American Water Works Association
Parti
Sunday Seminars Through
Poster Presentations
                                in
               Presented at the AWWA
               Water Quality Technology Conference .
               November 10-14,1991 .« Orlando, Florida'

-------
         PATHOGENS IN DRINKIBS MATER - ARE THERE ANY NEW ONES?

                       Donald J.  Reastmer, Chief
             Microbiological Treatment Branch,  DWRD,  RREL   •
                  U.S.  Environmental Protection  Agency
ABSTRACT

      Since 1976 three newly recognized human pathogens  have become
familiar to the drinking water industry as waterborne  disease agents.
These are:  the legionnaires disease  agent, Leoionella pnemnophila
and related species; and two protozoan pathogens, Giardia 1amblia
and CrvptosDO.rJLdj.un! parvuiB. both of which form highly  disinfectant
resistant cysts that are shed in the  feces of infected individuals.
The question frequently arises - are  there other emerging waterborne
pathogens that may pose a human health problem that  the  drinking water
industry will have to deal with?  This paper will review the current
state of knowledge of the occurrence  and incidence of  pathogens
and opportunistic pathogens other than Leglonella, Giardia and
Cryptosooridium in treated and untreated drinking water.  Bacterial
agents that will be reviewed include  Aergagnoas. Pseudomonas.
Campylobacter. Hvcobacterium. Yersjnja and Plesjomonas.   Aspects of
detection of these agents including detection methods  and feasibility
of monitoring will be addressed.

INTRODUCTION

      During the past 15 years three  specific human  pathogenic micro-
organisms, Leqionena. Giardia and Cryptosporidiunu  have been shown
to be waterborne and have caused waterborne disease  outbreaks.
Only Leqionella and Giardia are currently regulated  under the SDWA
Amendments of 1986.  These organisms  pose:  1) a potential health
threat to the general population and; Z) along with  several
opportunistic pathogens, may pose a significant waterborne health
threat to an estimated 30 million Americans who have reduced or
impaired immune function  (1)-  1° addition to these  microorganisms, a.
variety of enteric viruses are known  to be a major cause of human
waterborne disease outbreaks.  These  include Poliovirus, Coxsackievirus
A and B, Echovirus, Rotavirus, Adenovirus, and Hepatitis A virus.  In
most cases of waterborne viral disease, drinking water that received
complete treatment was implicated (53% of reported isolations), while
26% was associated with water that received only disinfection and 15%
was associated^with untreated water (2).  The identification of viral
agents that cause waterborne outbreaks is problematic  due to the fact
that some of these agents can't be propagated in the laboratory.  For
those that can be propagated, large sample volumes must  be examined
to detect the organism.  Propagation  and identification  of the organism
takes several days.  In addition, by  the time the outbreak has been
recognized, it is usually long after  the contamination event that
caused the outbreak and water containing the agent  is  no longer
available.  This problem  is common to all waterborne microbiological
agents.

      Legionella has already been mentioned, but what  about other
bacteria?  What other bacterial agents can cause human health problems
through exposure to water?  tfe are familiar with traditional waterborne
bacterial pathogens such as Vibrio cholera  (cholera  agent),  and
 REASONER    .                                                        S0g

-------
 Salmonella tvohi (typhoid fever agent), but these organisms are easily
 Inactivated by currently used treatment and disinfection practices.
 Are there other waterborne bacterial disease agents that we should
 be concerned with?  What about other viruses, protozoans, fungi and
 algae?  The following is a quick survey of non-traditional nricrobio-
 logical agents that have, or may have, an adverse impact on the water
 consumer's health.

 Bacteri al Haterborne Agents

       There are a variety of waterborne bacteria that are of concern
 from a human health perspective.  These are the opportunistic pathogens
 that can be found as part of the heterotrophic bacteria flora of
 aquatic systems.  The problens with most of these bacteria is that they
 are not obviously pathogenic to "healthy" humans and there is a
 tendency to discount their importance as agents of human disease.  In
 general, they cause apparent disease only in susceptible individuals
 who are immunologically compromised.  Also, as sanitary nicrobiologists
 we tend to be preoccupied with those disease agents that involve
 gastrointestinal tract upsets, rather than waterborne disease agents
 that are involved in wound infections; eye, ear, nose and throat
 infections; or generalized disseminated infections in the body.
 Finally, for many of the opportunistic pathogens, specific selective
 media for enumeration and isolation do not exist, making it difficult
 to directly and easily monitor for their presence and density in water.

       The bacteria generally considered opportunistic pathogens that
 are frequently found in drinking water are shown in Table 1.   Some
 organisms in this list may be considered frank or primary pathogens,
 meaning that they are capable of being the primary disease causing
 agent rather than & secondary invader.  Table 2 is a list of bacteria
 that includes some previously recognized, but still  emerging
 opportunistic organisms and some new candidates.  Table 3 shows new
 or emerging pathogens in the viral, protozoa and blue-gresn algal
 groups.

       Of the older/emerging opportunistic bacteria,  the organisms of
 most concern appear to be Aeromonas spp., Camoylobacter. 4-fnethylura-
 belliferyl-B-D-glucuronide (HUG) - negative Escherichia coll such as
 serotype 01S7:H7, and Hvcobacterimn spp.  However, it is difficult
 to assess the real human health risk of exposure, to these and other
 opportunistic organisms.   The following factors make it difficult to
 assess the health implications of these organisms,  })  The lack of data
 on the occurrence and densities of these organisms in U.S. water
 supplies.  2} The lack of data on infective dose required to establish
 infection.  3) The lack of data on the incidence of human disease
 caused by waterborne exposure to these organisms.  4) The interactive
 effects of exposure to mixed types and densities of these organisms.
 5) The range of susceptible individuals in the exposed population.
 6) The effectiveness of treatment processes and post-disinfection for
 control of these agents, individually and collectively.  7} The need
 for good detection methodologies that would permit adequate
 surveillance and monitoring for these organisms,

       Detection and enumeration methods for any of the waterborne
 pathogens and opportunistic pathogens remain a significant impediment
 to our ability to accurately assess the occurrence and-potential  health
 impact of these organisms.
510

-------
      Table 4 provides density estimates  obtained  from the literature
for selected opportunistic  pathogens  in drinking water.

      Table 5 summarizes  some of the  available  information on the
sensitivity of eaerging and raw candiate  opportunistic pathogens to
disinfection by chlorine.   Essentially no information  is  available on
the sensitivity of these  organisms  to alternative  disinfectants such as
chlorine dioxide,  and ozone.  Presumably  most of the organisms in Table
5 would be more sensitive to ozone  than they are to chlorine, chlorine
dioxide or chloramines.   However, such work remains to be done.

Aer omona.s_spp.

