Académique Documents
Professionnel Documents
Culture Documents
Researchers:
Honghong, Ma. Angelie T.
Jangad, Kristy Lou P.
Maus, Mae Ann J.
ABSTRACT
Bacteria are the simplest life form and are the most numerous organisms with
respect to the number of species and total biomass (Gerardi 2006). Not all bacteria are
pathogenic, some kinds of bacteria are very helpful to humans and a part of normal flora
in our bodies. However, pathogenic bacteria are able to lead sickness and possibly
even death, and usually can be found among rivers.
A study shows that in other countries specifically in North West province of South
Africa, there were cases of rivers that been impaired partly due to frequent
contamination of water sources with a number of pathogenic microorganisms (Meays et
al. 2004). Some rivers like the case when: Tiete River suddenly receives large water
volumes from its tributaries such as the Aricanduva River or the Pinheiros River, which
unload thousands of cubic meters in a few minutes. The resulting flood wave rapidly
raises the Tiete River to a new level, then transports large numbers of pathogenic
bacteria in the densely populated areas along the river (Abraham 2011).
Bacteria from the river can also transmit different kinds of diseases in its
surrounding areas; such as bacillary dysentery, viruses, fungi, protist and Animalia.
Evidences have shown that bacillary dysentery caused by Shigella bacteria found in
rivers alone is responsible for approximately 165 million cases of bacterial diarrheal
diseases annually. Of this, 163 million are in developing countries and 1.5 million in
industrialized ones accounting for an estimated 1.1 million death cases each year
(Sharma et al. 2010). There are a large number of bacteria, viruses, fungi, protist and
Animalia have been identified to be pathogenic for humans and the majority is water
borne which can be transmitted from rivers and a study from 2001 compiled 1415
pathogens (Abraham 2011).
Pathogenic bacteria that develops in rivers that can cause diseases can also be
the effect of poor sanitation and throwing waste products along the river which can also
cause fecal contamination in freshwater. Wastewater effluents play an important role as
sources of fecal contamination in freshwater environments. These non-point sources of
fecal pollution are broadly distributed in both urban and agricultural areas. Standard-
based water quality assessment is an essential component of monitoring programs for
the protection of human health. Microbiological indicators such as total coli forms, fecal
coli forms and fecal streptococci are widely used both in monitoring programs of this
kind and in human health risk assessment. These indicators may also point to the
presence of other non-bacterial pathogens such as enteric viruses and parasitic
protozoans (Dangelo and Pierdominici 2008). Pathogenic bacteria may include:
Klebsiella pneumoniae although found in the normal flora of the mouth, skin and
intestines, it can cause destructive changes to human lungs if aspirated (Ryan 2004). In
most cases, Klebsiella infections are seen mostly in people with a weakened immune
system. Most of the people that are affected is believed to have impaired respiratory
host defenses, including persons with diabetes, alcoholism, malignancy, liver disease,
chronic obstructive pulmonary diseases(COPD). The most significant source of patient
infection is the feces, followed by contact with contaminated instruments.
Escherichia coli which is commonly found in lakes, rivers, swimming pools and
soil from fecal sources, in most cases coliforms that do not cause illness. When E. coli
causes serious problems with the blood or kidneys symptoms may in includes: Pale
skin, fever, weakness, bruising and passing only small amounts of urine (May 2012).
Enteric pathogens in environmental surface waters originate mainly from effluent
discharge from sewage treatment plants (E.M Anastasi et al 2012).
Some bacteria may be introduced into the river water path through untreated and
treated waste waters. These are the predominantly Escherichia coli, and intestinal
enterococci, which are excreted by humans and warm blooded animals because these
bacteria pass sewage treatment plants to a great amount and survive for a certain time
in the aquatic environment. Good water quality will contribute to securing the drinking
water supply, as well as provide economic benefits by contributing towards the
protection of fish populations and preservation of biodiversity (Dragun and Kapetanovic
2011). The spread of potential pathogenic enterococci appears to be the manifestation
of a complex network of ecological processes and associated factors in the landscape
of the river Ganga (Lata et al. 2009).
