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Effects of pH, Temperature, Media Composition, Antimicrobial Agents and UV Light on Microorganism Growth

Effects of pH, Temperature, Media Composition, Antimicrobial Agents and UV Light on Microorganism Growth

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Published by AlJayT.Mejos
Microorganisms are open energy systems wherein the physical and chemical states of the environment affect the metabolic activities inside the cells.
This experiment was aimed to study the effects of a set of environmental factors on an aspect of microorganism performance as a function of its
growth. Five parameters were defined: pH, temperature, growth media composition, presence of antimicrobial agents and their effectiveness in
hindering microorganism growth and exposure to UV light. It was observed in the species used, E. coli, B. subtilis and S. aureus, that they grew best in
slightly alkaline environments and at room temperature. These were deemed their optimum growth pH and temperature, respectively. It was also
found that the presence/absence of nutrients in media greatly affect their growth that media allows for sufficient differentiation of bacteria between
species and that UV, antimicrobial agents are detrimental to their growth.
Microorganisms are open energy systems wherein the physical and chemical states of the environment affect the metabolic activities inside the cells.
This experiment was aimed to study the effects of a set of environmental factors on an aspect of microorganism performance as a function of its
growth. Five parameters were defined: pH, temperature, growth media composition, presence of antimicrobial agents and their effectiveness in
hindering microorganism growth and exposure to UV light. It was observed in the species used, E. coli, B. subtilis and S. aureus, that they grew best in
slightly alkaline environments and at room temperature. These were deemed their optimum growth pH and temperature, respectively. It was also
found that the presence/absence of nutrients in media greatly affect their growth that media allows for sufficient differentiation of bacteria between
species and that UV, antimicrobial agents are detrimental to their growth.

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Categories:Types, Research, Science
Published by: AlJayT.Mejos on Feb 17, 2010
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11/05/2012

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Effects of pH, Temperature, Media Composition, Antimicrobial Agents and UV Light onMicroorganism Growth
Al Jay Mejos, John Warner Carag, Jojiemar De Pano, Allison Vincent Labador, Erika Mari Macapagal
Institute of Biology, College of Science, University of the Philippines, Diliman, Quezon City 
Abstract
Microorganisms are open energy systems wherein the physical and chemical states of the environment affect the metabolic activities inside the cells.This experiment was aimed to study the effects of a set of environmental factors on an aspect of microorganism performance as a function of itsgrowth. Five parameters were defined: pH, temperature, growth media composition, presence of antimicrobial agents and their effectiveness inhindering microorganism growth and exposure to UV light. It was observed in the species used,
E. coli, B. subtilis and S. aureus 
, that they grew best inslightly alkaline environments and at room temperature. These were deemed their optimum growth pH and temperature, respectively. It was alsofound that the presence/absence of nutrients in media greatly affect their growth that media allows for sufficient differentiation of bacteria betweenspecies and that UV, antimicrobial agents are detrimental to their growth.
Introduction
Microorganisms are greatly affected by their environment.Different microbial populations respond differently to varyingconditions. There are certain conditions that favor their growth andothers that are detrimental for their survival. Understanding theirresponse to various physical and chemical conditions of theirenvironment, helps us to explain their distribution in nature, andmakes it possible for us to control the growth of beneficial bacteriaand cease the growth of the harmful ones. Effect of pH,temperature, culture media or energy source, chemotherapeuticagents like antimicrobials and UV light on different types of bacteriaare to be understood in this experiment.
Materials and Methods
Effect of pH 
Fourteen test tubes were prepared, seven each forNutrient (NB) and Glucose Yeast Peptone (GYP) broths, each testtube set-up having a different pH (1, 4, 5, 7, 9, and 11). The pH wasset using HCl or NaOH. NB tubes were inoculated with 0.1 mL or100
µ
L of
E. coli 
using a micropipette while GYP tubes wereinoculated with
S. cerevisiae 
. Both were incubated for 24 hours andthe resulting turbidity evaluated as a function of growth.
Effect of Heat Treatmen
Three Nutrient Agar (NA) plates was divided into fourquadrants. Each quadrant was inoculated with
E. coli 
,
S. aureus 
, and
B.subtilis 
. The last quadrant served as control. One plate was thenstored in the locker, refrigerator and incubator together with thetube of GYP inoculated with
S. cerevisiae 
for 24 hours.
Use of Differential or Selective Media 
Mannitol Salt Agar (MSA), MacConkey agar, and EosinMethylene Blue (EMB), were each divided into four quadrants.Leaving one quadrant as blank, each quadrant was inoculated with adifferent organism using cultures of
E. coli 
,
S. aureus 
, and
B. subtilis 
. Itwas incubated and growth was observed after 24 hours.
Effect of Antimicrobials/ Kirby Bauer Disc Diffusion Assay 
Six NA plates, two plates each for
E. coli 
,
S. aureus 
, and
B.subtilis 
, were inoculated using cotton swab plating. One cotton swabper plate was used, dipping the swab in the sample every half theplate. A sterile blank filter paper disc was placed at the center ofeach plate for control. Commercially available antimicrobial discs,Penicillin and Chloramphenicol, a disc with ethanol, and discsinfused with 20
µ
L of plant extracts Malunggay (
Moringa oleifera 
) and
Eucalyptus 
sp, were placed around the blank as shown below (fig. 1).Zones of inhibition as a function of antimicrobial capability of thetest substances were analyzed after 24-hour incubation.
Effect of UV Light 
100
µ
L 0.1%peptone water-100
µ
L
E. coli 
sample wasspread on three NA plates using a hockey stick. After the agarabsorbed the liquid, masking tape was suspended across the lidlessPetri plate without touching the surface of the agar. Each of the set-up was then exposed to 1 min. UV (lidless) then 1 min. sunlight (lidon & inverted), 1 min. UV (lidless) then 1 min. darkness (lid on andwrapped) and 5 min UV (lidless) then 24 hrs sunlight (lid on &inverted). All set-ups were then incubated at 26°C for 24 hours andobserved.
 
Fig. 1
. Placement of filter paper discs on plate surface.
Results
Table 1. Microbial growth in varying pH, temperature and UV exposure.
Group 1 Group 2 Group 3Temperatures4C – minimalgrowth20°C – intermediategrowth37°C – numerousgrowthpH (
NB
&
GYP
)1--4++5++7+++++++9+++++++++11+++++++UV lightGrowth undermasking tape andnumerousgrowth aroundarea surroundingmasking tape(1min)Growth undermasking tape andmedium growtharound areasurrounding maskingtape (5min)1 colony undermasking tape andlittle growtharound areasurroundingmasking tape(15min)
Table II.
Group 2
EMB
+ is a dark metallic color
E. coli
Theo + Actual +
B. subtilis
Theo - Actual +
S. aureus
Theo - Actual +
Mac Conkey
+ is a pink coloration
E. coli
Theo + Actual +
B. subtilis
Theo - Actual -
S. aureus
Theo - Actual -
MSA
+ is a yellow coloration
E. coli
Theo - Actual +
B. subtilis
Theo - Actual +
S. aureus
Theo + Actual +
Table III.
Chloramphenicol(25 units)
 
Penicillin
(10 units)
M.charantia Eucalyptus 
sp.EtOH
 
ControlEC1
27mm - - - - -
EC2
25mm - - - - -
BS1
28mm - 19mm 16mm - -
BS2
22mm - 17mm 17mm - -
SA1
37mm 49mm 23mm 18mm - -
SA2
39mm 52mm 21mm 3mm - -
Ethanol
Control
Penicillin
M. oleifera
Eucalyptus
sp.
 
Chloramphenicol

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