      For lactose  fermenting Aeromonas spp., detection methods are
similar to those for coliforas.  A  few additional  biochemical tests
are required to differentiate'and identify these organisms.   As a
general comment, it appears that Aeromonas spp. are not found in
significant densities in  treated and  disinfected drinking water,
although they have been found in higher numbers in water  that was
disinfected only.   They may gain entry, however, through  post-treatment
contamination events, or  some may survive treatment and disinfection
and regrow under suitable conditions  in some areas of  the distribution
system.  Aeromonads are adversely affected by copper concentrations
above 10 jsg/L (23). In the Netherlands,  limits on Aeromonas densities
in drinking water  have been set at maxi'mura values  of 20 CFU/1QO ml in
water leaving the  treatment plant, and 100 CFU/100 ml  in  distribution
waters.

      Non-lactose  fermenting aeromonads can be  isolated by the use of
selective media containing  starch related compounds and inhibiting
agents such as  ampicillin,  sodium lauryl  sulfate,  sodium  desoxycholate,
and others (5).    .

      For surface  water .supplies, water temperature is an important
factor in the occurrence  of Aeromonas  in  drinking  water since the
highest numbers reported  occurred during  the summer.   Aeroraonad
concentrations  may increase in the distribution water,  responding to
loss of disinfectant residual and to  assimilable organic  carbon in the
water.  Aeromonads can also utilize a  wide range of -biopolymers that
may be available in distribution biofilm  so control of biofilm in the
distribution systen may be  important  for  control of aeromonads.

      Hascher,  et  aj.. (32)  did not find seasonal variations  in
Aeroaonss spp,  in  well water and in the distribution systems.  They
concluded that  nearly 50% of the Aeromonas spp. they tested  could
be presumed to  be  enterotoxin - producers.

Campy!obacter

      Campylobacter jejuni  causes human diarrheal  disease throughout
the world and there is a  large reservoir  consisting of most  of man's
domestic animals.   Many outbreaks are  food and nilk related.

      Isolations of Campvlobacter from treated and disinfected,
unrecontaminated drinking water have not  been reported.   Interest in
the occurrence  of  these organisms has  been related to  their  occurrence
in surface waters  and, therefore, the  potential for their occurrence in
untreated or inadequately treated drinking water.  .Municipal  water
                                                                    511

-------
supplies  contaminated with C. .ie.iuni caused major outbreaks each
involving more than  2000 people (33,34) and an outbreak in Illinois
affected  78 persons  (62%) from 34 of 201 households (24).  Another
outbreak  that involved 94 people was due to Camovlobacter je.luni
contamination of a hospital water supply (35).

Escherichia coli 0157:H7

      This organism  is an atypical fecal coliform in that it does not
hydrolyze the fluorogenic substrate HUG used in recently developed
specific  substrate tests for total coliforms and E. coli.  £. coli
0157:H7  is better known as a foodborne (meat and milk) pathogen, but it
has been  implicated  in at least two waterborne disease outbreaks
(8,36,37).  It causes hemorrhagic colitis, hemolytic uremic syndrome,
and is a  leading cause of kidney disease in children (38).  In all
waterborne outbreak  cases due to E. coli 0157:H7, the water supply was
undisinfected ground or surface water that became fecally contaminated.

      Detection of E_. coli 0157:H7 cannot be readily done using the
standard  membrane filter procedure with m-FC medium and incubation
at 44.5°C for 24 h.  because the organism doesn't grow well at this
temperature.  Since  E. col i 0157:H7 does not ferment sorbitol, total
coliform  isolates that give a sorbitol negative reaction on MacConkey-
sorbitol  medium are  presumptive E. coli 0157:H7.  Final identification
is done by use of the API 20 E system and confirmed by serotyping
for 0:H antigens.

Plesiomonas shiqelloides

      This organism  is closely related to Aeromonas hvdroohila: both
are member of the family Vibrionaceae.  P. shiqelloides has been
found in  fresh water environments (lakes and rivers), fish and seafood
and animals (39).  It has been implicated in cases of traveller's
diarrhea  and consumption of raw oysters (40,41).  There appears to
be no consensus concerning whether or not Plesiomonas shioelloides
is a gastrointestinal pathogen for the normal host, but it is an
opportunistic pathogen for the immunocompromised host.  The organism
has also  been associated with extra-intestinal illnesses such as
bacteremia, meningitis, cholecystitis, osteomyelitis, and pseudo-
appendicitis.  In spite of this, the organism apparently has a low
pathogenic potential (42).   It's potential as a waterborne disease
agent has not been assessed and occurrence data in treated drinking
water are lacking.

      Detection of £. shiqelloides involves the same basic methodology
as for Aeromonas hvdroohila; £. shioelloides  is also oxidase negative.
Isolation can be accomplished using antibiotic supplemented agar media
such as  Salmonella-Shioella agar and KacConkey's agar.  Identification
of lactose non-fermenting, oxidase-positive, potential Plesiomonas
isolates  can be accomplished by use of a biochemical identification
system such as the API-20 E.  A medium  (KS5)  for selective isolation
of P. shiqelloides was reported by Sakata and Todaka (43).  To date,
however,  the use of this medium for direct recovery of P. shiqelloides
from water, including drinking water, has not been evaluated.

Hvcobacterium SDD.

      Hycobacteria have probably been 'emerging' as opportunistic
pathogens in potable water longer than any other genus or group of

512

-------
 bacteria.   Their emergence  has been accelerated  in recent years by
 their occurrence as disease agents in AIDS patients and the  Isolation
 of Hycobacterium spp.  from  chlorinated water sources  in patient care
 areas in hospitals. (11,26).  Diseases caused by  these organisms range
 from skin granulontas,  to .chronic pulmonary mycobacteriosis,  to
 disseminated disease  in AIDS patients or in patients  with underlying
 hematologic disorders.  In  the latter patients,  a case fatality rate up
 to 73% nay occur (11].

       The occurrence  and  incidence of mycobacteria in water,  including
 potable water,  is  well documented and was reviewed by Collins,  el al..
 (29).  In the time period  since that review additional work  has shown
 that potable hot water systems in hospitals contain higher concentra-
 tions of H. avium  than cold water systems (26).  Sixty-nine  percent of
 hot water {T = 52°C to S7°C) sites  sampled contained  H.  avium at
 densities that ranged from  1.0-500 CFU/100 ml (average 141 CFU/100 ml).
 By comparison only 17% of samples from cold water sites contained M.
 aviuB at densities 1.0-2.0  CFU/100 ml (average 1.5 CFU/100 ml).  In
; addition to the recovery  of H, avium serotype 4  from  distribution
 water, this serotype  has  been recovered from a majority of clinical
 specimens of patients with  AIDS in Massachusetts, indicating the
 likelihood of waterborne  transmission.  Aerosolized water  (showerheads,
 etc.) containing mycobacteria provides opportunity for inhalation
 exposure, in addition to  ingestion exposure.

       The use of low  level  chloranination (1 ng/L chloramine residual)
 before the first customer may select for mycobacteria because they
 have been found to be resistant to chlorine at this concentration (12).
 The mycobacteria are, in  general, more resistant to disinfection
 than other types of bacteria.  Havelaar, et al.  (25)  reported that a
 free available chlorine  concentration (hypochlorous  acid, HOCL) of 1
 mg/L was required  to  assure low mycobacteria levels  in bathing water.
 Susceptibility of  mycobacteria to disinfectants  has not been fully
 evaluated.  Pelletier, et aj,. (12) found that free chlorine  at 1.0
 rag/L gave 5 logs of inactivation of a variety of clinical  and
 environmental strains of mycobacteria in an 8 hour period,  but free
 chlorine at 0.15 mg/L had essentially no bactericidal effect on the
 same strains.

