Water and Reclaimed Water                                                                                                                July 2009
                                                                                                                      updated December 31, 2009

EPA's first problem was antibiotic resistance and chlorine resistant bacteria in sewage effluent and drinking water.
The second problem was to find they are created in these treatment plants. In an effort to get this pollutant out of
surface water Land Treatment was promoted. Land Treatment of sewage effluent is now classified as Reclaimed Water
and Biosolids without the Land Treatment controls.  The Bacterium coli test "
Fecal coliform"  was developed by the
German EIJKMAN to identify  thermotolerant bacteria from fresh human/animal feces in water, is now used on fecal
matter to make the claim that sewage sludge is safe. However, the elevated temperature test suppresses most gram
negative pathogenic bacteria, except E. coli -- the primary fecal colifom , growing at normal temperatures. Most coliform
gram negative bacteria" growing at normal temperature will cause serious illness and may lead to death. Other deadly
bacteria are ignored.

The present standards of the American Public Health Association (1925) have designated Gram-negative non-spore-
forming aerobic rods which ferment lactose with the production of gas as the organisms to be searched for to detect
fecal pollution. -- Bacterium coli is the organism which best fulfills the requirements for an index of fecal pollution. --is
seldom obtained except from feces or from substances recently polluted, -- B. coli itself, however, is occasionally
isolated from soil, is often found in feces of cold blooded animals, and occasionally occurs in streams free from
apparent human pollution. -- Eijkman (1904) has suggested a means of separating the B. coli originating in the feces of
warm blooded animals from the strains characteristic of cold blooded animals. This consists in incubating inoculated
glucose-peptone broth at 46°[C]. Gas formation is said to indicate the presence of B. coli from warm blooded animals.
B. coli from cold blooded animals, practically all B. aerogenes, (and B. cloacae), Bacterium proteus, and the various
fermenting anaerobic rods are said to be completely inhibited as far as gas production is concerned. -- however, -the
Eijkman test fails to detect all the fecal B. coli.  -- the lack of such gas formation does not prove the absence of
pollution. The absence of a positive presumptive lactose broth test at 37.5°[C] of course indicates the lack of fecal

Regrowth of coliforms and fecal coliforms in chlorinated wastewater effluent.
Observations made both in the field in chlorinated effluent, and in laboratory experiments show that coliforms are
capable of regrowth in chlorinated wastewater. Under field conditions regrowth of coliforms in chlorinated effluent held in
a storage reservoir for similar to 3 days appeared inversely correlated to (1) The residual chlorine in the storage
reservoir and (2) the number of coliforms surviving chlorination. In the laboratory experiments regrowth occurred after
initial doses as high as 11 ppm total chlorine even when there was no chemical inactivation of the chlorine. Fecal
coliforms were not detectable in 10-ml of samples after chlorination. Since coliforms and fecal coliforms are capable of
regrowth in chlorinated sewage effluent and admixtures of it, the sanitary significance of the number of coliforms after
storage or in receiving bodies of water is difficult to interpret. Thus standards might be based on the number of
coliforms, or fecal coliforms detected in effluent s immediately after chlorination. However, this would not be justified if in
addition to coliforms, pathogenic bacteria can regrow in chlorinated effluents.
Water Research [Water Res.]. Vol. 7, no. 4, pp. 537-546. 1973.

Factors That Influence Microbial Contamination of Fluids Associated with Hemodialysis Machines
These results were directly related to the design of recirculating systems which permits carbon- and nitrogen-containing
waste products dialyzed from the patient to accumulate, be used as nutrients by microorganisms, and subsequently
allow for 2- to 4-log increases in contamination levels during a dialysis treatment.
Appl Microbiol. 1974 November; 28(5): 822–830.

Escherichia coli: The Fecal Coliform
Over 90 percent of bacteria in raw sewage is E. coli.  However, in the treatment plant, E. coli only makes up about a
third to a fourth of the bacteria. The other 3/4 is 25% Klebsiella and 50% Enterobacter-Citrobacter
Dufour, A. P., Bacterial Indicators /  Health Hazards  Associated with Water  By A.W.Halley, Bernard J. Dutka,  Eds,
American Society for Testing and Materials, 1977, pp. 49-58
Limited preview - 1977 - 363 pages - Nature

Pure cultures of E. coli, Klebsiella and Enterobacter, obtained from hospital patients and from natural waters were
tested for their growth patterns by spread plate and membrane filtration procedures at the following temperatures; 35 °,
41.5 °, 43 °, 44.5 °, and 35°C for 4 h followed by 18 h at 44.5°C.
Results indicated that 44.5°C incubation produces the lowest population estimate and that the application of the
membrane filtration technique also reduced the potential population. Three water samples collected during June,
August and November were tested for fecal coliform and E. coli populations, with 11 different media (broth and agar)
and incubation temperatures of 35 °, 41.5 °, 43 °, 44.5 °, and 35°C for 4 h followed by 18 h at 44.5°C. During the study,
isolates were collected from all positive MPN tubes at each temperature and from each MF medium-temperature regime,
24 to 50 isolates were collected. From the isolate data corrected coliform (oxidase negative), fecal coliform and E. coli
population estimates were made.
A sample of feces was diluted in lake water and maintained at 20°C for 56 days. Samples were collected at
various times and tested for fecal coliform densities using five media and the same temperature regime as for
the lake water samples.
Data from these studies indicate that, depending on the age of the population being measured, the
temperature of the water sample, and the temperature-media-procedure combination used, fecal coliform
and E. coli population estimate techniques measure from 5 to 100% of the potential population.

