ENTERIC BACTERIA -- COLIFORM               36+ studies

E. coli (B.coli) has always been the primary coliform and has been a known pathogen since
1900. It has been known to infect wounds and cause meningitis since 1915. By 1903 the
theory was "virulent B. coli does not, of necessity, indicate faecal contamination."  One
hundred and ten years later, it is only an indicator that pathogens may be present. The new
industry science is even better. Fortunately, there are some good scientist who at least
report negative health effect.

The first two examples indicate the water and wastewater professionals are either lying about
the nature of coliforms or do not know what the coliform test is for. Furthermore, there is the
implication that E. coli 0157 is the only strain that will cause disease and fecal coliform is
something other than E. coli. Not only that but they claim the rest of the Family
Enterobacteriaceae (coliform) do not cause disease. The rest of the studies prove the lie or
lack of knowledge. Also see  
Health and Bacterial Terms , antibiotic resistance and regrowth
What Does E. coli in Water Really Mean?
What is E. coli?
E. coli (aka Escherichia coli) is a bacterium found in the feces of all warm-blooded animals.
First discovered by Theodor Escherich in 1885, E. coli is part of the normal gut flora and
benefits humans by producing vitamin K and competing with other pathogenic bacteria,
preventing their establishment in the intestine. Most strains of E. coli are completely
harmless and— assuming warm bloodedness—you are constantly exposed to the
bacteria and probably even bathing in it. A pair of underpants, on average, harbors 370,000
E. coli due to transfer from the skin. The bacteria were considered strictly commensal—
meaning naturally present and harmless—organisms of the large intestine until about 1935
when it was first associated with an outbreak of diarrhea in infants. A quick search of E. coli
on the web, however, usually leads one to sites describing illnesses such as diarrhea,
severe cramps, kidney failure and death. These reports are referring to the relatively rare
type of E. coli known as serotype O157:H7. Understanding that the species E. coli is further
divided into many different types of strains and serotypes is the first step in relating the
presence of the bacteria to specific health risks.

Coliform Bacteria and Drinking Water November 2007
Coliform bacteria are organisms that are present in the environment and in the feces of all
warm-blooded animals and humans. Coliform bacteria will not likely cause illness. However,
their presence in drinking water indicates that disease-causing organisms (pathogens) could
be in the water system.
Washington State DOH PUB. #331-181 (Revised)
The studies indicate the professional experts are wrong.

2009 -- Phylogeny and Disease Association of Shiga Toxin–producing Escherichia coli O91 -
Shiga toxin–producing Escherichia coli (STEC) infections are public health concerns
because of the severe illnesses they cause, such as hemorrhagic colitis and hemolytic
uremic syndrome (HUS) (1). STEC constitute a heterogeneous group of bacteria abundant in
the reservoir and in the environment (2). Transmission routes for human STEC infection are
numerous and include contact with animal excreta, person-to-person transmission, and
inadvertent ingestion of contaminated food and water. Many STEC serotypes have been
recovered from humans (3,4). Among them, STEC O91 is the most common serogroup
isolated from adult patients in Germany (5,6). The strains within this serogroup appear to be
transmitted predominantly by food, because 1) food vehicles have been identified as the
only risk factors for adults with sporadic STEC O91 infection in Germany (6); 2) O91 is the
second most frequently isolated STEC serogroup in routine food samples (5); and 3) O91 is
the only major STEC serogroup with no association between incidence of human infection
and cattle density (7).
EID Journal Home > Volume 15, Number 9–September 2009

2009 -- Enterobacteriaceae  -- instructional1.calstatela.edu/.../Lec%205%
Classification – more than15 different genera -- Some are enteric pathogens and others are
urinary or respiratory tract pathogens -- E. coli infections -- Neonatal meningitis – is the
leading cause of neonatal meningitis and septicemia with a high mortality rate. -- Usually
caused by strains with the K1 capsular antigen.  -- Gastroenteritis – there are several distinct
types of E. coli that are involved in different types of gastroenteritis: enterotoxigenic E. coli
(ETEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC) , enteroaggregative
E. coli (EAEC), and enterohemorrhagic E. coli (EHEC).

Bacteria belonging to the family Enterobacteriaceae are the most commonly encountered
organisms isolated from clinical specimens. --- Forty-four genera and over 130 species of
Enterobacteriaceae have been recognized. Some of the more common clinically important
genera of the family Enterobacteriaceae include: Salmonella; Citrobacter; Morganella;
Shigella; Enterobacter; Yersinia; Proteus; Serratia; Edwardsiella; Escherichia; Klebsiella;
Providencia -- They act as opportunistic pathogens when they are introduced into body
locations where they are not normally found, especially if the host is debilitated or
immunosuppressed. They all cause the same types of opportunistic infections, namely:
urinary tract infections, wound infections, pneumonia, and septicemia.

