| TOTAL COLIFORM = ENTEROBACTERIACEAE
In sewage sludge and sewage wastewater effluent
(Biosolids and Reclaimed water)
Fecal Coliform = thermotolerant version of coliform in sewage, sludge, biosolids, water.
Department of Health and Human Services adopted the simple colifom test in 1914 to indicate the presence of
fecal contamination in food and water. Coliform is not a taxonomic classification but rather a working definition
used to describe a family of 12 Gram-negative, facultative anaerobic rod-shaped bacteria that ferments lactose
to produce acid and gas within 48 h at 35°C. http://www.cfsan.fda.gov/~ebam/bam-4.html
by Jim Bynum
Would it surprise you to know that EPA's drinking water test does not require testing for pathogenic
E. coli strains, gram positive pathogenic bacteria, and other pathogenic organisms such as viruses,
protozoa or helminths? Both EPA and CDC claim the colifom test only reveal the presence of
nonpathogenic strains of E. coli.
For Nomenclature for Aerobic and Facultative Bacteria click on
What is Escherichia coli? E. coli is one of the coliform group and the primary marker for gaging the
safety of drinking water due to the ease of testing. However, the most pathogenic strains of E. coli
do not show up in the test as markers. The CDC explains it this way:
most strains of E. coli are harmless, others can make you sick. Some kinds of E. coli can cause
diarrhea, while others cause urinary tract infections, respiratory illness and pneumonia, and
other illnesses. Still other kinds of E. coli are used as markers for water contamination—so
you might hear about E. coli being found in drinking water, which are not themselves harmful,
but indicate the water is contaminated. It does get a bit confusing—even to microbiologists."
and inorganic substances, or a pathogenic organism that, after discharge and upon exposure,
ingestion, inhalation, or assimilation into an organism either directly from the environment or
indirectly by ingestion through the food chain, could, on the basis of information available to the
Administrator of EPA, cause death, disease, behavioral abnormalities, cancer, genetic mutations,
physiological malfunctions (including malfunction in reproduction), or physical deformations in
either organisms (humans) or offspring (children) of the organisms.
Family Enterobacteriaceae: (gram negative bacteria) are called total coliforms, and are defined by
their ability to ferment lactose within 24 - 48 hours incubation at human body temperature of 35-37°
C (95-99°F). EPA falsely claims that none of the bacteria included in the coliform group cause
diseases in humans.
EPA has made a little joke about its criminal activity of exposing public health to these deadly
pathogenic disease organisms: “When the news media announce a "boil water emergency,"
reporters often speak of a "total coliform violation." Coliforms are a group of bacteria, most of which
are harmless. At first glance, it might seem strange that a harmless group of bacteria such as
coliforms could cause such commotion. But like police tape and chalk outlines, coliform bacteria are
often found at the scene of a crime even though they are not themselves criminals.”
Mesophiles: Pathogenic micro-organisms that grow best at temperatures between 25 and 40 °C;
usually will not grow below 5 °C.
EPA claims, "Coliforms are a group of bacteria, most of which are harmless."
"total coliforms are common inhabitants of ambient water and may be injured by environmental
stresses (e.g., lack of nutrients) and water treatment (e.g., chlorine disinfection) in a manner similar
to most bacterial pathogens and many viral enteric pathogens." http://www.epa.
"Coliform Organism: Microorganisms found in the intestinal tract of humans and animals. Their
presence in water indicates fecal pollution and potentially adverse contamination by pathogens."
EPA: Organism: "Any form of animal or plant life."