Chlorine Resistance Patterns of Bacteria from Two Drinking Water Distribution Systems (1982)
Chlorine has also been shown to cause physiological injury of coliform microorganisms such as Escherichia coli,
resulting in underestimation of these indicator organisms in chlorinated waters.  -- Large numbers of viable
microorganisms, many of which have been shown to be human secondary opportunistic pathogens, can often be
recovered from potable water distribution systems maintaining free chlorine residuals (i.e., HOCl + OCI-) of 0.5 to 1.0
mg liter-' (21, 24).

Thus, specific mechanisms may exist for the survival of certain bacteria and viruses in waters containing relatively high
concentrations of chlorine (14, 16). Shaffer et al. (31) recently demonstrated that poliovirus isolates obtained from fully
treated, chlorinated drinking water were several orders of magnitude more resistant to free chlorine than two stock
laboratory strains which were used for comparison.

Bacterial colonization of the surfaces of drinking water distribution pipes has been recently documented by the use of
scanning electron microscopy (1, 28, 30). Studies in this laboratory have demonstrated that such microbial colonization
can occur even in highly chlorinated water systems (>1.0 mg of applied chlorine per liter). The cells comprising many of
these microcolonies are frequently covered with extracellular mucopolysaccharide or glycoprotein polymers,
which may contribute to their ability to survive and proliferate in such chlorinated environments.

The relative chlorine sensitivities of bacteria isolated from chlorinated and unchlorinated drinking water distribution
systems were compared by two independent methods. One method measured the toxic effect of free chlorine on
bacteria, whereas the other measured the effect of combined chlorine. Bacteria from the chlorinated system were more
resistant to both the combined and free forms of chlorine than those from the unchlorinated system, suggesting that
there may be selection for more chlorine-tolerant microorganisms in chlorinated waters. Bacteria retained on the
surfaces of 2.0-p.m Nuclepore membrane filters were significantly more resistant to free chlorine compared to the total
microbial population recovered on 0.2-p.m membrane filters, presumably because aggregated cells or bacteria attached
to suspended particulate matter exhibit more resistance than unassociated microorganisms. In accordance with this
hypothesis, scanning electron microscopy of suspended particulate matter from the water samples revealed the
presence of attached bacteria. The most resistant microorganisms were able to survive a 2-min exposure to 10 mg of
free chlorine per liter. These included gram-positive spore-forming bacilli, actinomycetes, and some micrococci.
The most sensitive bacteria were readily killed by chlorine concentrations of 1.0 mg liter-' or less, and included most
gram-positive micrococci, Corynebacterium, Arthrobacter, Klebsiella, Pseudomonas, Alcaligenes, Flavobacterium,
Moraxella, and Acinetobacter.

Aeromonas sobria in chlorinated drinking water supplies
Aeromonas species were recovered from over 27% of 183 chlorinated drinking water samples collected during an 18-
month period. Sixteen of 20 isolates tested elicited a cytotoxic response by Y-1 mouse adrenal cells. None of the strains
was either enterotoxigenic by the rabbit ligated ileal loop assay, exhibited piliation, or showed significant mannose
resistant adherence to human buccal cells. TheAeromonas isolates were further identified to beA. sobria and were
resistant to ampicillin and susceptible to chloramphenicol, kanamycin, streptomycin, and tetracycline. Total coliform
levels did not correlate with Aeromonas densities in distribution water. With 85% of the samplings,Aeromonas occurred
in distribution water when no coliforms were detectable by either the membrane filter or most-probable-number
techniques. A significant correlation (P<.001) existed between standard plate count levels and Aeromonas
Microbial Ecology,  Volume 8, Number 4 / December, 1982

Effect of Animal Grazing on Water Quality of Nonpoint Runoff in the Pacific Northwest
Total coliforms(TC) in runoff from grazed and ungrazed areas in the Pacific Northwest did not correlate with the
presence or absence of animals. Fecal coliform (FC) and fecal streptococal (FS) numbers were elevated in runoff from
the grazed are when cattle were present above that when they were absence from the area for several months. FC
numbers were elevated in the runoff above recommend levels of 200/100ml (FC) and 1,000 ml (TC) for primary contact.
In fact, FC and FS numbers appeared to increase from less than 100/100 ml to several thousand per 100 ml in the
runoff from grazed area for several months in the spring during warm, wet weather after animals were removed the
previous fall. Almost 3 years of cattle absence were required for FC numbers in runoff from the check water shed to be
consistently below the maximum recommended numbers for primary contact. The data indicate that the use of
conventional FC/FS ratios or FC numbers in runoff as a measurement of recent fecal pollution by cattle on grazed area
is of limited or no value. FC/FS ratios of 1, which indicate recent animal fecal pollution, were found after animals had
been removed for several months.

