Microbiology of milk

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Microbiology and Deterioration of Milk and Milk Products : 

Microbiology and Deterioration of Milk and Milk Products Mohammad Ashraf Pal Prof/chief scientist Division of LPT, FVSc & AH

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Milk constitutes an excellent medium for the growth of micro-organisms. Freshly drawn milk from healthy animals contains a small number of harmless micro-organisms, however, during milking process and storage the contamination takes place, the extent of which depends upon the hygienic measures taken before, during and after milking process and storage conditions observed thereafter .

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Significance of m.o . in milk Nearly all the changes that take place in the flavor and appearance of the milk after it is drawn form the cow, are the result of the activities of m.o ,therefore, it is very essential to control these m.o Microbial content serves as an indicator of production conditions and sanitary quality of milk. Prevention of spoilage. Prevention of milk borne illnesses. Production of dairy products with desired characteristics imparted via m.o . introduction

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Micro-organisms in Milk The types of m.o found in milk vary considerably Bacteria, yeasts, moulds and bacteriophages are commonly encountered. Viruses and protozoa are seldom observed in milk, except as occasional contaminants. Bacteria Most common and most numerous of m.o found in milk and milk products .

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They belong to four main groups: Gram + ive cocci Gram + ive non-spore forming rods Gram + ive spore-forming rods Gram - ive non-spore forming rods. LAB Normally present in milk and they are also used as starter culture for the production of cultured dairy products. They ferment lactose and yield lactic acid. + ++++

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LAB reclassified (older names in parenthesis) . Lactococci L.delbrueckii sub sps.lactis ( Str.Lactis ) L. Lactis sub sps . Cremoris (Str. Cremoris ) Lactobacili L. Casei L. delbrueckii sub.sps.lactis (L. Lactis ). L. delbrueckii sub Sps . bulgaricus ( L.bulgaricus ) Leuconostoc

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Coliforms Facultative anaerobes optimum growth @ 37 0 c indicator organism and are closely associated the presence of pathogens but not necessary pathogens themselves. They ferment lactose with prod n of acid and gas cause rapid spoilage of milk they are killed by pasteurization Their presence is an indication of PP contamination of milk.

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Yeast Most frequently encountered in raw cream during hot weather produce acid and co 2 . They are potential contaminants throughout the year. Moulds Their growth is visible as a fuzzy or fluffy growth on the surface of milk and milk products. They may be black, green, grey, blue or white. They discolor the product and often produce repulsive undesirable off odors Essential in production of certain varieties of cheese.

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Bacteriophages Particularly obnoxious in starter cultures used for the production of cultured dairy products. Phages kill bacterial culture and entire fermentation process fails (slow or dead vat). Factors affecting growth of m.o in milk Food supply ( H 2 o,energy,C,N Vit . &Mineral source ) Moisture ( a w ) Oxygen supply ( Obligate aerobes ,facultative ,micoaerophillic , Aerotolerant anaerobes,obligate anaerobes) Acidity and pH (Acidophilic)

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Preservatives Light ( phototrophic) Concentration (osmophillic yeasts) Temperature ( psychrotrophs - 20-30-07 , Mesophiles-30-40,Thermophiles-55-65 o c Antimicrobial constituents Products of microbial growth in milk ~Enzymes ~ Decomposition products of protein, fat & CHO etc. ~ Pigments , Toxins etc (mycotoxins & bacterial toxin )

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Result of microbial growth in milk (Spoilage) Principal cause are Psychrotrophs Most of these are destroyed by pasteurization Some may survive e.g. Pseudomonas fluorescens , Pseudomonas fragi Other species and strains that survive pasteurization and grow at Refrigeration temp. ,Produce heat stable proteolytic & lipolytic enzymes and cause spoilage : ~ Bacillus ~ Clostridium ~ Cornebacterium ~Arthrobacter ~ Lactobacillus ~ Microbacterium ~ Micococcus ~ Streptococcus

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Deteriorative changes: Souring- Lactose fermentation LA, VFA Souring & gassiness- coliforms  acid & gas Aroma production- starter culture  diacetyl Proteolysis – unpleasant odors- undesirable. controlled desirable –cheese prod n . Ropiness –Milk drawn into long threads (Alkaligenes viscolactis) Sweet curdling –due to prod n of rennin like enzyme which curdles without souring Stormy fermentation : Rapid fermentation by Clostridium perfringens Color changes Pseud.syncyanea (blue) ; Pseud.synxantha (yellow); Serratia marcescens (red)

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Pathogenic micro-organisms in m ilk Food borne illnesses occur as a result of : ~Ingestion of raw milk ~Improper pasteurization ~Poor handling ~Storage leading to PP contamination Measures to decrease the threat : ~Hygienic production practices ~ Proper handling and storage ~Pasteurization

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Bacterial Pathogens of current concern Bacillus cereus Listeria monocytogens Yersinea entrocolitica Salmonella spp. E.coli O 157:H 7 Compylobacter jejuni Coxeilla burnetii Moulds ( Aspergillus,Fusarium,Penicillium ) grow in milk and milk products & produce potentially hazardous mycotoxins

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Means of destruction of micro-organisms Heat - pasteurization , sterilization etc. Ionizing Radiations- UV., gamma rays High frequency sound waves- super & ultrasonic's Electricity - by the heat generated. Pressure - 600 x > atmospheric pressure Chemicals - Acids ,alkalis, halogens, H 2 0 2 etc.