In a 1984 STUDY, "A total of 1,900 lactose-fermenting bacteria were isolated from raw sewage
influent and chlorinated sewage effluent from a sewage treatment plant, as well as from chlorinated
and neutralized dilute sewage, before and after a 24-h regrowth period in the laboratory. Of these
isolates, 84% were resistant to one or more antibiotics. Chlorination of influent resulted in an
increase in the proportion of bacteria resistant to ampicillin and cephalothin, the increase being
most marked after regrowth occurred following chlorination. Of the other nine antibiotics tested,
chlorination resulted in an increased proportion of bacteria resistant to some, but a decrease in the
proportion resistant to the remainder. Multiple resistance was found for up to nine antibiotics,
especially in regrowth populations. Identification of about 5% of the isolates showed that the
highest proportion of Escherichia coli fell in untreated sewage. Some rare and potentially
pathogenic species were isolated from chlorinated and regrowth samples, including Yersinia
enterocolitica, Yersinia pestis, Pasteurella multocida, and Hafnia alvei. Our results indicate that
chlorination, while initially lowering the total number of bacteria in sewage, may substantially
increase the proportions of antibiotic-resistant, potentially pathogenic organisms."
Dr. Edward McGowan explains the problem:
problems----mainly that these are not reflective of actual pathogen levels. These coliform are
vegetative bacteria that are susceptible to low-level disinfection, yet there are many
pathogens that require high-level disinfection. These include viruses that are non-enveloped,
several robust pathogens such as the ones that cause multi-drug resistant tuberculosis, and
the bacillus group. In these we find spores but there are several other spore formers beside
these. The formation of toxins is also part of the issue and some toxins can withstand extreme
heat and thus are not destroyed by cooking. Their spores can survive harsh environmental
conditions. In one case, anthrax was tested on an island off Great Britain during WW II. The
small island remained quarantined for 50 years, see also Shackelton’s hut below.
In sewage sludge, a certain number of indicators are allowed to remain. If these low-level
vegetative bacteria survive, what of the more robust? Then we have antibiotic resistance and
on top of that, genetic material that does not show using a coliform test and viable but non-
culturable as well as certain pathogens that also do not show up well on lab tests---all thus
missed by relying on coliform. Further, several pathogens are chlorine resistant and certainly
the genetic material although being capable of transfer, is not affected by chlorine levels
used in water treatment.
These Enterobacteriacea (coliform) are estimated to be responsible for about 100,000 deaths each
year in the US, and account for about half of all the clinically significant bacteria isolated by hospital
[biosafety level 2] laboratories. Freezing does not destroy them -- whether in nature in the water, or
on frozen foods contaminated with the bacteria.
As you will see, there is nothing harmless about this Enterobacteriaceae family (coliform) of disease
organisms. The implication of EPA's statement is that just because they are found with necrotizing
fasciitis (flesh eating bacteria), doesn't mean they caused the infection.
Coliforms are Gram negative, non-spore forming, oxidase negative, fermentative rods belonging to
the Family Enterobacteriaceae. This group is widely distributed in nature, and many can live as
saprophytes in addition to being associated with the intestinal tract of most animals. Coliforms are
divided into two groups for convenience. The larger group, which includes all members of the
Family Enterobacteriaceae, are called total coliforms, and
are defined by their ability to ferment lactose within 24 - 48 hours incubation
at 35°C. The thermotolerant fecal coliform at (44.5 °C) are primarily E. coli and occasionally
Klebsiella. in the past, E. coli 0157:H7 was not thermotolerant and proper testing was done at low
temperatues. The tests takes 3 to 7 days, depending on the pathogen.
A recent study, July 2007, found thermotolerant 04 (Uropathogenic E. coli, UPEC), 025
(Enterotoxigenic E. coli, ETEC), 086 (Enteropathogenic E. coli, EPEC), 0103 (Shiga-toxin producing E.
coli, STEC), 0157 (Shiga-toxin producing E. coli, STEC), 08 (Enterotoxigenic E. coli, ETEC) and 0113
(Shiga-toxin producing E. coli, STEC) in drinking water systems.
These respond to very low levels of disinfection which may only injure some bacteria. Other
pathogenic bacteria (index) and viruses (index) will not be affected.
