Abid Khan Report (s2897438)

Q1.

Report Forms 1, 2 & 3.

Discuss which characteristics of the unknown samples allowed you to determine the
species of microbe present. Describe what each test indicates/suggests about the
unknown organism, how these results helped you in identifying the micro-organism
and ruling out other possible bacterial species, and if all of the test results correlated
with identification (4 marks).
1. Differential staining
Firstly differential staining was conducted to determine the gram
staining of the different bacteria which can be categorised into two
groups: gram-negative and gram-positive bacteria. In this
investigation, faecal and vaginal (white) bacteria were classified as
gram-negative which was determined by the characteristic pink colour.
The pink colour is due to the combination of the thin peptidoglycan
walls present in gram negative bacteria which causes crystal violet
stain to be easily washed/removed with alcohol and the addition of
safranin counter-stain. On the other hand, vaginal (cream) bacteria was
categorised as gram positive. The was established due to the deep
violet colour illustrated due to the characteristic thick peptidoglycan
walls of gram positive bacteria which allows the retention of the crystal
violet stain even after the addition of alcohol.
2. Isolation techniques to determine colony morphology and cellular
arrangement
Through isolation techniques (streaking to spread bacteria colonies
across a nutrient agar plate) colony morphology and cellular
arrangement of the bacteria could be determined. Faecal and vaginal
(cram) bacteria colonies were round gross morphology, flat in colony
height, smooth colony margins, small colony size and had a cream
colour/pigmentation. Conversely, vaginal (white) bacteria colonies
exhibited an irregular gross morphology, raised colony height, smooth
colony margins, pinpoint colony size and a white pigmentation.
3. Differential and selective media (blood agar, MacConkey Agar and
Mannitol Salt Agar
Differential and selective media were then utilised to conduct further
differentiation to understand the characteristics of each of the bacteria
colonies. Blood agar (BA), MacConkey agar (MCA) and Mannitol Salt
Agar (MSA) were used. BA recognises the haemolytic enzymes
secreted by bacteria which have the ability to lyse the red blood cells
present in the nutrient agar. Both faecal and vaginal (cream) were nonhaemolytic while vaginal (white) was beta-haemolytic. This
characteristic is an example of E.coli.

MCA was implemented only for gram negative bacteria (faecal and
vaginal white) to classify their lactose fermentation ability. Faecal
bacteria was classified as non-lactose fermenting due to the display of
colonies with no colour/pigmentation. Vaginal (white) was classified
as lactose fermenting exhibited by the red-pigmented colonies. This
characteristic is also another example of E.coli.
MSA is used to differentiate colonies according to their ability to
ferment mannitol. Only the gram positive bacteria (vaginal cream)
could ferment mannitol exhibited by the yellow colour surrounding
the colonies. This characteristic is an example of S.aureus.
4. Biochemical test- Catalase and Oxidase
The catalase and oxidase test were conducted to determine the bacteria
colonies capability of dealing with oxidative stress and determining
the presence of cytochrome oxidase respectively. All the bacteria is this
investigation tested negatively for this test
5. Microbact gram negative system
The microbact gram negative system is indicative of the identity of
aerobic and facultatively anaerobic gram-negative bacteria. For faecal
bacteria, the results correlated with the code number 7706 which
indicates that there is a 74% chance of the bacteria being Salmonella Spp.
For the vaginal (white bacteria), the code number produced was 6760
which indicates that there is a 95% chance of the bacteria being E.coli.
6. Sensitivity and resistance of antimicrobial test
Different antimicrobial agents were used as antibiotics to determine
their effects on the different bacteria. Clindamycin was the only
antibiotic to inhibit the growth of vaginal (cream) and faecal bacteria.

Q2.

Report Form 3.

Based on the results from Report Form 3, what could be a possible antibiotic
treatment regime for each of the patients? Was the sensitivity/resistance result
expected (2 marks)
According the results found determined in the antimicrobial agent/antibiotic
test, clindamycin is a possible antibiotic treatment for patients with the faecal
and vaginal cream bacteria but not with vaginal white bacteria. The other
antibiotics tested were all susceptible to growth from each of the three types
of bacteria.
This resistance result of clindamycin on faecal and vaginal cream bacteria is
expected as clindamycin is used to treat bacteria such as staphylococcus
aureus and salmonella (Ogbru, 2014).
Q3.

Report Form 4.

