Microbiology Presentation 31-3-2015

Result of Microbial
Identification
Name:
Nurul Maizura Binti Hashim
(GS 41438)
&
Mohammed Abdulrazzaq Jabbar
(GS 33846)
Supervisor:
Prof. Dr. Zailina Hashim
Introduction
The water-based MWFs were frequently been used in machining

industry .
The primary concern from the usage of these water-based
MWFs
is the presence of biological contaminants that
encourage the growth of bacteria and fungi.
Biological agents present in the MWFs liquids are emitted in

the formed of droplet bioaerosols during a rapid rotation of the
working metal tools which are inhaled by machine operator and
causing adverse health effects mainly on respiratory system
(Cyprowski et al., 2007).
A study shows that, some of the dominant bacteria species that

present in the MWFs
were Pseudomonas aeruginosa,
Burkholderia capacia, Acinetobacteria calcoatius (Bracker et.
al., 2003)
Methodology
Identification step
Isolation of pure colony
Gram-staining
Gram
negative
Gram positive
Oxidase Test
Catalase Test
API 20
E
Oxidase-positive
NonEnterobacteriacea
e.
API 20
NE
Oxidase-negative
Enterobacteriacea
e.
Catalasenegative
Catalasepositive
API 20 Staph
API 20 Strep
apiweb Softwere
Isolation of pure colony
A single colony of each morphology was picked from

the mixed culture medium.
Zigzag technique for isolation of pure culture on pure

solid media.
Gram Staining
A gram stain used in bacteriology to differentiate bacterial cells
into two major groups, gram-positive and gram-negative
(Cappuccino & Sherman, 2011).
The Gram stain reaction is based on the difference in the
chemical composition of bacterial cell walls.
The reagents required are Crystal violet, Grams iodine, 95%
ethyl alcohol and safranin.
Oxidase Test
The oxidase test is used to identify bacteria that produce cytochrome c oxidase (enzyme
of the bacterial electron transport chain).
All bacteria that are oxidase positive are aerobic (can use oxygen as a terminal electron
acceptor in respiration)
Bacteria that are oxidase-negative may be anaerobic, aerobic, or facultative; the oxidase
negative result just means that these organisms do not have the cytochrome c oxidase
that oxidizes the test reagent
(Cappuccino & Sherman, 2011)
Catalase Test
The catalase test is used to differentiate staphylococci (catalasepositive) from streptococci (catalase-negative).
The enzyme, catalase, is produced by bacteria that respire.
Catalase-positive bacteria include strict aerobes as well as
facultative anaerobes (respire using oxygen as a terminal
electron acceptor).
Catalase-negative bacteria may be anaerobes (do not respire
using oxygen as a terminal electron acceptor).
(Cappuccino & Sherman, 2011).
Catalase test
Results
Location
Bacterial Isolated in MWF bulk
Bacteria Isolated in Air at level 1.5 meters
SABB 1
5-Feb-15
1.
2.
3.
Pseudomonas aeruginosa
Klebsiella Pneumoniae
Candida Albicans
1.
2.
3.
E. Coli
Stenotrophomonas Maltophilia
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
E. Coli
Candida Albicans
E. Coli
Candida Albicans
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
1.
2.
1.
2.
1.
2.
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
1. Pseudomonas aeruginosa
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
3. Burkholderia cepacia,
1.
2.
3.
E. Coli
Burkholderia cepacia
Aspergillus
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
SABB 1 (Self Alignment Ball Bearing 1)

1.Pseudomonas aeruginosa.
2.Candida Albicans.
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
E. Coli
Candida Albicans
1.
2.
3.
1.
2.
1.
2.
1.
2.
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans

1.Klebsiella Pneumoniae
Location
SABB 1
5-Feb-15
SABB 2
9-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
E. Coli
Aspergillus
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans

1.Pseudomonas aeruginosa
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
DGBB 1 (Deep Groove Ball

Bearings 1)
1.E. Coli
2.Candida Albicans
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans

Bearings 2)
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans

Bearings 3)
1.E. Coli
2.Candida Albicans
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
SRB 1 (Spherical Roller

Bearing 1)
1.Candida Albicans
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
SRB 2 (Spherical Roller