       Detection and enumeration of mycobacteria in water  requires the
 use of membrane filtration and selective and inhibitory media.  After
 sample filtration, the membrane filter say or may not be  treated to
 reduce the background of other organisms.  Such treatment  usually
 involves the use of sodium hydroxide, hypochlorite or oxalic acid
 followed by neutralization before culturing the treated membranes.
 Alternatively, the water sample may be treated using  a selective agent
 such as'cetylpyridinium  chloride  {0.04% final concentration) for 24
 hour before membrane  filtration of the water sample  (44).

       The major problem  in examination of water for mycobacteria
 is that an extended time period may be needed to culture  the organisms
 (depending on the  species); up to 30 days at 37°C on  a medium such as
 Middlebrook-Cohn  7H10 agar.  The plates nust be sealed  in a container
 to prevent dehydration of  the medium during incubation.   Detailed
 methodology is available elsewhere  (44,45,46).

 New Candidate Organisms  -  Bacteria

       Anaerobiosoirillum succinciproducens  is an anaerobic or micro-

                                                                      513

-------
aerophillc, motile, spiral shaped rod that has been Isolated froa
diarrhea! feces,  the genus flnaerobiosoirillua was first described
in 1976 by Davis, et at.(47)  and there have been few reports of these
organisms producing disease in humans (3).  This organism has been
associated with human diarrhea and bactereraia.  Malnick, et al_. (3)
reported the  developaent of a median for the isolation of this organism
from diarrhea! feces and blood of humans.  On this medium, the organism
produces uniform, dark-blue, low convex shaped colonies, 1-2 mm
diameter, in  48 h. incubation.  The medium was used to survey human,
canine and feline feces for the presence of anaerobiospirillumlike
organisms,  Because these organisms are anaerobes, isolation requires
anaerobic incubation at 37°C for 48 h.  Previously,  the organisms
were isolated from Skirrow campylobacter medium.  Halnick, et al.
found no AnaerobiosoiriTlum spp. in 527 normal human fetal samples
and obtained  only one isolate from one of 100 selected diarrhea!
samples.  Anaeroblosplrillum spp. were isolated from rectal swabs
of 7 of  10 cats and 3 of 10 dogs.  These results clearly show that
Anaerobic- spirillum spp. are not part of the normal fecal flora of
humans.

      No data is available on the occurrence of these organisms in
water or on their susceptibility to water treatment disinfection
processes.  Currently used methods for bacterial analysis of potable
water would not detect these organisms up because the media used are
not appropriate, and anaerobic rather than aerobic incubation is
required.

Helicobacter  pylori

      He! 1cobacter pylori (formerly Campy!obacter pylori) is an
organism that causes the majority of human gastritis cases and is
associated with human duodenal ulcers.   Infection with the organism
may also contribute to an increased risk of gastric carcinoma.
Persons  at risk for gastric carcinoma have been shown to have a high
prevalence of H. pylon'  infection.  Little information is available
about reservoirs of the organism, routes of transmission, survival
in the environment, occurrence in water, and susceptibility to control
by water treatment processes.

      H- pvlori  infection is widespread  among Peruvian adults fay the
age of 30 and Klein, et  aj..,  {9} reported that municipal water supply
•appeared to  be an  important source of infection among Lima school
children.  Mai, et a],.,  (10)  reported that H. pylori may survive in
the environment  in a dormant  but viable  state.  Using stepwise
increases  in  nutrient concentration and  alteration of physico-chemical
factors, they demonstrated reversion of  H. pylori from the dormant
stage back  into dividing and  infective cells.  This information
suggests the possibility of water as an  environmental reservoir for
this organism. The fecal oral route of  infection is suspected but
not yet  proven.

      The detection of H. pylori by cultural Eethods is difficult
because  the  organism  is  fastidious and 3 or more days incubation are
required for isolation.   If 1t enters the dorsant state, recovery using
cultural  techniques may  be even more difficult.  The association of
H. pylori with the risk  of gastric carcinoma raises interesting
questions  concerning  the  possibility  of  other opportunistic organisms
having  some  association  with  gastrointestinal carcinomas.
514

-------
Vibrio fluvial 1s

      This organism is described in the literature as  a marine
bacterium.  Host cases of gastroenteritis caused  by V.  fluvial ii have
been associated with consumption of seafood.   The isolation of 1,
fluvial is from potable distribution waters from two systems is,
therefore, surprising (48).  Isolates of this organism were obtained
from m-Endo medium, apparently in the absence of  total  coliforms.
It is not known whether or not the identification of this organism
was serologically confirmed by appropriate serotyping  techniques.
V. fluv ial is is the name given' to organisms which were previously
designated as Group F or EF-6.                            .

      V. fluv ialls isolation from fresh surface water  and sediment was
reported by Venkateswaran, et a] . (49).  To isolate total vibrios from
the water, they used a membrane filter and pad preenrichment with
alkaline peptone water for 6 hr at 30°C.   This was followed by transfer
of the membrane filters to thiosul fate-citrate-bile salt-sucrose (TCBS)
agar and incubation at 30°C for 18-20 h.   Subsequent to isolation,  the
organisms were biochemically identified.

      Other than these two reports, very little is known of the
occurrence and distribution of V. fluv ial is in freshwaters that serve
as drinking source waters, or in potable waters.   Presumably, the
methodology of Venkateswaran, et al. (49) could be used for surveying
for the presence of these bacteria in surface and potable waters.
      This organism was not included in the tables because there have
been only a few episodes of waterborne disease attributed to these
organisms, primarily to X. enterocolitica.  These outbreaks have
involved untreated water and only a small number of individuals.  A
review of Y. enterocolitfca prepared by Schiemanri (50) summarized the
current state of knowledge about this organism.

Other Hicrobial Groups

Enteric Viruses

      Due to the variety of viruses that compose this group of micro-
organisms, and the concern about human disease caused by waterborne
exposure, there has been a significant research effort over the past
several years.  These efforts  ave been primarily directed at developing
information on inactivation of enteric viruses by drinking water
disinfectants.  Much less emphasis has been placed on surveying water
supplies for occurrences of enteric viruses, but some such studies have
been done.  A recent article by Hurst (51) reviewed the literature
for studies on the occurrence  of enteric viruses in natural freshwaters
and their removal by conventional drinking water processes.  Recently,
Payment (52) found no human enteric viruses in 100 L volumes of 162
finished drinking water samples.

