1

4TH COFFEE

Abnormal milk
 Mastitis 2

• Mastitis is the inflammation of udder (one or more

quarters), usually caused by bacterial infection
4TH COFFEE

• Mastitis milk is characterized by Most of the times it is

• Higher microbial count subclinical in nature
• Higher somatic cell count
• Altered physical and chemical composition
• Reduced yield
• High numbers of Somatic cells concentration

• Udder - swollen, hot and painful with edema

• Milk - discoloured, clots Can’t be eliminated but reduced by
Huge economic loss good management
 Mastitis 3

• Bacteria, yeast, fungi and viruses can infect

udder but bacteria are the main agents
4TH COFFEE

• Organisms penetrate teat duct and establish

infection

• Around 140 different microbes were isolated from

udder of a cow
 Classification 4

• Based on etiology
• Contagious
• Opportunistic
4TH COFFEE

• Environment
Based on symptoms
• Per acute:
• Acute or clinical:
• Sub acute or sub clinical: milk and udder appears
normal
• Chronic:
• Gangrenous:
 Microbes 5

• The udder gets infected with one or more of the

causative organisms.
4TH COFFEE

• Contagious mastitis transmitted during milking by hand

milkers, cloths, milking machine

• Opportunistic pathogens: induce pathogens when there

is any infection or wound in the udder/ teat and by
milking

• Environmental pathogens are the most common cause

of mastitis and they induce mastitis between milking
 Etiology 6

Based on causative agent

• Contagious mastitis: S. aureus, Strep. Agalactiae,
Corynebacterium bovis, Mycoplasma
4TH COFFEE

• Opportunistic mastitis: Coagualse neagtive S. aureus

• Environmental mastitis: Strep. uberis, Strep. dysagalactiae,

E. coli, Klebsiella, Enterobacter, Truperella

• Uncommon pathogens: Nocardia, Pasterurella, Yeasts, Fungi

Mycobacterium, Bacillus cereus, pseudomonas, serratia
marcescens,

• Summer mastitis: Corynebacterium pyogenes

 Risk factors 7

• Animal risk factors

• Age
• Stage of lactation - early and late
4TH COFFEE

• Condition of the teat

• High producing animals are more
susceptible
• Vit E and Selenium deficiency
• Environmental factors
• Poor housing and management
• Bedding • Pathogens factors
• Ability to survive in
environment
• Toxin production,
virulence
 Clinical signs
8

• Gross abnormalities in milk

• Abnormalities in udder – swelling, pain, redness,

4TH COFFEE

gangrene

• Systemic response - anorexia, toxemia, fever,

ruminal stasis, recumbency
 Changes in milk composition
9

• Increase in number of pathogenic bacteria

• Marked increase in somatic cell count.

4TH COFFEE

• Impairment of synthetic ability of the secretary

tissue
• Reduced yield

• Alternation in major and minor milk constituents

• Increased infiltration of blood constituents like

serum proteins, into milk.
 Changes in milk composition
10

• Increased in concentration:
• Total whey proteins (BSA, Immunoglobulin)
• Sodium
4TH COFFEE

• Chloride
• Cu, Fe, Zn
• Various enzymes
• pH
 Changes in milk composition
11

• Decreased in concentration:
• Level of lactose

• Fat
4TH COFFEE

• Total casein (α & β decreases but γ increases)

• Potassium and other Minerals Ca, Mg, P

So mastitic milk has lower SNF, Fat, casein and

lactose

Higher serum proteins, chloride ions and pH

 Somatic Cells
12

• The animal body cells are called as somatic cells

• Normally present in milk

4TH COFFEE

• But their number increases steeply in mastitis

milk.

• There are two types of somatic cells in milk

• Epithelial cells derived from the secreting tissue of the
udder.
• Leucocytes derived from blood, higher than
epithelial cells.
 Somatic Cells
13

• SC are numerous in colostrum compared to

normal milk but it again increases in late lactation
4TH COFFEE

• In normal milk SCC varies from 4000 to 1

million / ml.

