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111 Microbiology of Drinking Water The principal microbiological contaminants found in drinking water of the United States are bacteria, viruses, and pathogenic protozoa. Each is considered in a separate section of this chapter. Helminths are included along with the protozoa. Little information is available on mycoplasma, pathogenic yeast, and pathogenic fungi in drinking water. Microbiologi- cal contaminants, such as fungi and algae, do not seem to be important causes of waterborne disease, although they are sometimes associated with undesirable tastes and odors. EPIDEMIOLOGY The average annual number of waterborne-disease outbreaks in the United States reported since 1938 is shown in Figure III-1 (Center for Disease Control, 1976b). There was a decrease in the number of outbreaks during the late 1930's and 1940's, but this trend was reversed in the early 1950's. There has been a pronounced increase in the outbreaks reported by the Center for Disease Control (CDC) in Atlanta, Georgia, since 1971. The reason for this apparent increase is not entirely clear, but it could be either the result of improved reporting or an overloading of our treatment plants with source water of increasingly lower quality. Since 1971, the CDC, the Environmental Protection Agency (EPA), state epidemiolo- gists, and engineers in state water-supply surveillance agencies have cooperated in the annual reporting of outbreaks. The purposes of such 63
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64 DRINKING WATER AND H"LTH 50 _ CC m 40 in ~ 30 of LU 20 I: I: - o 1938- 1941- 1946- 1951 - 1956- 1961 - 1966- 197 1 1940 1945 1950 1955 1960 1965 1970 1974 YEARS FIGURE III-l Average annual number of waterborne disease outbreaks, 1938-1975. reports are to control disease by identifying contaminated water sources and purifying them, and to increase knowledge of disease causation. The roles of many microbial agents, including, for example, Yersinia enterocolitica and mycoplasma, remain to be clarified. The most important waterborne infectious diseases that occurred in 1971-1974 are listed in Table III-1. The etiologic agent was determined in only 53% of 99 disease outbreaks that involved 16,950 cases (Craun et al. 1976~. The remainder were characterized as "acute gastrointestinal illness of unknown etiology." Shigellosis was the most commonly identified bacterial disease (2,747 cases) in 1971-1974. Most of the cases were associated with non-municipal water systems. Four typhoid fever outbreaks affected 222 people and involved semipublic and individual water systems. In 1974, 28 waterborne-disease outbreaks, comprising 8,413 cases, were reported to the Center for Disease Control (1976a). The largest was an outbreak of giardiasis that occurred in Rome, N.Y., with an estimated 4,800 cases. The second largest involved about 1,200 cases caused by Shigella sonnet. In the third largest, which involved 615 cases of acute gastrointestinal illness, the etiologic agent was not definitely determined, but Yersinia enterocolitica was suspected. The fourth largest was caused by Shigella sonnet and involved 600 persons. Nineteen states reported at
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Microbiology of Drinking Water 65 least one outbreak. Craun et al. (1976) stated that "this probably reflects the level of interest in investigating and reporting in different states rather than the true magnitude of the problem within the state." Semipublic water systems were associated with 55% of the outbreaks and accounted for 32% of the total cases in 1971-1974. Municipal systems accounted for 31% of the outbreaks, but 67% of the cases. Individual systems accounted for 14% of the outbreaks and only 1% of the cases, but outbreaks associated with individual systems probably are under-report- ed, as opposed to those associated with municipal and semipublic systems. Deficiencies in treatment and contamination of groundwater were responsible for a majority of the outbreaks (onto) and cases (onto) in 1971- 1974. Inadequate or interrupted chlorination was involved in 31% of the outbreaks and 44% of the cases. Craun et al. (1976) have drawn attention to the large number of waterborne disease outbreaks involving travelers. In 1971-1974, 49 (onto) of the 68 outbreaks that occurred in connection with semipublic and individual systems affected travelers, campers, visitors to recreational areas, or restaurant patrons; and 86% of the 49 outbreaks occurred during April-September. Outbreaks on cruise ships are excluded from the above tabulations, but they are of interest and should be mentioned because they involve the traveling public. For