Bovine Mastitis

Sukolrat Boonyayatra DVM, MS Clinic for Ruminants

Whats mastitis ?
Inflammation of one or more quarters of the udder

Mammae = breast -itis = Latin suffix for inflammation

Normal

Inflamed

Swelling pain warm redness

Causes

Intramammary Infection (IMI):

Bacterial infection Mycoplasmal infection Mycotic (fungal) infection Algal infection

Mechanical trauma Thermal trauma Chemical insult

Predisposes the gland to IMI

Economic Losses

Mastitis accounted for 26% of the total cost of all dairy cattle diseases. Losses from mastitis were twice as high as losses from infertility and reproductive diseases. Sources of loss

Reduced milk production Discarded milk Early cow replacement costs Reduced cow sale value Drugs Veterinary services labor

Determinants of Mastitis

Timing of infection and stage of lactation

Active involution

High Pressure in the gland Bacteria inside the gland Teat dipping ceases. Phagocytic efficiency Increasing of immunoglobulins and lactoferrin cannot override the problems noted above. Dry cow treatment can not reduce coliform IMI during active involution. Reducing the period of active involution by infusing colchicine (disrupts milk secretion mechanisms) decreases IMI during the active involution phase.

Timing of infection and stage of lactation

Peripartum period

Fluid volume in the gland increases Citrate concentration rises and lactoferrin is low Phagocytic cells efficiency
High immunoglobulin concentrations in the gland at this time are not effective in preventing new IMI. IgG1 is not normally an effective opsonin in the mammary gland. Antibiotic concentration Teat dipping

Timing of infection and stage of lactation

Early lactation

Metabolically stressed Mastitis is sometimes associated with high concentrate feeding which accompanies early lactation.

Nutrition and Mastitis

Micronutrient
Se

Observation
Decreased efficiency in neutrophil funtion Improved bactericidal capabilities of neutrophils Decreased severity and duration of mastitis Increased neutrophil bactericidal activity Decreased incidence of clinical mastitis In combination with Se, decreased prevalence of IMI at calving

Vit E

Vit A -carotene
Cu Zn

Decreased SCC Moderated glucocorticoid levels Increased bactericidal function of phagocytes Increased mitogen-induced proliferation of lymphocytes
Deficiency decreased neutrophil killing capability Deficiency increased susceptibility to bactericidal infection Deficiency decreased leukocyte function Deficiency increased susceptibility to bacterial infection

Inflammation of Mammary gland

1. Multiplication of bacteria in mammary gland 2. Vasodilation 3. Increased vascular permeability 4. Swelling

5. Diapedesis 6. Phagocytosis and destruction of bacteria 7. Tissue repair

Development of mastitis and the cows defense against the infection

The major routes of bacterial transmission

Mastitis Clinical Syndromes

Categorized based on Severity of Immune Response

Peracute Mastitis: sudden onset, severe inflammation of the udder, and serous milk-Systemic illness often precedes the symptoms manifested in the milk and mammary gland. Acute Mastitis: sudden onset, moderate to severe inflammation of the udder, decreased production, and occurrence of serous milk/fibrin clots, Systemic signs are similar but less severe than for the peracute form.

Mastitis Clinical Syndromes

Subacute Mastitis: mild inflammation, no visible changes in udder, but there generally are small flakes or clots in the milk, and the milk may have an off-color. There are no systemic signs of illness. Chronic Mastitis: Chronic mastitis may persist in a subclinical form for months or years with occasional clinical flare-ups. Treatment usually involves treating the clinical flare-ups, or culling the cow from the herd.

Mastitis Clinical Syndromes

Subclinical Mastitis: the most common form of mastitis, 15x40 X more common than clinical mastitis, no gross inflammation of the udder and no gross changes in the milk, decreased production and decreased milk quality Elevated Somatic Cell Count

Abnormal Milk

Abnormal Udder

Somatic Cell Count

~98-99% White Blood Cell + 1-2% Epithelial cells from milk-secreting tissue Cows natural defense mechanism Normal or uninfected cow: 50,000-200,000 cells/ml >200,000 cells/ml: the likelihood of infection increase Prevalence of subclinical mastitis in Chiang Mai may be exceed 80%. 1 clinical mastitis : 15-40 subclinical mastitis

Effects on Milk Quality

Subclinical mastitis results in INCREASES in undesirable milk components and DECREASES in the desirable components. Pasteurized milk that is processed from raw milk with a somatic cell count below 250,000 has a significantly longer shelf-life than products made from milk with a somatic cell count above 500,000.