• In abnormal milk it may be up to 368 millions /

ml.

• As per the American Public Health Association

somatic cell count should not exceed
5x105 somatic cells / ml in normal milk.
 Milk constituents Normal Milk Mastitis milk 14

1. Fat % 3.45 3.2

2. Protein % 3.61 3.56
3. Lactose % 4.5-5.3 3.3-4.9
4TH COFFEE

4. BSA mg/ml 0.82-1.29 4.3-21.5

5. Lactoferrin mg/ml 0.2 6.2
6. Sodium mg/100ml 57 104.6
7. Potassium mg/100ml 172.5 157.3
8. Chloride mg/100ml 80-103 7250
9. pH 6.65 6.9-7.0
10. Somatic cell 102 / ml 20 - 1000 100 - 5000
 Milk constituents Normal Milk Mastitis milk 15

11. Catalase (µM 0.08 1.84

O2 produced/min/ml)
12. Lactic dehydrogenase (m 300 – 500 Up to 5525
units/ml)
13.
4TH COFFEE

Alkaline phosphatase 191 712

(units/ml)
14. Acid phosphatase (µ M 0.063 0.081
units/ml)
15. Lysozyme (µg/ml) 0.0 5.6

Protease, lipase, aldolase, sorbitol,

dehydrogenase etc also get increased
 Significance
16

• Mastitis affects the yield & quality

4TH COFFEE

• Unsuitable for the preparation of fermented products

• Yield reduced, starter cultures growth affected, aroma &
flavour affected

• Some microbes are also pathogenic to human beings.

• Staphylococci - cause food poisoning in humans
Produce enterotoxins, which are not inactivated even
during pasteurization and spray drying of milk.
• Salmonella and enteropathogenic strains of certain
coliforms
 Detection of Mastitis Milk
17

Based on compositional changes

• pH of milk: is higher than that of normal milk.
4TH COFFEE

• 5 ml of milk mixed with 1ml of 0.04% - 0.13%

bromothymol blue solution, the appearance of blue
green colour indicates mastitic milk (pH 6.8 or more)
as against grass green colour for normal milk (pH 6.6).

• Mastrip – indicator paper based

 Detection of Mastitis at herd level
18

• Bulk tank milk SCC

• Culture of bulk tank milk – S. aureus, S.
agalactiae
4TH COFFEE

• Sting sampling or milk line sampling – group of

cattle
 Detection of Mastitis at individual 19

level
• Abnormalities in udder and milk
• Culture
• CMT
4TH COFFEE

• SCC – automated electronic cell counters (<1L cells/ml)

• Electrical conductivity of milk - increases
• NAGase – cell associated enzyme - a lysosomal enzyme
derived from damaged mammary epithelium
• Infrared thermography
• Biopsy
• Ultra-sonography of mammary gland

• Indirect test detect inflammation not infection

 Detection of Mastitis Milk
20

• Based on compositional changes

• Chloride content:
• Normal milk - 0.08 to 0.14%
4TH COFFEE

• Abnormal milk - more than 0.14%.

• The chloride content of milk can be estimated by
addition of 5 ml of 0.1345% silver nitrate solution
and add 2-3 drops of 10% potassium chromate
indicator. A yellow colour indicates more than 0.14%
chloride content.
 Detection of Mastitis Milk
21

• Based on compositional changes

• Catalase test:
• The presence of catalase as examined by evolution
4TH COFFEE

of oxygen on adding hydrogen peroxide.

• Take 9 ml of milk to that add 1ml of H2O2,

presence of air bubbles in the sample indicates
mastitis.

• Place a drop of milk on a slide & a drop of H2O2 in

methylene blue put a cover slip immediately - air
bubbles under cover slip is positive
 Detection of Mastitis Milk
22

• Based on compositional changes

• Somatic cell count:

4TH COFFEE

• An increase in the number of somatic cells in

milk is indicative of mastitis.

