New Strategies for Staphylococcus aureus Mastitis

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New Strategies for Staphylococcus aureus
Mastitis

• Philip Sears, DVM, PhD
• Department of Large Animal Clinical Science
• College of Veterinary Medicine - Michigan State
  University
• East Lansing, Michigan USA
• www.cvm.msu.edu (extension)

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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
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Milk Cultures

• Infection level in herd
• Type of infections
• Source of infections
• Control and Prevention
• Treatment

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Milk Culturessample consideration

• Cow samples
• Composite cow samples for herd survey
• quarter samples - clinical cases, high SCC

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Milk Sampling
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Milk Culturessample consideration

• Individual samples
• composite sampling for herd survey
• quarter sampling - clinical cases, evaluate
  individual cows
• Bulk tank samples
• screen herd for contagious pathgens
• monitor herds for Strep ag, S. aureus, Mycoplasma
• String samples
• large dairies - treat each string as herd

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Culture 100
Culture 10
Staphylococcus aureus isolation Sampling
Culture positive 1
75 2
94 3 98
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Determine EtiologyBacteriologic

• False Positive
• environmental bacterial isolates
• False Positive are less likely when
• Staph aureus for subclinical and clinical cases
• single pathogen culture in large numbers
• multiply culture from the same quarter
• 2-3 consecutive or duplicate samples

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Determine EtiologyBacteriologic

• False Negatives
• cyclic shedding of S. aureus
• after antibiotic therapy
• freezing (? S. aureus)
• composite sample (quarter sample)
• reduce FN by multiple samples (consecutive or
  duplicate at the same time

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? ?
Staphylococcus aureus
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Staphylococcus aureus
Gangrene mastitis
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Staph MastitisStaphylococcus aureus
Contagious mastitis

• Lives in infected quarters infected skin
• injured teat ends (mechanical, chemical, weather)
• Spread from cow to cow(milking, hands, common
  cloth)
• Nonclinical with high SCC reduce production
• Clinical (mild to severe) with repeated flare-ups
• Treatment
• Lactation 10 - 60
• Dry off 40 - 70
• less 10 with chronic, long standing infections

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Efficacy for Gram-Positive Pathogens
Pathogen qrts treatment cure Staph aureus
121 ND 24.8 Strep agalactiae 31
ND 51.6 other Streps 111
ND 36.0 Cure (no-Tx) Staph aureus 1141
Clox 26 (56) 20 Strep agalactiae 762
Clox 85 (16) 19 Strep uberis 797
Clox 77
Craven, 1991
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Treatment of S.aureus IMIAntibiotics
Field Studies Product Quarters
Cures Clox/florfenicol 33 1
3, Wilson 1996 Amoxicillin 12 1
8, Sears 1994 Pirlimycin 68
12 18, Sears 1998 Total 104 14 13
Product Quarters Cures ND 121
24.8 Cloxocillin 1141 26.6 No
treatment 56 20.0 (5-6)
Craven,1986
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Treatment of S.aureus IMIAntibiotics
Field Studies Product Glands
Cures Clox/florfenicol 33 1
3, Wilson 1996 Amoxicillin 12 1
8, Sears 1994 Pirlimycin 68
12 18, Sears 1998 Total 104 14 13
Staphylococcus aureus isolation Sampling
Culture positive 1
75 2
94 3 98
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Antimicrobial susceptibilitytesting using
micro-broth dilutions
ug/ml 64 32 16 8 4 2
96 well microtiter plate
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Focus on Gram Positives (MIC)
Susceptibility Testing

• 10 random samples that cultured coagulase
  negative Staph spp. were submitted for MIC
• Based upon the results, the MIC50, MIC75 and
  MIC90 for each group of organisms were
  calculated
• (antibiotic susceptibility levels for 50, 75
  and 90)