2009 -- Chapter 16. Enteric Gram-Negative Rods (Enterobacteriaceae)
Excerpt: "The Enterobacteriaceae are a large, heterogeneous group of gram-negative rods
whose natural habitat is the intestinal tract of humans and animals. The family includes many
genera (Escherichia, Shigella, Salmonella, Enterobacter, Klebsiella, Serratia, Proteus, and
others). Some enteric organisms, eg, Escherichia coli, are part of the normal flora and
incidentally cause disease, while others, the salmonellae and shigellae, are regularly
pathogenic for humans. The Enterobacteriaceae are facultative anaerobes or aerobes,
ferment a wide range of carbohydrates, possess a complex antigenic structure, and produce
a variety of toxins and other virulence factors. Enterobacteriaceae, enteric gram-negative
rods, and enteric bacteria are the terms used in this chapter, but these bacteria may also be
called coliforms...."

2008 -- Microbiological Criteria
The Indicator Concept (Fecal Coliform)
• Difficult to test for specific pathogens
• Coliforms as surrogate for fecal contamination
• von Fritsch used Klebsiellae
• Escherich (1892) described Bacillus coli [E. coli}
• Schardinger suggested E. coli as indicator of fecal pollution.
• Eijkman (1904) differentiated “fecal coliforms” from environmental coliforms by growth at

2007 -- A comparison of ten USEPA approved total coliform/E. coli tests
Since 2002, the United States Environmental Protection Agency (USEPA) has approved ten
enzyme-based total coliform and E. coli detection tests for examination of drinking water.
These tests include: Colilert®, Colilert-18®, Colisure®, m-Coli Blue 24®, Readycult®
Coliforms 100, Chromocult®, Coliscan®, E*Colite®, Colitag™ and MI Agar. The utility of the
enzyme based test systems is based on both the ability of the test to detect the target
organisms at low levels and
the ability of the test system to suppress the growth of
non-target organisms that might result in false positive results
. Differences in the
ability of some of these methods to detect total coliform and E. coli, as well as suppress
Aeromonas spp., a common cause of “false positive” results, have been observed. As a
result, 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

2006 -- Method 1680: Fecal Coliforms in Sewage Sludge (Biosolids) by Multiple-Tube
Fermentation using Lauryl Tryptose Broth (LTB) and EC Medium  Fecal coli form Sampling
Appendex F

2005 --  Method 1681: Fecal Coliforms in Sewage Sludge (Biosolids) by Multiple-Tube
Fermentation using A-1 medium

2005 --
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

2005 -- Recovery of Chlorine-Exposed Escherichia coli in Estuarine Microcosms
Laboratory microcosm experiments were performed to determine whether chlorine-exposed
Escherichia coli are capable of recovery (i.e., increase in numbers of culturable cells) in
estuarine waters and if so what water-quality parameters are responsible for this recovery.
Suspensions of E. coli were exposed to 0.5 mg L-1 of chlorine for 5 min followed by
dechlorination with sodium thiosulfate. The chlorine-exposed bacteria were introduced into 2-
L microcosms containing estuarine water collected from the Seacoast region of New
Hampshire. Culturable cells in the microcosms were enumerated at 0, 10, 24, 48, and 74 h.
In all estuarine microcosms the number of culturable cells increased by factors ranging from
2.8 to 50 over the 74-h incubation period. Multiple linear regression analyses indicated that
ammonium and salinity were most significantly correlated with the recovery of E. coli over the
74-h incubation period; however, ammonium concentrations were strongly correlated with
dissolved organic carbon and total dissolved nitrogen, making it impossible to determine with
any degree of certainty the unique effect nitrogen or carbon had on recovery. The extensive
recovery observed in our study indicates that following exposure to concentrations of
chlorine that cause cell injury rather than death, numbers of culturable E. coli may increase
significantly when discharged into estuarine waters. Thus, depending on the effectiveness of
the chlorination process, the regular monitoring of chlorinated wastewater treatment effluent
may underestimate the true impact on water-quality and public health risks
Environ. Sci. Technol., 2005, 39 (9), pp 3083–3089