      Regarding the occurrence of emerging or new human enteric viruses
that may be waterhorne, rotaviruses are a common cause of acute
non-bacterial gastroenteritis  in infants and children (535-  Rotavirus
has been detected in contaminated drinking water and wastewater in
Mexico  (54).
                                                                     515

-------
      Bloch, et  al (55) reported the cell culture isolation of
hepatitis A virus  (HAV) from contaminated well water that was
implicated in a  coimon source outbreak that resulted in 35 cases of
'hepatitis A.  This study provided new information on the stability of
HAV in untreated water since the water samples from which the virus was
isolated were obtained three months after the onset of symptoms in the
index case.  In  addition, water concentrates were stored 8 months at
4°C before they were inoculated into cell  culture.

      The effectiveness of conventional water treatment processes and
disinfection in  the removal and inactivation of enteric viruses
suggests that very little enteric waterborne disease occurs in people
who consume water  that consistently receives full treatment.  However,
when breakdowns  occur in the treatment and/or disinfection processes,
waterborne illness risk is increased.

      A considerably greater risk exists for those people who consume
untreated groundwaters that are subject to surface water contamination.
This is, and will  continue to be, a topic of major concern because of
the potential for  viral contamination of groundwaters.  Because of the
low infectious doses for many viruses (1-2 infectious units) and the
fact that dissemination of infectious viruses may occur without"
detection, it is probable that there is much greater occurrence
of waterborne viral  disease in the population than is presently
recognized.  Efforts currently underway to use genetic probes to detect
viruses in water may provide the means to more easily and reliably
detect enteric viruses in all types of water, including drinking water.
The key question,  however, is - when viruses are detected by gene probe
methods, what does it mean?  How can one tell from probe detection
methods whether  the viruses were viable and infective, or if they were
inactivated, but the UNA or RNA was intact?

Pathogenic Protozoa

      The two enteropathogenic protozoa that have been in the limelight
as waterborne agents for the past several years are Giardia and Crypto-
sporidium.  It is  likely they will continue to receive attention
because of their resistance to inactivation by disinfection and the
recent waterborne  outbreaks attributed to them.  What about other
protozoa that are  enteropathogenic?  Are there any and are they
waterborne?  The answer is maybe!

BlastocYstis hominis

      The history  of this organism has been one of conflicting
descriptions of  it nature and pathogerncity.  This protozoon has proven
difficult to classify taxonomically.  It has been described as an
intestinal yeast and was first named Blastocvstis enterocola in 1911;
in 1912 the name was changed to £. hominis.  However, it has many
characteristics  that are protozoan in nature, and it was reclassified
in 1976.' It may eventually be placed in its own niche within the
protozoa.  This  is based on the diversity of its shapes and sizes,
unique physiology  and some of its reproductive modes.  Also, shared
RNA sequences with other eucaryotic cells confirm that B. hominis is a
protozoan.  Since  reclassification in 1S76, it has been placed in the
family Sporozoa, but several minor characteristics indicate a better
assignment would be to the Sarcodina.  A complete description of the
organism, its characteristics and current classification was given by
Zierdt (15).

516

-------
      In fecal samples,  this complex protozoan appears in a variety of
cell forms that range from yeast size (ca,  7 /m) to giant cells of
20-40 im.  The predominant hollow spherical  form of this organism has
been easily confused with unrelated cells from many animals.  Many
degenerated tissue cells, vegetable cells,  2nd many artifacts resemble
B. hominis.  Hence,  it will be extremely difficult to detect in en-
vironmental samples.  The organism is strictly anaerobic and has no
cyst form, but may have  glycocalyx (capsule).  This suggests that it
may be much more susceptible to disinfection than are Giardia cysts and
Cryptosporl.dium oocysts,

      The most frequent  complaint of blastocystosis patients is intense
abdominal discomfort accompanied with pain.   Diarrhea is not standard
and constipation is common; a variety of other symptoms such as vomit-
ing and fatigue may occur.  Many mild cases  of the disease resolve in
about 3 days without treatment and asymptomatic cases are often seen,

      The occurrence and distribution of fi.  hominis in water and the
environment is unknown.   Isolation of B. hominis has been confirmed
only in humans, monkeys,  apes,  pigs and possibly guinea pigs.  The
incidence rates of B.  hotninis in humans range from 5.3% in Saudi
Arabia (56) to 16% in New York (57).

Hicrosporidia

      Another group of pathogenic protozoa  that may bear watching are
the microsporidia.  These are obligate intracellular parasites found in
many vertebrates including man.   Of the microsporidia two forms are
enteric parasites, Encephalitozoon cuniculi  and Enterocytozapn biemisi.
These organisms have ellipsoid shaped spores approximately 2.5 by
1.5 pi for £. cuniculi and 1.5 by 0.5 pn for E. bienusi.  There are
known fecal-oral transmission mechanism cycles for these organisms
similar to Giardia and Crvptosporidium.  These organisms may be latent
parasites in many people, which would explain why, with highly immuno-
compronrised people (AIDS patients),  the microsporidian parasites are
being seen more frequently.  At present, there is no effective drug
therapy.

      There is no information available regarding their occurrence and
distribution in water or other environmental samples.  Likewise, the
effectiveness of conventional  treatment and  disinfection processes for
removal  and inactivation of the cysts of these organisms is unknown.

Blue fireen Alcrae fCyanobecteria)

      An alga has been associated with traveler's diarrhea in travelers
to the Carribean, Hexico, South America, India, or Southeast Asia,
and several AIDS patients (17).   The organism observed in stool
specimens was described  as resembling a coccidian oocyst or fungal
spore, or a "large CryptosDoridium".  Electron microscopy revealed cell
characteristics that indicate the organism  may be a species of blue
green algae similar in structure to ChloreTla.  .The immunologically
competent patients with  traveler's diarrhea  complained of nausea,
vomiting, anorexia,  weight loss and explosive, watery diarrhea.  In
most patients, the symptoms cleared within  two weeks along with a
decrease in the number of cyst!ike bodies in the stools.  The
The organisms may have been acquired by consumption of improperly
filtered water.
                                                                    51?

-------
      An outbreak of diarrhea!  illness occurred in 21 people who lived
or worked  in  a  hospital  dormitory in Chicago, Illinois (58).  The
outbreak occurred following  transient failure of a building water tank
pump.  Examination of patient stools for parasites resulted in the
observation of  coccidian-like bodies in 9 of 17 stools.   Light
microscopy examination of water tank sediment revealed occasional
coccidian-like  bodies.   The  Centers for Disease Control  identified the
organism as a cyanobacterium or cryptosporidium-like organism.

      The  specific identify  of  the organism, its occurrence and distri-
bution in  water and its  susceptibility to removal and inactivation by
water treatment process  is unknown,

      There have been numerous  reports of wildlife, livestock and
pet deaths due  to algal  toxins  in water (59).  Cyanobacteria (blue
green algae)  are known to produce several toxins, including neuro-
toxins, hepatotoxins and contact irritants.  The unusual  aspect of the
diarrheal  illness outbreaks  cited above is that the diarrhea] illnesses
were caused apparently by cyanobacterium infection rather than by
ingestion  of  toxins elaborated  and released into the water by the
cyanobacteria,  but this  is by no means certain.

