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Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
Hemofilus+bordetela+gardnerela
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Hemofilus+bordetela+gardnerela

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  • The vagina is a dynamic ecosystem that normally contains approximately 10 9 bacterial colony forming u/gm of vaginal fluid The normal flora is dominated by lactobacilli, but include a variety of other organisms, some potential pathogens, at lower levels Other common bacteria streptococcal species, Gm neg bacteria, G. vaginalis and anaerobes Lactic acids and other organic acids are metabolized from glycogen by the lactobacilli maintaining the vaginal pH between 3.8 and 4.2 The acidic environment inhibits the overgrowth of bacteria and other organisms with pathogenic potential The normal vaginal discharge is clear to white, odorless, and of high viscosity.
  • Lactobacillus-hydrogen peroxide producing, normally present in sufficient amounts Prevotella (genera, Bacteroides genus) commonly assoc with oral cavity. Prevotella species also indicated in women with BV. Mix of Gram-variable coccobacilli. Mobiluncus is an anaerobic bacteria, 2 subspecies: curtisii and mulieris. More often recovered from blacks and always occurs with other orgs assoc with BV. Gardnerella normally present in vaginal flora (looks the same as Bacteroides on Gram stain) Mycoplasma found more often in women w/ BV than w/o BV and may have a symbiotic relationship w/ Gardnerella or act alone.
  • -- So what is the causation? -- Why is the number of recurrences so high? -- Not sufficient data to conclude whether it is sexually transmitted or not. -It remains unclear whether recurrence is due to the use of antibiotics that do not eradicate infection or to reinfection from sex partners. 7 -- Important connections between characterization factors (see picture) - Initiation factor of BV remains a mystery, although an important inter-relation between lactobacilli, hydrogen peroxide production, vaginal pH, and overgrow of BV associated bacteria is present. 6 --In particular, let’s look further into the aspect of OVERGROWTH OF BV ASSOCIATED BACTERIA 6) Wilson, J. Managing recurrent bacterial vaginosis. Sex Transm Infect 2004;80:8-11. 7) Bradshaw CS, Morton AN, Hocking J, Garland SM, Morris MB, Moss LM, Horvath LB, Kuzevska I, Fairley CK. High recurrence rates of bacterial vaginosis over the course of 12 months after oral metronidazole therapy and factors associated with recurrence. J Infect Dis 2006;193:1478-86.
  • Gram stain and screening. Quantification of bacterial morphotypes (Lactobacillus, Gardnerella and Bacteriodes spp (Prevotella), and Mobiluncus. Not reliant on sypmtoms so it is a more sensitive test, better at diagnosing asymptomatic cases. More specific than wet mount for finding clue cells. Gram stain is best way to see Mobiluncus. 0 6 10
  • Transcript

    • 1. Haemophilus influenzae Dr Kamran Afzal Classified Microbiologist
    • 2. HAEMOPHILUS
      • H. influenzae
      • H. influenzae biogroup aegyptius
      • H. ducreyi
      • H. parainfluenzae
      • H. aphrophilus
      • H. paraphrophilus
      • Haemophilus= Blood-loving
    • 3. Haemophilus influenzae type b
    • 4. EPIDEMIOLOGY
      • Reservoir Human Asymptomatic carriers
      • Transmission Respiratory droplets
      • Temporal pattern Peaks in Sept-Dec and
      • March-May
      • Communicability Generally limited but higher in some circumstances (over crowding)
    • 5. SPECIMENS
      • Pus
      • Sputum
      • Naso-pharyngeal swab
      • Throat swab
      • CSF
      • Blood
      • Joint/ Pleural/ Pericardial aspirates
        • Specimens should not be refrigerated
        • Transport within 2 hrs
    • 6. MORPHOLOGY
      • Gram negative rods
      • Cocco-bacillus to filamentous (pleomorphic)
      • Capsulated
    • 7. CULTURAL CHARACTERISTICS
      • Grows aerobically
      • 5-10% CO 2 enriched moisturized atmosphere
      • Optimum temp 37 0 C, 20-42 0 C
      • Selective media
      • - Bacitracin for sputum
    • 8.
      • Differential media (Robert’s media)
      • - Bacitracin + Sucrose + Phenol red
      • - HI does not ferment Sucrose: Transparent colonies
      • - HP ferments Sucrose: Yellow colonies
      • Chocolate agar
      • - Contains both factors X and V
      • - During preparation, when heated it
      • inactivates NADase, releases extra
      • V from red cells
    • 9.