2004 -- E. coli History

2003 --
Microbiology of Milled Cereal Grains: Issues in Customer Specifications -- Cargill
coliform/enteric bacteria—commonly found in human and animal intestines, are sometimes
used as an indication of fecal contamination. Thirty genera of coliform/enteric bacteria are
classified in the family Enterobacteriaceae (7).
Four of these genera—Escherichia, Salmonella, Shigella, and Yersinia—can be pathogenic
for humans. --- Some of the enteric bacteria— Enterobacter, Erwinia, Klebsiella, Pantoea,
and Serratia—grow predominantly on plants.
In fact, some of these are involved in the
spoilage of produce, and are typically not found in fecal matter

2001 -- Indicators of microbial water quality -- WHO
The concept of ‘coliform’ bacteria, those bacteria resembling B. coli [E. coli], was in use in
Britain in 1901 (Horrocks 1901). Hence, the total coliforms can best be described as a range
of bacteria in the family Enterobacteriaceae varying with the changing composition of the
The use of bacteria as indicators of the sanitary quality of water probably dates back to 1880
when Von Fritsch described Klebsiella pneumoniae and K. rhinoscleromatis as micro-
organisms characteristically found in human faeces (Geldreich 1978).
Faecal indicator: A group of organisms that indicates the presence of faecal contamination,
such as the bacterial groups thermotolerant coliforms or E. coli. Hence, they only infer that
pathogens may be present.
The range of non-faecal bacteria represented in the coliform group and the environmental
growth of thermophilic (faecal) coliforms Klebsiella spp. and E. coli (Ashbolt et al. 1997;
Camper et al. 1991) have concerned bacteriologists and sanitary engineers since the 1930s
(Committee on Water Supply 1930).
The arbitrary definitions adopted for E. coli and the related coliforms were all based upon
cultural characteristics, including the ability to produce gas from lactose fermentation (HMSO
1969). Hence, the thermotolerant coliforms include strains of the genera Klebsiella and
Escherichia (Dufour 1977), as well as certain Enterobacter and Citrobacter strains able to
grow under the conditions defined for thermotolerant coliforms (Figureras et al. 1994;
Gleeson and Gray 1996). This phenotypic approach has also resulted in E. coli or a
related coliform being ignored simply because they failed to ferment lactose,
failed to produce gas from lactose or were indole-negative at 44.5°C. The approach had
been repeatedly questioned (Waite 1997), and was only resolved in the UK in the 1990s
(HMSO 1994).
It has long been recognised that artificial culture media lead to only a very small
fraction (0.01–1%) of the viable bacteria present being detected (Watkins and
Xiangrong 1997).

2000 -- Public health aspects connected to the use of sludge on land
Like other urban wastes, sewage sludge may contain different kinds of pathogens
that are infectious for different species of animals and plants as well as for humans.
The origin and nature of organic wastes such as different types of sludge always
causes a hygienic risk in storage, collection, processing, handling and utilisation.
These risks exist both when the organic wastes are generated during the treatment of
industrial or municipal wastewater, and when the sludge results from industrial
processing of organic material. Therefore hygienic principles must be followed in
processing, storage, transport and distribution of such materials. Recycling of organic
material to agriculture is a desirable aim from the point of view of saving raw
materials which are of limited availability such as phosphorous, but this aim may
conflict with the necessity to protect humans, animals and plants from undesired
infections as well as with general aims of environmental protection. This report
covers only the hygienic aspects of the use of sludge on land. However, undesired
organic and inorganic pollutants can also cause risks and must be kept in mind.
Hygienic risks due to sludge and related products will be discussed here. This includes the
direct transmission of pathogens to humans or animals and plants of agricultural importance
as well as the introduction of these pathogens into the biozoenosis and
environment by the application of such material as organic fertilizers.
Primary pathogene:
Salmonella spp.; Shigella spp. ; Escherichia coli; Pseudomonas aeruginosa;
Yersinia enterocolitica; Clostridium perfringens; Clostridium botulinum; Bacillus anthracis;
Listeria monocytogenes; Vibrio cholerae; Mycobacterium spp; Leptospira spp;
Campylobacter spp; Staphylococcus; Streptococcus
Secondary pathogene:
Escherichia; Klebsiella; Enterobacter; Serratia; Citrobacter; Proteus; Providencia;
Multiresistant bacteria

