Food Microbiology Discussion Responses

Tutorial Class/TA46
Tuesday, 7st March 2017
Lecturer: Mrs Inggrid Suryanti Surono

Group 1:
Ronald Horison /1901492375
Kristanto Rianto /1901467221
Putu Aditya Ragatama /1901517806
Johanes Evan Andjaya /1901476996
Joshua Firdaus /1901460732
Joshua /1901478995
Jason Purnama /1901500736
Ferry Octavian Sulaiman /1901522794
Janitra Saraswati Dewi /1901461874
Said Naufal Hibaturrahman /1901519540
Michelle Muliawidjaja /1901502022

Question:
1. In which phase, gram staining could be inducted? (2) {Putu, Tam}
2. In which phase we do the viable count?(2){Johanes,Joshua Firdaus)
3. Why do cells enter stationary phase?(2) {Joshua, Ferry}
4. How are the correlation of microbial growth characteristics related to food
preservation?(3) dengan contoh{Said, Janitra, Michelle}
5. How are the correlation of microbial growth characteristics related to food
safety?(2){Jason,Ronald

Responses:

1. There are 4 phases of the growth of bacteria, lag phase (adaptation phase), log phase,
stationery phase, and death phase. Gram staining could be inducted in log phase because the
shape and the conformation of bacteria, especially the cell wall, are more stable than another
phase. The numbers of bacteria are enough and stable. If use the old culture in gram staining,
may the positive gram stain becomes negative gram stain because the death of bacterium
which cause changes of permeability. In this phase the cells divide at a constant rate depending
upon the composition of the growth medium and the conditions of incubation. The rate of
exponential growth of a bacterial culture is expressed as generation time, also the doubling time
of the bacterial population. Typically, the increase is exponential. The bacteria are growing and
dividing at their maximum speed. The explosive growth of bacteria cannot continue forever in
the closed conditions of a flask of growth medium. Nutrients begin to become depleted, the
amount of oxygen becomes reduced, and the pH changes, and toxic waste products of
metabolic activity begin to accumulate. The bacteria respond to these changes in a variety of
ways to do with their structure and activity of genes.
 Figure 1. Growth Curve of Bacteria

2. In which phase we do the viable count?

During the stationary phase, if viable cells are being counted, it can not be determined
Whether some cells are dying and an equal number of cells are dividing, or the population of
cells has simply stopped growing and dividing (Todar,
http://textbookofbacteriology.net/growth_3.html). In the lag phase, viable count can be done,
because Viable count measurements are the only accurate way to determine population during
lag times (Swinnen IA, Bernaerts K, Dens EJ, Geeraerd AH, Van Impe, 2004)
Identification of Standard Plate Count bacteria, selected colonies of all morphological
types were picked from Standard Plate Count Agar plate. Isolates were purified by streaking on
nutrient agar. Pure cultures were maintained on NA slants at 5oC. The cultures were identified
according to Bergeys Manual of Systematic Bacteriology, the most important source of
information for the identification of unknown bacteria. According to it, the first approach to
bacterial identification involve preliminary microscopic examination of the gram-stained
preparation for its categorization into two broad groups. After knowing the gram reaction (gram
positive or negative), colony and cell morphology (rods or cocci) of the bacterium, the
identification was done with the help of various key charts [2, 3, 4] so as to confirm the identity
of bacteria (Chouhan, 2015).
Method of Viable Counting, the usual plating technique was employed, the culture
sample being diluted in sterile M/30-phospate buffer solution. Eight replicate plates on Difco
nutrient agar were made from a suitable dilution of each sample and these were incubated at
the temperature of the experiment concerned, expect when this was 15 oC ,when an incubation
temperature of 20oC was used in order to accelerate growth of the colonies (Jordan & Jacobs,
1946).

3. Stationary phase is attained when a culture exhausts nutrients and its OD remains constant.
Starved microorganisms need to respond to a wide range of stresses. such as nutrient
limitation, high concentrations of organic acids, osmotic and oxidative stress, and pH changes,
depending on the specific conditions that have led to stationary phase. (Gefen, et al. 2014).
During this phase, the number of living cells remained constant for the period different,
depending on the bacteria, but eventually towards a period of decline population. In some
cases, the cells contained in a culture contained in a cultured cell populations do not grow, can
be elongated, swollen abnormally, or the irregularities, a manifestation of the growth that is not
balanced (Kusnandi, dkk. 2003). The stationary phase may occurs in bacteria because breeding
began to decrease and some cells die. If the breeding rate equal to the rate of death, then the
overall number of cells remains constant. This can be due to reduced nutrient or the formation
of metabolic products which tend to accumulate may become toxic to the bacteria in question. In
this phase, the cells become more resistant to extreme conditions such as heat, cold, radiation
and chemicals

4. Microorganisms grow or multiply in numbers when exposed to a favorable environment, such

as food. Their growth is associated with food spoilage and foodborne diseases. In order to
design methods to control their growth in food as a way to control spoilage, a good
understanding of microbial growth characteristics is needed.
The growth of microorganisms is reflected by the increase in the number or mass of vegetative
cells of bacteria, yeasts, and molds. Normally, a food harbors a mixed population of
microorganisms that can include different species and strains of bacteria, yeasts, and molds.
The growth characteristics of a mixed population differ in several respects from that of a pure
culture. Depending on the environment, both the food environment (intrinsic) and the
environment in which the food is stored (extrinsic), some of the species or strains can be in an
optimum or near-optimum growth condition. Thus, in a mixed population, the intrinsic and
extrinsic environments determine which microorganisms in the mixed population will become
predominant and produce specific changes in a food. These aspects are very important in the
control of microbial growth. (Ray and Bhunia, 2013)