Effects of Livestock Pasturing on Nonpoint Surface Runoff
Microbiological analysis for total coliform (TC), fecal coliform (FC), and fecal streptococci (FS were conducted. The
number of bacteria exceeded water quality criteria for nonpoint source runoff, even from a non-pastured control

Injury and the Improved Recovery of Coliform Bacteria in Drinking Water
The importance of stressed indicator bacteria in assessing water quality has been gaining attention as microbiologist
have discovered the inaccuracy of present methods and media for recovering indicator bacteria in water. -- Recoveries
of fecal coliforms on m-T7 were 3.1 times greater that the standard m-FC method and 1,7 times greater than the two
layer enrichment, temperature acclimated procedure. -- Laboratory and field data indicated that copper at levels
normally found in drinking water could injury coliform bacteria. Study results showed that 90% injury could occur at
copper concentrations  of 0.050 mg/L. -- during the last decade a pronounced increase has occurred in both the
number of cases  and outbreaks of waterborne diseases -- At the same time, some experts believe that as many as
90% of the outbreaks are unreported, sine many people fail to associate their illness with contaminated drinking water.
--- Injury to indicator bacteria can result from a variety of sources, including chlorine and other disinfectants, heat,
freezing, sunlight, pH, transition metals, antagonistic plate count organisms, and possibly other undefined chemical and
physical parameters. Injured bacteria exist in every aquatic environment examined thus far.

Retrospective Epidemiological Study of Disease Associated with Wastewater Utilization
Lack of Data made outcome unclear

Viruses in Water and reclaimed Wastewater
Cultures incubated at 36C for 10 to 14 days.
Methods not capable of determining viruses in disinfected reclaimed water

Inactivation of Microbial Agents by Chemical Disinfectants (1986)
Two factors that can influence disinfection results are the relative resistance of laboratory-grown cultures versus that of
natural organisms and protection of bacteria be particulate matter. Cell cultures grown in the laboratory are more easily
inactivated. Bacteria that are within particles of feces or other organic matter or that are attached to activated carbon
particles are not inactivated as readily as bacteria that are not associated with such particulate matter. -- differences
exist in relative resistance to various chemicals disinfectants. Whereas organism A might be more resistant to chlorine
than organism B, the opposite might be observed for chloramine or chlorine dioxide.

Though pathogenic bacteria are among the target organism for disinfection, little information is available on their
inactivation. Most of the research related to bacteria has focused on indicator organisms. Studies in the 1940s did not
reveal substantial differences in disinfection resistance between bacterial pathogens and members of the coliform
group. Thus data for E. coli should indicate the degree of disinfection needed for the pathogenic bacteria.

Methods for laboratory growth and enumeration of pathogenic viruses are difficult or not yet available. Most of the
disinfection data presented in this report are for poliovirus.

Phenotypic Characteristics of Coliform and Noncoliform Bacteria from a Public Water Supply Compared with Regional
and National Clinical Species (1986)

The FC test was developed to differentiate typical and atypical E. coli from other members of the family
Enterobacteriaceae. The mechanism of temperature tolerance is not known. The basic premise is that E. coli will grow
and metabolize at 44.5°C while other Enterobacteriaceae will not.

In our study, Klebsiella and Enterobacter isolates, which other evidence suggested were of environmental origin,
demonstrated positive FC tests, suggesting that temperature tolerance may not conclusively
identify E. coli or FCs.

Identification to the species level of coliforms confirmed the presence of Klebsiella and Enterobacter species
but not Escherichia coli. Investigation suggested that the elevated coliform counts were due to biofilm growth and
sloughing and not to ongoing fecal contamination (8).

During the summer and fall of 1984, elevated total coliform counts were observed in the distribution system
of a public water supply serving 350,000 people in south central Connecticut. As part of an investigation of
possible health risks associated with the presence of bacteria in the water supply, bacterial isolates from the
distribution system were compared with bacterial isolates of the same species obtained from a large regional
teaching hospital and from a national compendium of clinical isolates. Characteristics analyzed included
phenotypic metabolic activity, antimicrobial susceptibilities to clinically utilized antibiotics, temperature
tolerance at 44.5°C, and beta-glucuronidase activity in single-test form and on a selective medium. Environmental
isolates lacked known plasmid-mediated characteristics, with the exception of one Escherichia coli isolate which showed
some antibiotic resistance. Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, and Enterobacter
agglomerans from all sources were temperature tolerant and yielded positive fecal coliform tests. Only E. coli showed
beta-glucuronidase activity (both in a single biochemical test and on a selective medium). No single characteristic
analyzed was sufficient to establish an organism as either environmental or clinical in origin.