To verify the coliform facts I went to EPA's expert, Mark Meckes.
In a message dated 7/16/2007 10:44:11 A.M. Pacific Daylight Time,
[email protected] writes:
The total coliform group consists of several genera of bacteria belonging to the family
According to Meckes 1982 study, the Family Enterobacteriaceae include bacteria such as E. coli,
Salmonella typhi, Shigella dysenteriae, Aeromonas, Yersinia species, Pseudomonas aeruginosa,
and Vibrio cholerae.
In 1984, researchers from Clinical Microbiology, Department of Pathology, University of California at
Los Angeles Medical Center, evaluated the rapid 18-24 hour Eiken system, a 21-
biochemical-test battery tray stored at 25°C and inoculated in one step. A total of 345 coliform
organisms in 16 (out of about 30) genera from the family Enterobacteriaceae and 2 members of the
genus Aeromonas were tested. They found, although the Eiken system correctly identified only
79.5% of the 347 isolates tested, the low accuracy was the result of an inadequate computer code
book data base.
In 1991, CDC verified the accuracy of the 24 hour rapid strip test for 291 coliform strains of 16
genera of these bacteria likely to be routinely isolated in a clinical microbiology laboratory
The results were about the same.
The Introduction To Clinical Microbiology, University of Texas - Houston Medical School, describes
coliform as the "Enterobacteriaceae family have earned a reputation placing them among the most
pathogenic and most often encountered organisms in clinical microbiology. They are the causative
agents of such diseases as meningitis, bacillary dysentery, typhoid, and food poisoning."
Furthermore, according to Kenneth Todar, University of Wisconsin-Madison Department of
Bacteriology, "The enterobacteriaceae include agents of food poisoning and gastroenteritis,
hospital-acquired infections, enteric fevers (e.g. typhoid fever) and plague. They also cause
infections in domestic, farm and zoo animals and include an important group of plant pathogens.
Their host range includes animals ranging from insects to humans, as well as fruits, vegetables,
grains, flowering plants, and trees." [now they are human pathogens]
Other gram negative rod bacteria in sludge/biosolids and reclaimed water
TOTAL COLIFORMS MOST OFTEN SEEN IN HOSPITAL SETTINGS IN THE PAST
Coliforms -- Gram- negative rods are usually associated with intestinal infections which may spread
to other parts of the body. The term EPA uses for all infections is gastroenteritis.
Aeromonas (1985) University of California at Los Angeles Medical Center,
Suppressed in Fecal Coliform Test
Cedecea strains (1984) The name Cedecea was proposed in 1980 for a new genus.
In 1982, Farmer and co-workers reported a case of bacteremia caused by a species of Cedecea
designated as Cedecea neteri (2).
Citrobacter: C. freundii is suspected to cause diarrhea and possibly extraintestinal infections. C.
diversus has been linked to a few cases of meningitis in newborns.
Edwardsiella tarda: E. tarda produces hydrogen sulfide. This bacterium is usually found in aquatic
animals and reptiles. However, it has been known to cause gastroenteritis and wound infections in
Enterobacter: several species cause opportunistic infections of the urinary tract as well as other
parts of the body. E. aerogenes and E. cloacae are two such pathogens that do not cause diarrhea,
but that are sometimes associated with urinary tract and respiratory tract infections.
ESCHERICHIA COLI: Besides being the number one cause of human urinary tract infections, E. coli
has been linked to diseases in just about every other part of the body. Pneumonia, meningitis, and
traveler's diarrhea are among the many illnesses that pathogenic strains of E. coli can cause.