Bearing 2)
1.Candida Albicans
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Roller 1
1.Pseudomonas Aeruginosa
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
E. Coli
Candida Albicans
E. Coli
Candida Albicans
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
1.
2.
1.
2.
1.
2.
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
E. Coli
Aspergillus
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Roller 2
Location
SABB 1
5-Feb-15
1.
2.
3.
Candida Albicans
1.
2.
3.
E. Coli
SABB 2
9-Feb-15
1.
2.
1.
2.
Aspergillus
SABB 3
11-Feb-15
DGBB 1
13-Feb-15
1.
2.
3.
1.
2.
1.
2.
3.
1.
2.
1.
2.
1.
2.
DGBB 2
16-Feb-15
DGBB 3
18-Feb-15
SRB 1
20-Feb-15
SRB 2
20-Feb-15
Rollers 1
25-Feb-15
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Candida Albicans
Candida Albicans
Rollers 2
25-Feb-15
Rollers 3
27-Feb-15
2. E. Coli
3. Staphylococcus
1. E. Coli
2. Staphylococcus
1.
2.
3.
1.
2.
1.
2.
1.
2.
3.
1.
2.
3.
1.
2.
3.
E. Coli
Aspergillus
Candida Albicans
Staphylococcus
Candida Albicans
Staphylococcus
E. Coli
Candida Albicans
E. Coli
Candida Albicans
E. Coli
Candida Albicans
Roller 3
Pathogenicity of Identified
Microorganisms
Pseudomonas Aeruginosa is pathogenic only when pass the natural

defensive layers of the body such us skin or mucous membrane by
direct tissue damage. A large number of enzymes and toxins are
liberated by this organism as lipase, proteolytic enzyme, endotoxins,
and haemolysins. Several clinical conditions which are highly
correlated with this organism and the respiratory system is the most
frequent body site of infections such as cystic fibrosis, and
pneumonia. Also skin can cause skin infection especially when there
is irritation to the skin. This microorganism is highly resistant to
antibiotics (Bhatia & Lal Ichhpujani, 2008).
Pathogenicity of Klebsiella Pneumoniae, which can cause

variety of diseases such as bronchopneumonia, wound
sepsis, bacteremia, and meningitis (Bhatia & Lal Ichhpujani,
2008).
E. Coli can cause wound infections, septicemia,

pneumonia and other infections (Bhatia & Lal
Ichhpujani, 2008).
Pathogenicity of Candida Albicans, Under normal conditions is

not pathogenic. However, with predisposing factors like skin
irritants as chemical, mechanical, biological irritations, with more
others the fungus can produce bronchopulmonary candidiasis,
septicemia, granuloma, cheilitis (infection of lips), intertrigo
(irritant dermatitis mainly on skin folds), , and many skin diseases
(Bhatia & Lal Ichhpujani, 2008).
Conclusion
Total 8 microorganisms were isolated and identified in both

MWFs bulk and air samples from 11 work production sections.
Only 4 microorganisms are selected for further immunological
studies because they are the dominant in MWFs bulk samples
and also exist in the air samples which are
E. Coli
Candida Albicans
The results are consistent with other studies (burton et al, 2012
& Bracker et al, 2003)

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Result of Microbial

The water-based MWFs were frequently been used in machining

Biological agents present in the MWFs liquids are emitted in

A study shows that, some of the dominant bacteria species that

Isolation of pure colony

A single colony of each morphology was picked from

Zigzag technique for isolation of pure culture on pure

(Cappuccino & Sherman, 2011)

(Cappuccino & Sherman, 2011).

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

SABB 1 (Self Alignment Ball Bearing 1)

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

SABB 2 (Self Alignment Ball Bearing 2)

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

SABB 3 (Self Alignment Ball Bearing 3)

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

DGBB 1 (Deep Groove Ball

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

DGBB 2 (Deep Groove Ball

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

DGBB 3 (Deep Groove Ball

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

SRB 1 (Spherical Roller

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

SRB 2 (Spherical Roller

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

Bacterial Isolated in MWF bulk

Bacteria Isolated in Air at level 1.5 meters

Pseudomonas Aeruginosa is pathogenic only when pass the natural

Pathogenicity of Klebsiella Pneumoniae, which can cause

E. Coli can cause wound infections, septicemia,

Pathogenicity of Candida Albicans, Under normal conditions is

Total 8 microorganisms were isolated and identified in both

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