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Microbiological Standards for milk & Milk Products Suggested references: Food Safety & Standards Act(2006) Food Safety & Standards Authority Of India

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Starter Cultures Starter cultures are those microorganisms that are used in the production of cultured dairy products such as yogurt and cheese. The natural micro flora of milk is either inefficient, uncontrollable and unpredictable, or is destroyed altogether by the heat treatments given to the milk.

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A starter culture can provide desired characteristics in a more controlled and predictable fermentation. The primary function of lactic starters is the production of lactic acid from lactose. Other functions of starter culture may include the following : Flavour ,aroma, and alcohol production proteolytic and lipolytic activities inhibition of undesirable organisms

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There are two groups of lactic starter cultures: Simple or defined : single strain, or more than one in which the number is known. Mixed or compound : more than one strain each providing its own specific characteristics Starter cultures may be categorized as mesophillic or thermophillic : Mesophilic Lactococcus lactis subsp. Cremoris L. delbrueckii subsp. Lactis

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L. lactis subsp. Lactis biovar diacetylactis Leuconostoc mesenteroides subsp. Creamoris Thermophilic Streptococcus salivarius subsp. thermophilus (S. thermophilus ) Lactobcillus delbrueckii subsp. bulgaricus L. delbruekii subsp. Lactis L. helveticus L. plantarum L.casei

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Mixtures of mesophillic and thermophillic microorganisms can also be used as in the production of some cheeses.

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Starter culture preparation Commercial manufacturers provide starter cultures in lyophilized ( freez-dryed ), frozen or spary - dried forms. The dairy product manufacturers need to inoculate the culture into milk or other suitable substrate. There are a number of steps necessary for the propagation of starter culture ready for production: Commercial culture Mother culture- first inoculation; all cultures will originate from this preparation

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Intermediate culture - in preparation of large volumes of prepared starter Bulk starter culture - this stage is used in dairy product production

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HACCP Raw and end-products may be tested for the presence, level, or absence of microorganisms. Traditionally these practices were used to reduce manufacturing defects in dairy products and ensure compliance with specifications and regulations, however, they have many drawbacks: Destructive and time consuming Slow response

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Small sample size Delays in the release of food In 1960’s the Pillsbury company, the U.S. Army, and NASA introduced a system for assuring pathogen free foods for the space programme. This system, called Hazard Analysis and Critical Control Points (HACCP), is a focus on critical food safety areas as part of TQM . Involves a critical examination of entire food manufacturing process to determine every step where there is a possibility of physical, chemical, or microbiological contamination which would render food unsafe or unacceptable for human consumption .

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These identified points are critical control points (CCP). Seven principles to HACCP : analyze hazards determine CCPs establish critical limits establish monitoring procedures establish deviation procedures establish verification procedures establish record keeping procedures Before these principles can be put into place, a prerequisite programme and preliminary setup is necessary.

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Prerequisite Programme Premise control receiving and storage control equipment performance and maintenance control Personnel training sanitation recall procedure Preliminary setup assemble team describe the product identify intended use construct flow diagram and plant schematic verify the diagram on site

Terms and definitions : 

Terms and definitions Food Safety - concept that food will not cause harm to the consumer when it is prepared and/or eaten according to its intended use Food chain - sequence of the stages and operations involved in the production, processing, distribution, storage and handling of a food and its ingredients, from primary production to consumption Food safety hazard- biological, chemical or physical agent in food, or condition of food, with the potential to cause an adverse health effect Food safety policy - overall intentions and direction of an organization related to food safety as formally expressed by top management End product - product that will undergo no further processing or transformation by the organization

Terms and definitions : 

Terms and definitions Critical control point CCP - at which control can be applied and is essential to prevent or eliminate a food safety hazard or reduce it to an acceptable level Critical limit - criterion which separates acceptability from unacceptability Monitoring - conducting a planned sequence of observations or measurements to assess whether control measures are operating as intended Correction - action to eliminate a detected nonconformity Corrective action - action to eliminate the cause of a detected nonconformity or other undesirable situation

Terms and definitions : 

Terms and definitions Validation - obtaining evidence that the control measures managed by the HACCP plan and by the operational PRPs are capable of being effective Verification - confirmation, through the provision of objective evidence, that specified requirements have been fulfilled Updating - immediate and/or planned activity to ensure application of the most recent information

Example of processing for safety: 

Example of processing for safety RAW MATERIAL Milk

Potential hazards: 

Potential hazards Salmonella Campylobacter

Control measure: 

Control measure Heating

Critical Control Point: 

Critical Control Point Boiling in pan

Monitoring: 

Monitoring Observation of foaming

Verification: 

Verification Observation of foam residues

Use: 

Use Drinking while still hot ( This prevents recontamination and growth which may lead to hazards )