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EFFECTS OF ANTIBIOTICS ON LACTIC ACID FERMENTATION

(Statistical Analysis of Data: Cochran Q Test)

Paul Gerald Albao, Caren Joy Amoto, Rosenette Azarcon, Margaux Barcelona, Margie Luz Rose
Borres, Chaina Eunice Caladiao, Kimberly Iligan, Tynna Eunice Pielago

INTRODUCTION
Based on our research, the internet says that Yoghurt is a dairy product consumed
extensively in Turkey. Lactobacillus delbrueckii subsp. Bulgaricus and S. salivarius subsp.
thermophilus cultures are used in its production. It has been revealed that the antibacterial effect
of bacteria is used in yoghurt production against various food pathogens. It has been reported that
antibacterial effect of yoghurt results from the lactic acid produced by cultures (Kilic, 1990;
Gülmez et al., 2003). In addition to the positive impact of yoghurt on the pathogenic
microorganisms, there are also some studies on the ability of reducing the negative effects of
antibiotics on intestinal flora (Coşkun, 2006).
Kefir is an acidic milk product in which some bacteria and yeasts can live in harmony and
weak alcoholic fermentation occurs. Bacteria and yeasts have a symbiotic relationship in kefir and
it has been proved that compounds they produce have an antibacterial effect on pathogenic bacteria
such as Salmonella, Helicobacter, Shigella, Staphylococcus and E. coli (Ulusoy et al., 2007; Raja
et al., 2009). For this reason, it has been thought that kefir can be used alternatively for therapeutic
purposes in food borne pathogens infections (Schneedorf and Anfiteatro, 2004).
Although probiotic microorganisms are widely known as lactic acid bacteria, some yeasts
and bacteria that do not produce lactic acid has been accepted as probiotic bacteria (Young and
Huffman, 2003; Senok et al., 2005). Because they inhibit the growth of other microorganisms in
the environment, lactic acid bacteria are used in the production of safe food in terms of
pathogenicity. Moreover, low pH, other organic acids, bacteriocins, hydrogen peroxide (H2O2),
ethanol, low oxidation-reduction potential are among the other inhibiting factors (Turantaş, 1998).
Bacteriocins produced by lactic acid bacteria are proteins showing bactericidal effects. Although
the control mechanisms in the synthesis of bacteriocins are unknown, these are thought to be
synthesized as mechanisms to continue the life under the conditions of ecological stress (Aymerich
et al., 1996). It is thought that bacteriocins change the potential of membranes by corrupting K+
ion and ATP and cause cells fail to balance the intracellular pH of cell (Sezer and Guven, 2009).
the antibacterial effects of homemade yoghurt, commercial yoghurt, commercial kefir and
probiotic yoghurt on viability of Salmonella typhimurium (ATCC 14028), Escherichia coli (ATCC
25922), Pseudomonas aeruginosa (ATCC 27853) and Staphylococcus aureus (ATCC 25923) was
investigated by using disc diffusion method. Antibaterial effects of the samples were tested at 24,
48 h and the 7 day intervals. Homemade yoghurt showed the maximum antibacterial effect against
the pathogens. It was determined that the most sensitive pathogenic bacteria to dairy products were
Salmonella typhimurium whereas the least sensitive pathogen was Pseudomonas aeruginosa.
Comparing to the antimicrobial effect of 0.9% lactic acid which was used as positive control, our
results concluded that the bacteria found in fermented dairy products have an antibacterial activity
and this activity is not caused by only lactic acid.

MATERIALS

*antibiotic capsules (500mg) sterile beaker (cover foe milk can)


30bsterile test tubes *yogurt
pipet and aspirator 3 test tube racks
distilled water *evaporated milk
disposable syringe cotton plugs

METHODOLOGY
I. Preparation of Antibiotic Solutions
The antibiotic solutions should be arranged in four different concentrations by diluting
the stock antibiotic solution with distilled water. Calculate the concentration using the
dilution formula.
II. Inoculation of 25 Samples of Milk with Bacteria
Sterilize the milk can for at least 2 minutes. Prepare and disinfect the 25 test tubes in blue
flame. Provide cotton plugs and put 3 mL of milk in the sterilized test tubes. Remove the cotton
balls before filling and replace them with new ones after.
III. Addition of Antibiotic Solutions
Each test tube should be labeled with proper numbers (1-25). The five control tubes should
be set aside. 5 drops of assigned antibiotic solution should be added to the remaining test tubes.
IV. Incubation of the Samples
Place the test tubes in randomized complete block design in the test tube block. Leave it
for three days, and after the period of three days, inspect the experiment and dispose all the
samples and experiments in the trash can for wet garbage.
V. Analyze the Data Using the Cochran Q test
The analysis should be done on single data set consisting of inspections and observations
done after the period of three days. Make sure to set up the null and alternate hypothesis. Add
the scores for each replicate, and the addition of data should be done in each row. Square the
sums, and calculate the Q using the given formula. Lastly, study and interpret the results.
DATA AND RESULTS
The observation is recorded using the following code:
 Write “0” if there is no curdling
 Write “1” if an appreciable degree of milk curdling was observed.

CONCENTRATION LEVEL
Replicate Numbers Results

1(control) 0

2(control) 0

3(control) 0

4(control) 0

5(control) 0

6 0

7 0

8 0

9 0

10 0

11 0

12 0

13 0

14 0

15 0

16 0

17 0
18 0

19 0

20 0

21 0

22 0

23 0

24 0

25 0

CONCLUSION
In this fermentation of milk experiment we observed that increasing amount of antibiotic
(insert name here) is inversely proportional with the amount of milk curdling. This behavior has
yielded the conclusion that the antibiotic has affected the growth of the organisms that is typically
seen to be a catalyst for fermentation hence the curdling or yellowing and "bunching" up of milk
because if the changes in the protein structure of the milk. The mechanism of the fermentation
reaction is anaerobic respiration of sugars in the milk. The sugars gets processed by anaerobic
respiration into acids which in turn de-synthesizes the protein hence the formation of "milk-
curdles"

REFERENCE
Antibacterial Effects of Some Fermented Commercial and Homemade Dairy Products and 0.9%
Lactic Acid against Selected Foodborne Pathogens. (n.d.). Retrieved from
https://scialert.net/fulltext/?doi=ajava.2011.189.195
Zenaida T. Nucum, Ed. D, (2010) (pages 46-47)