Microbial flora of milk

and milk products

-Dr. Shekhar Pokhrel

• A variety of microbes both saprophytic and
pathogenic are associated with milk and milk
products because these foods provide all the
nutrients required for microbial growth and
multiplication.
• These microorganisms may be
– bacteria,
– yeast,
– Moulds
– virus.
• Among these bacteria are the most important and
common group followed by moulds, yeasts and
viruses.
 A. BACTERIA
• Bacteria have the ability to utilize various milk
constituents to grow and multiply.
• While growing at the expenses of milk constituents
these microorganisms may release certain metabolites
like lactic and other organic acids, gasses, enzymes,
flavouring compounds, pigments, toxins etc. in the
system which may be useful and harmful and thus
directly effects the quality.
• Generally these metabolites lead to various spoilage
conditions in the products and make them unfit for
human consumption.
 Categorization of dairy-related bacteria
1. Shape wise
Cocci: cells are spherical
• Diplococci: Cells divide in one plane & remain attached in
pairs Ex. Neisseria
• Streptococci: Cells divide in one plane & remain attached
after some divisions, in the form of chains . Ex. Lactococcus
• Tetrads: Cells divide in two planes & form 4 cells. Ex.
Pediococci
• Sarcinae: Cells divide in 3 planes & form a cuboidal
arrangement. Eg. Sarcinaventriculi, Sarcinaureae,
• Staphylococci: Cells divide in 3 planes in an irregular pattern
producing bunches of cocci .eg. Staphylococcus aureus
Bacilli: cylindrical or rod like cells
• Diplobacilli: Pairs.
• Streptobacilli: Chains
• Ends may be rounded as in Lactobacillus delbruckeii ssp.
bulgaricusor squamosed as in Bacillus anthracis

Spirilla: spiral or helical shaped cells

• Single curves: Vibrio
• Many curves: Spirochetes
• Few curves: True spirilla

Pleomorphic: cells appearing in different shapes or lack of

uniform shape. Ex. Arthrobacter
 2. Temperature wise
• Mesophillic:
– Microorganisms capable of growing between 20 & 40°C with the
optimum growth temperature (OGT) of 37°C are termed as
“Mesophiles”.
– All pathogenic organisms are mesophillic in nature. Eg. S. aureus,
E. coli

• Psychrotrophic:
– Organisms capable of growing at or below 7°C (refrigerated) but
the OGT is between 15 & 20°C are termed as psychrotrophs.
– These are the significant spoilage organisms of refrigerated milk
& milk products. Eg. Pseudomonas sp.; Alcaligenes sp.
• Thermophilic:
– Organisms capable of growing over 50°C with O.G.T of 55°C
are termed as thermophiles.
– They are the important organisms causing outbreaks in heat
processed milk & milk products.
– Some are capable of growing between 40-85°C.
– Organisms produce enzymes at rapid rate, so that enzymes
are replaced quickly. Eg. Bacillus stearothermophilus,
Streptococcus thermophilus.

• Thermodurics:
– Organisms capable of withstanding pasteurizing
temperatures of 63°C/30 min. with O.G.T of 35-37°C are
termed as thermodurics.
– They form important flora of pasteurized or heat processed
foods. Eg. Micrococcus varians
 3. Oxygen requirement
• Aerobic:
– Organisms capable of growing in the presence of oxygen are termed as
aerobic organisms.
– They can grow in a standard air atmosphere of 21% oxygen.
– They are more efficient in utilization of available nutrients. Eg Bacillus
species
• Anaerobic:
– Organisms which cannot grow in the presence of oxygen but can grow in
the presence of CO2 are termed as anaerobic organisms.
– They don’t use O2 for energy yielding reactions.
– They are ever poisoned by O2.
– Some tolerate low concentrations of O2.
– They produce catalase & peroxidase enzymes.
– High tolerance –Clostridium perfringens and Moderate tolerance –
Clostridium tetani
• Facultative:
– Organisms which can grow either in the presence or
absence of oxygen are termed as facultative organisms.
Eg. .E. coli, Lactococcus lactis spp. lactis

• Microaerophillic:
– Organisms which grow best at 1-15% of O2 levels.
– They can use O2 for energy yielding reactions but
cannot withstand high levels of O2 i.e, 21% of O2. Eg.
Campylobacter jejuni
 4. Chemical properties
• Acid producers: Capable of fermenting lactose to form
lactic acid. Lactic acid coagulates milk by producing
precipitation of Casein at 4.6 pH
– Homofermenters: Lactococcus, some Lactobacilli
– Heterofermenters: Some Lactobacilli, Lueconostocsp.,

• Gas producers: Capable of producing CO2\&H2 from

lactose fermentations. Eg. E. coli, Clostridium species.