Lactose (good) Total proteins (good) Casein (good) Immunoglobulins (bad) Solids not fat (good) Total solids (good) Fat (good) Lipase (bad) Sodium (bad) Chloride (bad) Calcium (good) Phosphorus (good) Potassium (good) Trace minerals (bad) Cheese (good) Heat stability (good)

Decreased 5 to 20% Decreased slightly Decreased 6-18% Increased Decreased up to 8% Decreased 3 t0 12% Decreased 5 to 12% Increased rancidity Increased Increased Decreased Decreased Decreased Slight increase Decreased curd strength, fat and yield Reduced

What are the health concerns of mastitis ?

Animal health

Loss of functional quarter Lowered milk production Death of cow

Poor quality milk antibiotic residues in milk

Human health

How severe can mastitis be ?

Subclinical Mastitis

Clinical Mastitis

~ 90 -95% of all mastitis cases Udder appears normal Milk appears normal Elevated SCC (score 35) Lowered milk output (~ 10%)
Longer duration

~ 5 - 10% of all mastitis cases Inflamed udder Clumps and clots in milk

Acute type major type of clinical mastitis bad milk loss of appetite depression

prompt attention needed

Chronic type bad milk cow appears healthy

What causes mastitis ?

Bacteria ( ~ 70%) Yeasts and molds ( ~ 2%)

Unknown ( ~ 28%)

physical trauma weather extremes

Where do these organisms come from ?

Infected udder

Environment bedding soil water

manure Replacement animals

How does mastitis develop ?

Cow

Predisposing conditions Existing trauma (milking machine, heat or cold, injury) Teat end injury Lowered immunity (following calving, surgery)

Nutrition Organisms Environment

Process of infection
Organisms invade the udder through teat canal

Migrate up the teat canal and colonize the secretory cells

Colonized organisms produce toxic substances harmful to the milk producing cells

The cows immune system send white blood cells (Somatic cells) to fight the organisms

recovery

clinical

subclinical

Organisms

Contagious microorganisms

Staphylococcus aureus Streptococcus agalactiae Mycoplasma bovis Corynebacterium bovis Environmental streptococci Coliform Staphylococcus spp. (CNS) Pseudomonas aeruginosa Actinomyces pyogenes Nocardia Species

Environmental microorganisms

Opportunistic microorganisms

Others

Bacterial Infection: Streptococci

Environmental

Contagious

S. uberis S. dysgalactiae S. equinus

S. agalactiae

More subclinical mastitis Environment Predominant early and late lactation

Clinical mastitis Resides in the milk and on the surface of the milk channel Cannot invade the tissue Accumulate Neutrophils Ducts and acinar epithelium damage Inter-alveolar tissue fibrosis loss of secretory function Treated easily with penicillin

Bacterial Infection: Staphylococci

Staph aureus

Other staph

Gangrenous mastitis: alpha toxin Spread by milking equipment and milkers hands Fibrous tissue replacement low production Poor response to ABO Dry cow therapy Persistent, difficult to eliminate

Found normally on skin Lowers milk yield Elevated SCC Easily responds to antibiotics Relapse frequently seen

Fig. 1. Mammary parenchyma from which coagulase-negative Staphylococcus was isolated, showing the presence of mononuclear cells. HE. 660

Fig. 2. Mammary parenchyma from which coagulase-negative Staphylococcus was isolated, showing the presence of neutrophils within the alveolar lumen. HE. 660 .

Fig. 3. Mammary parenchyma from which Prototheca sp. was isolated, showing the micro-organisms within the alveolar lumen. HE. 660 .

The cocci in the lesions of the mammary glands show a positive reaction to antibody against Staph.aureus (ABC X 200)

a. The bacteria were round or oval in shape, showing a thick cell wall, characteristic of gram-positive bacteria (TEM. X 40,000) b. Fibrous material (arrows) stained by ruthenium-red, around the bacterial cell wall, which forms a capsule (TEM.X 250,000)

Severe necrosis of interlobular and intralobular ducts The lesions affected the intralobular duct, intralobular duts and alveoli (Azan x 30).

Bacterial clumps(arrows) surrounded by alveolar epithelial cells undergoing necrosis Thrombus(*)is seen in the blood vessel(He x 100).

