VPM 201-Lab 6 Bovine Mastitis, Bacillus & Mastitis (2012)

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1 Exercise 1. Bovine mastitic milk sample A. Note relevant images are on next page Sample A is Staphylococcus aureus Moderate size (1.0 mm), circular, convex, cream-to-light yellow, opaque Double-zone (target) hemolysis Gram +ve cocci Clumps, pairs, Catalase +ve Coagulase +ve MAC No growth - - Ed No growth - -

2 Staph. aureus on Staph. aureus is coagulase positive

3 Exercise 1. Bovine mastitic milk sample B. Note relevant images are on next page Sample B is Streptococcus uberis Pinpoint to small (<0.5 mm), circular, translucent at 24 hours. Opaque, ~ 1mm at 48 hours Strains are (weak) alpha or nonhemolytic Recall - hemolysis shows zone of greening around colonies Gram +ve Chains of cocci Catalase -ve CAMP -ve MAC No growth - - Ed Growth - grey to brown colonies Due to hydrolysis of Esculin and formation of Fe precipitate - Esculin +ve

4 Streptococcus uberis at 48 hrs, Strep. uberis on Edwards agar note dark/silvery/grey colonies

5 Exercise 1. Bovine mastitic milk sample C. Note relevant images are on next page Sample C is Escherichia coli 2-3 mm greyish, single narrow zone of hemolysis Gram-negative rods, singly and in pairs Oxidase ve Indole +ve Citrate & Urease -ve MAC Growth, Lactose Fermentor (LF) intensely pink with pink in agar around colonies Ed No growth - -

6 E. coli showing hemolysis on Urease Citrate E. coli E. coli E. coli E. coli on MAC

7 Exercise 1. Bovine mastitic milk sample D. Note relevant images are on next page Sample D is Corynebacterium bovis Small, white, dry colonies nonhemolytic MAC No growth - Ed No growth Gram +ve Coryneform - rods & coccobacilli - L & V forms Palisades Catalase +ve Urease -ve

8 Note: we don t have a good picture of C. bovis - This is a high mag shot of C. pseudotuberculosis - C. bovis is more off-white, less glossy & no hemolysis Catalase test

9 Exercise 1. Bovine mastitic milk sample E. NOTE there are no images for CoNS Sample E is a Coagulase Negative Staphylococcus spp. (CoNS or CNS) Moderate size (1.0 mm), circular, convex, off-white, opaque Non- or single-zone hemolysis MAC No growth - Ed No growth - Gram +ve cocci Clumps, pairs, occasional short (3-4) chains Catalase +ve Coagulase -ve

10 Exercise 1. Bovine mastitic milk sample F. Note relevant images are on next page Sample F is Streptococcus dysgalactiae Small translucent colonies, Strains are typically alpha-hemolytic Recall - hemolysis shows zone of greening around colonies MAC No growth - Ed Growth hemolytic zone is often Visible Gram +ve cocci Short chains - Smears from milk sample tend to show chaining better - Catalase ve CAMP -ve Esculin -ve

11 Streptococcus dysgalactiae 48 hour plate Close-up of -hemolysis: zone of greening

12 Exercise 1. Bovine mastitic milk sample G. Note relevant images are on next page Sample G is Klebsiella pneumoniae Large (5-10 mm), light grey, coalescing, mucoid colonies Nonhemolytic MAC Large, mucoid, LF- pale pink - Ed No growth - Gram-negative Short, plump rods occurring singly or in pairs. A large, unstained halo may be evident particularly with Fuchsin counterstain this is the capsule Oxidase ve Indole ve Citrate & Urease +ve

13 Klebsiella pneumoniae on Urease Citrate Klebsiella pneumoniae on MAC K. pneumoniae K. pneumoniae K. pneumoniae

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