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Sirindhorn International Institute of Technology Thammasat University GROWTHS, ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF KEFIR GROWN IN DIFFERENT CULTURE.

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Presentation on theme: "Sirindhorn International Institute of Technology Thammasat University GROWTHS, ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF KEFIR GROWN IN DIFFERENT CULTURE."— Presentation transcript:

1 Sirindhorn International Institute of Technology Thammasat University GROWTHS, ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF KEFIR GROWN IN DIFFERENT CULTURE MEDIA Sathika Jariyatamkitti, Somrutai Kitchalao, Pattarawan Wuttisan, Rachnarin Nitisoravut*, Issara Poljungreedand and Siwarutt Boonyarattanakalin

2 Sirindhorn International Institute of Technology Thammasat University Background Statement of Problem Objectives Methodology Results and Discussion Conclusion 2

3 Sirindhorn International Institute of Technology Thammasat University Kefir Fermented drink Natural prebiotic: rich in enzymes and beneficial live bacteria or microbial Benefits Nutrition Additive product Therapeutic effects 3 selling.html

4 Sirindhorn International Institute of Technology Thammasat University Diameter: 3 to 20 mm Grain matrix Polysaccharide Protein (dry weight) Cellular debris and unknown components Microorganisms: A complex symbiotic relationship Mostly lactic acid bacteria and yeasts 4

5 Sirindhorn International Institute of Technology Thammasat University Exopolysaccharides (EPS) Mainly produced from bacteria Provide… Specific rheology and texture of fermented milk products Biological Properties -Antimicrobial and antifungal -Antitumor o Fernando Lopitz-Otsoa, A. R., Natalia Elguezabal and Javier Garaizar, Rev Lbaroam Micol, (2006) 23, 67. o Hsi-Chia Chen, S.-Y. W., Ming-Ju Chen, Food Microbiology, (2008) 25, Anti-inflammatory -Antimutagenic and antioxidant

6 Sirindhorn International Institute of Technology Thammasat University Can milk kefir grown in media other than cow milk Would kefir grown in other media possess health benefits Alternative culturing liquid… Tropical country Plenty of coconut trees Relatively Cheap Growth of kefir grains grown in three different culture media Microorganism profiles of kefir and kefir grains Compare antibacterial and antifungal activities of milk, coconut milk and coconut water kefir Coconut milk Coconut water 6

7 Sirindhorn International Institute of Technology Thammasat University To see the effects of coconut milk and coconut water, as a culturing medium for kefir, on kefir culture growth To classify microorganism profiles in kefir and kefir grains To examine antibacterial and antifungal activities of kefir products 7

8 Sirindhorn International Institute of Technology Thammasat University METHODOLOGY 8

9 Suspend grains in phosphate buffer and homogenize using mortar Perform serial dilutions for enumerations Isolate bacteria using MRS agar, DECA agar, and YM agar and incubate Count and observe the colonies

10 Sirindhorn International Institute of Technology Thammasat University Inoculum Preparation Inoculation of Test Plates Interpreting Results Application of Disc to Inoculated Plates 10

11 Sirindhorn International Institute of Technology Thammasat University Inoculum Preparation Inoculation of Test Plates Interpreting Results Application of Disc to Inoculated Plates 11

12 Sirindhorn International Institute of Technology Thammasat University Select isolated colonies and transfer them into a broth tube Incubate the broth culture Compare the turbidity with 0.5 McFarland Standard 3_43_287_688/10171/Potato-Dextrose-Broth.html 12

13 Sirindhorn International Institute of Technology Thammasat University Inoculum Preparation Inoculation of Test Plates Interpreting Results Application of Disc to Inoculated Plates 13

14 Sirindhorn International Institute of Technology Thammasat University Streak the swab over the entire agar surface 14

15 Sirindhorn International Institute of Technology Thammasat University Inoculum Preparation Inoculation of Test Plates Interpreting Results Application of Disc to Inoculated Plates 15

16 Sirindhorn International Institute of Technology Thammasat University Place on inoculated plates Incubate plates antimicrobial-effect.html Soak ø 5 mm no.1 filter paper by fermented solutions and positive control solutions 16

17 Sirindhorn International Institute of Technology Thammasat University Inoculum Preparation Inoculation of Test Plates Interpreting Results Application of Disc to Inoculated Plates 17

18 Sirindhorn International Institute of Technology Thammasat University Measure the diameter of clear zones Sliding calipers A ruler k%20diffusion/ 18

19 Sirindhorn International Institute of Technology Thammasat University RESULTS AND DISCUSSION 19

20 Sirindhorn International Institute of Technology Thammasat University Coconut milk and coconut water can be used as an alternative culturing medium for kefir culture 20 Growth data for kefir grains grown in three different culture media (50 mL) at 25 °C 4.7g/100g Sugar level

21 Sirindhorn International Institute of Technology Thammasat University Coconut milk and coconut water can be used as an alternative culturing medium for kefir culture 21 Growth data for kefir grains grown in three different culture media (50 mL) at 25 °C 2.8g/100g Sugar level

22 Sirindhorn International Institute of Technology Thammasat University Coconut milk and coconut water can be used as an alternative culturing medium for kefir culture 22 Growth data for kefir grains grown in three different culture media (50 mL) at 25 °C 2.6g/100g Sugar level