example, in June 1973, about 90% of 655 passengers and 35% of 299 crew were affected by an outbreak of acute gastroenteri- tis. An epidemiological investigation identified Shigella flexneri type 6 among early cases, and contaminated water and ice aboard the ship were implicated as vehicles of transmission (Center for Disease Control, 19731. In 1975, outbreaks of diarrhea on 8 ships affected between 9% and 61% of the passengers. In most of these outbreaks the causal agents and vehicles TABLE III-1 Etiology Of Waterborne Outbreaks and Cases, 1971-1974 Disease OutbreaksCases Gastroenteritis 467,992 Giardiasis 125,127 Shigellosis 132,747 Chemical poisoning 9474 Hepatitis-A 13351 Typhoid fever 4222 Salmonellosis 237 TOTAL 9916,950
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66 DRINKING WATER AND H"LTH of transmission were unknown; water was identified as the vehicle in one of them (Center for Disease Control, 1976b). In 1975, 24 waterborne disease outbreaks involving 10,879 cases were reported to the Center for Disease Control (1976b). No etiologic agent was found for the two largest outbreaks (Sewickley, Pa. 5,000 cases and Sellersburg, Ind. 1,400 cases). The third largest outbreak, involving over 1,000 persons, occurred at Crater Lake National Park, Oreg. Enterotoxi- genic E. cold was isolated from residents of the park who became ill, and from the park's water supply. Seventeen of the 24 outbreaks and about 9070 of the cases reported to CDC were designated as "acute gastrointestinal illness." This category includes cases characterized by gastrointestinal symptoms for which no specific etiologic agent was identified. Cases resulting from water treatment deficiencies (2,695) or deficiencies in the water distribution system (6,961) accounted for almost 89% of the total cases in 1975. As in the past, most of the cases occurred in the spring and summer. The reported numbers of outbreaks and illnesses represent only a portion of the true totals. Craun et al. (1976) called attention to the outbreak at Richmond Heights, Fla., in 1974 as an example of why good disease surveillance is necessary and of the way in which many illnesses may go unnoticed. Initially, only 10 cases of shigellosis in this outbreak were recognized by authorities. An epidemiologic investigation revealed that approximately 1,200 illnesses actually occurred. This large outbreak might not have been detected if local health authorities had not been conducting shigellosis surveillance. In another outbreak, some 1,400 residents of Sellersburg, Ind. (31% of the town's population) experienced gastroenteritis. The high attack rate, rapid onset of the outbreak, review of water sampling data, and the town-wide survey suggested that the illness was waterborne, but no bacterial or viral pathogens or chemical toxins were found in the town water supply. Until improved detection and reporting systems are in use, the available epidemiological data will represent only a small fraction of the waterborne-disease problems in this country. BACTERIA The principal bacterial agents* that have been shown to cause human intestinal disease associated with drinking water are: Salmonella typhi, *Nomenclature in this report follows the 8th edition of Bergey's Manual of Determinative Bacteriology (Buchanan and Gibbons, 1974).
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Microbiology of Drinking Water 67 typhoid fever; Salmonella paratyphi-A, paratyphoid fever; Salmonella (other species and a great number of serotypes), salmonellosis, enteric fever; Shigella dysenteriae, S. pexneri, and S. sonnet, bacillary dysentery; Vibrio cholerae, cholera; Leptospira sp., leptospirosis; Yersinia enterocoli- tica, gastroenteritis; Francisella tularensis, tularemia; Escherichia cold (specific enteropathogenic strains), gastroenteritis; and Pseudomonas aeruginosa, various infections. Several other organisms have been associated with gastroenteritis, such as those in other genera of the Enterobacteriaceae: Edwardsiella, Proteus, Serratia, and Bacillus. Number of Cells Required to Infect In attempting to assess the hazards in drinking water, it is important to know how many viable pathogenic cells are necessary to initiate an infection. McCullough and Eisele (19Sla,b,c,d) found that a dose of 106-108 salmonellae per person was necessary for most strains, although 105 cells of some strains could infect. More recent studies by Dupont, Hornick, and associates on selected enteric bacterial pathogens are summarized in Table III-2. Some enteric pathogens are highly virulent, causing infection when relatively few cells are administered (e.g., Shigellaflexneri and S. dysenteriae), whereas others require