Microbes can grow in their optimum condition. There are physical requirement and chemical
requirement for microbial growth. The physical requirement consist of temperature, pH and
osmotic pressure. The chemical requirement consist of availability of useable carbon source,
useable sources of nitrogen, sulphur and phosporus, presence (or absence) of oxygen gas, and
availability of trace elemental nutrients (such as Fe, Mg, etc). One can effectively inhibit the
growth of a microbes of concern by controlling its physical and chemical environment.
Food preservation is the process of treating and handling food to stop or slow down food
spoilage, loss of quality, edibility, or nutritional value and thus allow for longer food storage.
Preservation usually involves preventing the growth of bacteria, fungi (such as yeasts), and
other microorganisms, as well as retarding the oxidation of fats which cause rancidity.
Preservative food additives can be antimicrobial. These inhibit the growth of bacteria or fungi,
including mold, or antioxidant, such as oxygen absorbers, which inhibit the oxidation of food
constituents. Common antimicrobial
preservatives include calcium propionate, sodium nitrate, sodium nitrite, sulfites (sulfur dioxide,
sodium bisulfite, potassium hydrogen sulfite, etc.), and disodium EDTA. Antioxidants include
BHA and BHT. Other preservatives include formaldehyde (usually in solution), glutaraldehyde
(kills insects), ethanol, and methylchloroisothiazolinone.

To reduce or stop the bacteria growth, first, we have to know about the factors of microbe
optimun growth, so we can use or choose proper food preservation. For exmple, Clostridum
botolinum will not produce its toxin below pH 4.5, and consequently hight acid products with a
pH below 4.5 do not need as severe a heat treatment (Campbell-Platt, 2012). In other words, to
prevent the growth of Clostridum botolinum makes the pH condition lower than 4.5. Other
exmple is, fruits. As we know, fruits products like juices have high water activity, which allows
microbial growth that may be easily spoiled the products. Therefore, to prevent the microbial
growth of this products, we have to preserved with combination of hurdle that include low pH,
suppretion of water activity by solute addition, added preservatives, and heat treatment (Theron
dan Lues, 2010).

5.To know about the correlation of microbial growth characteristics related to food safety better
we know about the growth of bacteria. When bacteria grow , they increase in numbers not in
size. This process is called doubling. Under ideal conditions, the number of bacteria can double
every 30 minutes. For example, if raw meat is left out at room temperature, after five hours there
could be more than 10,000 bacteria in the rice. This is more than enough bacteria to cause
foodborne illness. Many factors affect bacterial growth but the most important ones are food,
water, pH, oxygen, and temperature. The important factor about microbial growth is time and
temperature. (Anonym, online).

When the total of bacteria come at lethal dossage, the can cause foodborne illness. The
best way to prevent foodborne illness caused by bacteria is to implement food safety policies
that promote good personal hygiene like washing hands, preventing food cross contamination
by storing foods properly and only use cleaned and sanitized utensils and surfaces to store,
prepare, and serve food. The last one for controlling of the growth, it can be doing by keep food
out of the temperature danger zone (Washington State Department of Health, 2013).

References

Anonym. Online. Food Safety.

(http://www.foodsafetysite.com/resources/pdfs/EnglishServSafe/ENGSection2.pdf).
Accesed at 3 March 2017.

Boundless. Food Preservation. Boundless Microbiology Boundless, 26 May. 2016. Retrieved

05 Mar. 2017 from https://www.boundless.com/microbiology/textbooks/boundless-
microbiology-textbook/industrial-microbiology-17/food-preservation-202/food-preservation-
1016-7700/

Campbell-Platt, Geoffrey. 2012. Fod sciene and technologies. Wiley-Blackwell : Oxford

Chouhan, Sonu. 2015. Enumeration and Identification of Standard Plate Count Bacteria in Raw
Water Supplies. India : Paher University.

Gefen, Orit., Ofer Fridman., Irine Ronin., Nathalie Q. Balaban. 2014. Direct observation of
single stationary-phase bacteria reveals a surprisingly long period of constant protein
production activity. PNAS: Vol. 111 no. 1.
Jordan,R.C. & Jacobs, S.E. 1946. The Effect of Temperature on the Growth of Bacterium coli at
pH 7.0 with Constant Food Supply. London : Physiology Institute, University College of
South Wales and Monmouthshire, Cardiff, and the Bacteriological Laboratory, Imperial
College of Science and Technology.

Kusnandi., dkk. 2003. Mikrobiologi. Bandung: Universitas Pendidikan Indonesia.

Los Angeles Mission College. Lecturer Note, Chapter 6: Microbial Growth. Department of Life
Science, Los Angeles Mission College.

Ray, Bibek., Arun Bhunia. 2013. Fundamental Food Microbiology: Fifth Edition. New Delhi. CRC
Press.

Swinnen IA, Bernaerts K, Dens EJ, Geeraerd AH, Van Impe JF Int J Food Microbiol. 2004 Jul
15; 94(2).
Theron, Maria M. dan J.F. Rykers Lues. 2010. Organic Acid and Food Preservation. CRC
Press: London
Todar, Kenneth. Online Textbook of Bacteriology
http://textbookofbacteriology.net/growth_3.html

Waluyo,Lud.Drs.M.Kes.2004.Mikrobiologi Umum.Universitas Muhammadiyah Press : Malang.

Washington State Department of Health.2013. Washington State Food & Beverage Workers
Manual. Washington State Department of Health: Washington.

Volk and Wheeler. Mikrobiologi Dasar. Jilid 2 edisi V. Diterjemahkan oleh Sumarto
Adisumartono. Penerbit Erlangga. Jakarta. 1990