Conclusions

      A variety of microbial opportunistic pathogens and  primary
pathogens  may be found in water used as drinking source water.  Many of
these organisms have also been  isolated from treated drinking water,
particularly  the bacterial opportunistic pathogens.  Essentially all of
the other  microbial opportunists have been found in potable water that
was contaminated due to  inadequate treatment or breakdowns in the
treatment  processes.

      Perhaps none of the organisms covered herein pose a significant
or high health  risk to most  exposed healthy individuals.   However,
accumulating  evidence indicates that all of these organisms can be
severe health risks for  imraunocoiBprotaised individuals. The population
of inmunocompromised individuals in the U.S. population is steadily
increasing, partially due to the aging of the population, and partially
due to the increasing number of people receiving medical  treatment for
long term  diseases such  as AIDS and cancer.  It seems likely that we
will be seeing  an increasing emergence of diseases due to waterborne
opportunistic bacterial  pathogens in people less iraunologically com-
promised than are AIDS patients.

      He usually focus on single species of organisms when talking
about determining the causative agent in a disease outbreak.  It is
possible,  perhaps probable,  that in evaluating human health risks due
to drinking water, we should be considering the collective impact of
simultaneous  and repeated exposures to variable levels of several
opportunistic bacteria,  rather  than focusing on individual organisms.
The collective  adverse health impact »ay be far greater than the
impacts of individual organisms would suggest.

      In a recent study  by Payment, gt al_. (60), excess gastro-
intestinal illness was found in consumers of treated drinking water
compared to consumers who drank the sane water that was further
treated by reverse-osmosis.  However, it was also suggested that
heterotrophic bacteria which grew in drinking water produced by
reverse osmosis filtration units, and were enumerated on  R2A medium

518

-------
incubated at 35°C, may be associated with low numbers  of gastro-
intestinal illnesses  (61).  Heterotrophic bacteria?  densities in the
treated distribution  water ranged from  5.2  x 10l  to  4.3  x 102 CPU/ml
(35°C)  in one system  and from 2.2 x 10l to 9.5 x  if CFU/ml (35°C) in
the other system.  Heterotrophic bacterial  densities in  the R-0 water
ranged from 0 to  107  CFU/ml, but'most contained between  i04-105 CFU/ml.

      Significant differences between the exposure experienced by the
consumers of treated  distribution water versus  consumers of R-0 treated
water were:  1} treated distribution water  contained lower densities of
organisms/ml, but a greater diversity of types  of bacteria based on
organisms isolated and identified (14); and 2}  R-0 water contained high
densities of organisms, but of very limited diversity, sometimes only
one bacterial type or species at a time (48).   Overall,  data from
studies by Payment, et aj.. suggest that they saw  increased illnesses
in R-0 water consumers caused by ingestion  of water containing high
densities of one .or a few types of bacteria,  and  in  consumers of
treated distribution  water by ingestion of  water  containing low
bacterial densities but having higher diversity of bacterial  types.

      An alternative  possibility is that undetected  enteric viruses may
have been responsible for the observed  or reported illesses,  not the
heterotrophic bacteria.

      Thus, the collective impact of exposure to  many kinds of
bacteria, some of which are opportunistic pathogens, in  drinking water
may result in measurable disease, just  as exposure to an infective
dose of a single  species can result in  disease.

      Are there any new pathogens in drinking water?  The answer is
yes!  So-e are new in the traditional usage of  the term  new.   Others
are new in the sense  of dawning recognition of  pathogenic potential of
organisms that, individually and collectively,  have  not  been considered
pathogenic in the past.              :

ACKNOWLEDGEMENTS

      I thank Dr. Eugene W, Rice and Mr. Terry  Covert for their review
and comments on this  manuscript, and Mrs. Elizabeth  Creamer for her
able assistance in the preparation of this  manuscript.

                             DISCLAIMER

      This paper has  been reviewed in accordance  with  the U.S.  Environ-
mental  Protection Agency's peer and adminsitrative review policies and
approved for presentation and publication.

REFERENCES

1.    Olson,  B. H.  1989.  The Safety of Our Drinking  Water -  Reason
      for Concern But Not Alarm.  New England J.  Hed.  320(21): 1413-1414
      {Kay 25).

2.    Rose, J. B.  1990.  Emerging Issues for the Microbiology of
      Drinking Water.  Water/Engrg. Management  p. 23-26, 29.

3. *   Ralnick, H., K. Williams,  J. Phil-Ebosie  and A.  S. Levy.   1990.
      Description of  a Medium for Isolating Anaerobiospiri11 urn spp.,
                                                                    519

-------
      A Possible Cause of Zoonotic Disease,  from  Diarrhea!  Feces
      and Blood of Humans and Use of the Hediua. in  a  Survey of
      Human, Canine, and Feline Feces.  J. Clin.  Hicrobiol. 28(6):
      1380-1384.

4.    Schubert, R. H. W.  1991.  Aeromonads and Their Significance  as
      Potential Pathogens in Water.  J. App1.  Bacteriol.  Symp. Suppl.
      70:131S-135S.

5.    Van der Kooij, D.  1988.  Properties of Aeromonads  and Their
      Occurrence and Hygienic Significance in Drinking Water.  Zbl.
      Bakt. Hyg. B 187:1-17.

6.    Blaser, J. H., 0. 1.  Penner and J. G. Wells.   1982.  Diversity of
      Serotypes in Outbreaks of Enteritis Due to  Campy!obacter
      .ie.luni.  J. Infect. Dis. 146(6) :826.

7.    Skirrow, H. B.   1982.  Campy!obacter Enteritis  - The First Five
      Years.  J. Hyg.  (Camb.) 89:175-184.

8.    Dev, V. J., H. Main and  I. Gould.   1991. Waterborne Outbreak of
      Escherichia coll 0157.   Lancet  337:1412 (June 8).

9.    Klein, P. D., Gastrointestinal  Physiological  Working Group,
      D. Y. Graham, A. Gaillour, A. R. Opekun and E.  0.  Smith.
      1991.  Water Source as  a Risk Factor for Helicobacter pylori
      Infection in Peruvian Children.  Lancet 337:1503-1506
      (June Z2).

10.   Mai, U.  E. H., M. Shahamat and  R.  R. Colwell.  1990.  Survival
      of Helicobacter  pylori  in the Environment in  a Dormant but
      Viable Stage.  Revista  Espenola de Enfermedades Digestivas.
      78  (Suppl.  1); 17 Abst.  P-18, November.

11.   Du Moulin,  G.  C.,  I.  H.  Sherman, D. C. Hoaglin and K. D.
      Stottmeier.   1985.  Hycobacterium  aviuin Complex, An  Emerging
      Pathogen  in Massachusetts.  J.  Clin. Hicrobiol. 22(1):
      9-12.

12.   Pelletier,  P.  A., G.  C.  du Moulin  and K. D. Stottmeier.   1988.
      Hycobacteria  in  Public  Water Supplies: Comparative Resistance
      to  Chlorine.   Hicrobiol.  Sci.  5(5):147-148.

13.   Van  Loon,  F.  P.  L., 2.  Rahim,  K. A. Choadhury, B. A. Kay  and S.
      A.  Rahman.   1989.   Case Report  of  Plesiomonas  shiqelloides -
      Associated  Persistent Dysentery and Pseudomembranous Colitis.
      J.  Clin.  Hicrobiol.  21(8):1913-1915.