      • Media should contain :
      • - Haemin/ Fe containing Porphyrin (Factor X )
      • - NAD/P (Factor V )
      • Factor X : used by HI to produce essential resp enzymes Catalase, Peroxidase, Cytochrome c
      • Factor V : electron carrier in oxidation reduction system of the organism
    • 10. SATELLITISM
      • Blood agar: inoculate a streak/ spots of Staphylococcus aureus that provides V in excess of its own
      • May also be shown by
      • Streptococcus pneumoniae , Neisseria sp, Diphtheroids, other Streptococci
    • 11. IRIDESCENCE
      • When sub-cultured on transparent media (Filde’s or Levinthal’s agar) colonies of HI show iridescence of different colors
      • Non-capsulated: Blue
      • Capsulated: Red, orange, green, blue
      • Seen when a strong concentrated light is passed from beneath and looked obliquely
      • Due to: optical properties of capsular layers
    • 12. BIOCHEMICAL TESTS
      • Catalase +
      • Oxidase +
      • Nitrate reduction +
      • Glucose fermentation +
      • Antigen detection
        • CSF, urine, fluids, serum
      • 8 Biotypes (I-VIII) on the basis of
      • -Indole
      • -Urease
      • -ODC
    • 13. SEROLOGICAL TYPES
      • 6 Sero-types (a-f) on the basis of capsular antigens
      • Type ‘b’ is invasive and capsulated
      • Type ‘b’ causes:
      • Meningitis, Pneumonia, Ac. epiglottitis, Cellulitis, Osteomyelitis, Septic arthritis
    • 14. PATHOGENESIS
      • Organism colonizes nasopharynx
      • In some persons organism invades bloodstream and cause infection at distant site
      • Antecedent URI may be a contributing factor
      • Normal carriage
      • -Humans: upper resp tract
      • -2-10% of healthy people are carriers of type ‘b’
      • -25-80% carriers of non-capsulated HI
    • 15.
      • Virulence factors
      • -Polysacharide capsule
      • -Fimbriae
      • attachment
      • -IgA proteases
      • colonisation
      • -OMP
      • invasion
    • 16. ANTIBIOTICS
      • Ampicillin/ Amoxicillin
      • Chloramphenicol
      • Tetracycline
      • Cefuroxime
      • Cefotaxime/ Ceftriaxone
      • Co-amoxiclav
      • Ciprofloxacin
      • Azithromycin
      • Clarithromycin
    • 17. CONJUGATE VACCINE
      • For active immunization
      • Polysaccharide vaccine: poorly immunogenic
      • Polysaccharide is coupled to various proteins
      • (TT, N. meningitis , DT)
    • 18. Bordetella pertussis
    • 19. THE GENUS BORDETELLA
      • Very thin, ovoid or small rod shaped
      • Gram negative bacilli
      • One of the most common and serious bacterial respiratory disease
      • All species cause severe childhood respiratory infection
      • SPECIES
        • Bordetella pertussis
        • Bordetella parapertussis
        • Bordetella bronchiseptica
    • 20. Bordetella pertussis
      • CHARACTERISTICS
      • Most fastidius
      • Bordet-Gengou medium (BGM-Selective medium)
        • Charcoal and blood to absorb toxins
      • Colonies are shiny, grey and convex
    • 21.
      • Strict aerobe
      • Optimum temp 35 – 36 0 C, takes minimum 3 days
      • Non-motile, non-sporing
      • Three human pathogenic serotypes
        • Type 1,2, type 1,3 and type 1,2,3
    • 22. PATHOGENESIS
      • WHOOPING COUGH
      • Non-invasive infection of the respiratory mucosa
      • Humans – only natural hosts
      • Incubation period
        • 1-2 weeks
    • 23. VIRULENCE FACTORS
      • Agglutinogens
        • 1,2 and 3
      • Tracheal cytotoxin
        • Paralyses tracheal cilia
      • Pertussis toxin
        • Lymphocytosis
    • 24. Tracheal cytotoxin
    • 25. Pertussis toxin
    • 26. CLINICAL FEATURES
      • Per – tussis
        • Severe cough
      • Child suffers bouts of paroxysmal coughing each day
      • During paroxysm -> no pause for air intake, the tongue is protruded fully, fluids stream through nose and eyes, face becomes cyanotic. When death seems imminent, a final cough clears the secretions and, with a massive inspiratory effort through the narrow glottis, a long, high pitched ‘whoop’ -> WHOOPING COUGH
    • 27. CLINICAL PHASES
    • 28. Cough Plate Method
      • Culture plate held at 10-15 cm infront of the mouth when the patient is coughing spontaneously or induced
      • Droplets of respiratory secretions impinge on the media
        • Helpful as a bed side investigation
    • 29. LAB DIAGNOSIS
      • Bacterial culture
        • Specimen – Per-nasal swab is better
        • BGM or Charcoal-Blood agar with Penicillin
        • Incubate aerobically for 7 days
        • Advantage -> isolate can be serotyped / genotyped
    • 30.
      • Detection of bacterial antigen or DNA
        • Specimens – serum, urine
        • Bacteria in NP secretions -> labelled with fluorescein -> UV microscopy
        • PCR detects Bordetella antigen in NP secretions
      • Detection of Bordetella antibody
        • ELISA
        • Florescent Antibody method
      • Differential blood count
        • Lymphocytosis
    • 31. TREATMENT
      • Antimicrobial drugs
        • Erythromycin, Clarithromycin – 14 days
        • Penicillin is not useful
      • Other measures
        • Cough suppressants, corticosteroids
          • Retention of symptoms
      • Pertussis vaccine – supreme importance
    • 32. CONTROL
      • Vaccination
        • Safe, >90% effective and strongly recommended
        • Suspension of whole bacterial cells, killed by heat or chemicals
        • 1-2 months, 1 st injection I/M
        • Herd immunity – breaks cycle of transmission as the organism dies before finding a new host
    • 33. Gardnerella vaginalis
    • 34. Normal Vagina
      • Dynamic ecosystem
      • Lactobacilli metabolize glycogen -> lactic acids that maintain pH 3.8 - 4.2
      • The normal vaginal discharge is clear to white, odorless, and of high viscosity
    • 35.