Pathogenic strains of E. coli can cause severe cases of diarrhea in all age groups by producing a
powerful endotoxin. [Central America Shigella strain Toxin] Treating E. coli infections with
antibiotics may actually place the patient in severe shock which could possibly lead to death. This is
due to the fact that more of the bacterium's toxin is released when the cell dies. (Primary Fecal
E. coli O157:H7 is markedly different from other pathogenic E. coli, as well. In particular, the O157:H7
serotype is negative for invasiveness (sereny test), elaborates no colonization factors (CFA/I or
CFA/II), Doesn't produce heat stable or heat labile toxins and is non-hemolytic. In addition, E. coli
O157:H7 is usually sorbitol negative whereas 93% of all E. coli ferment sorbitol. E. coli O157:H7
also lacks the ability to hydrolyze 4-methylumbelliferyl-β-D-glucuronide (MUG) and does not grow
at 45 °C in the presence of 0.15% bile salts. Because of the latter characteristic this serotype cannot
be isolated by using standard fecal coliform methods that include incubation at 45 °C.
Ewingella (2003) A fatal case of Waterhouse-Friderichsen syndrome resulting from infection in a
previously healthy 74-year-old woman is reported. The patient died suddenly within 14 hours after
presentation. The diagnosis of Waterhouse-Friderichsen syndrome as the cause of death was
established post mortem based on autopsy findings, microscopic examination, measurement of
serum procalcitonin concentration (113 ng/ml), and outcome of postmortem bacteriologic cultures
that grew in heart and spleen blood samples. Since the introduction as a new group in the family in
1983, more recent case studies have established its clinical significance and pathogenic potential
to cause severe, life-threatening bacteremia and sepsis. It is a rare pathogen that should be added
to the list of unusual bacteria causing Waterhouse-Friderichsen syndrome.
Hafnia alvei strains (1994-1998), ) Three patients had hospital-acquired urinary tract infections.
Hafnia alvei also caused community-acquired cholangitis, cholecystitis, appendicitis, psoas abscess
and prosthetic endocarditis. In (2000--2005) A total of 138 patients with Hafnia alvei isolates were
identified [in canada] (2.1/100,000/year) and two-thirds were of community onset. Older age and
female gender were important risk factors for acquisition. The most common focus of isolation was
urine in 112 (81%), followed by lower respiratory tract in 10 (7%), and soft tissue in 5 (4%), and the
majority (94; 68%) were mono-microbial.
Klebsiella: Klebsiella's pathogenicity can be attributed to its production of a heat-stable
enterotoxin. K. pneumoniae is second only to E. coli as a urinary tract pathogen. Klebsiella
infections are encountered far more often now than in the past. This is probably due to the
bacterium's antibiotic resistance properties. Klebsiella species may contain
resistance plasmids (R-plasmids) which confer resistance to such antibiotics as ampicillin and
carbenicillin. To make matters worse, the R-plasmids can be transferred to other enteric bacteria
not necessarily of the same species. (sometimes show in Fecal Coliform Test)
Kluiyvera strains (2003) The spectrum of disease due to Kluyvera infection in children includes
central venous catheter infection and/or sepsis, urinary tract infection, enteritis, and, in one
instance, fatal peritonitis. It is clear on the basis of our case report that uncommon, opportunistic
organisms such as Kluyvera can be significant pathogens. four isolates were from blood (including
our case), four were from urine, and one each were from peritoneal fluid, endo-tracheal aspirate,
autopsy lung tissue, and stool (Table 2)....