1999 -- Comparison and Recovery of Escherichia coli and Thermotolerant Coliforms in
Water with a Chromogenic Medium Incubated at 41 and 44.5°C
The significance of various coliform organisms in water has been and remains an extensively
studied subject. Since fecal coliforms are not defined taxonomically, Escherichia coli is the
only member species for which standardized data exists. According to Leclerc et al. (16), the
coliform species of fecal origin and their isolation frequency in human feces (15) are as
follows: E. coli, 100%; Citrobacter diversus, 70%; Citrobacter amalonaticus, 70%; Citrobacter
freundii, 70%; Klebsiella pneumoniae, 49%; Klebsiella oxytoca, 49%; Enterobacter cloacae,
9%; and Enterobacter aerogenes, 9%. The following species will probably be of nonfecal
origin (16): Klebsiella trevisamii, Enterobacter agglomerans, E. gergoviae, E. sakazakii,
Hafnia alvei, Serratia marcescens, S. liquefaciens, S. marinorubra, and S. odorifera.
Unfortunately, the specificity of fecal coliforms as indicators of fecal pollution varies
considerably depending on the environmental conditions and the presence of industrial
effluent (3). Some authors (7, 19) have suggested that the term "fecal coliforms" should be
excluded from microbiology. "Thermotolerant coliforms" (TTC) is considered to be a more
appropriate description of these organisms (4). The acronym KEC is introduced in this study
to describe the -galactosidase positive thermotolerant coliforms other than E. coli.
Applied and Environmental Microbiology, August 1999, p. 3746-3749, Vol. 65, No. 8

1995 --

1989 --
Ammonia-induced injury in pure cultures and natural populations of coliform bacteria.
Ammonia-induced injury was investigated in pure cultures of Escherichia coli and
Enterobacter aerogenes, and in natural coliform populations obtained from the oligotrophic
Luxapallila and the eutrophic Sunflower Rivers in northern Mississippi. Pure cultures were
affected by ammonia exposure as indicated by changes in the injury ratio (IR) of CFU on m-
T7 agar/CFU on m-Endo agar. Ammonia concentrations between 0 and 20 (mg NH3-N/1)
had little or no effect and concentrations between 40 and 80 caused the greatest injury.
Natural coliform populations from the oligotrophic river were more prone to ammonia-induced
injury than those from the eutrophic river. The results stress the need for the routine use of
m-T7 media and the enumeration of injured cells when using the membrane filter procedure
to ascertain domestic water quality.
FEMS Microbiol Lett. 1989 Nov;53(1-2):65-9.

1987 -- Endotoxin Promotes the Translocation of Bacteria From the Gut
Experiments were performed in mice to determine whether endotoxin could cause bacteria
normally colonizing the gut to spread systemically, a process termed bacterial translocation.
Endotoxin given intraperitoneally promoted bacterial translocation in a dose-dependent
fashion from the gut to the mesenteric lymph node (MLN). The incidence of bacterial
translocation to the MLN was similar whether the endotoxin was administered intramuscularly
or intraperitoneally, although the number of bacteria colonizing the MLN was greater with
intraperitoneal endotoxin. The incidence and magnitude of endotoxin-induced bacterial
translocation were similar between CD-1 and C3H/HeJ (endotoxin-resistant) mice, indicating
that bacterial translocation is not prevented by genetic resistance to endotoxin. Thus, it
appears that the gut may serve as a reservoir for bacteria causing systemic infections during
Arch Surg. 1987;122(2):185-190.

1984 -- Evidence for the role of copper in the injury process of coliform bacteria in drinking
Low levels of copper in chlorine-free distribution water caused injury of coliform populations.
Monitoring of 44 drinking water samples indicated that 64% of the coliform population was
injured. Physical and chemical parameters were measured, including three heavy metals
(Cu, Cd, and Pb). Copper concentrations were important, ranging from 0.007 to 0.54
mg/liter. Statistical analyses of these factors were used to develop a model to predict
coliform injury. The model predicted almost 90% injury with a copper concentration near the
mean observed value (0.158 mg/liter) in distribution waters. Laboratory studies with copper
concentrations of 0.025 and 0.050 mg/liter in an inorganic carbon buffer under controlled
conditions of temperature and pH caused over 90% injury within 6 and 2 days, respectively.
Studies of the metabolism of injured Escherichia coli cells indicated that the respiratory chain
is at least one site of damage in injured cells.