23 Sirindhorn International Institute of Technology Thammasat University Species of bacteria and yeasts in kefir and kefir grains can be found 23 Selective platesGECAMRSYM Shape of microorganisms BacillusCoccusYeastBacillusCoccusYeastBacillusCoccusYeast Milk kefir Milk kefir grain Coconut milk kefir Coconut milk kefir grain Coconut water kefir Coconut water kefir grain *+ indicates the presence of the microbial where – indicates the absence of the microbials Table 1*: Microscopic examination of kefir and kefir grains in three different culturing media

24 Sirindhorn International Institute of Technology Thammasat University Species of bacteria and yeasts in kefir grains can be found 24 Selective platesGECAMRSYM Shape of microorganisms BacillusCoccusYeastBacillusCoccusYeastBacillusCoccusYeast Milk kefir Milk kefir grain Coconut milk kefir Coconut milk kefir grain Coconut water kefir Coconut water kefir grain *+ indicates the presence of the microbial where – indicates the absence of the microbials Table 1*: Microscopic examination of kefir and kefir grains in three different culturing media

25 Sirindhorn International Institute of Technology Thammasat University Kefir grown in coconut milk and coconut water have antibacterial and antifungal activities 25 Strains Antibiotic AgentsKefir Products FlucozoleAmpicillin Chloram- phenicol PenicillinMilk Coconut milk Coconut water E. Coli (-) S. Typhi (-) S. Aureus (+) B. Subtilis (+) C. albican ** The results represent the mean of clear zone diameters (mm). The non fermented media (milk, coconut milk and coconut water) show no inhibition zone Table 2**: Antibacterial and antifungal activities using disk diffusion method of 24 h kefir fermentation in various culturing media

26 Sirindhorn International Institute of Technology Thammasat University Kefir grown in coconut milk and coconut water have antibacterial and antifungal activities 26 Strains Antibiotic AgentsKefir Products FlucozoleAmpicillin Chloram- phenicol PenicillinMilk Coconut milk Coconut water E. Coli (-) S. Typhi (-) S. Aureus (+) B. Subtilis (+) C. albican ** The results represent the mean of clear zone diameters (mm). The non fermented media (milk, coconut milk and coconut water) show no inhibition zone Table 2**: Antibacterial and antifungal activities using disk diffusion method of 24 h kefir fermentation in various culturing media

27 Sirindhorn International Institute of Technology Thammasat University Kefir grown in coconut milk and coconut water have antibacterial and antifungal activities 27 Strains Antibiotic AgentsKefir Products FlucozoleAmpicillin Chloram- phenicol PenicillinMilk Coconut milk Coconut water E. Coli (-) S. Typhi (-) S. Aureus (+) B. Subtilis (+) C. albican ** The results represent the mean of clear zone diameters (mm). The non fermented media (milk, coconut milk and coconut water) show no inhibition zone Table 2**: Antibacterial and antifungal activities using disk diffusion method of 24 h kefir fermentation in various culturing media

28 Sirindhorn International Institute of Technology Thammasat University Coconut milk and coconut water can be used as the alternative to cow milk Bacillus and Yeasts  All kefir and grains Coccus  Milk kefir and grains Antibacterial activities - Coconut milk kefir - Coconut water kefir Antifungal activities - Only milk kefir 28

29 Sirindhorn International Institute of Technology Thammasat University Semih Otes, a. O. C., Pakistan Journal of Nutrition, (2003) 2, 54. Fernando Lopitz-Otsoa, A. R., Natalia Elguezabal and Javier Garaizar, Rev Lbaroam Micol, (2006) 23, 67. Hsi-Chia Chen, S.-Y. W., Ming-Ju Chen, Food Microbiology, (2008) 25, 492. Zhou Jianzhonga, L. X., Jiang Hanhub, Dong Mingshengb, Food Microbiology, (2009) 26, 770. Chujian Chen, McGill University Montreal, (2005), p 222. Prof.Dr.Lalitha Kameswaran, Manual on Antimicrobial Susceptibility Testing, In (2004). (Retrieved September 26,2012) Kamila Leite Rodrigues, L. R. G. C., Jose Carlos Tavares Carvalho, Joao Evangelista and Jose Mauricil Schneedorf, International Journal of Antimicrobial Agents, (2005) 25, 404. Michel Andre Wattiaux, Milk Composition and Nutritional Value, In Dairy Essentials - Lactation and Milking. Muhammad Majeed, P. D., Lakshmi Prakash, Ph.D., Cococin TM : The Nourishment Factor TM Freeze-Dried Coconut Water Solids, In (2007). g=en (Retrieved September 15, 2012) 29

30 Sirindhorn International Institute of Technology Thammasat University QUESTIONS? 30

31 Sirindhorn International Institute of Technology Thammasat University Cow milk - 4.7g/100g sugar level Coconut milk - 2.8g/100g sugar level Coconut water - 2.6g/100g sugar level 31

32 Sirindhorn International Institute of Technology Thammasat University Inoculum Preparation Inoculation of Test Plates Interpreting Results Application of Disc to Inoculated Plates 32

33 Sirindhorn International Institute of Technology Thammasat University Select isolated colonies and transfer them into a broth tube Incubate the broth culture (35 °C for 2-6 hr) Compare the turbidity with 0.5 McFarland Standard 3_43_287_688/10171/Potato-Dextrose-Broth.html 33

34 Sirindhorn International Institute of Technology Thammasat University Place on inoculated plates Incubate plates (35 °C for hrs) antimicrobial-effect.html Soak ø 10 mm no.1 filter paper by fermented solutions and positive control solution 34


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