Survival of Coliforms and Bacterial Pathogens within Protozoa during Chlorination (1988)
The traditional indicator of bacterial pathogens in drinking water has been the presence of coliform bacteria. The
increasing spontaneous occurrence of coliforms in chlorine treated drinking water has generated concern because
presently accepted standard chlorine levels are unable to eliminate these and other microorganisms (9, 25, 31).
Parallelling the increases in reported cases of coliform occurrence in chlorinated drinking water has been an increase in
the outbreaks of waterborne diseases in the United States (11, 12, 44).

The susceptibility of coliform bacteria and bacterial pathogens to free chlorine residuals was determined
before and after incubation with amoebae and ciliate protozoa. Viability of bacteria was quantified to
determine their resistance to free chlorine residuals when ingested by laboratory strains of Acanthamoeba
castellanii and Tetrahymena pyriformis. Co cultures of bacteria and protozoa were incubated to facilitate
ingestion of the bacteria and then were chlorinated, neutralized, and sonicated to release intracellular bacteria.
Qualitative susceptibility of protozoan strains to free chlorine was also assessed. Protozoa were shown to survive and
grow after exposure to levels of free chlorine residuals that killed free-living bacteria.
Ingested coliforms Escherichia
Citrobacter freundii, Enterobacter agglomerans, Enterobacter cloacae, Klebsiella pneumoniae, and
Klebsiella oxytoca
and bacterial pathogens Salmonella typhimurium, Yersinia enterocolitica, Shigella sonnei,
Legionella gormanii, and Campylobacter jejuni had increased resistance to free chlorine residuals. Bacteria could be
cultured from within treated protozoans well after the time required for 99% inactivation of free-living cells. All bacterial
pathogens were >50-fold more resistant to free chlorinc when ingested by T. pyriformis. Escherichia coli ingested by a
Cyclidium sp., a ciliate isolated from a drinking water reservoir, were also shown to be more resistant to free chlorine.
The mechanism that increased resistance appeared to be survival within protozoan cells. This study indicates that
bacteria can survive ingestion by protozoa. This bacterium-protozoan association provides bacteria with increased
resistance to free chlorine residuals which can lead to persistence of bacteria in chlorine-treated water. We propose
that resistance to digestion by predatory protozoa was an evolutionary precursor of pathogenicity in bacteria and that

Qualitative Pathogen Risk Assessment for Ocean Disposal of Municipal Sludge
It is difficult to assess the risk

Factors promoting survival of bacteria in chlorinated water supplies.
Results of our experiments showed that the attachment of bacteria to surfaces provided the greatest increase in
disinfection resistance. Attachment of unencapsulated Klebsiella pneumoniae grown in medium with high levels of
nutrients to glass microscope slides afforded the microorganisms as much as a 150-fold increase in disinfection
resistance. Other mechanisms which increased disinfection resistance included the age of the biofilm, bacterial
encapsulation, and previous growth conditions (e.g., growth medium and growth temperature). These factors increased
resistance to chlorine from 2- to 10-fold. The choice of disinfectant residual was shown to influence the type of
resistance mechanism observed. Disinfection by free chlorine was affected by surfaces, age of the biofilm,
encapsulation, and nutrient effects. Disinfection by monochloramine, however, was only affected by surfaces.
Importantly, results showed that these resistance mechanisms were multiplicative (i.e., the resistance provided by one
mechanism could be multiplied by the resistance provided by a second mechanism).
Appl Environ Microbiol. 1988 March; 54(3): 649-654

Susceptibilities to antibiotics and antiseptics of new species of the family Enterobacteriaceae.
One hundred and sixty-nine strains of new species of the family Enterobacteriaceae, isolated mainly from the
environment, were tested to determine their susceptibilities to 13 antibiotics and 4 antiseptics or disinfectants. All the
species were susceptible to aminoglycosides, doxycycline, and trimethoprim but were resistant to chloramphenicol.
Susceptibility to beta-lactams varied more among the strains. However, all the strains were cefotaxime susceptible, apart
from some Buttiauxella agrestis strains for which MICs were greater than 256 micrograms/ml. The antiseptic MBCs were
similar to those published elsewhere for species of the Enterobacteriaceae of clinical origin. No resistance to
chlorhexidine was observed. On the other hand, the environmental strains presented a greater resistance to active
chlorine than did the reference strains.
Antimicrob Agents Chemother. 1988 June; 32(6): 873-876

Total Coliform Rule
Control of coliform bacteria, including fecal coliforms and Escherichia coli (E. coli 0157:H7)
[The implication is that thermotolerant E. coli is something other than a coliform or a fecal coliform]

Nutrients for Bacterial Growth in Drinking Water: Bioassay Evaluation
Bacteria regrowth in drinking water systems occurs on the walls of the systems and as free living cells attached to
suspended solids. It is attributed to heterotrophs bacteria which may be opportunistic pathogens that interfere with
detection of coliforms.
[All animals are heterotrophic, as well as fungi and many pathogenic bacteria]