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Results of Susceptibility TestingEnvironmental
Staph spp.
MIC50 MIC75 MIC90 Max MIC
Ampicillin 0.12 0.12 0.133 0.12
Pirlimycin 0.50 0.50 1.1 2.00
Pen/Nov 0.50 0.50 0.50 0.50
Penicillin 0.12 0.12 0.21 0.12
Tetracycline 0.25 0.25 16 1.00
Cephalothin 1.0 1.0 1.0 1.0
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Decisions

• 90 of Coagulase Negative Staph spp.
• Were susceptibile to Cephalothin,
  Penicillin/Novobiocin and potentially Pirlimycin,
  Ampicillin (Amoxicillin) and Penicillin

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Clinical Mastitis Treatment Protocol

• Staphylococcal Mastitis
• 5-7 days of Cefa-Lak therapy intramammary BID
• 5-7 days of Penicillin therapy systemically BID

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Treatments

• Treatment 1 No IMM or systemic antimicrobial
  therapy
• Treatment 2 IMM antimicrobial therapy only
• Strep spp. Amoximast ?
• Staph spp. and Staph aureus Pirsue?
• once a day for five days.
• Treatment 3 IMM Systemic antimicrobial
  therapy
• Polyflex ? (30cc IM) once a day for five days.

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Results
Cows (Quarters) cured 27 days post treatment Cows (Quarters) returned to production
Strep spp Control 6/27 (22) 14/27 (52)
Amoximast 11/28 (39) 20/28 (71)
Amox Ampicillin 18/27 (67) 21/27 (78)
Staph spp Control 3/15 (20) 10/15 (67)
Pirsue (IMM) 5/15 (33) 8/15 (53)
PirsueAmpicillin 4/16 (25) 14/16 (88)
S. aureus Control 0/10 (0) 4/10 (40)
Pirsue 0/9 (0) 7/9 (78)
PirsueAmpicillin 0/7 (0) 7/7 (100)
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Results
Cows (Quarters) cured 27 days post treatment Cows (Quarters) returned to production
Strep spp Control 6/27 (22) 14/27 (52)
Amoximast 11/28 (39) 20/28 (71)
Amox Ampicillin 18/27 (67) 21/27 (78)
Staph spp Control 3/15 (20) 10/15 (67)
Pirsue (IMM) 5/15 (33) 8/15 (53)
PirsueAmpicillin 4/16 (25) 14/16 (88)
S. aureus Control 0/10 (0) 4/10 (40)
Pirsue 0/9 (0) 7/9 (78)
PirsueAmpicillin 0/7 (0) 7/7 (100)
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Results
Cows (Quarters) cured 27 days post treatment Cows (Quarters) returned to production
Strep spp Control 6/27 (22) 14/27 (52)
Amoximast 11/28 (39) 20/28 (71)
Amox Ampicillin 18/27 (67) 21/27 (78)
Staph spp Control 3/15 (20) 10/15 (67)
Pirsue (IMM) 5/15 (33) 8/15 (53)
PirsueAmpicillin 4/16 (25) 14/16 (88)
S. aureus Control 0/10 (0) 4/10 (40)
Pirsue 0/9 (0) 7/9 (78)
PirsueAmpicillin 0/7 (0) 7/7 (100)
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Results
Cows (Quarters) cured 27 days post treatment Cows (Quarters) returned to production
Strep spp Control 6/27 (22) 14/27 (52)
Amoximast 11/28 (39) 20/28 (71)
Amox Ampicillin 18/27 (67) 21/27 (78)
Staph spp Control 3/15 (20) 10/15 (67)
Pirsue (IMM) 5/15 (33) 8/15 (53)
PirsueAmpicillin 4/16 (25) 14/16 (88)
S. aureus Control 0/10 (0) 4/10 (40)
Pirsue 0/9 (0) 7/9 (78)
PirsueAmpicillin 0/7 (0) 7/7 (100)
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Staph Mastitis Therapy
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Therapy
New thoughts towards treatment regime has lead
to Immune modulators Timed treatment -
drugs immune modulation
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Staphylococcus aureus Cross Section
Capsule Type 2, 5 8
Cell membrane
Cytoplasma
Experimental Trial Vaccination Treatment
Peptidoglcan-techoic acid
Cell wall protein (Protein A)
Using vaccination to produce antibodies to help
cure S. aureus mastitis with antibiotic therapy
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Vaccination and TherapyCombination of Two
Immuno-modulators???