1984 -- Human Pathogenic Viruses at Sewage Sludge Disposal Sites in the Middle Atlantic
Human enteric viruses were detected in samples of water, crabs, and bottom sediments
obtained from two sewage sludge disposal sites in the Atlantic Ocean. Viruses were isolated
from sediments 17 months after the cessation of sludge dumping. These findings indicate
that, under natural conditions, viruses can survive for along period of time in the marine
environment and that they may present potential public health problems to humans using
these resources for food and recreation.
The isolation of viruses in the absence of
fecal indicator bacteria reinforces previous observations on the inadequacy of
these bacteria for predicting the virological quality of water and shellfish.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Oct. 1984, p. 758-763 Vol. 48, No. 4

1980 -- Bacteria Associated with False-Positive Most-Probable- Number Coliform Test
Results for Shellfish and Estuaries
Aerobic and facultatively anaerobic bacteria isolated from false-positive, presumptive,
total coliform, most-probable-number tests of Chesapeake Bay oyster,
water, and sediment samples were characterized and then classified by numerical
taxonomy. A total of 538 bacterial strains clustered into 17 phena, the predominant
groups of which were Enterobacteriaceae (including Escherichia coli),
Aeromonas spp., and Bacillus spp. Bacillus spp. were recovered most frequently
from sediment samples. Gas-producing strains which were not members of the
Enterobacteriaceae were not isolated during this study. However, disproportionately
large numbers of atypical and anaerogenic lactose-fermenting strains were
encountered. We concluded that no single, specific bacterial group can be identified
as being responsible for the false-positive reaction in the presumptive coliform
test. Instead, the false-positive reaction is a result of complex interactions among
various genera, representing predominantly bacteria other than coliforms.

1980 -- Enumeration of Potentially Pathogenic Bacteria from Sewage
The paucity of quantitative data concerning the content of pathogenic and
potentially pathogenic bacteria in sewage sludge has been cited by Carrington (4)
as being a major obstacle in determining the impact pathogens from sludges may
have on the environment. Such sludges could pose a significant health risk
through the contamination of vegetables, surface waters, and groundwaters.
Hess and Breer (18) found Salmonella species in 90% of the sludges they examined
and observed that
the organisms could survive for up to 72 weeks in sludges that
had been applied to land.
Furthermore, they reported that neither aerobic nor
anaerobic digestion significantly reduced the contamination of sludge with
Salmonella. McKinney and co-workers (27) reported that the survival of Salmonella
in sludge in seeded bench-scale digesters was dependent on the density of the
initial population, available nutrients, and detention time. Foliguet and Doncoeur
(11) have urged the routine disinfection of digested sludges because retention
times and temperatures in field digesters varied so that consistent killing of
Salmonella was not obtained.

1980 -- Two antibiotic-resistant strains of Escherichia coli for tracing the Sewage in
Using laboratory cultured bacteria which are not pathogenic
Journal of Hydrology (NZ), 1980

1979 -- Chlorine injury and the enumeration of waterborne coliform bacteria.
Reports in the literature indicate that injury of a reversible nature occurs in cells that
have been exposed to water (4), freezing in foods (25), and sanitizers (31). Other potential
sources of injury may also be present in aquatic environments. The subject of chlorine-
induced injury has been controversial, with some investigators stating that enumerated cells
were not injured (7, 14), whereas others suggested that certain cells were capable of
repairing the injury incurred (23, 22). It has also been proposed that the addition of certain
metabolites aided in the reversal of injury (12) and that reducing agents would inhibit or
reverse the oxidation caused by chlorine (16, 27). Studies have shown that the membrane
filtration procedure for enumerating coliform bacteria with the selective M-FC medium does
not enumerate chlorine-injured cells
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Mar. 1979, p. 633-641, Vol. 37, No. 3

1979 -- Hemagglutination Patterns of Enterotoxigenic and Enteropathogenic Escherichia coli
Determined with Human, Bovine, Chicken, and Guinea Pig Erythrocytes in the Presence and
Absence of Mannose - Texas Medical School

1972 --
Enteric Bacterial Growth Rates in River Water
Enteric bacteria, including stocked strains of pathogenic species and organisms naturally
present in the stream, were capable of growth in a chemostat with autoclaved river water
taken 750 m below a sewage outfall. Maximal specific growth rates for all organisms occurred
at 30 C, whereas culture generation times ranged between 33.3 and 116 hr. Of the six
laboratory strains of enteric species used, Escherichia coli and Enterobacter aerogenes
grew at generation times of 34.5 and 33.3 hr, respectively, while the remaining Proteus,
Arizona, Salmonella, and Shigella spp. reproduced at a rate two to three times slower than
the coliforms. Little or no growth occurred in the water at incubation temperatures of 20 and
5 C, and death was observed for Salmonella senftenberg at 20 and 5 C and for E.
aerogenes and Proteus rettgeri at 5 C. When enteric bacteria naturally present in the river
water were employed in similar experiments, coliform bacteria demonstrated a generation
time of approximately 116 hr, whereas fecal coliforms failed to grow. Growth of the bacteria
from the river demonstrated a periodicity of approximately 100 hr, which suggests that much
of the growth of these organisms in the chemostat may be on the glass surfaces. This
phenomenon, however, was not observed with any of the stocked enteric species. Neither
the stock cultures nor the aquatic strains were capable of growth in autoclaved river water
taken above the sewage outfall at the three temperatures tested.
Appl Microbiol. 1972 August; 24(2): 168–174.