What characteristics of the normal flora could be determined? Were the characteristics
of the normal flora expected for that region of the body? Can you make suggestions as
to the possible identity of your normal flora? (1 mark)
The gram stain reaction for skin behind the ear and oral cavity bacteria were
both gram positive. They both displayed cocci cellular morphology and
staphylococcus cell arrangement. Additionally their colony morphology were
similar apart from the colony size and pigmentation which was small and
cream for skin behind the ear and medium and yellow for oral cavity. These
characteristics were expected for the flora presented in these regions of the
body. The possible identities of the flora in the skin behind the ear are
staphylococcus epidermis, staphylococcus aureus or staphylococcus warneri. The
possible flora for the oral cavity could be staphylococcus aureus or
staphylococcus epidermis.

Q4.

Report Form 5.

Discuss the results you observed following washing of your hands with the different
cleaning agents? Which chemical agent was the most effective at reducing microbial
growth and why? Did your results reflect the class results? (1 mark)
The chemical agents used for washing hands were soap and hand wash and
to compare the effectiveness of them, the difference between the growth of
bacteria in unwashed and washed hand must be found. The growth of
bacteria in unwashed hands in the class sample of soap and handwash was
1.86 and 0.73 respectively while for washed hands with soap and handwash
was 1.21 and 0.46 respectively. The difference in the growth of bacteria in the
soap and hand wash samples is 0.64 and 0.267 respectively. Thus it can
concluded that soap is more effective in reducing microbial growth.
The individual results correlated with this conclusion as the unwashed result
for soap and handwash was 3 and 2 respectively. The growth of bacteria after
washing hands with soap and handwash were 1 and 1 respectively thus
correlating with the class results.
Q5.

Report Form 6.

Was an increase or decrease of turbidity observed in the tube for each of the chemical
agents? Was an increase or decrease of microbial growth in the tube observed with the
serial dilution of your chemical agent? Was this increase or decrease in microbial
growth reflected in growth on the plates? (1 marks)
The chemical agent used in this investigation was bleach. There was a slight
increase in the bacterial growth in the plates but no bacterial growth on the
tubes. However in the class results, floor cleaner, surface spray and vinegar
exhibited increase in turbidity across dilutions. Bleach, NaCl and ethanol had
slight increase to no increase in turbidity across dilutions.

Q6.

Report Form 6.

What effect did the dilution series of the chemical agent have on microbial growth?
Which was the most effective chemical agent overall and how did you determine this?
What is the MIC for each of the chemical agents (the highest dilution that prevents
appearance of turbidity AND subsequent growth on agar plates)? (1 marks)
The effect of the dilution series was variable due to inconsistencies in
microbial growth. Bleach appeared to be the most effective chemical agent
overall as the average turbidity in the tube was 1 and the growth on the
microbial plate was classified as 2 when the highest dilution was observed.
This allows the conclusion to be made that bleach is effective against bacteria
even at low concentrations. Dilution 1 was the MIC for bleach and floor
cleaner which resulted from the prevention of a turbid solution and growth
on nutrient agar plates. The MIC for ethanol, vinegar, NaCl and surface spray
were greater than the first dilution. At dilution 1 either turbidity or microbial
growth or both were exhibited. This leads to the conclusion that the dilution
of those chemical agents decreases its effectiveness at inhibiting or destroying
microbial growth.
Q7.

Report Form 6.

Which of the chemical agents are bacteriocidal and which are bacteriostatic? How did
you determine this? (1 mark)
Bacteriocidal chemical agents are known as bacteria-killing while
bacteriostatic agents are bacteria inhibiting (News-Medical, 2004).
Bacteriodical agents induce less growth on the nutrient agar plate when
compared to growth in the tube (ie. growth on the plate is smaller when
compared to the turbidity of the tube). Bacteriostatic agents hinder the growth
growth/replication of bacteria in the tube but cannot inhibit growth on the
nutrient agar plates. According to the results found in this investigation, floor
cleaner and bleach are bacteriocidal agents. Surface spray, ethanol, vinegar
and NaCl are bacteriostatic agents.

References
News-Medical. (2004). Bactericidal (bacteria-killing) versus bacteriostatic (bacteria-inhibiting) drugs
for treating infections. Retrieved September 21, 2014, from http://www.newsmedical.net/news/2004/10/21/5741.aspx
Ogbru, O. (2014). clindamycin, oral (Cleocin). Retrieved September 20, 2014, from
http://www.medicinenet.com/clindamycin-oral/article.htm