OBJECTIVES:
To determine the amount of anti-microbial peptide production
by Staphylococcus warneri under various conditions when 2L and 10L
Fermented. To Test the effects of one uncontrolled parameters sush
as pH, Temperature or dissolved Oxygen and compare findings. To
produce anti-microbial activity from Staphylococcus warneri.
INTRODUCTION:
Staphylococcus warneri is a member of bacterial genus
Staphylococcus, consisting of Gram-positive bacteria with spherical
cells appearing in clusters. Colonies of S. warneri are usually
tan, yellow and about 2-4mm in diameter after 48 hours incubation
at 35°C.
It is commonly found as part of the skin flora on humans and
animals. S. warneri rarely causes disease, but may occasionally
cause infection in patients whose immune system is compromised. S.
warneri is known to produce antimicrobial peptide activity in the
form of Nisin. The optimum conditions for this to occur are pH 7.
Nisin is a polycylic antibacterial peptide with 34 amino acid
residues used as a food preservative. It is produced by bacterium
and which contains antimicrobial activity and which is known as a
bacteriocin. Nisin has been found to have properties that can
control spoilage caused by lactic acid bacteria.
It is used in processed cheese, meats, beverages, etc. during
production to extend shelf life by suppresing Gram-positive
spoilage and pathogenic bacteria. In food it is common to use Nisin
at levels depending on the food type regulatory approval. Nisin
cannot be produced chemically therefore it has to be synthesised
using fermentation. During fermentation various stages of growth
occur and as a result different conditions can occur during this
fermentation process, eg pH, most organisms produce acid as they
grow and therefore in the Lag phase ( a period of adptation for the
cells to their new environment, new enzymes are ynthesized) and in
the lag phase can produce alkaline substances and therefore pH
plays an important role in efficient fermentation. As acid is
produced alkaline substance needs to be added to the process to
maintain the optimum pH of 7 and likewise in the lag phase when
alkaline substances are produced, acidic substance needs to be
added to maintain the pH, temperature, and oxygen. III. MATERIALS
AND METHODS:“As per manual. ”
RESULTS:
All parameters are controlled using sensor probes in the
vessels connected to a data logging software system. The vessels 4
and 5 are controls where the optimum environmental growth
parameters for the strain are kept. To determine the results
obtained in each vessels are as follows: Vessel 1: No antimicrobial
peptide activity seen at any of the time intervals. This indicates
that when temperature is not controlled the temperature can
increase significantly. As shows in TABLE 1. 1 History Plot Vessel
1 – 2L NO Temperature control. Vessel 2: No antimicrobial activity
seen at 13:00.
However antimicrobial activity seen in both neat and 1:2
sample at 14:00 and 15:00. Antimicrobial activity seen in neat
sample at 16:00 and 09:00. When air flow is not controlled the
reduced air content reduces the rate of fermentation, As Oxygen is
required for cell growth and when air is in reduced quantity this
slows down rate of cell reproduction as shows in TABLE 1. 2 History
Plot Vessel 2 – 2L NO Air Flow. Vessel 3: Antimicrobial activity
seen in neat sample at all time intervals. Antimicrobial activity
seen in 1:2 sample at 14:00, 16:00 and 09:00.
Activity seen in 1:4 for the first time at 16:00 and 09:00.
There is greater anti-microbial peptide activity with temperature
and air controls which shows that the pH does not have significant
effects as the other two parameters. The fermentation was not
affected to the same extent by pH as shown in TABLE 1.
History Plot Vessel 3 – 2L NO pH control. Vessel 4:
Antimicrobial activity seen in neat sample at all time intervals.
Activity seen in 1:2 sample at 14:00, 15:00, 16:00 and 09:00. For
the first time see antimicrobial activity in 1:8 sample at 09:00.
This shows the three uncontrolled vessels has greater
anti-microbial peptide, where in fermentation took place on its
fastest rate as all conditions are maintained at most favourable
for the organism to grow and reproduced as shown in TABLE 1. 4
History Plot Vessel 4 – 2L Optimum conditions. Vessel 5: (In error
no result recorded for 14:00 time interval) Antimicrobial activity
seen in neat and 1:2 sample at all time intervals. Activity seen in
1:4 and 1:8 (for the first time) at 15:00, 16:00 and 09:00. The
effects produce the highest level of anti-microbial peptide
activity of all the system.
The organism has greater supply of oxygen and nutrients and
temperature and pH has a lesser effect due to the larger volume as
shown in TABLE 1.
History Plot Vessel 5 – 10L Optimum conditions.
CONCLUSIONS:
In this practical the results was successfully determined
that Temperature is the most important parameter to control in
relation to microbial growth. Therefore, if temperature was not
controlled, NO amount of anti-microbial peptide activity produced
by Staphylococcus warneri. While in Oxygen level and pH level if
NOT controlled S. warneri will still grow and produced the
anti-microbial peptide.