45% chance of not getting sick! 0 0 10 20 30 Number of pathogens What is probability of k < (dose)? Let dose = 15 Find cumulative probability for k=14 Find probability that k>0 0! 0 e CV e CV Pk 0 1 e CV 0.4 1 0.3 0.8 0.6 0.2 0.4 0.1 0.2 0 0 0 2 Cumulative probability Pk 0 e CV k! CV CV=1 Probability of ingesting exactly k pathogens Pk CV k 4 Number of pathogens For CV = 1, Pk>0 = 0.63 For CV = 0.001, Pk>0 = 0.001 (converge for small CV) Effect of Pathogen Dose For CV = 0.001, Pk>0 = 0.001 What happens if the pathogen dose is 10 rather than 1? Let’s assume that the concentration of this new pathogen is 10 times as great (CV=0.01) What is the probability that you ingest 10 or more? For CV = 0.01, Pk≥10 = 3x10-27 Pathogens with an infectious dose of 1 are potentially quite harmful even at very low concentrations! Pathogens with an infectious dose>1 are not dangerous at low concentration! Waterborne Disease Outbreaks in the US (1985) G. lamblia was the most frequently identified pathogen for the seventh consecutive year, causing three (20%) of 15 waterborne outbreaks. In each of the outbreaks, as in well-characterized waterborne outbreaks of giardiasis in the past, water chlorination had been maintained at adequate levels to make outbreaks of bacterial diseases unlikely, but the lack of an intact filtering system capable of filtering Giardia cysts, distribution system problems, and mechanical deficiencies allowed drinking water to become a vehicle of giardiasis. Waterborne Disease Outbreaks (1993) The majority of outbreaks (68%) during 1991-1992 were classified as AGI of unknown etiology Water sampling showed the presence of coliforms and/or deficiencies in chlorination for 91% of these outbreaks 24 outbreaks (71%) were associated with contaminated untreated or inadequately treated groundwater Two outbreaks were associated with treatment deficiencies in water systems using UV light for disinfection Three protozoal outbreaks during 1991-1992 occurred in systems that were equipped with chlorine disinfection and met EPA coliform standards but were not equipped with filtration Waterborne Disease Outbreaks (1993) Four of the six surface water systems associated with WBDOs were equipped with filtration. In three of these outbreaks, raw water quality had deteriorated because of sewage effluents that were not appropriately diluted as a result of low stream flows during dry weather. During the outbreaks associated with these systems, filtration deficiencies were noted, with elevated turbidity in finished water. Decreased filtration efficiency combined with deterioration in raw water quality also contributed to the WBDO in Milwaukee (1993). It appears that chlorination was unable to provide an effective barrier when filtration failed A fatal waterborne disease epidemic in Walkerton, Ontario An estimated 2,300 people became seriously ill and seven died from exposure to microbially contaminated drinking water in the town of Walkerton, Ontario, Canada in May 2000 The Walkerton operators were asked to provide a chlorine residual (majority to be free chlorine) of 0.5 mg/L after 15 min. Evidence at the Inquiry revealed that chlorine dosage practice at Well #5 was insufficient to achieve a 0.5 mg/L residual even in the absence of any chlorine demand. Although the evidence did not allow for an estimate of the chlorine demand at the time Well #5 was contaminated, it was reasonable to assume that the contamination causing this outbreak was accompanied by a chlorine demand sufficient to consume entirely, or almost entirely, the low chlorine dose thereby allowing inadequately disinfected water into the distribution system Chlorine must have been consumed because there was an outbreak Chlorine Sources On Site Production (electrolysis) Chlorine gas (Cl2) Liquid Bleach (NaOCl) Calcium hypochlorite (Contains 65% available chlorine) Ca(OCl)2 