1964 - Sanitary Significants of coli form and fecal coli form -- Public Health Service _ Cin
THE SCIENCE of sanitary water bacteriology began in 1880 when von Fritsch described
Klebsiella pneumonia and K. rliinoscleromatis as organisms characteristic of human fecal
contamination. A short time later Escherich identified Bacillus coli as an indicator of fecal
pollution. Both observers considered human feces as dangerous pollution while the feces of
other warm-blooded animals were not so regarded. From this origin the current "coliform
group" developed to include numerous micro-organisms of diverse biochemical and
serologic characteristics, natural sources and habitats, as well as controversial sanitary
Public Health Reports, Vol. 79, No. 1, January 1964

1958 -- The Coliform Group II -- Public Health Service
Escherichia coli has been suggested by several research workers as a more accurate
indicator of fecal contamination than the more inclusive "coliform group." Media and
procedures for the enumeration of E. coli have been proposed by Eijkman (1904), Leiter
(1929), Perry and Hajna (1933, 1944), Hajna and Perry (1939, 1943), Vaughn et al. (1951),
and Levine et al. (1955). Some of these procedures apparently yield satisfactory results with
certain samples of water, milk, or food products; however, there is no universal agreement
as to their criteria of application and significance of results in water bacteriology. There are
also uncertainties regarding the significance of E. coli when assayed by rapid tests as
compared to the traditional  IMViC classification.
This investigation was undertaken to determine the reactions of 12 IMViC types of coliform
bacteria in EC medium after 24 hr incubation at 45 C. It is believed the resultant data will be
of value in choosing the procedure and in the interpretation of test results for E. coli and the
coliform group.
Public Health Department Report.
Appl Microbiol. 1958 September; 6(5): 347–348.

1957 The Coliform group I
Public Health Department Report, VOL. 5
Several methods have been suggested for the accurate, rapid, and convenient detection
and enumeration of Escherichia coli in potable waters, stream samples, sewage, and in
certain food products. The first of these procedures was described by Eijkman who
proposed a "fecal coli" test that he believed differentiated between coliforms from the gut of
warm-blooded animals and strains originating from cold-blooded ones. The method was
based on the assumption that only fecal types of coliforms from warm-blooded animals
could grow in glucose broth at 46 C with the production of gas. Twenty-five years later a
close relationship between the positive Eijkman reaction and indole production was observed
by Leiter (1929) and he suggested that the combination of the two positive tests was
practically specific for E. coli.
Many procedures have been proposed which claim to
have provided a more suitable environment for the growth of E. coli and at the
same time to have suppressed to a large degree the growth of other coliform types
along with the majority of noncoliform bacteria.

1955 -- The Use of Serology in the Classification of Micro -organisms
A further important example of the use o€ serology in elucidating broad relationships is
provided by the large group of bacteria classified (Bergey’s MaNuaZ, 1948) as the family
Furthermore, the sharing of some antigens between genera, e.g. Escherichia and Klebsiella
(Kauffmann, 1949), supports the view that not only are the members of this
family closely related but they do in fact form a continuous series
J . gen. Microbial. 12, 367-374

S Afr Med J. 1954 May 22;28(21):439-41.

1952 -- The Status of Serologic Typing in the Family Enterobacteriaceae -cdc
Salmonella, Shigella, Escherichia coli group, Proteus group, Klebsiella-Aerogenes group,
Aerobacter- aerogenes,
The value of definitive typing of E. coli cultures is well illustrated by results recently obtained
in the study of infantile diarrheas in England, Holland, the Scandinavian countries, and the
United States. Due largely to the work of Bray,24 Bray and Bea anG,25Giles and Sangster,
2" Giles, Sangster, and Smith,27 Smith, Galloway, and Speirs,28 Taylor, Powell, and Wright,
29 Rogers, et al.,30-33 Taylor,34 and Kauffmann and Dupont,3' it has become evident that
certain E. coli types play a prominent role in the causation of infantile enteritis. While only
two serologic types of E. coli have as yet been definitely incriminated, it seems likely that
additional types will be found which have a definite relationship to the disease.
Am J Public Health Nations Health Edwards and Ewing 42 (6):

