BOVINE MASTITIS: AN UPDATE

By Dr S.V. Singh, Assistant professor, Deptt. of Vet. Clinical Medicine College of Veterinary Sciences N.D.U.A. & T.

MASTITIS
Inflammation of mammary gland parenchyma Characterised by: abnormalities of milk & abnormalities of udder

Development of mastitis
A B

Organisms remain on .the udder surface.

Organisms enter teat canal and disrupt the teat canal barrier.

Organisms move further and affect the udder immune barrier.

Organisms establish and multiply in the udder.

Somatic cells engulfing bacteria

Inflammation and damage to the udder

PREDISPOSING

Interaction of bacterial, host and environmental factors leading to mastitis

FACTORS

Bacterial Factor Host Factor Environmental factor

Bacterial Factors
Ability to survive in the immediate environment of the animal Ability to colonise the teat duct. Ability to adhere to mammary epithelium and not be flushed out with the milk flow The degree of invasiveness. For example, streptococci cause little pathological change to secretory cells but staphylococci initiate degenerative changes. Ability to resist phagocytosis and antibacterial substances in udder, including resistance to antibiotics.

Host Factors
Genetic predisposition to mastitis - teat shape, sphincter tone, anatomy of the teat canal and susceptibility to weakening of suspensory ligament. (Pendulous udder) Age: older cows, especially after four lactations. Stage of lactation: more susceptible just after calving and for the following 2 months. Presence of lesions on the teats that may predispose to inadequate milking or may harbour
mastitis producing bacteria.

Environmental factors
Presence of large numbers of potential pathogens in the immediate environment of the animal. (poor hygiene ) General management of animals. For example, Coliform mastitis is much more frequent in housed cattle. Milking machine malfunction or inadequate design. Milking shed environment including poor milking techniques and hygiene. External traumas such as that arising from rough, middy approaches to the milking shed or with cause, the sucking of large vigorous lambs.

CLASSIFICATION
Per acute Acute Sub-acute Chronic Subclinical

 Peracute form: hot, swollen and painful quarter, abnormal milk, systemic disturbance - fever, depression, anorexia, weakness and a rapid weak pulse Acute form: severe inflammation, but systemic reaction is not so marked.

 Chronic from: Absence of systemic signs, very few external signs of change in the udder, recurrent episodes of mild inflammation and slight change in milk. Subclinical form: No visible abnormality in udder and milk , milk appears normal unless tested, positive in laboratory.

Gangrenous mastitis
Cracked skin of teats/udder Reddish to bluish black discoloration Hard, cold and painful with honey like discharge from fissures Sloughing off

Gangrenous mastitis in goat

Sloughed off teat

Udder abscess

Udder edema

DIAGNOSIS
STEPS INVOLVED
1.Thorough examination of udder, 2. Examination of individual quarters 3. Examination of milk.

EXAMINATION OF UDDER
Inspection, Palpation, Inspection of the udder secretion

INSPECTION
The size and shape of the udder, individual quarters and the teats should be inspected by viewing it from front, on each side, and from behind. Preferably the udder should be examined from the back with hind legs held apart and tail lifted upwards. This position allows view of whole udder.

PALPATION
The teat: Palpated with the finger tips, The quarters: Palpated (preferably after milking) with the flat of the hand, first feeling the surface, then the deeper tissue. A convenient procedure is to start at the tip of the teat and proceed upwards.

 The teat (streak canal) normally feels firm when rolled between the fingers and all four teats should feel the same. Abnormalities include swelling, injury and neoplasia of the tip and teat canal: also abnormal tenderness and increased warmth. Patency of the teat canal is tested by drawing off a few streams of milk (into a stripping cup-not onto the ground).

 Udder skin is palpated for

Surface temperature (exceptionally warm in mastitis accompanied by cellulitis; cold in gangrenous mastitis), Tenderness, Thickening, Hardening and Elasticity. The skin of milked out udder should be more so elastic that a fold can be pulled out.

EXAMINATION MILK

OF

THE

Broadly classified under two categories i.e. (i) The detection of alteration in milk as a consequence of pathophysiological changes due to an inflammatory response and (ii) The direct detection of microorganisms in milk by culture.

Sample transport

collection,

storage

and

Storage- Sterilized glass or plastic tubes of 10-15 ml capacity with appropriate tight fitting caps Udder - washed properly & dried with the help of a muslin cloth. Collection -just prior to the milking particularly from the fore milk which contains more mastitis organisms.

Sampling- individually.
Herd testing :a composite sample from all the quarters.

 Teats - scrubbed thoroughly within a cotton ball soaked in 70% alcohol. Hands- gloved, rinsed in a germicidal solution and thoroughly dried or change the gloves after taking the sample, from proceeding to another animal. Examinationimmediately after the collection. Transport to laboratory - in ice. If stored for 15-30 minutes it should be stored at 4-5C.