      • 1. Epithelial Cells
      • 2. Lactobacilli
      • - 5 to 15 µ
      • 3. WBCs
      • - Few = NL
      • - Never > Epi’s
      • - Many = Inflammation
      • ( Parabasilar Cell) >
      • Normal Epithelial Cells with Sharp Borders
      • Normal Lactobacilli - 5 to 15 µm
      • WBCs
      • - Few = Normal
      • - Many = Inflammation
    • 36.
      • Three main types of vaginitis
        • Bacterial vaginosis
          • About 50% of vaginitis cases
          • Gardnerella or a mixed anaerobes
        • Trichomoniasis
          • About 20% vaginitis cases
          • Trichomonas vaginalis
        • Candidiasis
          • About 25% of vaginitis cases
          • C. albicans (80-92%)
    • 37.  
    • 38. Vaginitis Differentiation Normal Trichomoniasis Candidiasis Bacterial Vaginosis Symptom presentation Itch, discharge, 50% asymptomatic Itch, discomfort, dysuria, thick discharge Odor, discharge, itch Vaginal discharge Clear to white Frothy, gray or yellow-green; malodorous Thick, clumpy, white “cottage cheese” Homogenous, adherent, thin, milky white; malodorous “foul fishy” Clinical findings Cervical petechiae “strawberry cervix” Inflammation and erythema Vaginal pH 3.8 - 4.2 > 4.5 Usually < 4.5 > 4.5 KOH “whiff” test Negative Often positive Negative Positive NaCl wet mount Lacto-bacilli Motile flagellated protozoa, many WBCs Few WBCs Clue cells ( > 20%), no/few WBCs KOH wet mount Pseudohyphae or spores if non- albicans species
    • 39. What is bacterial vaginosis?
      • Shift in normal vaginal flora
      • Lactobacillus
      • Anaerobic bacteria
        • Prevotella sp., Mobiluncus sp.
      • Gardnerella vaginalis
      • Mycoplasma hominis
    • 40. Microbiology
      • Lactobacillus acidophilus
      • Gardnerella vaginalis
      • Mycoplasma hominis
      • Mobiluncus species
      • Anaerobes
        • Bacteroides (Porphyromonas)
        • Peptostreptococcus
        • Fusobacterium
        • Prevotella
      Mobiluncus mulieris Mycoplasma hominis
    • 41. Gardnerella vaginalis
      • Normal vaginal flora
      • Gram negative or Gram variable
      • Non capsulated
      • Non motile
      • Facultative anaerobe
      • Catalase negative
      • β - haemolytic
    • 42. Clue cell Clue cell on Gram stain and saline wet mount of vaginal discharge (on >20% cells) Bacteria adhered to epithelial cells; most reliable single indicator
    • 43. Etiology and Pathogenesis
      • Not fully known
      • Sexually transmitted?
      Reduction of lactobacilli Reduction in H 2 O 2 production Overgrowth of BV associated bacteria Raised pH
    • 44. Specimen Collection
      • Swab the lateral or posterior fornices of the vagina
      • Place the swab in a sterile tube containing
      • ~ 6 drops of saline
      • Transfer to the lab ASP
    • 45. Methods for Diagnosis of Bacterial Vaginosis
      • Clinical Diagnosis, Amsel’s criteria
      • – Used to clinically manage patients
      • Gram stain, Nugent score
      • – Centralized and standardized lab based method
      • – Not used for clinical management
    • 46. Amsel’s criteria (3 of 4)
      • Thin, grey/white, homogenous, malodorous discharge
      • Clue cells on microscopy
      • pH of vaginal fluid > 4.5
      • Release of fishy odor on adding alkali (10% KOH)
        • Positive “whiff” test (Amine Test)
        • Absence of inflammation is the basis of the term &quot;vaginosis&quot;
        • Pruritus is absent
    • 47. Bacterial Vaginosis testing
      • Diagnosis by Gram Stain
      • Nugent Method: Scores 0-10
      Normal (0-3) Intermediate (4-6) BV (7-10)
    • 48. Treatment Recommendations
      • Oral Metronidazole 500 mg bid x 7 days
        • 84-96% cure rate
      • Oral Clindamycin 300 mg bid x 7 days
        • Less effective
      • Topical treatments (higher recurrence rates)
        • Metronidazole gel (0.75%)
          • 70-80% cure rate
        • Clindamycin cream (2%)
          • Less effective
    • 49.  
    • 50.  
    • 51.  
    • 52.  

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