Koserella (1994) Yokenella regensburgei (Koserella trabulsii) was isolated from a 74-year-old male
with a septic knee and from a 35-year-old immunocomprised female whose transient bacteremia
occurred without overt signs of sepsis. Neither strain was correctly identified by laboratories using
a variety of techniques
Leminorella (1999 ) The site of isolation was urine in 6, wound in 3, and blood, peritoneal fluid, and
sputum in 1 each. Lm was the only isolate from the site of infection in 7 cases. Based on clinical data
Lm was classified as a definite pathogen in 6 cases, probable in 2, and a possible pathogen in 3;
causing UTI in 5 patients, surgical site infection in 2, bronchitis, soft tissue infection, primary
bacteremia and secondary peritonitis in 1 each. Only in one case did Lm isolate have no clinical
significance. All 12 isolates were resistant to second gen. cephalosporins. 5/12 isolates were
susceptible to penicillins, third gen. cephalosporins and ciprofloxacin. 7/12 of isolates were
resistant to penicillins, penicillin/inhibitor combinations, and to cefotaxime. 11/12 isolates were
susceptible to ceftazidim, and all were susceptible to imipenem and amikacin. We conclude that
Leminorella spp. should be considered as a significant nosocomial pathogen capable of causing a
variety of clinical syndromes. Clinical isolates of Leminorella are often resistant to multiple antibiotic
Morganella morganii can cause urinary tract and wound infections, as well as diarrhea.
Providencia species have been associated with nosocomial (hospital acquired) urinary tract
infections. P. alcalifaciens, has been associated with some cases of diarrhea in children.
Proteus, can cause urinary tract infections and hospital-acquired infections. P.mirabilis, a cause of
wound and urinary tract infections. most strains of P. mirabilis are sensitive to ampicillin and
cephalosporins. P. vulgaris is not sensitive to these antibiotics.
Rahnella (2005) A 76-year-old male with prostatic hyperplasia presented with acute pyelonephritis.
Blood cultures yielded Rahnella aquatilis. Treatment with intravenous followed by oral levofloxacin
resulted in cure. Important characteristics of this organism include its biochemical similarities to
Enterobacter agglomerans, its apparent ability to cause bacteremia from a renal focus, and its
response to quinolone therapy.
Salmonella: S. typhimurium and S. enteritidis are the two leading causes of salmonellosis
(inflammation of the intestine caused by Salmonella). S. typhi is unique because it is only carried by
humans. This intracellular parasite can cause typhoid fever (enteric fever) which is characterized by
fever, diarrhea, and inflammation of the infected organs.
Serratia genus were once known as harmless organisms that produced a characteristic red
pigment. Today, Serratia marcescens is considered a harmful human pathogen which has been
known to cause urinary tract infections, wound infections, and pneumonia. Serratia bacteria also
have many antibiotic resistance properties which may become important if the incidence of Serratia
infections dramatically increases
Shigella is also an invasive pathogen which can be recovered from the bloody stool of an infected
host. Invasive pathogens colonize the host's tissues as opposed to growing on tissue surfaces.
Genetically, Shigella and E. coli are virtually identical. Were they to be discovered and named today,
they would be grouped into one species. However, Shigella has been set apart traditionally for its
consistent non-motility, its failure to ferment lactose and its cause of bacillary dysentery,
characterized by severe abdominal pain and bloody diarrhea. Only a few ingested cells are needed
to result in disease.
Tatumella strains (2000) Tatumella ptyseos was mostly reported to be found in the specimens
obtained from respiratory tract, with an isolation ratio of 68 % in the sputum of these cases.
Besides, it was isolated in the blood cultures of neonatal sepsis cases. Newborn and elderly are
also among the vulnerable groups
Yersinia genus: Y. enterocolitica and Y. pestis. Y. enterocolitica is the most often encountered
species of Yersinia in the lab. This bacterium is an invasive pathogen which can penetrate the gut
lining and enter the lymphatic system and the blood. Infection, which is usually through ingestion of
contaminated foods, can cause a severe intestinal
inflammation called yersiniosis. Release of its enterotoxin can cause severe pain similar to that
found in patients with appendicitis.
Y. pestis is included here because it causes the bubonic, pneumonic, and septicemic plagues.
Human contraction of bubonic plague is usually through flea bites. Once inside the body, Y. pestis
releases a toxin which inhibits electron transport chain function. Swelling of the lymph nodes, skin
blotches, and dilerium are sometimes observed within a few days of infection. Untreated infections
usually result in death within a week of initial infection.
A comparison of ten USEPA approved total coliform/E. coli tests