large numbers to infect (e.g., Salmonella typhosa and Vibrio cholerae). Virulence is a genetic trait and can vary markedly from strain to strain (Meynell, 19611. Phenotypic variation in virulence can occur within a given clone. A small percentage of the cells in a population may be unusually virulent (Meynell, 1961; Meynell and Meynell, 1965~. Thus, it does not always follow that because large numbers of cells are required for infection in feeding trials, that large numbers in drinking water are necessary to cause infection. Some few individuals may become infected by small numbers of unusually virulent cells. Recent evidence also indicates the possibility of genetic transfer of virulence from invading microbes into the resident intestinal population, providing another me.ans by which small numbers of organisms might initiate a disease state. The consequences of an increasing prevalence in livestock and their excrete of coliform organisms containing infectious plasmids and giving rise to clinical conditions were not examined in detail because of time constraints and their lack of immediate relevance to standard setting. Similarly, the consequences of adding antibiotics to animal and poultry feed and the enhanced hazards-of spreading drug-resistant organisms were not examined. The infecting dose also varies with the age and general health of the
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68 DRINKING WATER AND H"LTH TABLE III-2 Infective Doses For Man of Bacterial Enteric Pathogens Subjects Infected/Total Tested Enters Pathogen Dose: Viable Cells 10i 102 103 104 105 106 107 108 109 Shigella dysenteriae Strain M131 1/10 2/4 7/10 5/6 Strain A-1 1/4 2/6 Shigella flexr~eri Seam 2A# 6/33a 33/49b66/87 15/24 Strain 2A# # 1/4 3/4 7/8 13/19 7/8 Salmonella typhi Strain Quailes 0/14 32/116 16/32 8/9 40/42 Vibrio cholerae Strain Inaba With NaHCO3 11/13 45/52 2/2 No NaHCO3 0/2 0/4 0/4 2/4 1/2 Enteropathogen~c E. cold Strain 4608 0/5 0/5 4/8 SOURCES: Shigella dysenteriae: Levine et al., 1973; Shigella flexneri: Dupont et al., 1972b; Dupont et al., 1969; Salmonella typhi: Cornice et al.. 1970; Vibrio cholerae: Cash et al., 1974; Enteropathogen~c E. coli: Dupont et al., 1971. aD.ose 1.8 x 102. bDose: 5 x 103. host population (MacKenzie and Livingstone, 1968~. Infants and the aged may be particularly susceptible. Previous exposure to a given pathogen is important, in that coproantibodies may prevent infection with a strain that is generally present in the population, whereas a new serotype introduced into the water supply may present an increased hazard. Not all strains of Shigella are highly virulent. Shaughnessy et al. (1946) determined infecting doses of four strains of Shigella while studying immunization in volunteers. They found that infectivity in mice could not be directly correlated with infectivity in humans and that doses of 109 organisms or higher `.vere needed to produce human infection. In their extensive studies to develop a Shigella vaccine, Hornick, DuPont, and associates observed the infective dose for several strains. With S. flexneri 2A, 30 of 39 volunteers became ill from a dose of 105-108 organisms
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Microbiology of Drinking Water 69 (DuPont et al., 1969~. They showed that Shigella must penetrate the intestinal mucosa to produce symptoms of classic dysentery and that addition of bicarbonate facilitated this process. Two vaccine strains of S. exneri 2A, a hybrid of a Shigella mutant, E. colt, and a streptomycin- dependent strain, could be safely administered orally in doses of 10~° organisms or higher (DuPont et al., 1972a). A virulent strain could cause symptoms in doses of as few as 180 organisms (DuPont et al., 1972b). With Shigella dysenteriae 1 (Shiga strain)-an organism that has two pathogenic modes, invasiveness and enterotoxin elaboration the infecting dose in man was shown to be as low as 10 organisms (Levine et al., 19731. With such high infectivity of Shigella, why are waterborne outbreaks not more common? One possibility is that Shigella survives poorly in water. Wang et al. (1956) pointed out that, in a number of bacillary dysentery outbreaks involving water, the organism was not, or could not be, isolated. Over several years of studying irrigation water in Colorado, Wang et al. (1956) and Dunlop et al. (1952) were not successful in isolating shigellae, although salmonellae were frequently isolated. The survival of shigellae in water appears to be shorter than that of many other bacteria; Dolivo-Dobrovolskiy and Rossovskaya (1956) found Shigella survival times of only 0.5-4.0 h during the warmest time of the year. However, enteric pathogens may survive