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GRAM-POSITIVE BACTERIA
Clinical Importance, Diagnosis, Antimicrobial Susceptibility test, Control and Prevention
Universiti Malaysia KelantanFaculty of Veterinary Medicine
Course coordinator: Dr. Erkihun AkliluLecture II (24 September 2013)
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STAPHYLOCOCCI
Occurs worldwide
Colonize nasal cavity, skin, mucous membrane, GIT (transient)
Infections are mainly endogenous
Indirect transmission is possible
Can survive in the environment for prolonged period
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STAPHYLOCOCCI
Pathogenecity
Staphylococci are pyogenic (abscess formation and suppuration) Botryomycosis (Chronic staphylococcal wound) Granulomatous lesion with pockets of pus
Pathogenic strains produce various enzymes and toxins
Enterotoxins (A-E) cause food poisoning – emetic
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STAPHYLOCOCCI
Exfoliatin causes staphylococcalscalded skin syndrome (SSS) ininfants and possiblby in dogs
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STAPHYLOCOCCI
TSS-1: causes toxic shocksyndrome (TSS) in humans
Epidermolytic toxins: porcineexudative epidermatitis
The alpha toxin (haemolysin)produces gangrenous mastitis
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STAPHYLOCOCCI
Leukocidin: kills neutrophils and macrophages
Protein A: binds to the FC region of IgG
Staphylokinase: activates plasminogen
Coagulase: causes plasma coagulation in vitro
Hyluronidase: Spreading factor
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ANIMAL DISEASES CAUSED BY PATHOGENIC STAPHYLOCOCCI
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ANIMAL DISEASES CAUSED BY PATHOGENIC STAPHYLOCOCCI
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STAPHYLOCOCCAL HAEMOLYSINS
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STAPHYLOCOCCUS
Laboratory Diagnosis
Specimen: includes, exudate, pus from abscess,mastitic milk, skin scrapings, urine,affected tissues
Direct Microscopy: Examination of Gram-stainedsmears (pus/exudate) may reveal Gram-positive cocci “Grape-like”
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STAPHYLOCOCCUS
Isolation and Identification
Media: Ox or sheep blood agarSelective media: Sheep/ox blood agar with 15 mg
nalidixic acid and 10 mg colisitin sulphateper liter of media
For food microbiology: Mannitol salt agar (MSA) and Baird-Parker medium.
Incubated aerobically at 370C for 24 – 48 hours
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STAPHYLOCOCCUS
Isolation and Identification
Colonial characteristics:
Colonies appear in 24 hours Round, smooth, glistening On BA appear larger and opaque compared to
smaller, colonies of haemolytic streptococci Golden-yellow pigment (human, domestic animals) Non-pigmented (white): dogs S.aureus and S.intermedius : produce double
haemolyis (alpha and beta) S. hyicus: non-haemolytic
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STAPHYLOCOCCUS
Isolation and Identification
Haemolysis caused by S. aureus
Non-haemolytic colonies (S. hyicus)
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STAPHYLOCOCCUS
Test for pathogenicity of staphylococcal isolates
Coagulase test DNAase test Test for protein A
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STAPHYLOCOCCUS
Biochemical tests
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STAPHYLOCOCCI
Differentiation of staphylococci, micrococci and streptococci
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STREPTOCOCCUS
Habitat
Indigenous respiratory tract microbial flora ofanimals and humans
Certain species are also found in the gastrointestinaland urogenital tracts of humans and animals
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STREPTOCOCCUS
General characteristics
Streptococci are facultative anaerobic Gram-positive organisms that often occur as chains or
pairs Catalase-negative
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STREPTOCOCCUS
General characteristics
Hemolytic Reactions Beta Hemolysis
Complete hemolysis Clear zone around
colonies
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STREPTOCOCCUS
Alpha hemolysis A greenish discoloration around the area
surrounding the colony due to incompletehemolysis of the red blood cells.