1951 -- The Anaerogenic Effect of Nitrates and Nitrites on Gram-Negative Enteric Bacteria
W HILE estimating the Escherichia-Aerobacter densities (most probable numbers) of meat
curing brines,  difficulties were encountered because of the almost invariable appearance of
' skips" or false-negatives in the lowest dilutions (10 ml. portions), while gas was often formed
in the higher dilutions (1 ml. and 0.1 ml. portions). The standard 3-dilution fermentation tube
method was employed, using 5 tubes in each dilution and double lactose broth for the 10 ml.
portions.1 It became apparent that some factor in the curing brine, or "pickle," was inhibiting
the gas production of the coliform organisms.

1944 -- Evaluation EC Medium -- State of Maryland
AT the St. Louis meeting of the American Public Health Association in October, 1942, the
authors had the pleasure of discussing their experience with a new EC medium (buffered
tryptose lactose bile salt t) for the isolation of coliform bacteria at 370 C. and of Escherichia
coli at 45.50 C.1 In the examination of 147 samples of drinking water of various types, not a
single false presumptive was encountered among 1,176 gas tubes while 58.5 per cent of
false presumptives were obtained with standard lactose broth. At the same time 14.2
per cent more of the EC tubes were positive for coliforms. The medium was found to have
comparable sensitivity to Mallmann and Darby's 2 LST (lauryl sulfate tryptose) medium when
tested with 33 stool specimens and 25 samples of raw sewage.
Am J Public Health Nations Health. 1944 July; 34(7): 735–738.

1937 -- Escherichia-Aerobacter Intermediates from Human Feces
THE phrase, "Escherichia-Aerobacter intermediates," has been variously employed in the
literature of the past decade. At present it customarily refers to coliform bacteria which differ
from typical Escherichia or Aerobacter strains in one or more of the major differential
reactions, such as the Voges-Proskauer, methyl-red and citrate utilization tests. Recent
discussion has tended to confine the designation to those organisms which are Voges-
Proskauer-negative, methyl-red-positive, and capable of multiplying in a medium containing
citrate as the sole source of carbon.
Am J Public Health Nations Health. 1937 August; 27(8): 822–827.

The Escherichia-Aerobacter group comprises one of the important groups of bacteria in
dairy products. Milk, even under careful conditions of production; practically always contains
some of these organisms. The fact that the Escherichia type generally represents organisms
coming from the intestinal tract of man and animals and the Aerobacter type organisms
from soils and grains makes a distinction between the two types useful. The growth of
Escherichia-Aerobacter organisms in dairy products is undesirable for, in addition to forming
acid and gas from lactose, they produce undesirable flavors and aromas. Some of the
defects which have been reported as due to this group of organisms are ropiness in milk and
cream, and gassy fermentations in cottage cheese, cheddar cheese, milk, and
sweetened condensed milk.

' -- Dept. of Health, Chicago
Recently developed differential plating methods of enumerating colon-aerogenes organisms
are in need of further study from the standpoint of interpretation of findings. Such a method,
originally devised by Noble (1928) and later revised and simplified for use in routine water
analyses (Tonney and Noble (1931a), has been used by us in studying the sanitary
significance of Bact. coli and Bact. aerogenes in water supplies (Tonney and Noble
(1931b)), and also for studying the relative persistence of the two types of organisms in
nature (Tonney and Noble (1931c)). The data of these studies seem to indicate that the
Bact. coli count in this medium has a much closer and more consistent relationship to fecal
sources of pollution than has the Bact. aerogenes count.

NATURE -- Depart of Health, Chicago
A growing question for practical consideration in water control is the relative sanitary
significance of Bact. coli and Bact. aerogenes as criteria of fecal pollution. It is generally
agreed that Bact. coli is strictly of fecal origin, and its presence in water therefore is
accepted as evidence of dangerous pollution. Bact. aerogenes, however, occupies a
doubtful position as an organism of fecal origin. Some observers regard it as of the same
fecal significance as Bact. coli, while others consider it of little, if any, value as
indicating contamination from fecal sources. There appears to be growing evidence in
support of the latter view (Chen and Rettger (1920); Koser (1927); Hinman (1925); Tonney
and Noble (1930)). Certainly, the great predominance of Bact. aerogenes over Bact. coli at
large in nature needs some explanation other than the assumption that both are of direct
fecal origin, since in feces itself the reverse relationship is true. In previous studies
we have found the ratio of Bact. coli to Bact. aerogenes, by the differential plate count, to be
about 100 to 1 in feces of both human and animal origin, and on the other hand about 1 to
20 in soils and vegetation (Tonney and Noble (1930)).
J Bacteriol. 1931 December; 22(6): 433–446.