"A Randomized Trial to Evaluate the Risk of Gastrointestinal Disease Due to Consumption of Drinking Water Meeting
Current Microbial Standards,
" American Journal of Public Health, vol. 81, no. 6, June 1991, pp. 703-708. study found
that as many as one in three gastrointestinal illnesses -- often chalked up to "stomach flu" -- are caused by drinking
water contaminated with microorganisms

Improved Membrane Filtration Method Incorporating Catalase and Sodium Pyruvate for Detection of Chlorine-Stressed
Coliform Bacteria
It is well recognized that coliforms subjected to chlorine may become stressed, resulting in a reduced ability of these
injured bacteria to proliferate on selective media employed for their detection (7, 10, 24, 33). Failure to detect coliforms
can lead to an overly optimistic estimate of the safety of a water when, in essence, pathogens may still exist. -- The
inability to recover chlorine-injured coliforms on recommended M-fecal coliform (M-FC) and M-Endo media has been
attributed, in part, to various surface-active ingredients
in each medium -- Without exception, all [3] E. coli strains exposed to chlorine exhibited significant (P < 0.001)
reductions in the specific activity of catalase compared with activity in their respective controls (Fig. 1). E. coli SWEI was
the most sensitive to chlorine, as indicated by a difference in the specific activities of control and chlorine-exposed cells
of 20.3 U/mg of protein. --  Exposure to chlorine often renders a portion of the bacterial population injured, resulting in
the inability of these indicator organisms to be recovered on recommended media -- The existence of injured coliform
bacteria in chlorinated waters presents an ongoing challenge to microbiologists attempting to detect these indicator
organisms with currently accepted methodologies. -- Subsequent analysis of colonies from plates containing
compounds which enhanced recovery indicated coliform verification percentages of >80% on M-FC, >90% on mT7, and
>94% on M-Endo media. These data suggest that the addition of peroxide-degrading compounds to various standard
recovery media may improve detection of both coliform and heterotrophic bacteria in chlorinated waters.

Control of Biofilm Growth in Drinking Water Distribution Systems
Researchers realized that water treatment and disinfection systems were merely inactivating bacteria in raw water; the
microorganism were surviving the treatment process. The current definition of a biofilm is an organic or inorganic
surface deposit consisting of microorganism, microbial produces, and detritus (Marshall, 1976; Characklis, 1981;
Characklis and Marshall, 1990).

A Focus on EPA's Research
Waterborne Disease
Currently, it is estimated that there will be over 100,000 violations of the SDWA annually. Nearly half of these will be
MCL violations. Of these, the majority will be microbiological violations by small systems.

Investigations into Biofouling Phenomena in Fine Pore Aeration Devices

Centers for Disease Control, "CDC, EPA Issue Drinking Water Guidance for People with Weakened Immune Systems --
Press Advisory," June 1995. in 1994 and 1995, 45 million Americans drank water from water systems that fell short of
SDWA standards

Legionella: Human Health Criteria Document
Symbiotic existence with algae and other bacteria in biofilms effect survival. They infect protozoans  and can reproduce
in th organisms which protect them from disinfection. They are prevalent in anthropogenic water such as potable water,
cooling towers reservoirs and whirlpools

Mycobacteria: Health Advisory
Primarily cause tuberculosis and leprosy. Environmental sources include aerosols, water, biofilms, soil, dust, food
products and contaminated medical equipment. Causes pulmonary infections, Lymph infection, ear infections, skin and
soft tissue infections, Catheter-associated infection and disseminated infection.

Mycobacteria: Drinking Water Fact Sheet
20 of the 90 species of Mycobacteria are known to cause disease in humans. They have been isolated from
wastewater, surface water, recreational water, ground water and tap water. Biofilms may serve as a reservoir for these
opportunistic pathogens.

How Does Time-Dependent Dental Unit Waterline Flushing Affect Planktonic Bacteria Levels?
"The reported list of bacterial genera isolated from DUWLs [dental unit water lines] is long and varied. For example, a
variety of opportunistic pathogens belonging to the following genera have been identified:
Legionella, Klebsiella,
Staphylococcus,  Enterococcus, Mycobacterium,  Pseudomonas, Moraxella, Sphingomonas, Brevindimonas, and
others." According to the
World Health Organization (WHO), [see HPC-2003]"The cooling water of dental units, which is
sprayed into the patient’s mouth, is also often heavily contaminated with Pseudomonas aeruginosa.

Heterotrophic Plate Counts and Drinking-water Safety
The Significance of HPCs for Water Quality and Human Health
Public health aspects of the role of HPC
2.3.3 Risks from bacteria detected in water
A number of studies have yielded virtually the same characteristic spectrum of heterotrophic bacterial strains. The
predominant species in this spectrum are
Acinetobacter spp., Aeromonas spp., Alcaligenes spp., Comamonas spp.,
Enterobacter spp., Flavobacterium spp., Klebsiella spp., Moraxella spp., Pseudomonas spp., Sphingomonas spp.,
Stenotrophomonas spp., atypical Mycobacterium spp., Bacillus spp. and Nocardia (Burlingame et al. 1986; LeChevallier
et al. 1987; Payment et al. 1988; Payment 1989; Reasoner et al. 1989; Manaia et al. 1990; Edberg et al. 1997; Rusin et
al. 1997; Norton and LeChevallier 2000).