Bacterial Infection: Coliforms

Groups of organisms

E. coli, Klebsiella, Enterobacter

Environmental source (manure, bedding, barns, floors and cows) Coliforms cause acute clinical mastitis

Multiply rapidly with low SCC Endotoxin releasing High temp, and inflamed quarter Watery milk with clots and pus Toxemia The udder can be gradually return to normal without fibrosis

Bacterial Infection: Other organisms

Pseudomonas aeruginosa

Out breaks of clinical mastitis or subclinical mastitis Similar pathogenesis to coliform mastitis Severe endotoxaemia can occur. Out breaks of clinical mastitis Most common in Europe Actinomyces pyogenes + Peptostreptococcus indolicus Non-lactating heifers and cows at pasture in the summer months and more common during wet weather Fly borne ?? Severe systemic reaction and Loss function Abcess develop

Serratia

Summer mastitis

Bacterial Infection: Other organisms

Mycoplasma mastitis

Clinically severe mastitis Rarely systemic involvement All ages & all stages of lactation Post calved cows show more severe signs. Long-term persistence in udder (up to 13 mths) Some cows can shed the organism without clinical signs. Normal secretion in the early stage of infection Flaky material settles out leaving a turbid Whey-like supernatant fluid Very high SCC

How is mastitis diagnosed ?

Physical examination

Cultured Analysis

Signs of inflammation Empty udder Differences in firmness

Cowside tests California Mastitis test

The most reliable and accurate method

Unbalanced quarters

Treatment

Clinical mastitis

Strip quarter every 2 hours Oxytocin valuable high temp, give NSAIDs Seek veterinary assistance
Subclinical mastitis Questionable

Treatment with penicillins

1/8 1/4

Partial Insertion

Standardized Milking Procedures

Stanchion/ Tie stall

Parlor

Ware gloves Wipe off excess dry manure, straw and bedding Strip each teat into a stripcup Dip teats with an approved pre-dip Allow the pre-dip to react for at least 30 sec.

Wear Gloves Wipe off excess dry manure, straw and bedding Strip each teat into a stripcup Dip teats with an approved pre-dip Dip 3-4 cows Allow the pre-dip to react for at least 30 sec.

Standardized Milking Procedures

Stanchion/ Tiestall

Parlor

Clean teat and teat ends using single paper towel or individual towel clo Return to the first cow and The teats must be dried for at least 15 clean sec teat and teat ends Attach milking machines immediately after teats are dried using a single paper towel Dip teats with post-dip immediately after or milking individual towel cloth The teats must be dried for at least 15 sec Attach milking machines immediately after teats are dried Dip teats with post-dip immediately after milking

Detection of Mastitis
Visualization Detection

and palpation of the udder

of Somatic Cells California Mastitis Test

N-acetyl--D-glucosaminidase (NAGase) - a lysosomal enzyme which increases in milk when mastitis is present

Indirect chemical tests to detect mastitis

Electrical conductivity: Sodium and Chloride ions A radial immunodiffusion test : Serum albumin concentration increases if epithelium damage is present. An anti-trypsin test: Anti-trypsin activity tends to naturally high at the beginning of a lactation the values are high only if serum anti-trypsin has leaked through damaged mammary epithelium.

California Mastitis Test (CMT)

The CMT reagent reacts with genetic material of somatic cells present in milk to form a gel. A plastic paddle having four shallow cups marked A, B, C and D for easy identification of the individual quarter. Approximately 1/2 teaspoon (2 cc) of milk is. An equal amount of the CMT reagent is added to the milk. A circular rotating to thoroughly mix the contents. Score in approximately ten seconds while still rotating. Read the test quickly as the reaction tends to disintegrate after about 20 seconds. Rinse the paddle thoroughly with water and it is ready for the next test.

Advantages of CMT

Fairly accurate in measuring SCC in milk Primarily developed for sampling quarters, it can also be used on "bucket" and "bulk tank" milk samples. Foreign material does not interfere with the test. It is inexpensive, simple, and little equipment is needed. Easy clean-up after each test--simply rinse with water. Environmental temperature changes have little effect on the CMT as long as the milk has been refrigerated and is not over two days old. Herd mastitis levels can be estimated from tank CMTs. A CMT of 2 or 3 on tank milk indicates a probable high percent of infected cows.

Disadvantages of CMT

Scoring the test may vary between individual testers. It is necessary to be as consistent as possible to insure uniform results. Scores represent a range of leucocyte content rather than an exact count. False positive reactions occur frequently on cows that have been fresh less than ten days, or on cows that are nearly dry. These animals should be tested closer to the middle of the lactation. Occasionally, acute clinical mastitis milk will not score positive due to the destruction of leucocytes by toxins (poisons) from the infecting organism.