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. From time to time other
organisms have been proposed wholly or partially to replace these bacteria, but, as stated
so definitely and convincingly by Levine (1921), Bacterium coli is the organism
which best fulfills the requirements for an index of fecal pollution. It is constantly present in
the feces of warm blooded animals, is seldom obtained except from feces or from substances
recently polluted, is slightly more resistant than intestinal bacterial pathogens, but is not
sufficiently resistant to remain long in water after the water has been purified enough to kill
J Bacteriol. 1930 August; 20(2): 139–150.

1929 -- Eijkman medium in examination of oysters and crab meat -- state of Maryland
T HE Eijkman test for Bacillus coli has been used and studied in the laboratories of the
Bureau of Bacteriology of the Maryland State Department of Health since 1929. One of the
authors (Perry, 1929), first applied the test to a number of cultures of Bacillus
coli recovered from oysters and oyster bearing waters. He found " such cultures
invariably produce indol from a suitable medium and were able to ferment
the dextrose of Eijkman broth with gas at 46' C." He also found that " only 11 . 2 per cent of
223 cultures of lactose-fermenters (coli-aerogenes group) from oyster and water samples
from four areas were Bacillus coli. The four areas differed geographically, topographically,
and in respect to their sanitary features. The presence of Bacillus coli was found in close
agreement with probable fecal pollution."
Am J Public Health Nations Health Perry and Hajna 25 (6): 720

The author has felt for some time that before much light will be thrown on the true relative
incidence of B. coli and B. aerogenes in water and in feces, etc., it will be necessary first, so
to modify our preliminary enrichment media, or other conditions,
as to enable the
investigator to isolate or suppress either B. coli or B. aerogenes at will.
Am J Public Health (N Y). 1921 January; 11(1): 21–23.

1911 -- Classification of the B. Coli Group
The term B. coli as an indication of fecal contamination in water and milk has been so often
misapplied that the result has been much confusion and frequent misinterpretation of
bacterial examinations. It has been the custom of many bacteriologists to throw out of
sanitaryconsideration all bacteria which do not absolutely conform to the so-called "typical"
B. coli. There are many known varieties, all of fecal origin and closely related to typical B.
coli, which will be described in this paper, and there probably exist many more varieties which
will be discovered in the future. Any of these varieties, when they occur in water or milk,
have a sanitary significance, and because of their close relationship, all
should be included in the B. coli group. The fermentative reactions have been chosen as a
means of classification, not only because of the ease with which these organisms are thus
separated from those of other groups, but because of the facility with which each variety may
be separated from the others...
After the identification of this variety, a vaccine was made which was specific for this
particular variety of infection, whereas the vaccine previously made from B. communis B
apparently had no curative effect.

1903 -- The Pathogenicity of B. coli in Relation to the Bacteriological ...by WG Savage -  
virulent B. coli does not, of necessity, indicate faecal contamination. His points of
identification for his B. coli are however very inadequate. ... In considering the question of
virulence of B. coli in water supplies ..... For the two B. coli isolated from milk, rabbits were
used for testing the virulence:- ...

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Jim Bynum, VP
Help for Sewage Victims

Coliform exhibit optimum
growth in  the 20-37° C range
(68 - 98.6°F).  Total Coliform
are gram negative members of
the Family Enterobacteriaceae
that ferment lactose at 98.6°F
to produce gas and/or acid
within 48 hours. Other members
of the Family
take a longer time to ferment

Fecal coliform exhibit minimal
growth at an elevated
temperature. They are a few
thermotolerant members of the
Family Enterobacteriaceae
that ferment lactose at
112.1°F  to produce gas
and/or acid within 48 hours.

E. coli and Klebsiella are the
primary fecal coliform that
exhibit less than 5% growth rate
at  the elevated temperature.
They act as opportunistic
pathogens when they are
introduced into body
locations where they are not
normally found, especially if
the host is debilitated or

While there may be some
non-pathogenic strains, the
International Escherichia and
Klebsiella Centre (WHO) has a
collection of approximately
60,000 E. coli strains, most of
which are clinical isolates.
Clinical isolates means they are
pathogens associated with
disease.  About half of the
hospital acquired infections are
caused by the coliform bacteria