much longer times in lake or river sediment than in free waters, and resuspension of such pathogen-loaded sediments at a later time may introduce a "slug" of bacteria into the waters that is not completely removed by treatment systems. Estimation of Disease Potential by Direct Quantitation of Bacterial Pathogens The detection of bacterial pathogens in water polluted with human or animal fecal matter is relatively easy when large numbers of organisms are present (American Public Health Association, 1975~. Pathogenic bacteria have been isolated from relatively clean reservoirs, rivers, streams, and groundwater; large samples, concentration techniques, and often elaborate laboratory procedures are used. However, detecting the presence of these pathogenic organisms in processed and disinfected water is far more difficult. Scientific literature presents a vast array of media and methods for direct pathogen detection in finished water (Geldreich, 1975~. The greatest emphasis has been on the Salmonella-Shigella group of enteric organisms. Numerous modifications of well-known media are used for
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70 DRINKING WATER AND H"LTH pre-enrichment, enrichment, selective inhibition, and isolation, and there are many recommended modifications of incubation temperature and time. Some methods use the classic most-probable-number (MPN) procedure for quantification; others use membrane filtration. Reviews of proposed procedures may be found in the Journal of the Water Pollution Control Federation (Geldreich, 1968, 1969, 1970b; Van Donsel, 1971; Reasoner, 1972, 1973, 1974, 1975~. A recent review appeared in the fourteenth edition of Standard Methods (American Public Health Association, 1975~. There are serious limitations to the use of direct isolation of specific pathogenic bacteria for evaluating water quality. First, there is no single procedure that can be used to isolate and identify all these microorgan- isms. Second, only for salmonellae are the available procedures sufficiently accurate; the methods for other major pathogens such as Shigella, Vibrio, and Leptospira-are inadequate. Third, none of the available procedures is applicable to quantitative isolation of small numbers of pathogens in drinking water. Fourth, even if procedures could be recommended, it is doubtful whether laboratories doing routine bacteriologic studies of water would have the expertise to carry out the procedures reliably. In outbreaks caused by gross contamination, the standard procedures would be of value. Recently, Reasoner and Geldreich (1974) reviewed several of the rapid-detection methods proposed for water and concluded that the cost per test, although perhaps higher than for conventional procedures, must be tolerated for potable-water quality assessment in emergency situations created by natural disasters, treatment breakdown, or rupture in the distribution network. None of these procedures would provide protection to the public as great as that provided by the currently used indicator organism, the coliform. Indicator Organisms The term "indicator organism," as used in water microbiology, means: a microorganism whose presence is evidence that pollution (associated with fecal contamination from man or other warm-blooded animals) has occurred. Indicator organisms may be accompanied by pathogens, but do not necessarily cause disease themselves. As noted above, pathogens are usually more difficult to grow, isolate, and identify than indicator organisms, and often require special media and procedures. Indicator organisms, rather than the actual pathogens, are used to assess water quality because their detection is more reliable
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Microbiology of Drinking Water 71 and less time-consuming. Pathogens appear in smaller numbers than indicator organisms and are therefore less likely to be isolated. An indicator organism should have the following characteristics: · Applicable to all types of water. · Present in sewage and polluted waters when pathogens are present. · Number is correlated with the amount of pollution. · Present in greater numbers than pathogens. · No aftergrowth in water. · Greater survival time than pathogens. · Absent from unpolluted waters. · Easily detected by simple laboratory tests in the shortest time consistent with accurate results. · Has constant characteristics. · Harmless to man and animal. No organism or group of organisms meets all these criteria, but the "coliform group" of organisms fulfills most of them. ESCHERICHIA COLI AND THE COLIFORM GROUP Escherichia cold is commonly found in the human intestine. It is not normally a pathogen, although