• CMT
• Contains a detergent and Bromocresol purple indicator
(pH 6.6)
 Detection of Mastitis Milk
23

• Somatic cell count:

• Sodium lauryl sulphate test: It is a presumptive test,

4TH COFFEE

based on the increase in viscosity of milk on adding

sodium lauryl sulphate solution 4% SDS in a 15%
teepol solution (pH to 12.0)

• Two ml of milk is mixed with 2ml of reagent by

shaking gently for 20 seconds in a tube and observed
for coagulation.
 Detection of Mastitis Milk
24

• Somatic cell count:

• Sodium lauryl sulphate test:

4TH COFFEE

Observation Leucocyte count/ml Mastitis condition

No viscous layer Less than 1,00,000 No mastitis condition
Slight viscous layer 1,00,000-5,00,000 Sub-acute condition
Central viscous cone 5,00,000-50,00,000 Acute condition
disappears after
stopping rotation
Central cone persists More than 50,00,000 Acute condition
 Detection of Mastitis Milk
25

• Somatic cell count:

• Direct leukocyte count:

4TH COFFEE

The presence of more than 5,00,000 somatic

cells/ml of milk is indicative of mastitis.

It is performed in a similar way as DMC by

staining with Newman’s stain.
 Detection of Mastitis Milk
26

• Somatic cell count:

• Resazurin rennet test:

4TH COFFEE

• This test is based on the disturbance in the salt balance

and increase in leucocytes content in mastitic milk.

• Coagulation of milk by rennet is slowed down due to

disturbed salt balance and leucocytes reduce resazurin
dye faster.

• Done similar to RRT, - mastitic milk shows delayed

coagulation but faster resazurin reduction
 Detection of Mastitis Milk
27

• Based on the causative microorganisms

Microscopic examination:
• This is helpful even in detecting the admixture of
4TH COFFEE

mastitic milk with herd milk.

• Presence of long chains of streptococci is indicative of

mastitis due to [Link],

• Cells in grape like bunches - Staphylococcal mastitis.

 Detection of Mastitis Milk
28

• Based on the causative microorganisms

Hotis test:
This test is meant for differentiating [Link] from
4TH COFFEE

[Link].

Here 9 ml of milk should be mixed with 0.5ml of 0.5%

bromocresol purple solution and incubated at 37 0C for
24-72 h in a culture tube.

Appearance of canary yellow colour colonies along the

walls and at the bottom of the tube is diagnostic of
[Link] mastitis whereas [Link] gives rusty
brown colour colonies
 Detection of Mastitis Milk
29

• Based on the causative microorganisms

Blood agar test or use of selective agars:

4TH COFFEE

• Mastitic milk sample should be streaked on blood agar.

• The causatives of mastitis can be detected based on the

following observations.
 Detection of Mastitis Milk
30

Observation Organism
Small colonies, alpha or beta haemolysis [Link]
will occur or no haemolysis will occur in
some cases
4TH COFFEE

Alpha haemolysis (small zone around [Link]

colonies and green discoloration)
No reaction [Link]

Large colonies than streptococci, beta Staph. aureus.

haemolysis (a wide zone of clearance
around colonies)
 Detection of Mastitis Milk
31

• Based on the causative microorganisms

CAMP Test (CHRISTIE, ATKIN, MUNCH & PETERSON TEST)

This test is meant specifically to confirm Str. agalactiae from
4TH COFFEE

mastitis milk.

A standard culture of Staph. aureus should be streaked

vertically across the center of a blood agar plate.

The suspected streptococci should now be cross streaked

horizontally at an angle taking care not to touch
[Link] streak.

After incubation at 370C over night Str. agalactiae will

produce a clear zone between its streak and that of S.
aureus.
4TH COFFEE 32
 Prevention and control
33

• Proper milking management

• Proper management of milking equipments

4TH COFFEE

• Dry cow therapy- drying off period

• Early Diagnosis and management

• Culling of infected animals

• Hygienic maintenance of shed

• Routine udder health maintainence

4TH COFFEE

Thanks
34