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VPM 201
Veterinary Bacteriology and Mycology
Laboratory Review-Pt 2: 2010
Lab 6a/b: Respiratory Infections
P. multocida
Exercise 1. feline pneumonia, Abx (Clavamox) unresponsive : TTA: Enterobacter
cloacae: No pix, LF, motile, Indole/Urease –ve, Citrate +ve
Exercise 2. BRDC – mixed culture Mh/Pm . Ex. 3 . Cat bite wound : P. multocida
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Lab 6b: Respiratory Infections
A. pleuropneumoniae
Morse code Demo 4: Kennel cough
“honking cough” ITB
Lab 7: Canis Otitis Externa
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Lab 7. Felis with a felis x 2
Staph. albus !?
- 12 week old cat new in household
- Came from short stay at SPCA
- About week ago one eye was “tearing”
- Now both affected , muco-purulent
Chlamydophila felis
Giemsa stain
EB
Conjunctival epithelial cell
Lab 7: Pinkeye
• BA – β-hemolytic
• No MAC growth
• Gram -ve, coccobacillus
• Oxidase +ve, nonmotile
• Intervention
– Vaccination – several
– Face fly (M. autumnalis)
– Abx – Oxytetracycline
– Optic corticosteroids
– Zoonotic
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Lab 8a/b: Urinary Tract Infections
Inte rpre tation of Quantitative Urine Culture Results in Dogs
Method ofCollection
Infection(cfu/mL)
Equivocal (cfu/mL) Contamination(cfu/mL
Cystocentesis >1,000 100-1,000 <100
Catheterization >10,000 (D) 1,000-10,000 (D) <1,000 (D)
Voided (Midstream) >100,000 (C) 10,000 - 100,000 (C) <10,000 (C)
cfu = colony form ing units ( ie. the number of bacteria)
Note: most UTI’s caused by single bacter ial species and are not the result of mixed
infections.
UTI = urethritis, cystitis, ureteritis, pyelonephritis
Lab 8a/b: UTI • Dog UTI. Cystocentesis plated on BA/Mac.
No Mac Growth. I.d. – BA – double zone, Catalase positive, Coag Positive:
gram-positive cocci
• S. pseudintermedius
– Quantitative plating: Cysto > 1,000 CFU/mL
– Counts/purity
• Radiology/Ultrasonography • Other Agents of K9 UTI
– E. coli, P. mirabilis, K. pneumoniae, Strep/Enterococci spp., P. aeruginosa
– Cats different – Staph felis case ,
• Goal of UTI Abx: 3 – 5 x MIC
– If Baytril MIC is 0.5 ug/mL then need to be able to achieve 2.5 ug/mL in bladder (pharm/tox)
1 bug / 10 HPF = 105
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Morse code
Lab 8a: UTI: Demo’s
• Kidney of 4-day old foal. P.M. shows micro-abscess in kidneys.
• A. equuli Sleepy Foal Disease
• BA: viscid, +/- hemolysis
• LF on MAC
• Cow with UTI: C renale
– Coryneform; Palisade, acute angles, Urease +ve
• Recall: Enterococcus spp.
• Actinobaculum suis
Lab 9/10: Clostridium/Mycoplasmas
• No peptidoglycan, highly pleomorphic
• Colony: “Fried Egg” , microcolonies
• Specialized media (10-20 % serum – cholesterols)
– Hayflicks, Freys, Friis
• Disk Growth Inhibition test(s)
– Digitonin sensitivity (Biochemical) genera
– Growth Inhibition with Specific Antisera - species
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Mycoplasma Genus/Species
Q/A
• M. gallisepticum: CRD (Air Sacculitis) Chickens, IS in Turkeys
• M. bovis
– Caseonecrotic bronchopneumonia in feedlot: arthritis/tenosynovitis ; contagious agalactia mastitis dairy
• M. felis
– Conjunctivitis, Feline Resp. Dis. Complex, polyarthritis in immunodeficient cats
– Resp. disease in immunocompromised humans and foals
• M. hyopneumoniae : Enzootic pneumonia in swine
• M. haemofelis : Feline Infectious Anemia
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Lab 10: Misc. Demo cases
• A cow presents with extensive dermatitis on the dorsal aspects of the face and body. Large scabs “paintbrush” were removed from the lesions. A Gram-stain prepared from the scab material is provided.
• Can you make a diagnosis with information provided ? Why?
• If yes, what organism might this be?
• Vaccines? Penicillin?
Lab 10: Demo Cases
• Sheep, exudative dermatitis – P. aeruginosa
– BA: large, metallic sheen, hemolytic,
– NLF on MAC, Ox +ve
– Green wool or fleece rot – moisture/feces/blow flies (wool strike)
– MHA shows pigment
• Equine (stiff, sawhorse stance hyperexcitable) – puncture wound exudate
– Gram positive –
– drumstick/tennis racket morphology
– Vaccines?
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Lab 11: Mycology
• Mycological Examination of tissue
• Wet Mount - 10-20% KOH
• Stains - Giemsa, PAS, Methenamine Silver, India
ink, LPCB
• Culture – BHI +, Sabourauds
• Dermatophytosis, Aspergillosis, Zygomycosis
• Yeast
• Dimorphic Fungi
Zygomycetes
• Culture
– Mucor spp.
• Microscopic
– Aseptate hyphae
– Sporangium
– Rhizoids
• Silver stained placenta
– Brown hyphae
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Aspergillus fumigatus
• Dog: chronic sinusitis
• Culture
• Microscopic morphology
– Septate hyphae
– Vesicle
– Conidiospores
• Nasal Aspergillosis
– Radiology, Rhinoscopy
• Guttural Pouch Mycosis
• Brooder pneumonia
• Abortion in ruminants
Dermatophytes
• Cultures
– M. canis (Cats)
– M. gypseum
– T. mentagrophytes
– T. verrucosum
• Microscopic
• KOH/PAS
– Ectothrix arthrospores
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Yeasts
• Candida albicans
• Malassezia pachydermatis
– “footprint”, lipophilic
– Bigger than Staph/Strep
– Otitis externa,
– Seborrheiac dermatitis
Malassezia
Systemic Mycoses
• Cryptococcus neoformans
Encephalitis: PAS
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Thermally Dimorphic Fungi
• Histoplasma capsulatum
• Endemic Miss., Ohio, St. Lawrence
• Bat/bird feces
– Intracellular Yeast
– Mold phase (tuberculate chlamydospores)
• Blastomyces dermatitidis
• Endemic as per H. capsulatum
• Terminal chlamydospore (lollypop)
Histo – 37 oC
Blasto – 37 oC
Lollipop Chlamydospores
• Coccidioides immitis
• S.W. U.S.
• 37 oC “Spherule” phase
• Mold Phase
– Barrel shaped arthrospores
Thermally Dimorphic
Barrel arthrospores
Spherule – 37 oC
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• Sporothrix schenckii
– Ulcerative lymphangitis
• Horses, cats, cattle, people
– Mold phase (septate)
• Microconidia (flowerettes)
– Body Temp.
• Cigar bodies (cats)
• Plants and mastitis?
SubQ Thermally Dimorphic + Algae