14.    Payment,  P.,  F.  Gamache and  G.  Paquette.   1989.  Comparison of
      Data from Two Water Filtration Plants and  Their Distribution
      System.   Water Sci.  Tech.  21(3):287-289.

15.   Zierdt,  C.  H.   1991.   Blastocvstis hominis - Past  and  Future.
      Clin.  Hicrobiol. Rev. 4(l):61-79.

16.    Ashkenazi,  S.  and L-  K. Pickering.  1991.  New Causes  of
       Infectious Diarrhea.   Eur.  J. Clin. Hicrobiol.  Infect.  Dis.
 520

-------
 17.    Long,  E.  G.,  A.  Ebrahimzadeh,  E.  H.  White,  B.  Swisher and C. S.
       Galloway.   1990.   Alga Associated with Diarrhea in Patients with
       Acquired  Immunodeficiency  Syndrome and in Travelers.  J. Clin.
       Hicrobiol.  2J(6): 1101-1104.

 18.    Burke, V.,  J, Robinson, H. Gracey, D.- Peterson and K. Partridge.
       1984.   Isolation of Aeromonas  hydrophi'la from the Metropolitan
       Hater Supply:  Seasonal Correlation with Clinical Isolates.
       Appl.  Environ. Microbiol.  48:361-366.

' 19.    Picard, B,  and Ph. Goullet.   1987.  Seasonal Prevalence of
       Nosocourial  Aeromonas hydrophila Infection Related to Aeromonas in
       Hospital  Water.   3.  Hosp.  Infect. 10:152-155.

 20.    Havelaar,  A.  H., J.  F. H.  Versteegh and M.  During.  1990.  The
       Presence of Aeromonas  in Drinking Water Supplies in the
       Netherlands.   Zbl. Hyg. 190:236-256.

 21.    LeChevallier, M. H., T. H. Evans, R, J. Seidler, 0. P. Daily, B.
       R. Herrel,  0. M. Rollins and S'. W. Joseph.   1982.  Aeromonas
       sgbria in Chlorinated Drinking Water Supplies.  Microb. Ecol.
       8:325-333.

 22.    Havelaar, A.  H,, H.  During and J. F. H. Versteegh.  1987.
       Anipicillln Dextrin Agar Medium for the Enumeration of Aeromonas
       Species in Hater By Hembrane Filtration.  J. Appl. Bact.
       62:279-287.

 23.    Versteegh,  J. F. M., A. H. Havelaar, A. C. Hockstra and A.
       Visser.  1989.  Coraplexing of Copper in Drinking Water Samples
       to Enhance Recovery of Aerpmonas and Other Bacteria.  J. Appl.
       Bacteriol.  67:561-566.'

 24.   Taylor, D.  N., H. Brown  and K. T. KcDermott.   1982.  Haterborne
       Transmission of Campy!obacter Enteritis.  Hicrob. Ecol. 8:
     '  347-354.

 25.   Havelaar, A. H.» 1. 6. Berwsld, D. J. Sroothuis  and J. G.
       Baas.  1985.  Hycobacteria in Semi-Public Swimming-Pools
       and Whirlpools.  Zbl. Bakt.  Hyg., I. Abt. Orig.  B 180:
       505-514.

 26.   Du Moulin, 6. C., K. D.  Stottmeier, P. A. Pelletier, A. Y.  Tsang
       and J. Hedlcy-Whyte.  1988.   Concentration of  Hvcobacterium
     •  avium by Hospital Hot Water Systems.  J. Amer. Med. Assoc.
       . 260(111:1599-1601.

 27.   Blaser, H. J., P. F. Smith,  W.-L. W. Wang and  J. C. Hoff.   1986.
       Inactivation of Campvlobacter .ie.iuni by  Chlorine and Chloraraine.
       Appl.  Environ. Hicrobiol.  51:307-311.

 28.   Hegraud, F.  and R. Serceau.  1990.  Search  for Caapylobacter
       Species in the Public Water Supply  of  a  Large  Urban Community.
       Zbl.  Hyg.  185:536-542.

 29.   Collins, C.  H., J. H. Grange and  H. D. Yates.   19S4.
       Hvcobacteria in Water.  J. Appl.  Bacteriol. 57:193-211.
                                                                      521

-------
30.   Carson,  L. A., N. J. Peterson, H. S. Favero and S.  H.  Aguero.
      1978.   Growth Characteristics of Atypical  Hycobacteria in Water
      and Their Comparative Resistance to Disinfectants.   Appl.
      Environ. Hicrobiol. 36(6):839-846.

31.   Sobsey,  M. 0.  1989.  Inactivation of Health Related Micro-
      organisms in Water by Disinfection Processes.  Water Sci.
      Techno!. 21(3):179-195.

32.   Hascher, F, F. F. Reintholer, D. Stunzner and B. Lamberger.
      1988.   Aeromonas Species  in a Municipal Water Supply of a
      Central  European City:  Biotyping of Strains and Detection of
      Toxins.   Zbl. Bakt. Hyg.  B 186:333-337.

33.   Mentzing, L. 0.  1981.  Waterborne Outbreaks of Canrovlobacter
      Enteritis in Central Sweden.  Lancet 11: 352.

34.   Vogt,  R. L., H.  E. Sours, T. Barrett, R. A. Feldman, B. S.
      Dickinson and L. Witherell.  1982.  Campy!obacter Enteritis
      Associated with  Contaminated Water.  Ann. Internal. Med. 96:
      292-296.

35.   Rautelin, H., K. Koota, R. von Essen, M. Jahkola, A. Siitonen
      and T.  U. Kosunen.   1990.  Waterborne Carapylobacter .ie.iuni
      Epidemic  in a Finnish Hospital for Rheumatic Diseases.
      Scand.  J.  Infect. Dis.  22:321-326.

36.   Geldreich, E. E., K. R. Fox, J.  A. Goodrich, E. W.  Rice, R. H.
      Clark and D. Swerdlow.  (in press).  A Water Supply Connection
      in the  Cabool, Missouri Outbreak.  Water Res.  (in press).

37.   HcGowan, K.,  F.  Wickersham  and N. Strockbine.   1989.
      Escherichia coli 0157:H7  from Water.   Lancet 1:967-968.

38.   Anonymous.   1990.   Food Safety and Toxicology.  Health Environ.
      Digest  4(Aug./Sept.):l-6.

39.   Wadstrom, T.  and A.  Ljungh.   1991.  Aeromonas  and Plesiomonas
      as Food -  and Waterborne  Pathogens.  Intn'l. J. Food Hicrobiol.
      12:303-312.

40.   Holmberg,  S.  D., I.  K.  Wachsmuth, F. W. Hickman-Brenner, P. A.
      Blake and  J.  J.  Farmer  III.   1986.  Plesiomonas Enteric
      Infections  in the  United  States. Ann.  Intern. Med. 105:
      690-694.

41.   Centers for  Disease Control.   1989.  Aquarium  - Associated
      Plesiomonas  shiqelloides  Infection  - Missouri.  Morbidity
      and Mortality Weekly Report 38(36):617-619.

42.   Abbott, S.  L.,  R.  P. Kokka and J. M. Janda.  1991.  Laboratory
      Investigations  on  the Low Pathogenic Potential  of Plesiomonas
      shigelloides.   J.  Clin. Hicrobiol.  29(1):148-153.

43.   Sakata, T.  and  K.  Todaka.  1987.  Isolation  of Plesiomonas
      shiqelloides  and It's Distribution  in  Fresh  Water Environ-
      ments.   J.  Gen.  Appl.  Microbiol. 33:497-505.
522

-------
44.   Du Moulin,  G.  C.  and K.  D.  Stottmeier.  1978.  Use of Cetyl-
      pyridiniun Chloride in the  Decontamination of Hater for
      Culture of Mycobacteria. Appl.  Environ. Microbiol. 36(5):
      771-773.

45.   Songer, J.  G.   1981.  Methods for Selective Isolation of
      Mycobacteria from the Environment.  Can. J. Microbiol.
      27:1-7.

46.   Engelbrecht, R. S. and C. N.  Haas.  1977.  Acid Fast Bacteria
      and Yeasts as Disinfection  Indicators:  Enumeration Methodology.
      Proc. AVft/A Water Quality Tech.  Conference, Dec. 1976,
      Kansas City, MO.

47.   Davis, C. P.,  D.  Cleven, J. Brown and E. Balish.  1976.
      Anerobiosoirill urn, A New Genus  of Spiral-Shaped Bacteria.
      Int. J. Syst.  Bacteriol. 26:498-504.

48.   Payment, P.  1989.  Bacterial Colonization of Domestic
      Reverse-Osmosis Water Filtration Units.  Can. J. Microbiol.
      35:1065-1067.

49.   Venkateswaran, K., C. Kiiyukia, M. Takaki, H. Nakano, H.
      Matsuda, H. Kawakami and H. Hashimoto.  1989.  Appl. Environ.
      Microbiol. 55(10):2613-2618.

50.   Schieeann, D. A.  1990.  Yersinia enterocolitica in Drinking
      Hater,  pp. 322-339.  In:  McFeters, G. A. (ed.) Drinking
      Water Microbiology.  Springer-Verlag, N.Y.

51.   Hurst, C. Jt   1991.  Presence of Enteric Viruses in Freshwater
      and Their Removal by the Conventional Drinking Water Treatment
      Process.  Bull. World HHh. Org. 69(1):113-119.

52.   Payment, P.  1991.  Fate of Human Enteric  Viruses, Coliphages,
      and Clostridium perfringens During Drinking-Water Treatment.
      Can. J. Microbiol. 37:154-157.

53.   Tyler, J.   1985.  Occurrence in Hater of Viruses of Public
      Health Significance.  J. Appl.  Bacteriol.  (Symp. Supplement)
      37S-46S.

54.   Keswick, B. H., C.  P. Gerba, H. L. Dupont  and J. B. Rose.
      1984.  Detection  of Enteric Viruses  in  Treated Drinking  Water.
      Appl.  Environ. Microbiol.  47:1290-1294.

55.   Bloch, A.  B.,  S.  L. Stramer, J. D. Smith,  H. S. Margolis,
      H.  A.  Fields,  T.  H. McKinley, C.  P.  Gerba, J.  E. Maynard,  and
      R.  K.  Sikes.   1990.   Recovery of  Hepatitis A Virus from  a
      Hater Supply Responsible for a Common Source Outbreak  of
      Hepatitis  A.   Amer. J.  Public Health  80:428-430.

56.   Qadri,  S.  M. H.,  G. A.  Al-Okaili  and  F. Al-Dayel.   1989.
      Clinical Significance of Blastocvstis hominis.  J. Clin.
      Micro&iol.  27(11):2407-2409.

57.   Sheehan, D. J., B.  J. Raucher  and J.  C. McKitrick.   1986.
      Association of Blastocvstis  hominis  with  Signs  and Symptoms
      of Human Disease.   J. Clin. Microbiol.  24:548-550.
                                                                     523

-------
58,   Wurtz, R.,  F.  E, Kocka,  C.  ICallick,  C. Peters and E,
      Oacumos.   1991.  Blue Green Algae Associated with a
      Diarrheal  Outbreak.   Abst.  C-21,  Abstracts 91st Gen. Mtg.
      Araer. Soc.  Microbiol,, Kay  5-9,  1991, Dallas, TX.

59.   Repavich,  M. M., M.  C, Sonzogni,  J.  H. Standridge, R. E.
      Wedepol and L. F. Melsner.   1990.  Cyanobacteria (Blue  -
      Green Algae) in Wisconsin Haters:  Acute and Chronic
      Toxicity.   Water Res.  24(2):225-231.

60.   Payment, P., L, Richardson, J.  Siemiatycki, R. Oewar, H.
      Edwardes and E. Franco.   1991.   A Randomized Trial to Evaluate
      the Risk of Gastrointestinal  Disease Due to Consumption of
      Drinking Water Meeting Current  Microbiological Standards.
      Anier. J. Publ. Health.   81:703-708.

61.   Payment, P.  E. Franco, L. Richardson and J. Siemiatycki.
      1991.  Gastrointestinal  Health  Effects Associated with the
      Consumption of Drinking  Water Produced by Point-of-Use
      Domestic Reverse- Osmosis Filtration Units.  Appl. Environ.
      Hicrobiol.  57(4):945-948.
524

-------
         Table 1.  Opportunistic Bacterial  Pathogens Isolated
                              from Drinking Water
Acinetobacter spp.
Achromobacter xvlosoxidans
Aeromonas hvdrophila*
Bacillus spp.
Camovlobacter spp.*
Citrobacter spp.
Enterobacter aeroqenes
E. aoglomerans
E. cloacae
Flavobacterium meningosepticum
Hafm'a alvei
Klebsiella pneumoniae*
LeoioneTIa pneumoohila*
Horaxella spp.
Hvcobacterium spp.
Pseudomonas aeruginosa*
Pseudomonas spp. (non-aeruginosa)
Serratia fonticola
S. Tiouifaciens
S. marcescens
Stachvlococcus spp.*
Vibrio fluvial is*

* indicates the organism may be a primary (frank) pathogen
                                                                     525

-------
Table 2.  Summary of Emerging and New Potentially Waterborne
                        Opportunistic Pathogens
Organism Old/Emerging New Candidates
Anaer_QblospJ_rJ lltmt sued n 1 c |producens ?
Aeroffionas IPJK X
CampYloba^ter. spp. X
EjjchgrJchla c.o.1,1 0157;H7 X
He.11<;ftha.c.t.C-C Mlfid. ' ? •
Mycopacterium spp. X
Plesjomonas shlgelloldes ?
Vibrio fluvial is ?
Reference
3
4
5
6
7
8
9
10
11
12 •
13
14

-------
               Table 3,   Summary of New or Emerging Potential Waterborne Mieroblal  Pathogens
Viruses


Protozoa
               Rotawiruses (Group A)


               Blastoc-vstls homj.nls.