• Proteolytic: Degrade milk proteins into soluble

components by enzymes known as proteinase or
proteases. Eg. Bacillus species, Pseudomonas species.
• Lipolytic: Organisms capable of attacking milk fat by
enzymes known as lipases liberating glycerides & fatty
acids. Eg. Pseudomonas species, Achromobacter
lipolyticum

• Sweet curdling: Organisms capable of causing curdling

of milk by the enzyme known as rennin like enzyme
before the development of sufficient acidity. Eg. B.
subtilis, B. cereus, Enterococcus liquifaciens.

• Ropiness: Causing change in the viscosity of milk or

forming threads when the milk is poured from one
container to other, due to production of gums, mucins
etc. Eg. Alcaligenes viscosus
• Flavour producing
– Fruity: Pseudomonas fragi
– Malty: Lactococcus lactis var. maltigenes
– Fishy: Proteus icthyosmius

• Colour fermentations
– Yellow coloration: Pseudomonas synxantha
– Blue coloration : Pseudomonas cyanogenes
– Green coloration: Penicillium roqueforte
– Black coloration : Pseudomonas nigrifaciens
– Red coloration :Serratia marcescens
 B. YEAST
• Yeasts are unicellular, ovoid or elliptical cells, gram positive,
non-motile and larger in size when compared to bacteria.
• Reproduction is by ‘budding’.
• Grow well at 25-400C and can tolerate pH 3.5 and are
strictly aerobic.
 
• Saccharomyces cerevisiae
– Used in the brewing and baking industry and in Kefir production.
– Cells are cylindrical to ellipsoidal, occurring singly or in pairs,
short-chains or clusters.
– Lactose is not fermented.
• Candida kefir
– Associated with kefir, buttermilk and cheese.
– Produces ‘pseudomycelium’; some strains ferment lactose.

• Candida lacticondensi
– Isolated from condensed milk.
– Lactose not fermented.

• Candida pseudotropicalis ( false yeast)

– Ferments lactose with the production of alcohol and carbon
dioxide.
– Associated with “yeasty” or “gassy” cream and Kefir grains.
• Debaromyces hansenii
– Has been isolated from cheese, occurring in short chains and
propagation by ‘multipolar budding’.
– Does not ferment lactose.

• Kluyveromyces marxianus var. marxianus

– Previously known as Kluyveromyces fragilis, associated with
manufacture of kefir and kumis.
– Cells occur as sub-globose and singly or in pairs.

• Kluyveromyces marxianus var. lactis

– Previously known as Kluyveromyces lactis.
– Associated with yoghurt and has been isolated from milk, gassy
cheese, buttermilk and cream.
– Cells are ‘amoeboid’ and will ferment lactose.
 C. MOULDS
• Moulds are complex, multicellular
organisms, aerobic, growing over a wide
range of pH.
• The colonies appear cottony or wooly and
are generally white, creamy, green or black.
• The individual mould is composed of an
aggregate of branching protoplasmic
threads known as mycelium.
• Individual threads of the mycelium are termed
as hyphae. They are vegetative, concerned with
nourishment or fertile concerned with asexual
or sexual reproduction.

• Some produce sporangium, which in turn

produce sporangiospore and some, conidia,
which produce conidiospores (Penicillium sps).
MOULDS ASSOCIATED WITH MILK AND MILK
PRODUCTS
• Geotrichum candidum
• Scopulariospsis brevicaulis
• Sporendonema sebi
• Penicillium roqueforti
• Penicillium casei
• Penicillium camemberti
• Rhizopus stolonifer
 D. VIRUS
• A virus particle or virion consists of nucleic acid DNA or RNA
that is covered by a protein coat called capsid, are
symmetrically constructed. Two kinds of symmetry have been
recognized.
– Helical
– Cuboidal

• Hepatitis virus
– Causes hepatitis which is generally referred to as jaundice.
– It is due to the consumption of milk or food contaminated with faeces
or carriers of the disease.

• Cowpox virus
– Produces pustules and vesicles on the teats of cow udder
• Poliomyelitis virus
– Belongs to the group of enterovirus.
– It is readily killed by pasteurization.
– It has tendency to get localized in the CNS leading to paralysis.

• Tick borne encephalitis virus

– Belongs to the genus Flavivirus and is responsible for meningitis or
meningioencephalitis.
– Its vectors are the ticks Ixodes recinus and I. persulcatus.
– The disease may be caused due to consumption of unboiled goat
milk.

• Bacteriophages of lactic acid bacteria

– Bacteriophages are the viruses that infect bacteria.
– Many times in cheese industry starter cultures fails which is due to
bacteriophages