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Staphylococcus aureus Cross Section
Capsule Type 2, 5 8
Cell membrane
Cytoplasma
Peptidoglcan-techoic acid
Cell wall protein (Protein A)
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Trial Schedule
Vaccination

• 1st inoculation - 2 wks before treatment
• 2nd inoculation - 48 hours pretreatment
• 3rd inoculation - 7days after 2nd inoculation

Treatment
Lactation therapy (infected quarters) 3
sequential courses of pirlimycin - 36 hrs between
each course of treatment (total - 6 IMM
infusion) 1st infusion 48 hrs after 2nd
inoculation
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Treatment Results
Michigan
Vacc/treat Cleared Percent Lactation
Cow s 10 8 80 Quarters
19 17 89 Dry Cow Cow s 5
4 80 Quarters 11
10 91 50 cure for Treatment only
Sears, Proc NMC, 1999
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Vaccination-Treatment Trial
S. aureus infection

• Number of herds 11
• States 6
• Number cow tx 591
• Number quarters tx 869

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Clearance of S. aureus following vaccination and
treatment
100
90
80
70
60
Percent
Quarters
50
Cows
40
30
20
10
0
10
51
6
19
5
585
10
7
19
30
77
348
1
2
3
4
5
6
7
8
9
10
11
All
Herds (cows)
P. Sears A. Belschner, Proc. NMC, 1999
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Summary of Vaccination TrialCure Rates following
Treatment
90
79
80
70
70
Vaccination
60
Percent
Quarters
50
36
Cows
40
30
18
17
20
10
3
0
68
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190
132
146
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Antibiotic
Autogenous
Autocapsule
Only
Intramammary antibiotic
P. Sears A. Belschner, Proc. NMC, 1999
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Trivalent serotypes serve as markers

• T5 and T8 - Capsule
• 336 - Cell wall

Guidry et al., Am. J. Vet. Res., 1998
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Staphylococcus aureus serotypes ()
(n) Type 5 Type 8 336 NT
US 273 18 23
59 0
Europe 94 34 32
30 2
Korea 108 29 20
48 3
Guidry et al., Am. J. Vet. Res., 1998
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Farm-specific and Universal Staphylococcus aureus
Vaccine
Sears, Proc IMMQS, 2001
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Teichoic acid (cell wall)

• Present on all Staphylococcus aureus
• Elicits opsonizing antibodies
• Acts as an adhesion

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Antigen specific vaccine for Staphylococcus aureus
Sears, Proc IMMQS, 2001
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Sears, Proc IMMQS, 2001
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Treatment Protocol for Staphylococcal Mastitis
Clinical
Mastitis
Cow
Culture
Physical
Fever
No Fever
Exam

Mastitis Pen

Boxstall
Staph aureus
Staph sp
1. Await Culture
1. Banamine
2. Monitor
Intramammary
Intrmammary
2. Oral Fluids
antibiotic therapy
antibiotic therapy
3. Monitor
Flag records
Flag records
Cull?
Combination
Combination
Extented
Vaccination/
treatment
treatment
treatment
Treatment
Intramammary
Intramammary
Intramammary
Vaccination twice
antibiiotic
antibiiotic
pirlimycin
at 2 week intervals
Penicillin G IM
Penicillin G IM
6 to 8 treatments
Extented treatment
3 days
3 days
once daily
Outcome
Loss quarter
Lost quarter
Return to Herd
Milk Herd
Animal cull
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Animal Treatment Plan AABP record sheet - 2A

• Staphylococcal Mastitis (Treatment pathogen
  specific)
• 5-7 days of Cefa-Lak therapy intramammary BID
• 5-7 days of Penicillin therapy systemically BID

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