Fate of a representative pharmaceutical in the environment -- Lubbock’s Water Reclamation Plant
Texas Water Resources Institute, Audra Morse, Ph.D.; Andrew Jackson, Ph.D., P.E., 2003
The antibiotic resistance patterns of the LWRP varied monthly; heterotrophic bacteria were resistant to most of
the antibiotics investigated during the nine month study.

Contaminants in Well Water  (CDC Fact Sheet) (doesn't include but a few pathogens and chemicals)
Wondering what microorganisms (germs) and chemicals can be found in your well water, and what they can do to your
Bloody or non-bloody diarrhea, stomach cramps; little or no fever -- Can cause hemolytic uremic syndrome (HUS) and
kidney failure in young children or the elderly - Vomiting and stomach cramping may also occur --
“flu-like” symptoms with fever and muscle pains, or a rash --  Meningitis  and illnesses that affect the heart and brain
may occur, -- Jaundice (yellowing of eyes and skin) -- but can be serious or fatal for people with weakened immune
congestion of heart, lungs, and kidneys; low blood pressure; muscle spasms; weight loss; damage to adrenal glands --  
weight loss, cardiovascular damage, eye and muscle degeneration; cancer -- Stomach pain, nausea, vomiting, diarrhea,
numbness in hands and feet, partial paralysis, and blindness -- skin damage, circulatory system problems -- liver
problems -- Delayed physical and mental development in babies -- Shortened attention span, hearing, and learning
abilities of children -- increased blood pressures - stroke, kidney disease, -- Methemoglobinemia –( a blood disorder
that causes shortness of breath and blueness of skin, and can lead to serious illness or death -- Methmoglobinemia
mainly affects infants and pregnant women --  effects include increased urination and bleeding of the spleen) --
Drowsiness and decreased responsiveness -- Skin irritation

Validity of the Indicator Organism Paradigm for Pathogen Reduction in Reclaimed Water and Public Health Protection
The validity of using indicator organisms (total and fecal coliforms, enterococci, Clostridium perfringens, and F-specific
coliphages) to predict the presence or absence of pathogens (infectious enteric viruses, Cryptosporidium, and Giardia)
was tested at six wastewater reclamation facilities. Multiple samplings conducted at each facility over a 1-year period.
Larger sample volumes for indicators (0.2 to 0.4 liters) and pathogens (30 to 100 liters) resulted in more sensitive
detection limits than are typical of routine monitoring. Microorganisms were detected in disinfected effluent samples at
the following frequencies: total coliforms, 63%; fecal coliforms, 27%; enterococci, 27%; C. perfringens, 61%; F-specific
coliphages, 40%; and enteric viruses, 31%. Cryptosporidium oocysts and Giardia cysts were detected in 70% and 80%,
respectively, of reclaimed water samples. Viable Cryptosporidium, based on cell culture infectivity assays, was detected
in 20% of the reclaimed water samples. No strong correlation was found for any indicator-pathogen combination. When
data for all indicators were tested using discriminant analysis, the presence/absence patterns for Giardia cysts,
Cryptosporidium oocysts, infectious Cryptosporidium, and infectious enteric viruses were predicted for over 71% of
disinfected effluents. The failure of measurements of single indicator organism to correlate with pathogens suggests
that public health is not adequately protected by simple monitoring schemes based on detection of a single indicator,
particularly at the detection limits routinely employed. Monitoring a suite of indicator organisms in reclaimed effluent is
more likely to be predictive of the presence of certain pathogens, and a need for additional pathogen monitoring in
reclaimed water in order to protect public health is suggested by this study.
Applied and Environmental Microbiology, June 2005, p. 3163-3170, Vol. 71, No. 6

Ecology of Pseudomonas aeruginosa in the intensive care unit and the evolving role of water outlets as a reservoir of
the organism.
In spite of the significant changes in the spectrum of organisms causing intensive care unit (ICU)-associated infections,
Pseudomonas aeruginosa has held a nearly unchanged position in the rank order of pathogens causing ICU-related
infections during the last 4 decades. Horizontal transmissions between patients have long been considered the most
frequent source of P aeruginosa colonizations/infections. The application of molecular typing methods made it possible,
during the last ~7 years, to identify ICU tap water as a significant source of exogenous P aeruginosa isolates. A review
of prospective studies published between 1998 and 2005 showed that between 9.7% and 68.1% of randomly taken tap
water samples on different types of ICUs were positive for P aeruginosa, and between 14.2% and 50% of
infection/colonization episodes in patients were due to genotypes found in ICU water. Faucets are easily accessable for
preventive measures, and the installation of single-use filters on ICU water outlets appears to be an effective concept to
reduce water-to-patient transmissions of this important nosocomial pathogen.
American Journal of Infection Control. 33(5) Supplement 1:S41-S49, June 2005.