Bleach Concentration in terms of sodium hypochlorite (NaOCL) Bleach concentration in terms of Available Chlorine (As Cl2) Additional Information (estimated) Grams per liter Density of the solution (lb/U.S. gal) Specific gravity of the solution Wt. % Trade % Grams per liter Wt. % Trade % 5 5.4 53.9 4.8 5.1 51.4 9.0 1.08 10 11.6 115.8 9.5 11.0 110.4 9.7 1.16 15 18.6 185.7 14.3 17.7 177.0 10.3 1.24 Chlorine First large-scale chlorination was in 1908 at the Boonton Reservoir of the Jersey City Water Works in the United States Chlorine Widely used in the US oxidizes organic matter Typical dosage (1-5 mg/L) variable, based on the chlorine demand goal of 0.2 mg/L residual Trihalomethanes (EPA primary standard is 80 mg/L) Chlorine concentration is measured as Cl2 even when in the form of HOCl or OCl- Chlorine Reactions Charges 0 +1 -2 +1 -1 Cl2 + H2O H+ + HOCl + ClHypochlorous acid HOCl H+ + OCl- Hypochlorite ion The sum of HOCl and OCl- is called the ____ ______ residual _______ free chlorine HOCl is the more effective disinfectant Therefore chlorine disinfection is more effective at ________ low pH Dissociation constant is 10-7.5 HOCl and OCl- are in equilibrium at pH 7.5 Fraction of Free Chlorine Chlorine and pH HOCl H+ + OCl- 1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 HOCl OCl- pk 5 6 7 8 pH 9 10 Ammonia Reactions -3 +1 -3+1 +1 NH3(aq) + HOCl NH2Cl+ H2O Combined chlorine NH2Cl + HOCl NHCl2+ H2O Substitution reactions… The combined chlorine maintains its oxidizing potential Breakpoint Chlorination Removal of ammonia by chlorination -3 +1 0 -1 2NH3(aq) + 3HOCl N2+ 3Cl- + 3H2O Oxidizing equivalents of chlorine are consumed Does Chlorine Completely oxidize organic matter? NO! 4HOCl + CH4 CO2 + 2H2O + 4Cl- + 4H+ Oxidation states Carbon in organic matter (-4) Carbon in carbon dioxide (+4) Chlorine in HOCl (+1) Chloride (-1) Therefore should take 4 moles of chlorine (Cl2) per mole of organic carbon 23.6 g chlorine/g organic carbon Chlorine Demand (mg/L) Chlorine Demand vs. Total Organic Carbon 2.5 2 1.5 1 y = 0.51x - 0.16 R2 = 0.93 0.5 0 0 1 2 3 4 Total Organic Carbon (mg/L) 5 0.5 mg chlorine mg carbon Reaction with organic compounds with unsaturated linkages C H C H + HOCl → Cl C H OH C H Chlorine doesn’t oxidize the organic carbon Chlorine maintains its oxidation number 0.5 mg Cl2 mole Cl2 mole Cl 1 12 g C mole Cl 1 mg C 70.9 g Cl2 mole Cl2 mole C 12 mole C Disclaimer – There is no solid connection between chlorine demand and turbidity But – organic matter is often associated with particulate matter Chlorine Demand (mg/L) Chlorine Demand vs Turbidity If using the standard dose of 2 mg/L, then no residual above 15 NTU! 2 1.5 1 Based on 6 watersheds in western Oregon 0.5 0 0 5 10 Turbidity (NTU) 15 The Case for Chlorine It kills stuff Residual Recontamination Regrowth Effect of Chlorination on Inactivating Selected Bacteria Bacteria Cl2 Concentration (mg/l) Time (min) Ct Factor (mgmin/l) Reduction(%) Campylobacter jejuni 0.1 5 0.5 99.99 Blaser et al, 1986 Escherichia coli 0.2 3 5 99.99 Ram and Malley, 1984 Legionella pneumophila 0.25 60-90 18.75 99 Kuchta et al, 1985 Mycobacterium chelonei 0.7 60 42 99.95 Carson et al, 1978 Mycobacterium fortuitum 1.0 30 30 99.4 Pelletier and DuMoulin, 1987 Mycobacterium intracellulare 0.15 60 9 70 Pelletier and DuMoulin, 1987 0.5-1.0 5 3.75 99.6-100 Salmonella typhi 0.5 6 3 99 Shigella dysenteriae 0.05 10 0.5 99.6-100 Staphylococcus aureus 0.8 0.5 0.4 100 Bolton et al, 1988 Vibrio cholerae(smooth strain) 1.0 <1 <1 100 Rice et al, 1993 Vibrio cholerae (rugose strain) 2.0 30 60 99.999 Rice et al, 1993 Yersinia enterocolitica 1.0 30 30 92 Paz et al, 1993 Pasteurella tularensis Reference Baumann and Ludwig, 1962 Korol et al, 1995 Baumann and Ludwig, 1962 Effect of Chlorination on Inactivating Selected Viruses Viruses Cl2 Concentration (mg/l) Time (min) Ct factor (mg-min/l) Reduction (%) Reference Adenovirus 0.2 40-50 sec 0.15 99.8 Clarke et al, 1956 Coxsackie 0.16-0.18 3.8 0.06 99.6 Clarke and Kabler, 1954 Hepatitis A 0.42 1 0.42 99.99 Grabow et al, 1983 0.5-1.0 30 22.5 -- Keswick et al, 1985 Parvovirus 0.2 3.2 0.64 99 Churn et al, 1984 Poliovirus 0.5-1.0 30 22.5 100 Keswick et al, 1985 Rotavirus 0.5-1.0 30 22.5 100 Keswick et al, 1985 Norwalk Effect of Chlorination on Inactivating Selected Protozoa Cl2 Concentration (mg/l) Time (min) Ct Factor (mg-min/l) Reduction (%) Cryptosporidium parvum 80 90 7200* 90 Korich et al, 1990 Entamoeba histolytica 1.0 50 50 100 Snow, 1956 -- -- 68-389 99.9 AWWA, 1999 0.5-1.0 60 45 99.99 de Jonckheere and van de Voorde, 1976 Protozoa Giardia lamblia Naegleria fowleri Reference The Case for a Residual Disinfect any recontamination Prevent bacteria growth in the treated water Do pathogens grow in water? “The real reason for maintaining residuals during treatment and distribution is to control microbiological growths when the water is biologically unstable.” Control those non-pathogenic slime-forming organisms “Current practice in North America tries to kill all microorganisms whenever possible” Protection Against Recontamination In order to be effective the following requirements must be met The amount of chlorine demand must not exceed the residual The pathogens must be dispersed (not associated with other particles) This is unlikely if the contamination is faecal The pathogens must be susceptible to chlorine Growth of Bacteria in Water Distribution Systems Consumption of dissolved oxygen Increased heterotrophic plate counts or coliform counts This does not imply a health risk Decreased hydraulic capacity of the pipes Formation of taste/odor compounds Geosmin, mercaptans, amines, tryptophans, sulfates Increased rates of pipe corrosion I don’t have any evidence that this biological growth has a significant public health impact Pathogen Growth in Distribution Systems (CDC) Biofilms are coatings of organic and inorganic materials in pipes that can harbor, protect, and allow the proliferation of several bacterial pathogens, including Legionella and Mycobacterium avium complex (MAC). Mycobacterium avium complex, MAC, is an opportunistic bacterial pathogen, is resistant to water disinfection (much more so than Giardia cysts), and grows in pipe biofilms I need more data here! Is this a real public health threat? WHO on Regrowth There is no evidence to implicate the occurrence of most microorganisms from biofilms (excepting, for example, Legionella, which can colonize water systems in buildings) with adverse health effects in the general population through drinking water, with the possible exception of severely immunocompromised people Water temperatures and nutrient concentrations are not generally elevated enough within the distribution system to support the growth of E. coli (or enteric pathogenic bacteria) in biofilms. Thus, the presence of E. coli should be considered as evidence of recent faecal contamination. http://www.who.int/water_sanitation_health/dwq/en/gdwq3_4.pdf WHO on Regrowth (2) Viruses and the resting stages of parasites (cysts, oocysts, ova) are unable to multiply in water. Relatively high amounts of biodegradable organic carbon, together with warm temperatures and low residual concentrations of chlorine, can permit growth of Legionella, V. cholerae, Naegleria fowleri, Acanthamoeba and nuisance organisms in some surface waters and during water distribution http://www.who.int/water_sanitation_health/dwq/en/gdwq3_7.pdf Where is the original research for these conclusions? Life without Chlorine Amsterdam stopped chlorinating in 1983 Recommendations for Chlorine as Sole Treatment Low turbidity (<30 NTU) and low chlorine demand for effective use; pre-treat turbid water (WHO) Pre-treat means using a particle removal technology first Low turbidity