Correlation between the California mastitis test result and the somatic cell count.
CMT Interpretation score 0 Negative Visible reaction Milk fluid and normal Total cell count (/ml) 0-200,000 0-25% neutrophils

T
1

Trace
Weak positive

Slight precipitation
Distinct precipitation but no gel formation

150,000-500,000 30-40% neutrophils

400,000-1,500,000 40-60% neutrophils

2
3

Distinct positive

Mixture thickens with a gel formation

800,000-5,000,000 60-70% neutrophils

Strong positive Viscosity greatly increased. >5,000,000 Strong gel that is cohesive 70-80% neutrophils with a convex surface.

Steps involved in employing HACCP-based concepts for establishing proper milking procedures

Step 1

Educate owners and milkers about implementing a standardized milking procedure (Benefits !!!!!!) IF a dairy farm initiates and shows sustained interest

Establish ground rules They will have to be proactive and adopt changes TEAM EFFORT !!!

STEP TWO

Establish a team ( owner, milkers, veterinarian, facilitator) Mission statement Goals and timeline Written Procedures

Protocols Critical Limits ( SCC > 250,000) Milking time/milking Bulk Tank Temp; end of 1 hr of milking Sanitation

Recording Keeping

Schedule team meetings to review the process

STEP THREE

Train milkers and owners in implementing the standardized milking procedure Monitor the application of the standardized milking procedure Floor tests (each step is a critical point !) Laboratory tests (SPC or BTSCC) Monitor records Establish corrective actions to be implemented if milk quality critical limits have exceeded.

STEP FOUR

STEP FIVE

Factors Affecting Somatic Cell Counts

1. Mastitis (Udder infection) 2. Teat or udder injury 3. Number of quarters with mastitis 4. Age of cow 5. Stage of lactation 6. Season 7. Stress 8. Day to day variation 9. Technical factors 10. Management factors

Uses of SCC on Individual Cows for Management Decisions

1. Milk Culture and Sensitivity Testing 2. Treatment During Lactation 3. Drying Cows Off Early 4. Culling

1. Milk Culture and Sensitivity Testing

High SCC >500,000 cells/ml Very useful when:

High SCC two or more tests Beginning of lactation

2. Treatment During Lactation

Strep. agalactiae infection Very few cases of subclinical mastitis High SCC vs culture vs sensitivity

3. Drying Cows Off Early

The best method of eliminating infection High SCC and relatively low production There is evidence to suggest that a repeat dry treatment 3 weeks after the first therapy could increase success rate. Teat dipping for 10 days after lasting milking and for 10 days prior to calving

4. Culling

Persistently high SCC from lactation to lactation Staph. aureus or Mycoplasma spp. Milk production

5. Milking Routine

High SCC cow could be milked last or the milking machines could be sanitized after milking.

Mastitis Bulk Tank Culture Report Interpretation

Type of Bacteria
Strep agalactiae

Usual Infection Cause

Infected udders of other cows in herd

Major Means of Spread

Cow-to-cow by contaminated udder wash

Mastitis Control
Use separate towels to wash/dry; Teat dipping; dry cow treatment; eradicate in special cases Use separate towels to wash/dry; Teat dipping; dry cow treatment; milk infected cow last, cull chronically infected cows

Staph aureus

Infected udders of other cows, contaminated bedding from milk of infected cows

Cow-to-cow by contaminated udder wash rag, milkers hands contaminated milking equipment , and improperly functioning equipment

Mycoplasma

Infected udders of other cows, often from infected purchased cows/heifers

Cow-to-cow by hands of milkers, equipment, and common towels. Aerosol transmission from animals with respiratory signs may also occur. Or the bacteria can move from a respiratory tract infection to the udder or joints.

Careful purchasing of replacement cattle, using bulk tank and cow culturing to monitor herd status and clinical cows. Use separate towels to wash/dry; teat dipping; dry cow treatment; milk infected cows last, cull any positive clinical case.

Mastitis Bulk Tank Culture Report Interpretation

Type of Bacteria Non-ag Streps Usual Infection Cause Environment of cow Major Means of Spread Environment of the cow by; wet dirty lots, contaminated bedding, milking wet cows, poor cow prep, milking machine air slips Environment of the cow by; wet dirty lots, contaminated bedding, milking wet cows, poor cow prep, milking machine air slips. Hot humid weather. Poor teat dip coverage, poor cow prep, old bedding. Mastitis Control Improve stall and lots sanitation; milk clean dry cows, avoid air leaks and liner slips, changes bedding frequently. Keep cows standing after milking. Improve stall and lots sanitation; milk clean dry cows, avoid air leaks and liner slips, changes bedding frequently. Keep cows standing after milking. Consistent teat dipping, adequate cow prep, and more frequent bedding change.

Coliforms

Environment of cow

Staph species

Environment of cow

Good Milking Procedures

1. Provide Cows with a Clean, Stress-Free Environment 2. Check Foremilk and Udder for Mastitis 3. Wash Teats and Lower Surface of the Udder with a Warm Sanitizing Solution 4. Use a Premilking Teat Dip (Optional) 5. Dry Teats Thoroughly 6. Attach Teat Cups within 1 min. 7. Adjust Milking Units as Necessary 8. Shut Off Vacuum Before Removing Teat Cups 9. Dip Teats with a Safe and Effective Teat Dip 10. Disinfect Teat Cups Between Cows (Optional)

Problem Herd Handling

Problem Solving Techniques

One or more specialists: vet, fieldman, extension agent or milking machine dealer A visual inspection of the general environment Good detectives Specific approach

Problem Herd Handling

High Incidence of Clinical Mastitis and High SCC

Detection and discarding of visibly abnormal milk. Not milking fresh cows with cows that have clinical mastitis Collection of milk samples and culturing in a diagnostic laboratory. Treatment of selected cows, especially those infected with Streptococcus agalactiae. Culling of cows with chronic infections, particularly those caused by Staphylococcus aureus, environmental streptococci, Nocardia asteriodes, and Mycoplasma species. Drying off of selected cows and dry treating. Correction of deficiencies in management and environment.

Problem Herd Handling

(Continued) Upgrading of milking equipment Correction of deficiencies in milking hygiene. Improvement in the manner in which milking machines are used. Initiation of predipping. Strengthening of postmilking teat dip procedures. Arranging for fresh feed to be available when cows exit the milking parlor or barn so they will be encouraged to stand for at least 1 hour after milking to provide time for the teat canal to close tightly. Segregation of infected cows. Initiation of backflushing, particularly if the problem is caused by contagious microorganisms such as Staphylococcus aureus, Streptococcus agalactiae, or Mycoplasma species.

Problem Herd Handling

High Bacteria Counts

>10,000 /ml, Streptococci >75% Infected Udder Streptococci < 25% Improper Cleaning of Milking Equipment, Poor Udder Preparation and Poor Cooling of Milk High Streptococci & High Staphylococci + Coliforms + Spore Formers + Other Organisms A dual problem of infected cows and poor udder preparation. >15,000 /ml of Staphylococci Poor cooling of milk High coliform counts Broken teat cup liners, low water temperature, milkstone on milk-contact surfaces and failure to use correct chemicals for cleaning milking equipment Large number of coliforms, staphylococci, and environmental streptococci Faulty cooling of the milk

Mastitis Prevention Principles

1. Milk cow with clean, dry teats and teat ends.

Impact: Milk quality, environmental mastitis, liner slips, milk out and parlor throughput

2. Prevent transfer of pathogens from cow to cow during milking.

Impact: Contagious mastitis, milk quality

Impact: Mastitis, milk out, parlor throughput

3. Prevent injury to the teats during milking.

Mastitis Prevention Principles

4. Provide an environment that allows the cows to remain clean between milking.

Impact: Environmental mastitis, milk quality, parlor throughput, cow comfort

5. Early detection of new infections (clinical and subclinical).

Impact: Response to treatment, chronic infections, culling Impact: Success of treatment, cost control, residues in milk and meat

6. Proper use of medications.

Mastitis Prevention Principles

7. Control duration of infections.

Impact: Decreased prevalence, decreased culling Impact: Prevent outbreaks, culling information Impact: Permit culling for production, reduced herd prevalence

8. Monitor mastitis status.

9. Raise mastitis free replacements.

10. Assume all purchased replacements are infected.

Impact: Control introduction of new pathogens

Mastitis Prevention Principles

11. Provide adequate nutrition to preclude increased susceptibility to mastitis.

Impact: Control new infection rate

Impact: Teat end injury, new infection rate Impact: All areas of mastitis prevention and control, milk quality

12. Fly control.

13. Provide routine milker training

14. Assigned responsibilities for all areas of mastitis prevention.

Impact: Job knowledge, shared responsibility, improved compliance