pathogenic strains are known. Physiologi- cally, E. cold and members of the genera Salmonella and Shigella are quite similar. All are classified as enteric bacteria of the family Enterobacteria- ceae (Cowan, 1974~. They are facultatively anaerobic, and are able to ferment sugars with the production of organic acid and gas. These three genera carry out a type of fermentation called "mixed-acid fermenta- tion," but differ in a number of physiological characteristics. Many physiological differences between various enteric bacteria are known (Ewing and Martin, 1974), but at the beginning of the twentieth century this was not so. In the early days of water bacteriology, some simple operational distinctions were necessary. The lactose-fermentation test became the prime diagnostic tool: E. cold ferments lactose with the formation of acid and gas; Salmonella and Shigella do not ferment lactose. One source of confusion is the necessity to distinguish between E. cold and the "coliform group" of bacteria. Although the taxonomy of bacteria is constantly undergoing revision (see Buchanan and Gibbons, 1974, for the latest version), the genus Escherichia is well defined. It is distinguished from other mixed-acid fermenters of the Enterobacteriaceae primarily on
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72 DRINKING WATER AND H"LTH the basis of sugar-fermentation reactions, motility, production of indole from t~yptophan, lack of urease, inability to utilize citrate as sole carbon source, and inhibition of growth by potassium cyanide. However, the "coliform group" is not so precisely defined. The "coliform group," as defined in Standard Methods (American Public Health Association, 1975), comprises all "aerobic and facultative anaerobic, gram-negative, non- spore-forming, rod-shaped bacteria which ferment lactose with gas formation within 48 hr at 35 C." This is not a taxonomic grouping, but an operational one that is useful in water-supply and sewage-treatment practice. It includes organisms in addition to E. colt, most importantly Klebsiella pneumoniae and Enterobacter aerogenes, which are not m~xed- acid fermenters. The entry of the term "coliform" into sanitary bacteriology was associated with a policy established by H. E. Jordan when he became editor of the Journal of the American Water Works Association; he stated that he would substitute "coliform" for "E. colf' in papers submitted to him (Jordan, 1937~. Although most isolates classifiable as Escherichia by modern methods ferment lactose, about 5-9% of them do not (Ewing and Martin, 1974~. No isolates of the genus Salmonella, either in the species S. typhi or in other species, produce gas from lactose (Ewing and Martin, 1974~; therefore, a water sample containing Salmonella and a lactose-negative E. cold would be negative on the coliform test and would probably be discarded without further examination, because of the definition of"coliform." Even if glucose were substituted for lactose in a coliform analysis, a significant fraction of organisms would be missed, inasmuch as about 9% of isolates of Escherichia do not form gas from glucose (Ewing and Martin, 1974~. Because there are two procedures the multiple-tube-dilution or most- probable-number (MPN) technique, and the membrane-filter (MF) technique-the coliform group of organisms requires two definitions (American Public Health Association, 1975~. On the basis of the MEN technique, the group consists of all aerobic and facultatively anaerobic, gram-negative, non-spore-forming, rod-shaped bacteria that ferment lactose with formation of gas within 48 h at 35°C. On the basis of the ME technique, the group consists of all organisms that produce a dark colony (generally puIplish-green) with a metallic sheen within 24 h of incubation on the appropriate culture medium; the sheen may cover the entire colony or appear only in a central area or on the periphery. These two groups are not necessarily the same, but they have the same sanitary . ·^ slgnlucance. If the coliform group is to be used as an indicator of fecal pollution of water, it is important to know that the coliforms do not lose viability in the water environment faster than pathogenic bacteria, such as salmonel