The treatment of hospital waste: and appraisal
Hospitals discharge considerable amounts of chemicals and microbial agents in their wastewater. Problem chemicals
present in hospital wastewater belong to different groups, such as antibiotic, X-ray contrast agents, disinfectants and
pharmaceuticals.  Many of these chemical compounds resist normal wastewater treatment. They end up in surface
waters where that can influence the aquatic ecosystem and interfere with the food chain. Humans are particularly
exposed by drinking water. Microbial agents of special concern are multiresistant microbial strains. The latter are
suspected to contribute to the spread of antibiotic resistance.

Aeromonas: Human Health Criteria Document
Aeromonads are found in foods: fresh produce, seafood, raw meats, packaged ready to eat meats, cheese and milk.
They are present in high numbers in sewage before and after treatment. They colonize drinking water systems and
produce biofilms. They cause diarrheal illness, wound infections, septicemia, meningitis, ophthalmitis, endocarditis,
aspiration pneumonia, and biliary tract infections.

Economic Analysis for the Final Ground Water Rule
Office of Water (4606-M) EPA 815-R-06-014 October 2006 www.epa.gov/safewater

Hepatitis has a long incubation period and the virus remains viable in the environment and especially ground water for
months. Thus, the infection source is often obscure. Ground water HAV outbreaks have been identified (Georgetown,
TX (Hejkal et al, 1982), Racine, MO (Missouri Department of Health, 1992); Lancaster, PA (Bowen and McCarthy,
1983); Quebec, Canada (De Serres et al., 1999); all in sensitive aquifers). HAV is not favored in typical
cell lines used in cell culture and so identification is difficult in environmental samples.

The E. coli O157:H7 ground water outbreak in Walkerton, Ontario was also a large outbreak of
campylobacterosis. Arcobacter (now a separate genus from Campylobacter) was responsible for a ground
water outbreak at a camp in Coeur d'Alene, Idaho (McMillan, 1996). The sensitive aquifer was
contaminated by a septic tank. Campylobacter were associated with the recent outbreak in South Bass
Island, Ohio due to widespread fecal contamination in a sensitive aquifer (Ohio EPA, 2005). Like all
bacterial pathogens, special enrichment methods are needed to identify Campylobacter and Arcobacter in
environmental samples and so no data are available on the occurrence of these ubiquitous pathogens in
PWS well water. Campylobacter, like Vibrio has a viable but non-culturable environmental form which
makes it difficult to detect at times in water (Rollins and Colwell, 1986; Koenraad et al., 1997). Limited
dose-response data are available for Camplybacter (but not for Arcobacter). Because no occurrence data
are available, EPA is unable to quantify the benefits associated with avoided camplybacterosis (and
arcobacterosis) and their chronic sequelae (Guillane-Barre paralysis and reactive arthritis).

A comparison of ten USEPA approved total coliform/E. coli tests
[A study of 10 EPA approved coliform tests]
this study was undertaken to elucidate the strengths and weaknesses of each method. Water samples were collected
from three geographically and chemically diverse groundwaters in Wisconsin. One-hundred milliliter aliquots were
individually spiked with both low concentrations (one to ten organisms) and high concentrations (fifty to one-hundred) of
each of five different total coliform organisms (Serratia, Citrobacter, Enterobacter, E. coli, & Klebsiella). These spiked
samples were used to test the capability of ten enzyme-based test systems to both detect and enumerate the spiked
organisms. In addition, 100 ml samples were independently spiked with two different strains of Aeromonas
spp. at six different levels, to assess the ability of each enzyme-based test to suppress Aeromonas spp.
Analysis of the data indicated that wide variability exists among USEPA approved tests to detect and quantify total
coliforms, as well as suppress Aeromonas spp.
Journal of Water and Health Vol 5 No 2 pp 267–282 © IWA Publishing 2007 doi:10.2166/wh.2007.008

Detection of Viable but Nonculturable Escherichia coli O157:H7 Bacteria in Drinking Water and River Water
A sensitive method for specific detection of viable Escherichia coli O157:H7 cells, including viable but nonculturable
(VBNC) cells, in water samples was developed. This method involved capture of the bacterial cells on a low-protein-
binding membrane and direct extraction and purification of RNA followed by reverse transcription-PCR and electronic
microarray detection of the rfbE and fliC genes of E. coli O157:H7. It detected as few as 1 CFU of E. coli O157:H7 in
diluted cultures, 3 to 4 CFU/liter in tap water, 7 CFU/liter in river water, and 50 VBNC cells in 1 liter of river water,
demonstrating the best limit of detection reported to date for VBNC cells in environmental water samples.
[A CFU is the assumption that each colony was started by one colony forming unit at the beginning of the test without
any indication of the number of bacteria in each colony at the end of the test]
Applied and Environmental Microbiology, March 2008, p. 1502-1507, Vol. 74, No. 5

Deaths of Premature Babies
An article in the Canadian Press on April 27, 2007 reported that the illnesses of some 50 premature babies, including
4 deaths at Ste. Justine Hospital in Montreal resulted from pseudomonas infection in tap water.
Pseudomonas is a common bacteria found in tap water and may cause infection in individuals with weakened or
underdeveloped immune systems. It is virtually impossible to eliminate pseudomonas and other environmental bacteria
from tap water and most people are not adversely affected by these organisms. Ste. Justine Hospital has stopped
using tap water in its pre-natal intensive care unit: sterile water is now being used.