Strip cup test

Routine examination of the milk using a filter cup Moak firstly made use of strip cup test for the detection of visible particles of milk Types of strip cups (Two)
Fine screen type : Filters the pus, clots and flakes. Black background type: Helps to detect the change in colour as well as presence of particles. NOTE: Milk that is hotter than normal may be a good indication of a Staphylococcus aureus infection

California

Method described by Schalm et al. (1971) Principle: reagents cause rupture of somatic cells and release cellular proteins, including the DNA. These proteins unite with the reagent causing it to precipitate. The reaction is best seen in alkaline pH and since pH of the mastitic milk is mostly alkaline, therefore, it is most likely to give the above reaction. The reagent Bromocresol purple imparts purple colour to the mixture.

Mastitis (CMT)

Test

CMT Reagent
The test reagent maybe prepared as follows Bromocresol purple 0.01 gm Sodium hydroxide 1.5 gm Teepol 0.5 ml Distilled water 100 ml

 2-3 ml of milk + equal amount of CMT reagent gentle circular motion Positive reaction : gel formation & change of colour When the mixture is distinctly deep purple colour : + (Alkaline milk)

 INTERPRETATION
Reaction No precipitate or colour change Slight precipitate Degree of test - (Negative) T (Trace)

A distinct precipitate without gel 1+ formation and slight purplish colour positive)
Viscosity of milk increased with moderate gel formation Immediate thick gel formation, sticking at the bottom

(Weak

2+ (Positive) 3+ (Strong positive)

Whiteside Test (WST)

Based on the reaction between 4% sodium hydroxide and milk. The leukocytes of mastitic milk gets disintegrated upon contact with sodium hydroxide. The nucleic acid forms a sodium salt in the presence of sodium hydroxide, producing a gelatinous mass to which serum solids and fat globules become adsorbed to produce characteristic precipitate. Procedure: Five drops of milk + 2 drops of reagent (Stirring for about 20 seconds )

REACTION Mixture remain opaque & free of particles No apparent reaction during stirring, inspection reveals finely dispersed particles close

DEGREE OF TEST - (negative)

Thickening during stirring with little or no tendency for the mass to adhere to the stick; on continued stirring the mixture separate into milky whey and white particles
Thickening, the coagulum follows the stick and finally separated into a clear whey and white, thread-like whorls A tenacious mass forms immediately adhering to the stick; the mass separates into whey and thready, clumped, opaque material

1+

2+

3+

A tenacious coagulum with little or no tendency to break down into whey and particulate material

4+

Mastrip Test (MST)

Cellulose based bromothymol blue strip impregnated with stabilized ion One drop of milk is placed on the mastrip The colour reaction is graded as Reaction Degree of test No change in colour (yellow) Light green + Dark green ++ Bluish green +++

Electrical Conductivity
The electrical conductivity of milk increases in mastitis owing to altered concentration of ions in mastitic milk. Measured by using electrical conductivity meter. Conductivity is measured in Siemens (S) and is calculated by dividing amphere by voltage. The electrical conductivity of normal milk ranges between 4.0 5.5 mS/cm.

Somatic Cell count

Interpretations..

Normal SCC in milk is generally below 200,000 per ml An SCC above 250,000 - 300,000 is considered abnormal and nearly always is an indication of bacterial infection causing inflammation of the udder.

TREATMENT
Antibiotics Systemic Intramammary Preferably the same antibiotic should be used Anti inflammatory drugs Anti histamines

Antibiotic therapy
Mastitis is the most common reason for antibiotic use in the dairy industry Properties of antibiotic therapy intended for mastitis therapy
The bioavailability from intravenous administration with low plasma protein binding and high milk, plasma ratio. The antibiotic should have low minimum effective concentration (MEC) against major causative pathogens with longer elimination half-life from the udder tissue.

 Drugs of choice Aminoglycosides neomycin, Framycetin, ampicillin & cloxacillin and third generation cephalosporins Best performance: Ampicillin, Amoxycillin, Erythromycin & Tylosin Medium performance: Penicillin G, Cloxacillin & Tetracyclines Poor diffusers which do have a longer T1/2 : Streptomycin & Neomycin

Trial reports (gangrenous mastitis)

Treatment group Ceftriaxone Ceftriaxone

% efficacy 66.6 93.75

Reference Singh, 2004 Singha et al, 2004 Sharma, 2004 Singh et al ,2008

Ceftriaxone + gentamicin Ceftriaxone + gentamicin

92
83.33

Failure therapy

of

antibiotic

Development of resistance:
indiscriminate use of antibiotics the ubiquitous nature of mastitogens.

The bacteria too sometimes become refractory to the effects of the drug.

Demerits of intramammary therapy

Makes animal susceptible to the injury to teat canal if sterile conditionsplaces microbes directly into the udder Causes the dilation of teat canal lumen, and enhancing bacterial penetration. The inflammation, edema or fibrosis of the mammary gland results in a poor distribution of the drug within the udder.

ALTERNATE THERAPY
ANTIOXIDANT THERAPY PHYTOTHERAPY HOMEOPATHY CYTOKINE THERAPY OXYTOCIN THERAPY

Entry of bacteria into mammary gland Mobilisation of PMN cells Engulf and kill bacteria through RESPIRATORY BURST Release of free radicals

Adequate antioxidant

Insufficient antioxidant

Allow a more active and more prolonged effect of a cows natural immune system on invading bacteria

Free radicals accumulate

OXIDATIVE STRESS

 Cells can tolerate a mild oxidative stress Antioxidants- scavange and minimize the formation of ROS, but they are not 100% effective. When ROS are not effectively and safely removed, oxidative stress may impair health in dairy cows both directly and indirectly Directly: per oxidative damage to important lipids and macromolecules Indirectly: by changes induced in cellular membranes and components Tissue injury contributes to the development of disease conditions, mastitis being one of them

Commonly used antioxidants

Vitamin E Vitamin A Vitamin C Selenium Zinc

VitaminE Supplementation
Most important lipid soluble antioxidant. An integral component of all lipid membranes and protects lipid membranes from attack by Reactive Oxygen Species, ROS. Plays a regulatory role in the biosynthesis of various inflammatory mediators and influences both cellular and humoral immunity. Increases intracellular killing of both Staphylococcus aureus and Escherchia coli (Hogan et al.,1993)

Vit E conc. during periparturient period

Declines around 7 to 10 days prior to calving, and first two to three weeks of lactation Large quantity of the vitamin E is secreted in the colostrum causing a 47% drop in maternal Vitamin E concentration Increased cortisol concentration in the blood prior to calving reduces vitamin E content in neutrophils. Therefore, Vitamin levels are lowest around calving, the time of maximum susceptibility.

 Nauriyal (1996)
Dose: 1000 IU of tocopherol during the dry period and during early lactation Result:
Supplemented group - 12.5% incidence Unsupplemented group -56.25% incidence

Singh et al. (2000)

80% efficacy in animals administered Vitamin E and Selenium injections (E care Se) and gentamicin 60% efficacy in gentamicin treated group.

Dose recommendations
Dry cows :1000 IU/day Lactating cows :500 IU/day Injection of selenium and vitamin E or feeding supplemental amounts of these nutrients during the 3 weeks prepartum may reduce the incidence of post lactational disorders.

PHYTOTHERAPY
Ingredients/ Product Findings References
in Saxena et al., 1995, and Madan et al., 1996. of Kamal et al.,1998 Pachauri et al,. 1999 Dash, 2001 Mastilep (Dabur Efficacious Ayurvet Ltd.) treatment prophylaxis SCM Bonmilk

100% recovery 79.55% cure rate

Oil extract of 88.88% Ocimum sanctum with Azaderachta Aqueous extract of 100% recovery Tinospora cordifolia

Advantages
Nontoxic nature Cost effectiveness The ingredients have antibacterial, antiinflammatory, analgesic and immunostimulatory properties Effective against not any specific organism but plethora of organisms, The immunopotentiating activity of these drugs enhance the bodys defence mechanism along with udder immunity.

HOMEOPATHY
Belladonna
1 m: For acute postpartum mastitis. Udder very hot and red, painful to the touch. Animal is hot, and pulse is quick and strong. 1 dose every hour. 4 doses. Bryonia Alba 30 c : udder swollen, hard, Pain less intense when pressed. Animal is often Iying down. Especially good for chronic cases with fibrosis. Acute cases: 1 dose every 4 hours. 4 doses. Chronic cases: 1 dose 2 times a week for one month. Phytolacca 30 c : Clinical cases with sour, coagulated milk. Chronic cases with small clots at mid-lactation. Silicea 200 c : Few flakes in thin yellow milk. Also useful for summer mastitis cases with purulent abscess. 2 doses a week for 4 weeks. Ipeca 30 c : Useful for treating internal bleeding that produces pink or bloody milk. 3 doses a day for 3 days. Calcaria florica with silicia: Chronic mastitis and fibrosis. Lachesis: Udder discoloration, fever and putrified curd like appearance of milk

PREVENTION & CONTROL

Contagious mastitis - break the chain of transmission
Eliminating the source of infection (the infected mammary gland, by isolation and quarantine) and With strict sanitation at the time of milking.

Environmental mastitis - restrict the contact of populations of environmental bacteria with the mammary gland.
Good hygiene

 Invasion state :
Avoid the animals to sit immediately after calving, approx. 10 min (prevents the contact of microbes to udder). Teat dipping

Infection stage :
Boosting udder immunity by supplementing different antioxidants

Inflammation stage :
immediate medical care animals should be segregated from the rest.

Satyavrat Singh, Sonu Jaiswal, Desh Deepak Singh, Harnam Singh