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Microbiology of Drinking Water 73 TABLE III-3 Comparative Die-Off Rates (Half-TimeJa of Fecal Indicator Bacteria and Enteric Pathogens Bacteria Half-t~me Number O of strains 1 Indicator Bacteria Coliform (avg.) 17.~17.5 29 Enterococci (avg.) 22.0 20 Streptococci (from sewage) 19.5 S. equines 10.0 1 S. bovis 4~3 1 Pathogenic bacteria Shigella dysenteriae 22.4 1 S. sonnet 24.5 1 S. pexneri 2S.8 1 S. enteritidis, paratyphi A&D 16.0 19.2 2 S. enteritidis, typhimurium 16.0 1 S. typhi 6.0 2 V. cholerae 7.2 3 S. enteritidis, paratyphi B 2.4 1 aTime required for 50~O reduction in the population. From McFeters et al. (1974). lee and shigellae. Little information exists on the survival of bacteria in finished water, and the data on other types of water are scattered and fragmentary. McFeters et al. (1974) recently reviewed previous work and presented their own data on die-o~ of intestinal pathogens in well water. As seen in Table III-3, die-o~ rates for pathogens and coliforms are approximately the same. Earlier work on the survival of salmonellae in water was reviewed by McKee and Wolf (1963~. Another factor to be considered is the relative sensitivity of coliforms and bacterial pathogens to disinfection. Although this subject has been studied little recently, the older work (Butterfield et al., 1943; Butterfield and Wattle, 1946; Wattle and Chambers, 1943) indicated that there was essentially no difference between these different organisms in sensitivity to disinfection. This is not true when the coliform group is compared with viral pathogens. Viruses survive longer than bacterial pathogens (Colwell and Hetrick, 1976~. SOME DEFICIENCES OF COLIFORMS AS INDICATOR ORGANISMS Coliforms meet many of the criteria for an ideal indicator organism previously listed; however, there are some deficiencies. There is after
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124 DRINKING WATER AND HEALTH Committee on the Enteroviruses. 1962. Classification of human enteroviruses. Virology 16:501-504. Cowan, S.T. 1974. Enterobacteriaceae. In R.E. Buchanan and N.E. Gibbons, eds. Bergey's Manual of Determinative Bacteriology, 8th ea., pp. 290 293. Williams & Willcins, Baltimore. Craig, C.F. 1934. Amebiasis and Amebic Dysentery. Charles C Thomas, Springfield, Ill. 315 PP Craun, G.F. 1975. Microbiology-Waterborne outbreaks. J. Water Poll. Control Fed. 47:1566-1581. Craun, G.F., and L.J. McCabe. 1973. Review of the causes of waterborne-disease outbreaks. J. Am. Water Works Assoc. 65:7484. Craun, G.F., L.J. McCabe and J.M. Hughes. 1976. Waterborne Disease Outbreaks in the U.S. 1971-1974. J. Am. Water Works Assoc. 68:420-424. Culp, G.L., R.L. Culp, and C.L. Hamann. 1973. Water resource preservation by planned recycling of treated wastewater. J. Am. Water Works Assoc. 65:641-647. Dahling, D.R., G. Berg, and D. Berman. 1974. BGM, a continuous cell line more sensitive than primary rhesus and African green kidney cells for the recovery of viruses from water. Health Lab. Sci. 11:275-282. Dalldorf, G., and G.M. Sickles. 1948. Unidentified, filterable agent isolated from the feces of children with paralysis. Science 108:61-62. Danielsson, D. 1965. A membrane filter method for the demonstration of bacteria by fluorescent antibody technique. 1. A methodological study. Acta Pathol. Microbiol. Scand. 63:597-603. Davis, B.D., R. Dulbecco, H.N. Eisen, H.S. Ginsberg, and W.B. Wood. 1967. Microbiology. Harper and Row, New York. 1464 pp. Denis, F. 1974. Les virus pathogenes pour l'homme dans les eaux de mer et dans les mollusques: Survie-Recherche-Bilan. Med. Mall Infect. 4-6 bis:325-334. DeBlanc, H.J., Jr., F. DeLand, and H.N. Wagner, Jr. 1971. Automated radiometric detection of bacteria in 2967 blood cultures. Appl. Microbiol. 22:846-849. Di Girolamo, R., J. Liston, and J.R. Matches. 1970. Survival of virus in chilled, frozen and processed oysters. Appl. Microbiol. 20:58-63. Dismukes, W.E., A.L. Bisno, S. Katz, and R.F. Johnson. 1969. An outbreak of gastroenteri- tis and infectious hepatitis attributed to raw clams. Am. J. Epidemiol. 89:555-561. Dolivo-Dobrovolskiy, L.B., and V.S. Rossovskaya. 1956. The problem of the survival of dysentery bacteria in reservoir water. Gig. Sanit. 1956(6): 52-55. Cited in Biol. Abstr. 34: 13898 (1958). Dougherty, W.J., and R. Altman. 1962. Viral hepatitis in New Jersey 1961) 1961. Am. J. Med. 32:704716. ~ Duff, M.F. 1967. The uptake of enteroviruses by the New Zealand marine blue mussel Mytilus edulis aoteanus. Am. J. Epidemiol. 85:486-493. Dulbecco, R. 1952. Production of plaques in monolayer tissue cultures by single particles of an animal virus. Proc. Nat. Acad. Sci. USA 38:747-752. Dulbeeco, R., and M. Vogt. 1954. Plaque formation, and isolation of pure lines with poliomyelitis viruses. J. Exp. Med. 99: 167-182. Dunlop, S.G., R.M. Twedt, and W.L.L. Wang. 1952. Quantitative estimation of Salmonella in irrigation water. Sewage Ind. Wastes 24:1015-1020. Dutka, B.J. 1973. Coliforms are an inadequate index of water quality. J. Environ. Health 36:39-46.
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Representative terms from entire chapter: