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Title: Trouble Shooting Mastitis Problems: Using laboratory culture diagnosis to make mastitis treatment de


1
Trouble Shooting Mastitis ProblemsUsing
laboratory culture diagnosis to make mastitis
treatment decisions
  • Philip Sears, DVM, PhD
  • Department of Large Animal Clinical Science
  • College of Veterinary Medicine - Michigan State
    University
  • East Lansing, Michigan USA

2
Herd Investigation Disease Control
Step 1 Define Problem Use Dairy Analysis
Workbook - page 24) List possible
problems Review records (health, DHIA)
Review farm monitors to identify patterns Step
2 Use Diagnostic Testing to Determine Problem
Diagnostic Laboratory results
Veterinarian Other laboratories Step 3
Generate and Select Solutions Step 4 Develop a
Farm Plan Protocols Set goals
Create Standard Operating Procedures (SOPs)
Monitor records
3
Clinical Mastitis Definition
  • Score 1 Mild (signs milk)
  • abnormal milk
  • Score 2 Moderate (signs milk gland)
  • abnormal milk
  • swelling, pain
  • Score 3 Severe (signs milk, gland cow)
  • abnormal milk
  • swelling, pain
  • systemic fever, dehydration shock

Sears, 2001
4
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5
Clinical Predictive Schemefor Gram-negative
Mastitis
  • White, et al Jones Ward Morin, et al
  • watery milk watery milk watery milk
  • swollen gland summer summer
  • fever fever ? milk protein
  • weakness gt10 dim ? rumen sounds
  • clear/white milk old cow
  • no abscesses
  • previous mastitis

watery milk more common in Strep mastitis than
coliform mastitis - Obritzhauser et al, 1995
6
Clinical Predictive Schemefor Gram-negative
Mastitis
  • 3 experienced practitioners correctly predicted
    21/36 (58) culture results
  • 4 less experienced practitioners correctly
    predicted 39/82 (48) culture results
  • For coliform mastitis
  • Sens .64, Spec .64
  • PPV .51, NPV .76 (prevalence 31)

White et al, 1986
7
Milk Cultures
  • Infection level in herd
  • Type of infections
  • Source of infections
  • Control and Prevention

8
Milk Cultures
  • Clinical quarters or cows
  • Herd analysis
  • all cows
  • HSCC cows
  • monitoring herd
  • bulk tank milk

9
Milk Culture
10
Milk Culturessample consideration
  • Cow samples
  • Composite cow samples for herd survey
  • quarter samples - clinical cases, high SCC

11
Milk Sampling
12
Mastitis Problem Solving
Step 1 Identify possible mastitis problems.
(review records)
Subclinical mastitis
Clinical mastitis
Bulk tank SCC gt250,000 cell/ml
DHIA - SCC gt15 with LSgt4.5
New clinical cases gt2 per month
Severe clinical cases gt1 per year
Step 2 Use diagnostic test to determine
causative organism.
Bacteriological Cultures
Environmental organisms (usually clinical)
Contagious organisms (usually subclinical)
Coliforms
Strep sp
Staph sp
Staph aureus
Strep ag
13
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14
Culture Results
  • Culture results from clinical mastitis and high
    SCC
  • Combine with SCC, milk prod, DIM and preg
    diagnosis
  • Cow and herd recommendation decision

15
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16
Mastitis Problem Solving
Step 1 Identify possible mastitis problems.
(review records)
Subclinical mastitis
Clinical mastitis
Bulk tank SCC gt250,000 cell/ml
DHIA - SCC gt15 with LSgt4.5
New clinical cases gt2 per month
Severe clinical cases gt1 per year
Step 2 Use diagnostic test to determine
causative organism.
Bacteriological Cultures
Environmental organisms (usually clinical)
Contagious organisms (usually subclinical)
Coliforms
Strep sp
Staph sp
Staph aureus
Strep ag
Step 3 Generate possible solutions.
Treatment Decision
Treatment Decision
Treat clinical cases Treatment Plan Treatment
Protocols
Culture high SCC Segregate Cull chronic
Culture herd Treat positive cows Reculture treat
/cull
Prevention
Prevention
Clean/dry bedding Clean udders Premilking udder
pre Proper milking Maintain milking
equip Nutrition Vit E Se
Premilking udder prep Proper milking Postmilking
teat dip Maintain milking equip Dry cow
therapy Culture purchased animals
17
Clinical Mastitis Therapy Decisions
  • Large dairy 3200 milking cows in Michigan
  • Culture all clinical mastitis cases
  • Treat mastitis based upon culture results

18
  • Standard Mastitis Protocol
  • Abnormal milk
  • Intramammary antibiotic for 3 days
  • Continue treatment for 2 additional days
  • Change antibiotics two days
  • Remove from production untreatable

19
METHODS
  • Culturing Clinical Cases
  • Culture every new clinical case as they enter
    pen
  • With hold treatment 24 hrs for culture results
  • Unless cow is systemic ill or fever

20
METHODS
New Mastitis Protocol Culture
and record in Dairy Comp 305 Protocol base on
culture results Gram-negative NO
TREAT Gram-positive TREAT
21
New Mastitis Protocol Protocol
Gram-negative NO TREAT Monitor both the cow
and the quarter at each milking until milk is
normal Fever or Illness supportive therapy
Banamine Fluid therapy
22
New Mastitis
Protocol Protocol Gram-positive
TREAT Monitor at each milking until milk is
normal Intramammary antibiotic treatment If
milk does not return to normal cease milking
qrts If chronic repeater and clinical cease
milking qrts
23
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24
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25
20 Tx
80 NT
Hess 2003
26
20 Tx
80 NT
Hess 2003
27
Results
  • New Treatment Protocol
  • With the 24 hour with hold for culture ,
    antibiotic treatment was reduced 80
  • Gram-negative bacteria 30
  • No growth (coliforms/strep sp) 50

28
Results
  • New Treatment Protocol
  • With the 24 hour with hold for culture
  • Gram-positive bacteria
  • no change in days out of tank
  • no change in number of treatments
  • no change in quarters loss

29
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31
CONCLUSION
E. coli and no growth did not benefit from
antibiotic treatment for salable milk and
clinical signs. Culture identified no growth and
Coliforms rapidly (24 hrs). Treatment delay
reduced antibiotic use by 80. Treatment delay
did not effect clinical outcome for Gram-positive
bacteria (Strep Staph, 20 of cases). Change
in treatment protocol did not jeopardize the
health or well-being of the animal.
Hess 2003
32
Milk Cultures
1. Select cows for culture (herd, HSCC, etc) 2.
Collect an aseptic milk sample 3. Collect
composite or quarter milk sample 4. Refrigerate
(or ice) sample at collection
33
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34
watery milk more common in Strep mastitis than
coliform mastitis - Obritzhauser et al, 1995
35
Culture milk on blood agar plates or selective
media.
36
Blood agar Selective media MacConkey
agar Blood-esculin agar
37
Major Mastitis Pathogens Isolation
Selective Media
Philip Sears, DVM, PhD Department of Large Animal
Clinical Science
38
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39
Selective Media - Bi-plates
1
Large grey colonies
arrow shape
2
3
40
Gram-negative rods KOH positive Large grey
colonies moists or mucoid
Coliforms
Klebsiella sp
E. coli
41
Gram-negative rods KOH positive Large grey
colonies moists or mucoid
Coliforms
Klebsiella sp
E. coli
42
Gram-negative rods KOH positive Large grey
colonies moists or mucoid
Coliforms
Klebsiella sp
E. coli
43
Gram-positive cocci KOH negative Small
translucent colonies
Streptococci
Streptococcus agalactiae
Selective media Blood agar-esculin
Streptococcus spp) Str. uberis, enterococci,
beta-hemolytic colonies
esculin reaction
CAMP Test
44
Gram-positive cocci KOH negative Small
translucent colonies
Streptococci
Streptococcus agalactiae
Selective media Blood agar-esculin
Streptococcus spp) Str. uberis, enterococci,
beta-hemolytic colonies
esculin reaction
CAMP Test
45
Gram-positive cocci KOH negative Small
translucent colonies
Streptococci
Streptococcus agalactiae
Selective media Blood agar-esculin
Streptococcus spp) Str. uberis, enterococci,
beta-hemolytic colonies
esculin reaction
CAMP Test
46
Gram-positive cocci KOH negative Small
translucent colonies
Streptococci
Streptococcus agalactiae
Selective media Blood agar-esculin
Streptococcus spp) Str. uberis, enterococci,
beta-hemolytic colonies
esculin reaction
CAMP Test
47
Gram-positive cocci KOH negative Opaque
colonies white to yellow colonies
Staph aureus
Staph sp
Staphylococci
Staph sp
Staphylococcus aureus
Staph aureus

? ?
-
Staph sp
Coagulate plasma
Double zone hemolysis
Coagulase Test
(? - ?)
48
Gram-positive cocci KOH negative Opaque
colonies white to yellow colonies
Staph aureus
Staph sp
Staphylococci
Staph sp
Staphylococcus aureus
Staph aureus

? ?
-
Staph sp
Coagulate plasma
Double zone hemolysis
Coagulase Test
(? - ?)
49
Gram-positive cocci KOH negative Opaque
colonies white to yellow colonies
Staph aureus
Staph sp
Staphylococci
Staph sp
Staphylococcus aureus
Staph aureus

? ?
-
Staph sp
Coagulate plasma
Double zone hemolysis
Coagulase Test
(? - ?)
50
Clinical Mastitis Pathogen Isolation
Sears, 2003
51
Clinical Mastitis Pathogen Isolation
Selective media
Selective media
Sears, 2003
52
Mastitis Problem Solving
Step 1 Identify possible mastitis problems.
(review records)
Subclinical mastitis
Clinical mastitis
Bulk tank SCC gt250,000 cell/ml
DHIA - SCC gt15 with LSgt4.5
New clinical cases gt2 per month
Severe clinical cases gt1 per year
Step 2 Use diagnostic test to determine
causative organism.
Bacteriological Cultures
Environmental organisms (usually clinical)
Contagious organisms (usually subclinical)
Coliforms
Strep sp
Staph sp
Staph aureus
Strep ag
Step 3 Generate possible solutions.
Treatment Decision
Treatment Decision
Treat clinical cases Treatment Plan Treatment
Protocols
Culture high SCC Segregate Cull chronic
Culture herd Treat positive cows Reculture treat
/cull
Mild mastitis - abnormal milk Intermediate/moderat
e - pain, swelling Severe mastitis - systemic
fever, dehydration shock
53
Clinical Mastitis Treatment Protocol
Remove cow
Quarter loss to
production
Cull
Return to Herd
Milk
Hess 2003
54
Treatment Decisions
In a recent article that used a clinical mastitis
model, antibiotic treatment is not profitable.
The cost of milk discard off-sets the gain in
production.
(Bennedsgaard, 2003) However, these models place
all pathogens together and do not separate out
staphylococcal and streptococcal mastitis that
can respond favorably to antibiotic therapy.
(Wilson, 1996)
55
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56
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57
Summary
  • Culturing milk from subclinical infected cows
    can aid in the development of mastitis treatment
    protocols and mastitis prevention plans.
  • Culturing milk from clinical mastitis cases can
    be useful in identifying cases responsive to
    antibiotic therapy and reducing antibiotic
    residues without jeopardizing animal health.

58
Susceptibility Testing Methods
Incubate plate 18-24 hr, 35 C Measure and record
zone of inhibition around each disk
Innoculate MH plate
Place disks on agar plate
59
Disk Diffusion Susceptibility Testing
60
Antimicrobial susceptibilitytesting using
micro-broth dilutions
ug/ml 64 32 16 8 4 2
96 well microtiter plate
61
Antimicrobial Gradient TestingE-test
Read plates after recommended Incubation
Read MIC where elipse intersects scale
62
Focus on Gram Positives (MIC)
Susceptibility Testing
  • 10 random samples that cultured coagulase
    negative Staph spp. (CN Staph) and 10 random
    samples that cultured environmental Strep spp.
    were submitted to Dr. Bolins lab at the Animal
    Health Diagnostic Lab at Michigan State
    University
  • The MIC for each sample was determined
  • Based upon the results, the MIC50, MIC75 and
    MIC90 for each group of organisms were calculated
    (antibiotic susceptibility levels for 50, 75
    and 90 of each group)

63
Results of Susceptibility TestingEnvironmental
Strep spp.
64
Results of Susceptibility TestingEnvironmental
Staph spp.
65
Decisions
  • 90 of Environmental Strep spp. Were susceptible
    to Ampicillin (Amoxicillin) and
    Penicillin/Novobiocin and potentially Penicillin
  • 90 of Coagulase Negative Staph spp. Were
    susceptibile to Cephalothin, Penicillin/Novobiocin
    and potentially Pirlimycin, Ampicillin
    (Amoxicillin) and Penicillin

66
Current Clinical Mastitis Treatment Protocol
  • Environment Strep
  • 5-7 days of Amoximast therapy intramammary BID
  • 5-7 days of Penicillin therapy systemically BID
  • Coag-negative Staph
  • 5-7 days of Cefa-Lak therapy intramammary BID
  • 5-7 days of Penicillin therapy systemically BID

67
Keep Health and Treatment Records
  • Calendar or notebooks
  • Daily Treatment Records (AABP records)
  • Health Programs (Dairy Comp 305, Dairy CHAMP)
  • DHIA - PC Dart ( health input summaries)

68
Animal Treatment Plan AABP record sheet - 2A
  • Disease diagnosis and treatment protocol
  • Milk and meat withdrawal time

69
Animal Treatment Plan AABP record sheet - 2A
70
Packet Page 2
Daily Treatment Record (Barn Sheet) Fresh Cows
Record AABP record sheet
  • Cow by calving and treatment activity
  • Disease diagnosis and treatment
  • Treatment record for each fresh cow

71
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72
Health Monitor - Number of cases
Detecting the Demons in your Dairy Diagnostic,
Protocols, and Monitoring Tools for Your Dairy
Time period ___________
Number cows ____
Number cases
1 2 3 4 5 6 7 8 9 10 11 12 13
14 15 16 17 18 19 20
RP
DA
Off feed
Downer
CMT 1-3
Diarrhea
Lameness
Milk fever
BCS lt2.5
Metritis
Ketosis
Udder edema
Mastitis-New
Mastitis-Severe
73
Health Monitor - Number of cases
Detecting the Demons in your Dairy Diagnostic,
Protocols, and Monitoring Tools for Your Dairy
Time period ___________
Aug - Oct, 1999
46
Number cows ____
(full)
43
33
22
Number cases
1 2 3 4 5 6 7 8 9 10 11 12 13
14 15 16 17 18 19 20
9
7
7
4
4
2
2
RP
DA
CMT 1-3
Downer
Off feed
Diarrhea
Lameness
Milk fever
BCS lt2.5
Metritis
Udder edema
Mastitis-severe
Ketosis
Mastitis-new
74
BEST MANAGEMENT PRACTICES
MASTITIS DECISION TREE
Bulk Tank Somatic Cell Count (BTSCC)
DHI-SCC
Cow DHI-LS 4 or greater
CMT quarters
Positive quarters (CMT 1 or greater)D
Staph aureus (SA) Mycoplasma Coag neg Staph
Strep agalactiae (Sag)
Environmental Strep sp Coliforms
Depopulate (Sag)
Treat IMM
Depopulate IMI cows Dry off lt80 days Sell
chronic cows Kill bad quarter(s)
Identify source Housing Bedding Teat dip
pre post Pasture
Reculture
Treat IMM
Segregate at milking Test all fresh cows
heifers Positive SA to SA group
Reculture
Cull
Culture all fresh cow heifers
Mellenberger, 1996
75
BEST MANAGEMENT PRACTICES for CONTAGIOUS MASTITIS
76
BEST MANAGEMENT PRACTICES for CONTAGIOUS MASTITIS
(cont)
77
BEST MANAGEMENT PRACTICES for ENVIRONMENTAL
MASTITIS
78
BEST MANAGEMENT PRACTICES for ENVIRONMENTAL
MASTITIS (cont)
79
BEST MANAGEMENT PRACTICES for ENVIRONMENTAL
MASTITIS (cont)
80
Mastitis Problem Solving
Step 1 Identify possible mastitis problems.
(review records)
Subclinical mastitis
Clinical mastitis
Bulk tank SCC gt250,000 cell/ml
DHIA - SCC gt15 with LSgt4.5
New clinical cases gt2 per month
Severe clinical cases gt1 per year
Step 2 Use diagnostic test to determine
causative organism.
Bacteriological Cultures
Environmental organisms (usually clinical)
Contagious organisms (usually subclinical)
Coliforms
Strep sp
Staph sp
Staph aureus
Strep ag
Step 3 Generate possible solutions.
Treatment Decision
Treatment Decision
Treat clinical cases Treatment Plan Treatment
Protocols
Culture high SCC Segregate Cull chronic
Culture herd Treat positive cows Reculture treat
/cull
Prevention
Prevention
Step 4 Develop Farm Plan Protocols
  • Set goals
  • Create Standard Operating Procedures (SOPs)
  • Monitor records
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