Some waterborne bacteria like in the rivers are anti-microbial resistance and this
resistance becoming commonly found in hospitals and communities and also the need
of antibiotics will continue to go higher and increase. A study shows that anti-microbial
resistance like E. coli revealed multiple drug resistance in this most deadly strain of
prevalent in low hygienic areas, slum areas and urban-rural interfaces. Basically,
bacteria borne, resistance is more dominant, and hospital and community acquired
resistance is becoming more common. The need for antibiotics will thus continue to be
high and will increase because of an ageing population (Lawrence and Jeyakumar
2013).
The review of literature suggests that pathogenic bacteria found in the rivers can
cause various diseases due to its surroundings, poor sanitation, throwing of waste
products and waterborne bacteria are also antimicrobial resistance. However, there are
different kinds of pathogenic bacteria present in the rivers that can transmit different
kinds of diseases and this bacterias having the largest population in the rivers have the
tendency to cause the diseases mentioned.
Theres no such study that has undergone Microbial Identification in Pala-o
Riverside, Iligan city. The study seeks to identify the possible microorganisms found in
the river of Pala-o Riverside Iligan City. Specifically it aims to identify the different
pathogenic bacteria and its implications to human health.
Methods and Design
Design
The study utilized the descriptive type of research since the researchers seeks to
identify pathogenic bacteria in the river of Pala-o Riverside of Iligan City. The research
was conducted at the laboratory since possible pathogenic bacteria cannot be seen by
the naked eye. The researcher used biochemical tests, gram staining method,
coagulase test and oxidase test.
Research Locale
This study was conducted at Pala-o riverside, located at the Barangay Pala-o
542. Pala-o is one of the barangays outlying the highly urbanized area of Iligan City.
The river has a length of 860 meters and a width of about 30 to 40 meters. The river
originates from Pugaan Iligan City.
Instruments
The following culture media used in the research study are; Blood agar,
McConkey agar, Salmonella Shigella agar, Thiosulfate Citrate and Blue Sucrose agar.
Biochemical methods such as; coagulase, oxidase and the gram staining method were
used to classify the specific microorganism. The different Biochemical test for the
identification of microorganism includes the Triple Sugar Iron (TSI), Lysin Iron Agar
(LIA), Methyl Red (MR), Motility Testing, Indole Production, Citrate Utilization, Urea
Hydrolysis, Decarboxylase and Ornithine Decarboxylase. Farther the bacterias were
identified through microscopy.
Specimen Collection
With the use of gloves, and mask, the specimens were collected in a 236
milliliters sterile bottle. Twenty-four water samples were obtained. Twelve from
the morning samples as well as from the afternoon. The river was divided into
three streams; upper, middle and down. The streams were divided into four
stations each measured 10 meters horizontally and four water samples were
collected. Water samples were brought at the Medical Technology laboratory of
Adventist Medical Center College for experimental procedure.
Bacterial Culture and Isolation Method
The following agar were used; MacConkey agar, Salmonella Shigella agar,
Thiosulfate Citrate Bile Salt Sucrose agar and Blood agar for isolation and
bacterial growth. Sterile micropipettor blue tip was used as the instrument for the
inoculation of 1000m of the water samples for the bacteria culture media then
applied the streaking method for pure culture so that only a single cell occupies
an isolated portion of the agar surface. This single cell will go through repeated
multiplication to produce a visible colony of similar cells, or clones. The culture
medias were then incubated for about 24 hours at 37 degree Celsius to enhance
bacterial growth.
Data Analysis
Data was analyzed by creating a table of observed pathogenic bacteria in each
stream stations, from morning to afternoon samples.
Results
Table 1 shows that there are different kinds of bacteria that are present in each
streams in the river. In the upstream, the biochemical results for the identification of
bacteria for the four stations in simplified form were Vibrio metschnikovii,
Pseudomonas, and Klebsiella pneumoniae. While in the middle stream, Vibrio
metschnikovii, Enterobacter agglomerans, Klebsiella pneumoniae and Acinetobacter.
For the downstream, Klebsiella rhinoscleromatis, Vibrio metschnikovii and
Acinetobacter were found.
Table 1. Bacterias identified in the morning from upstream, midstream and downstream
UPSTREAM
Study Site
Station 1 S S Sta
t t tio
a a n4
ti ti
o o
n n
2 3
M M
A A MA
Agar MAC C C C
G
r
a
m
G G G -
G r r r G
r a a a r
a m m m a b
m - - - m a
- b b b - Gr c
b a a a b am i
a c c c a - l
ci il il il ci ba l
Gram Staining Gram-bacilli lli li li li lli cilli i
Oxidase Test - + - - - - - -
Biochemical Tests
A A
K/ / A A / A A
N A / / A / /
TSI A/A C G A A G A A
- -
, ,
-, -, -, -, -, - -
-, -, -, -, -, , ,
SIM -,-,- + - - - - - -
MR - - + - - - - -
- - -
-/ -/ / -/ -/ / /
LIA -/- - + - - + - -
CITRATE - - - - - + - -
UREASE - - - - - + - -
ORN - - - - - - - -
V
.
V
V V .
. . M M
K M M K e e
P . e e . t t
s p t t p s s
e n s s n c c
u e c c e h h
d u h h u n n
o m n n m i i
m o i i o k k
o n k k n o o
n i o o i v v
a a v v a i i
RESULT V. Metschnikovii s e ii ii e i i
MIDDLESTREAM
Study Site
G G
r r
G a a
r m m
a - - G
m r
- a
b b b m
a a a -
c c c b
i i i a
l l l c
l Gram- l Gram- l Gram- il
Gram-bacilli i bacilli i bacilli i bacilli li
- + - - - - - -
Biochemical Tests
A A K
A / / /
/ A A N
A/A A A/AG G A/A G A/AG C
- - -
, , ,
- + + -,
, , , -,
-,-,- - -,-,- + -,-,- + -,-,- -
- - + - - - - +
- - -
/ / / -/
-/- - -/- - -/- - -/+ +
- - + + - + + -
- - - - - - + -
- - - - - - - +
A
V E E c
. . . i
M n
e e
t a a t
s g g o
c g g b
h l l a
n o o c
i m m t
k e e e
o r r r
v V. a V. a K. s
i Metschni n Metschn n pneumo p
V. Metschnikovii i kovii s ikovii s niae p
DOWNSTREAM
Study Site
G
r
a G G
m r r
G - a a
r m m
a - -
m b b b
- a a a
b c c c
a i i i
c l l l
il Gram- l Gram- l Gram- l
Gram-bacilli li bacilli i bacilli i bacilli i
- - - - - - - -
Biochemical Tests
K K
A A / /
/ / N N
A/A A A/A A A/A C A/A C
- - - -
, , , ,
- - - -
, , , ,
-,-,- - -,-,- - -,-,- - -,-,- -
+ + - - - + + +
-/- - -/- - -/- - -/- -
/ / / /
- - + +
- - - - - - - -
- - - - - - - -
- - - - - + - +
K A A
. V c c
r . i i
h n n
i e e
n M t t
o e o o
s t b b
c s a a
l c c c
e h t t
r n e e
o i r r
m k
a o
t V. v V. s K. s
i Metsch i Metschn p rhinoscle p
K. rhinoscleromatis s nikovii i ikovii p romatis p
Table 2 shows the different bacteria's that has been identified mainly in each streams
with its four study site/stations. In the afternoon, the pathogenic bacteria found in the
upper stream were; Pseudomonas, Enterobacter cloacae, Vibrio Metschnikovii and
Shigella sonnei. And in the middle stream pathogenic bacteria present were; Shigella
sonnei, Vibrio metschnikovii, Enterobacter agglomerans and Acinetobacter. The
bacteria present in the downstream were, Shigella sonnei, Acinetobacter, Vibrio
metschnikovii, Escherichia coli and Enterobacter agglomerans.
Study Site
Station 1 St St Sta
ati ati tio
on on n4
2 3
Agar MAC SS SS SS
G G
r r
a G G a
m r r m
- a a -
b m m b
a Gr - Gr - Gr a
c a b a b am c
i m- a m- a - i
l ba c ba c ba l
l cill il cill il cill l
Gram Staining Gram-bacilli i i li i li i i
Oxidase Test + + - + - + - -
Biochemical Tests
TSI K/K(LF) A A/ K A/ K A/ A
/ A( / A( / A(L /
A LF K LF K F) A
G ) ( ) ( (
( N N N
N
L L L L
F F F F
) ) ) )
- - - -
, , , ,
- - - -
, -,-, , -,-, , -,-, ,
SIM -,-,- + - - - - - -
MR - - - - - - - -
n
o
r
e
a
c
- - - t
/ / / o
LIA -/- + -/- - -/- - -/- n
CITRATE - + - - - - - -
UREASE - - - - - - - -
ORN - + - - - - - -
E
n
t
e
r
o S
b h
a i
c g
t e
e P P l
r s s l
e e a
V. u V. u
c M d M d
l et o et o V. s
o sc m sc m Me o
a hn o hn o tsc n
c ik n ik n hni n
a ov a ov a ko e
RESULT Pseudomonas e ii s ii s vii i
MIDDLESTREAM
Study Site
G
r
G G a
r r m
a G a -
m r m
- a -
b m b b
a - a a
c b c c
i a i i
l c l l
l Gram- il Gram- l Gram- l
Gram-bacilli i bacilli li bacilli i bacilli i
- - - - - - - -
Biochemical Tests
- - - -
, , , ,
- - - -
, , , ,
-,-,- - -,+,+ - -,-,- - -,+,+ -
- - - - - - - +
- - - -
/ / / /
no reacton - -/- - -/- - -/- +
- - + - - - + -
- - - - - - - -
- - - - - - - +
V V V A
. . . c
M M M i
e e e n
t t t e
s s s t
c c c o
h h h b
n n n a
i i i c
k k k t
o o o e
v E. v V. v E. r
i agglome i Metschni i agglom
Shigella sonnei i rans i kovii i erans
DOWNSTREAM
Study Site
G G G
r r r
a G a a
m r m m
- a - -
b m b b
a - a a
c b c c
i a i i
l c l l
l Gram- il Gram- l Gram- l
Gram-bacilli i bacilli li bacilli i bacilli i
- - - - - - - -
Biochemical Tests
K K A
/ A / /
N / N A
C A C G
( ( ( (
N N N N
L L L L
F F F F
A/A(LF) ) A/A(LF) ) A/A(LF) ) A/A(LF) )
-,-,- -,-,- -,-,- -,-,+
- - - -
, , , ,
- - - +
, , , ,
- - - +
- + - - - + - -
- - - -
/ no / / /
no reacton + reacton - -/- + -/- -
- - - - - - - +
- - - - - - + -
- + - - - + - -
A A
c V c
i . i E
n M n .
e e e a
t t t g
o s o g
b c b l
a h a o
c n c m
t i t e
e k e r
r o V. r a
Shigella v Metschni n
Shigella sonnei sonnei ii kovii E. coli s
Table 3 shows that V. metschnikovii is the most dominant in all streams more
especially in the upstream. While K. rhinoscleromatis is the most dominant in the
downstream then followed by the Acinetobacter. In the midstream, the least dominant
bacteria were, K. pneumoniae and Acinetobacter. While for the upstream were,
Pseudomonas, E. agglomerans and K. pneumoniae.
Table 3. Summary of the bacterias identified in the morning
Table 4 also shows the dominance of V. metschnikovii which is also present in all
streams but for the afternoon result it is more dominant in the midstream. The next
dominant bacteria is S. sonnei which is also present in all streams but more dominant in
the downstream. K. pneumoniae which is only present in the upstream followed by E.
agglomerans which is only present in the midstream and downstream same as to
Acinetobacter. The least dominant bacteria present are the Enterobacter cloacae which
is only present in the upstream while E. coli in the downstream.
Table 4. Summary of the bacterias identified in the afternoon
DISCUSSION
The following bacteria are identified in the morning and afternoon. In the
morning, the identified bacterias were, Vibrio metschnikovii, Pseudomonas, Klebsiella
pneumoniae, Enterobacter agglomerans, Acinetobacter and Klebsiella rhinoscleromatis.
While in the afternoon, the bacteria found were, Pseudomonas, Enterobacter cloacae,
Vibrio metschnikovii, Shigella sonnei, Enterobacter agglomerans, Acinetobacter and E.
coli. The most dominant bacteria present in both morning and afternoon sample is
Vibrio metschnikovii.
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