probably means good quality groundwater (from a spring or from a well) Surface water would generally not meet the NTU requirement Chlorine requires process control (feedback based on residual chlorine concentration) Hypochlorinator Float 1.0 m 1.5” PVC overflow tube Transparent flexible tube 1.78 m (0.5”) 1.05 m PVC needle valve 0.5” PVC tube Water in the distribution tank Example Conservation of Mass Changing Volume Float ¶ r V ×n ˆ dA = r dV ò ò ¶t cv cs Vor Aor Vor Aor ¶ =dV ò ¶t cv Ares dh dV dt dt Vor Aor Qor dh Ares K or Aor dt 1.0 m 1.5” PVC overflow tube Transparent flexible tube 1.78 m (0.5”) 1.05 m h0 PVC needle valve 0.5” PVC tube Water in the distribution tank Orifice in the needle valve 2 gh 0 Integrate to get h as f(t) Finding the chlorine depth as f(t) Ares K or Aor h 2g Ares K or Aor 2g h0 dh h t dt 0 2 h1/ 2 h01/ 2 t 1/ 2 tK or Aor 2 g h h0 2 Ares 1/ 2 Aor h h0 tK or Ares g 2 2 2 h h0 tK or Aor Ares g 2 Finding Q as f(t) Q K or Aor 2 gh h h0 tK or Q K or Aor 2g Aor Ares g 2 h0 tK or Aor Ares g 2 Find Aor as function of initial target flow rate Aor Q0 K or 2 gh0 Set the valve to get desired dose initially Surprise… Q decreases linearly! Q K or Aor Q 2g 2 gh0 K or Q0 K or 2g h0 tK or h0 tK or g 2 Aor Ares K or Ares 2 gh0 Q0 g Q Q0 Aor Q0 K or h0 Ares 2 h0 tQ0 Q 1 Q0 2 Ares h0 1 h0 2 gh0 tQ0 Relationship between Q0 and Ares? Flow at Q0 for 4 days (tdesign) would empty reservoir Q0tdesign Ares hres Q 1 t hres 1 Q0 2 tdesign h0 Q0 hres Ares tdesign CCl2 CCl2 0 1 t hres 1 2 tdesign h0 Hypochorinator Fix http://web.mit.edu/d-lab/honduras.htm Conclusions Reflections What is one explanation for why Biosand filters don’t perform as well as expected? Calicivirus - An Emerging Contaminant in Water Annual estimates among adults in the U.S. reveal approximately 267 million episodes of diarrhea leading to 612,000 hospitalizations and 3,000 deaths In the last 3 decades it has become increasingly clear that viral agents are responsible for much of this public health burden Human caliciviruses (HuCVs) have been estimated to cause 95–96% of nonbacterial gastroenteritis outbreaks (Fankhauser et al., 1998; K.Y. Green et al., 2000). These viruses are considered ubiquitous in nature and stable in the environment, thereby increasing their propensity to spread and cause disease Calicivirus Four genera Lagovirus Vesivirus Norwalk-like viruses (Noroviruses or NLVs) Sapporo-like viruses (Sapoviruses or SLVs) Single structural protein that makes up the viral capsid 27-40 nm in diameter Cause disease in humans Norovirus: Infectious Dose Human volunteer feeding studies have determined the number of viral particles needed to initiate Huffman, D. E., K. L. Nelson, et al. (2003). infection is 10 to 100 The infectious dose may well be the result of mass transfer limitations in the gut i.e. the virus needs to be transported to the villi in the gastric mucosa and attach to initiate infection Thus it is likely that 1 viral particle is sufficient to Weber-Shirk initiate infection Norovirus: Clinical Illness and Diagnosis The most common symptoms of NLV infections include mild to moderate diarrhea, abdominal cramps, and nausea (Adler and Zickl, 1969; Hedberg and Osterholm, 1993). Other symptoms may include headache, malaise, chills, cramping, and abdominal pain. Stools typically do not contain blood or mucus. The onset of illness is generally within 24 to 28 h of exposure with a relatively short duration of illness (12 to 60 h). Adult infection with NLV can be distinguished from bacterial pathogens such as Salmonella and Shigella due to its characteristic projectile vomiting (Adler and Zickl, 1969; Caul, 1996) Norovirus: Inactivation A 1-log reduction in the RTPCR signal of Norwalk virus was observed after treatment with 2 mg/L monochloramine at pH 8 for 3 h Norwalk virus appears to be fairly resistant to free chlorine and monochloramine