1 Faculty of Food Technology, Osijek

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PRODUCTION OF SHORT AND MEDIUM CHAIN FATTY ACIDS
DURING FERMENTATION OF GOAT AND COW MILK WITH
Bifidobacterium longum BB-46
Vedran Slacanac1, Jovica Hardi1, Hrvoje
Pavlovic1, Mato Vlainic4, Darko Curik2, Marko
Jukic1, Vitomir Cutic3

• 1 - Faculty of Food Technology, Osijek
• 2 - Faculty of Medicine, Osijek
• 3 - Health centre, Valpovo
• 4 - MEGGLE, Dairy Industry, Osijek

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Possibilities of manipulating the composition of
the gut microflora by foods 2nd generation of
functional food

• INTESTINAL MICROFLORA ? by a number of
  physiological function contribute to overall
  health
• PROBIOTICS ? living micoorganisms which (when
  ingested or locally applied) confer one or more
  demonstrated health benefits for consumer
• PRODUCTION OF ANTI-MICROBIAL SUBSTANCES ? one of
  the health benefits of LAB

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COULD BE RESPONSIBLE FOR ANTAGONISTIC EFFECT
AGAINST OPORTUNISTIC MICROORGANISMS
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Physiological functions of short (SCFA) and
medium (MCFA) chain fatty acids

• SCFA affect colonic epithelial cell transport,
  hepatic control of lipid and carbonhydrates
• SCFA provide energy to muscle, kidney, herth and
  brain
• MCFA (from the MCT) are rapidly absorbed from the
  small intestine, intact or the following
  hydrolysis, into the portal circulation
• MCFA (from the MCT) are easier to metabolize
• MCFA can promote long-chain fatty acid oxidation
• SCFA and MCFA had strong in vivo and in vitro
  antagonistic effect against many oportunistic
  organisms (Foster Call, 1990 Cherington et.
  al., 1991 El-Ziney et. al., 1995 Cresci et.
  al., 1999Kievit Iglewski, 2000 Kunte et. al.,
  2000.)

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Goat milk versus cow milk

• Better digestibility
• Higher buffering capacity
• Smaller dimeter of fat globules and better
  distribuition in milk emulsion
• Higher content of SCFA and MCFA in the milk fat
• Higher content of some minerals especially
  soluable calcium, magnesium and chlorides
• Better immunological and antibacterial
  characteristics

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The aim of this work was to determine 1. The
intensity of production of lactic and acetic
acid, SCFA and MCFA during the fermentation of
cow and goat milk with Bifidobacterium longum
Bb-46 2. Potential antagonistic activity of
fermented goat and cow milk against some
pathogens The basic hypothesis - Goat milk
fermented by the probiotic Bifido- bacterium
longum Bb-46 rise higher amounts of examined
substances in comparison to cow milk
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METHODS OF ANALYSIS
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• 1. Fermentation of goat and cow milk
• SUBSTRATE goat and cow milk with 3,2 of
  milk fat
• HEAT TREATMENT UHT
• STARTER Direct Vat Set Bifidobacte- rium longum
  Bb- 46 culture
• FERMENTATION 37 0C / 24 hours

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• 2. Measuring during the fermentation
• a) pH values (METTLER TOLEDO, MA 235, pHIon
  Analyzer)
• b) lactic and acetic acid content (GC/MS)
• c) fatty acids contents (GC/FID)
• a), b) after every hour of fermentation
• c) before the fermentation, after 4 and 8 hour
  of fermentation

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• MICROBIOLOGICAL ANALYSIS
• CFU of Bifidibacterium longum Bb-46
• Modificated Bifidobacterium medium
• Anaerobic conditions (ANAEROBIC JARS)
• Incubation on 37 0C from 48 hours

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Antibiotic assay

• Standard method for Antibiotic sensitivity test
  (1 colony in 5 mL of sterile physiological saline
  solution, Mueller-Hinton agar for detection)
• Tested antibiotics NORFLOXACIN, OFLOXACIN
  (kinolon), TETRACYCLIN
• Tested probiotics goat and cow milk fermented
  by the use of B. longum Bb-46
• (inoculated milk, after 12 hour of fermentation,
  after 24 hour of fermentation)
• Oportunistic microorganisms E. coli, Salmonela
  enteritidis D
• Dosing of probiotics antibiogram susceptibility
  discs (diameter 12.7 mm producer Schleicher
  Schuell, Germany)

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RESULTS
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Changes of Bifidobacterium longum Bb-46 CFU
during the fermentation of goat and cow milk
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Changes of pH values during the fermentation of
goat and cow milk with Bifidobacterium longum
Bb-46
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Changes of acetic acid content during the
fermentation of goat and cow milk with
Bifidobacterium longum Bb-46
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Changes of short chain (SCFA) and medium chain
(MCFA) fatty acid contents (molar percent) during
fermentation
Mean values followed by the same letter in the
same column and in the same row are not
significantly different (Plt0.05) - for all fatty
acid separately Mean ? standard deviation, n
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Tendency of changes of SCFA and MCFA contents in
goat and cow milk during the fermentation with
Bifidobacterium longum Bb-46
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Results of Antibiotic assay (E. coli)
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Results of Antibiotic assay (S. enteritidis D)
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CONCLUSIONS

• pH values decreased more rapidly and larger
  number of viable cells of B. longum Bb-46 were
  found in goat milk during the fermentation
  (statistically significant, Fcalculated
  Fcritical 34,605,05 14,675,05)
• Total increase of the analyzed SCFA and MCFA in
  fermented goat milk was 45,74.
• Total increase of the analyzed SCFA and MCFA in
  fermented cow milk was 25,36.
• Increase of all the examined SCFA and MCFA in
  fermented goat milk was statistically
  significantly higher than in fermented cow milk.

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CONCLUSIONS

• Only the content of myristic acid increased
  statistically significantly during the
  fermentation of cow milk.
• Bifidobacterium longum Bb-46 was able to produce
  acetic acid in both types of milk.
• Higher rate of production of acetic acid was
  noted in goat milk in comparison to cow milk
  (statistically significant, Fcalculated
  Fcritical 14,495,05).
• 8. Antibiotic assay show that E. coli was weakly
  sensitive to the presence of fermented goat and
  cow milk (higher degree of probiotic sensitivity
  to fermented goat milk).

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CONCLUSIONS

• Salmonella enteritidis D was sensitive to the
  presence of fermented goat and cow milk (stronger
  inhibitory of fermented goat milk was noted).
• Measured diameter zones of inhibition for S.
  enteritidis D were in correlation with increasing
  values of examined parameters (pH value, viable
  cells of B. longum Bb-46, concentration of acetic
  acid, concentration of SCFA and MCFA) during
  fermentation.

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Antibiotic assay S. enteritidis fermented
goat milk