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STREPTOCOCCUS
Gamma Reaction Absence of a hemolytic reaction No change around colonies on blood agar
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STREPTOCOCCUS
Lancefield Groups Based on serological groupings Can also be detected by biochemical testing or antibiotic
Sensitivity
Group A Streptococcus pyogenes The most virulent human pathogen of the genus Is beta hemolytic Often identified by rapid serological tests or by
antibiotic resistance
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STREPTOCOCCUS
Group B
Streptococcus agalactiae
Mildly to moderately virulent
Usually beta or alpha hemolytic (some strains are gamma)
Detected biochemically
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STREPTOCOCCUS
Group D
Includes the fecal streptococci (enterococci)
Normal colon flora in humans and other animals
Genus Enterococcus
Several species; eg. Enterococcus faecalis
Occasionally pathogenic (often in urinary tract infections)
Usually gamma reactive
Detected biochemically
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STREPTOCOCCUS
Pathogenesis
Pyogenic (associated with suppuration and abscess formation)
Enterotoxins: Haemolysins (Streptolysin O and S) Hyaluronidase Dnase NADase Protease Streptokinase Phage-coded pyrogenic toxin (Scarlet fever)
Polysaccaride capsules (S. pyogenes, S. pneumoniae, S. porcinus ---Antphagocytic
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Polysaccaride capsules (S. pyogenes, S. pneumoniae,S. porcinus ---Antiphagocytic
Polysaccaride capsules in S. equi sbsp. equifunctions as adhesin.
STREPTOCOCCUS
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STREPTOCOCCUS
Laboratory Diagnosis
Specimens: Exudates, pus, mastitic milk, skinscrapings, cerebrospinal fluid, urine, affectedtissues
Submit swabs in transport media
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STREPTOCOCCUS
Laboratory Diagnosis
Direct microscopy:
Fixed smears from pus, exudates, sediments from centrifuged milk or urine
Gram stains
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STREPTOCOCCUS
Laboratory Diagnosis
Isolation:
Routine media (BA), Edwardsmedia
Incubate at 37oC for 24 – 48 hours
Colonial morphology varies: small,translucent (hemolytic), mucoid (S.equi subsp equi & S.pneumoniae)colonies
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STREPTOCOCCUSLaboratory Diagnosis
Isolation:
Peptone water sugar withadded serum (fastidiousstreptococci)
Enterococci and Group Dstreptococci grow at 45oC and40% bile (hyrdolyse aesculin)
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STREPTOCOCCI
Streptococci identification tests
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STREPTOCOCCUS
Laboratory Diagnosis
Identification:
CAMP test:
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STREPTOCOCCI
Differentiation of equine group C streptococci
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STREPTOCOCCI
Differentiation of enterococci and group D streptococci
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STREPTOCOCCI
Differentiation of streptococci causing bovine mastitis
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CORYNEBACTERIUM ANDRHODOCOCCUS
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CORYNEBACTERIA Many corynebacteria are opportunistic pathogens
except for C. bovis (pyogenic infections)
Non-nitrate reducing biotypes of C. pseudotuberculosis cause caseous lymphadenitis in sheep, goat and rarely in cattle
Abscessation and enlargement of superficial or internal lymph nodes
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CORYNEBACTERIA
Mechanical injury (e.g. shearing, insect bite) can be predisposing factors
Incubation period – about 3 months
C.pseudotuberculosis can survive in the environment longer
Bacteria can survive intracellularly and causes chronic infections
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CORYNEBACTERIA
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CORYNEBACTERIA
Laboratory diagnosis
Smear from lesion showingcoryneform bacteria
Isolation and identification of C.pseudotuberculosis from abscessmaterial (confirmatory)
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RHODOCOCCUS
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RHODOCOCCUS
R. equi (formerly known as Corynebacterium equi)
Gram-positive, soil saprophyte
Some spp of R. equi are cocci, while others appear as rods
Opportunistic pathogen of foals of less than six months old
Causes suppurative bronchopneumonia of foals
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RHODOCOCCUS
Infection is usually through inhalation ofcontaminated dust particles
R. equi presents in large number in the feces of younghealthy foals and in the feces of older horses
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RHODOCOCCUS
Laboratory diagnosis
Sample: tracheal aspirates or pus from lesions
Grows on non-enriched media
Produces characteristic mucoid salmon-like colonies