EPA/600/R-07/128, February 2008
The classic measurement of viability is to culture an organism. Culture remains the most sensitive method, however,
culture methods are slow and not all target organisms can be cultured.

Waterborne pathogens continue to contaminate drinking water supplies and cause waterborne disease outbreaks
(WBDO) despite current regulations designed to prevent and control their spread. In the United States, from 1991-
2000, 173 waterborne outbreaks and 432,733 cases of illness were reported in public and individual water systems. In
1993, an estimated 400,000 people became ill in Milwaukee, Wisconsin after drinking water contaminated with
Cryptosporidium parvum, a waterborne pathogen that contaminates drinking water supplies. Annually, t
he CDC
estimates that pathogen infected drinking water results in about a million new cases of illness and about a
thousand deaths. Most cases, however, are not reported to health care providers because of their self-
limiting nature and that do not result in WBDOs.

Review of Zoonotic Pathogens in Ambient Waters
Some well-known waterborne pathogens were excluded from analysis because they are not zoonotic. Excluded
pathogens include bacteria that are generally found in the environment, free-living protozoa, viruses, and helminthes
that have cold-blooded hosts (e.g., snails, copepods). Some common zoonotic pathogens were also excluded because
they do not have well-documented waterborne transmission (i.e., primarily transmitted via soil, food, or drinking water).
E. coli O157:H7 can survive for at least several weeks in animal feces and slurries and has been demonstrated to
survive at least 500 days at -20 °C in frozen soil.
Campylobacter has been shown to survive in aquatic environments with low temperatures (4°C) up to 4 months.
Under suitable environmental conditions, Salmonella can survive for weeks in waters or years in soils.

DC Water and Sewage Authority Coliform Reports   [below table of coliform diseases]
EPA allows up to five percent of the samples collected in a month to be positive for total coliform. The graph below
shows the percent of monthly samples testing positive for total coliform over the past year.

More D.C. Kids Had Elevated Lead Than Stated
More than twice as many D.C. children as previously reported by federal and local health officials had high levels of
lead in their blood amid the city's drinking water crisis, according to congressional investigators, throwing into doubt
assurances by those officials that the lead in tap water did not seriously harm city children.

A Final PR piece
Antibiotic resistant bacteria in wastewater processed by the Metropolitan Water Reclamation District of Greater Chicago
The Metropolitan Water Reclamation District of Greater Chicago (District) initiated a research study to determine the
total numbers and percentages of antibiotic resistant fecal coliform (FC) bacteria in raw sewage (RS) entering and final
effluents (FE) discharged from its seven Water Reclamation Plants (WRPs). The density of FC was determined on m-
FC agar containing ampicillin (ampR-16 microg/ml), gentamycin (genR-8 microg/ml), tetracycline (tetR-8 microg/ml), or
all three antibiotics. The study was primarily undertaken to determine whether secondary sewage treatment at the
District WRPs adequately reduces the numbers and percentages of FC(ampR), FC(tetR), FC(genR), FC(amp/tet/genR)
in the FE. The numbers of ampR, tetR, genR, and amp/tet/genR FC observed in RS ranged from 2.0 x 10(5) to 1.1 x 10
(7), 9.5 x 10(4) to 2.2 x 10(6), 95 to 1.5 x 10(4) and 90 to 9.5 x 10(3) per 100 mL, respectively. Secondary sewage
treatment without disinfection was shown to reduce the number of antibiotic resistant FC by two-three orders of
magnitude. The numbers of FC(ampR), FC(tetR), FC(genR), and FC(amp/tet/genR) observed in non-disinfected FE
ranged from 2.0 x 10(2) to 6.4 x 10(3), 2.2 x 10(2) to 4.1 x 10(3), 9 to <20 and 9 to <20 per 100 mL, respectively. The
relative percentages of antibiotic resistant FC observed in FE followed the same trend observed in RS: FC(ampR) > FC
(tetR) > FC(genR) > FC(amp/tet/genR). Only one FC(amp/tet/genR) bacteria was found in this study indicating that
multiple-antibiotic resistant FC was virtually eliminated by secondary sewage treatment. The results of multivariate
regression analysis showed that the percentages of antibiotic resistant FC in the FE from all seven District WRPs were
lower than the percentages of these organisms in RS (p<0.01). These results support the conclusion that secondary
sewage treatment in the District effectively reduces the number of antibiotic resistant FC and that the environments of
the District's seven WRPs are not conducive to the propagation or survival of antibiotic resistant fecal coliform bacteria.

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