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Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
Honey as a Natural Preservative in Milk
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Honey as a Natural Preservative in Milk

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  • 1. HONEY AS A NATURAL PRESERVATIVE OF MILK By ARCHANA.NATARAJAN MSc.BT-09022
  • 2. HONEY
    • Introduction
    • Materials required
    • Methods
    • Result
    • Discussion
    • Disadvantages
  • 3. MILK
    • Milk and other dairy related products comprise a major chunk of food products for their nutritive values
    • A good growth medium of many micro-organisms like Klebsiella, Bacillus, Pseudomonas and Staphylococcus
    • Preservation of Milk – a major problem !
  • 4. HONEY
    • Natural food product produced by honey bees
    • Its usage since ancient times
    • Major use in Ayurveda, Cosmetics and Food products
    • Composed of sugars – fructose, glucose and with some amount of sucrose
  • 5. HONEY
    • Numerous studies on the anti-bacterial activity of honey on clinically significant bacteria have been reviewed
    • The anti-bacterial activity of honey can be grouped into peroxide component and non-peroxide component
    • The major anti-bacterial factor in honey is HYDROGEN-PEROXIDE
  • 6. MATERIALS
    • CHEMICALS – Microbiological media and individual components of agar and broths of Luria-Bertani and Nutrient Agar were from HiMedia laboratories,Mumbai,India
    • Common chemicals salts and sugars were also from HiMedia laboratories and Sisco Research Lab,Mumbai
    Luria-Bertani medium Nutrient agar
  • 7. Milk and Honey
    • Four commercially available pasteurized milk samples were obtained and refrigerated at 4 º C until further use
    • Samples stored from 1 to 4 days were serially diluted and the isolated pure cultures were used for further analyses
    • Honey was obtained commercially and dilutions of honey were made in sterile water with or without honey at 100 º C for 10 mins in water bath
    • Artificial honey was prepared and used as control
    • Sterile water was used as negative control for milk and honey samples
  • 8. Isolation of Bacterial Cultures
    • 1ml of refrigerated milk samples were serially diluted in sterile water
    • From this,200 µl of sample were plated onto nutrient agar plates and incubated at 37ºC/24 hrs
    • The randomly picked colonies were re-inoculated in 15 ml LB broth and incubated for 14-16hrs at 37ºC
    • Culture thus obtained was characterized by various biochemical tests
  • 9. DISC DIFFUSION ASSAY
    • Sterile filter discs (10mm,diameter) were immersed in 5 µl diluted honey solutions and air dried
    • The cells were harvested from the cultures grown to mid log phase and the pallet was suspended in 3 ml fresh LB medium
    • From this culture,200µl corresponding to 1x10 7 CFU/ml was plated on LB plates
    • The discs containing honey of different concentrations were placed on culture plates and incubated at 37ºC for 24 hrs
    • The diameter of zones of inhibition were studied and recorded.
  • 10. Honey inhibits the growth of both Catalase Positive and Negative Bacteria
    • Cultures of bacteria isolated from milk were grown with a density of 1x10 7 CFU/ml
    • 5% inoculated into 1ml Nutrient Broth
    • Each flask supplemented with honey of different concentrations
    • The change in absorbance at 550nm was assessed using a photoelectric colorimeter
  • 11. HONEY- as a Preservative
    • Monitoring the bacterial growth in 500 ml of milk samples that were stored with 100 µl of 500mg/ml solution of honey added
    • Stored at 4ºC for 3-6 days
    • Comparison with milk sample without broth(550nm) inoculated with 100µl milk sample supplemented with honey and a similar volume of the same milk sample without honey were quantitated for its bacterial growth
  • 12. RESULT Spoilage of Milk during Shelf life
    • Milk samples were evaluated for contaminating bacterial species by serially diluting the samples and plating them on NA plates
    • Sterile water used as negative control,did not yield any bacterial colonies
  • 13. Isolation and Characterization of Bacteria
    • Morphological characteristics, biochemical and microbiological tests
    • Most Common – Bacillus spp. , Staphylococcus spp. , Pseudomonas spp. and Klebsiella spp.
    • Broadly classified into Catalase positive and catalase negative based on the results of the catalase tests
  • 14. Disc Diffusion Assay
    • Zones of inhibition were measured
    • Diameter of inhibition zones dependent more on the bacterial species and the concentration of honey
    • Honey was found to inhibit catalase negative cultures while catalase positive cultures were not inhibited to the same extent
    • Major role of Hydrogen Peroxide
    • Artificial honey was used at various concentrations and had no inhibitory effect on the organisms
  • 15. Honey inhibits growth of both Catalase Positive and Negative Bacteria
    • The inhibition of growth increased in a concentration dependent fashion at 24 hrs incubation in catalase negative cultures containing honey with a percent inhibition of 50-60% in average
  • 16. HONEY – As a Preservative
    • Milk samples stored at 4 ºC from 3-6 days in the presence or absence of honey were assayed for their bacterial content and growth
    • The addition of honey at a final concentration of 50mg/ml had a considerable inhibitory effect on bacterial growth as compared to the samples devoid of honey
  • 17. DISCUSSION
    • Preservation and storage of milk
    • The Lactoperoxidase system –
    • Its natural role in preservation of milk
    • Mechanism
    • Properties
    • Chemical preservation discouraged by FAO/WHO
    • Honey as a safe,natural product
  • 18. DISADVANTAGES
    • Spores of C.botulinum have been found in about 10% of honey by the Centre for Disease Control and Preservation (CDC) in the US.
    • Known to cause Botulism in infants which can cause serious illness in the first year of life or can prove fatal
    • Many people are found to be allergic to Honey
  • 19. REFERENCES
    • Taornima PJ,Niemira B A & Beuchat P R , Inhibitory activity of honey against food borne pathogens as infuenced by the presence of hydrogen peroxide and the level of antioxidant power, Intl J Food Microbiol,69(2001),217
    • Roushdy A,Studies on the efficiency of hydrogen peroxide and methods for its detection and quantification. J Food Prot,59(1996),1223
    • White J W, Honey Adv Food Sci,24 (1978),287 etc
  • 20.  

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