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    Honey   Presentation ( Final) Honey Presentation ( Final) Presentation Transcript

    • HONEY AS A NATURAL PRESERVATIVE OF MILK By ARCHANA.NATARAJAN MSc.BT-09022
    • HONEY
      • Introduction
      • Materials required
      • Methods
      • Result
      • Discussion
      • Limitations
    • 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 !
    • 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
    • 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
    • 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
    • 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
    • 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
    • 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.
    • 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
    • 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
    • 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
    • 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
    • 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
    • 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
    • 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
    • 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
    • LIMITATIONS
      • 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
    • 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
    •