               Mlcrosporldia
New Candidate




       X

       X
15

16
Algae
               Blue Green Algae
               (Cyanobacteria)
                             17

-------
in
N
00
                                    Table  4.   Densities  of Selected Opportunistic Pathogens
                                                           in Drinking Mater
pjflatLjsfl
Aeromonas hydroohjla
Aerononas spp.
spbr1..a
ELaiiojMUHs liUaallaidas
CMfiKlfiMfitej: spp.
Hveobacterluni spp.
M- flprdgriaB, M- avlum. H. fortultum
DensltoJ-CFU/Muiiel
100 - 150/100 ml
10 - 80/100 ml (mains water)
300/100 ml (storage tank)
600 - 800/L (ground water)
0 - 100/L (deep ground water)
0 - 3,300/100 ml
1 - 1,900/100 ml
1 - >H6/100 ml
<1 - >100/100 ml
<1 - 16/100 ml
«. *
- HA **
1.5/100 ml (cold water)
111/100 ml (hot water)
200-500/100 ml (unchlor. ground water) '
1 - <80/L (tapwater)
1 - >500/100 ml
Reference
18
19
If
4
(1
20'
21
18
22
23
24
12
M
25
26
        * -,  no Information  available
       ** NA,  not  available,  detection  usually  by  enrichment,  not  direct isolation.

-------
                               Table  5.   Disinfectant  Sensitivity of Emerging and New Candidate
                                                   Waterborne Opportunistic Pathogens
Organism
An a ero b i o s pi r 11 1 urn syce 1 n_i c iproducen s
Cafflpjjflbacter, spp.
Disinfectant
Any
Free chlorine; >0.3 mg/L
Free chlorine; 0.1 mg/L
Free chlorine
Comments on
Survival 	 lime
Sensitivity
unknown.
Short, increases
with decreased
residual .
>4 logs removal in
5 win (? 4°C,
Sensitive; rapidly
Reference
18
5
27
28
       Eschierichia coll 0157:H7
       PAesiofflonaji shigelloides
       Hvcobacterium spp.
           Any
           Any
Freo chlorine; 1.0 mg/L
inactivated unless
high level of
organic contami-
nation present.

Sensitivity probably
similar to other
enteric bacteria.

Unknown, but suspected
to be more sensitive
than Aerojnonas.

3 Mvcobacterlum spp.
were eliminated in
fl hrsj.
12
01
8

-------
                    Table  5  (Cont'd.)  Disinfectant  Sensitivity  of Emerging  and  New Candidate
                                           Waterborne Opportunistic Pathogens
Hycobacterium spp. (Cont'd,}
llejicobacter pylori

Rotavlrus, human


Blastocvstis hoffiinis
      DJsJnfectant

         Free chlorine; 0.15 mg/L



         Chlorine; 0,1-1,0 mg/L
                                       Free chlorine; 0.3 and
                                                 0.7 mg/L
      Comments on
     Survi'vaLTlrne                    Reference,

       No bactericidal effect.     "
       Inverse relationship
       between disinfectant
       residua] and CFU/100 ml.
       3 species survived         29
       7 days.
       Survived 60 min.            30
      Any



      Any

Chlorine, free,  0.75 mg/L


      Any
Sensitivity unknown, but probably
similar to that of vegatatlve
enteric bacteria cells.

Sensitivity unknown.

Approx. 5% survival after
60 minutes,

Sensitivity unknown.
31*
Blue-Green Algae
 (Cyanobacteria)
      Any
Sensitivity unknown
* Excellent review of disinfectant inactivation of viruses and other health related microorganisms 1n water.

-------
                                   TECHNICAL REPOHT DATA
                            fftesjt nsdhstoacti&u on At nvtn? btfoft tomfttl?"
 1. REfORTNO.

  EPA/600/A-937022
2.
 I, TiTLt AND SUBTITLE
                                                           !. REPORT OATS
 Pathogens  In  Drinking Hater - Are There Any New Ones?
                             t. f£«f ORMSHQ ORGANIZATION COOS
7. AUTHORISJ

  Donald J. Reasoner
                             3. PERFORMING ORGAM5ZAT1CN REPORT NO.
 I. f ERFORMJNG ORGANIZATION NAME AND ADDRESS

   Chief, Microbiological  Treatment Br.
   Drinking Water Res.  Div.,  RREL
   U.S. EPA, Cincinnati, Ohio  45268
                             10. PROGRAM ELEMENT MO.
                             II. CON TRACT/GRANT WO.
 12. SPONSORING AGENCY NAME AND ADDRESS

   Risk Reduction Engineering Laboratory—Cincinnati,  OH
   Office of Research--and Development
   US  Environmental Protection Agency
   Cincinnati,  OH 45268
                             13. TYPE OF REPORT AND PERIOD COVERED
                              published paper
                             14. SPONSORING AGENCY CODE
                               EPA/600/14
 is. SUPPLEMENTARY NOTES project Officer = Donald Reasoner   (513)  569-7234
 Proceedings  of the American  Water Works Association, Water Quality Technology
 Conference,  Orlando, FA, 11/10-14/91, 5:509-530	
 12. ABSTRACT
      Since  1976 three newly recognized  human pathogens have  become familiar
      to the drinking water industry as waterborne disease agents.  These are:
      the  legionnaires  disease  agent,  Leqionella  pneumophila  and  related
      species; and two protozoan pathogens, Giardia lamblia and Cryptosporidium
      parvum, both  of  which form highly disinfectant  resistant cysts that are
      shed  in the  feces of infected individuals.   The question  frequently
      arises  -  are  there other emerging  waterborne pathogens  that  may pose a
      human  health  problem that the drinking water industry will  have to deal
      with?   This  paper will  review the  current state  of knowledge  of the
      occurrence and incidence of pathogens and opportunistic  pathogens other
      than  Leqionella, Giardia  and Cryptosporidium in treated and untreated
      drinking   water.    Bacterial   agents  that  will  be  reviewed  include
      Aeromonoas,   Pseudomonas,   Campvlobacter,  Hycobacteriunu  Yersinia  and
      Plesiomonas.   Aspects of detection of these  agents  including detection
      methods and feasibility of monitoring will  be addressed.
 7.
                               KEY WORDS AND DOCUMENT ANAS.YSSS
                  DESCRIPTORS
                                             b.lDENTIFlERS/OPEH ENDED TERMS
                                         c.  COSATi Fie!d/Croup
     protozoa
     viruses
     bacteria
                                             Drinking water quality,
                                             bacterial  occurrence,
                                             opportunistic pathogens,
                                             pathogens
 S. DISTRIBUTION STATEMENT
  Release to Public
CFA F«m 2220-1 {R«T. X-773   PHEVJOUS COITION is OBSOLETE
                                             IS. SECURITY CLASS [Tlia Report!
                                                 Unclassified
                                          21. NO. OF PAGES
                                            24
                20. SECURITY CLASS iTJiispsjr}

                    Unclassified
                                         22. PRICE

-------