Midterm Notes 3.pdf

MIDTERM - BACTE LECTURE NOTES # 3 - prepared by: MAM LIGHT GRAM-POSITIVE ACID-FAST BACILLI – GENUS MYCOBACTERIUM  Aerobic, non-motile;  Slow growers and requires whole egg for growth  Difficult to stain but once stained difficult to decolorize due hydroxy m...

MIDTERM - BACTE LECTURE NOTES # 3 - prepared by: MAM LIGHT GRAM-POSITIVE ACID-FAST BACILLI – GENUS MYCOBACTERIUM  Aerobic, non-motile;  Slow growers and requires whole egg for growth  Difficult to stain but once stained difficult to decolorize due hydroxy methoxy acid / mycolic acid M. tuberculosis complex MOTT/ Mycobacterium other M. leprae than tuberculosis/ Runyon’s Classification M. tuberculosis _______________________ PHOTOCHROMOGENS  a.k.a Hansen’s disease M. bovis _____________________________ SCOTOCHROMOGENS  Causes LEPROSY M. africanum _________________________ NON-PHOTOCHROMOGENS  Non-culturable In vitro but viability can be M. microti & M. canetti – can infect RAPID GROWERS maintains using MICE FOOT PADS or FOOT those who are immunocompromised PADS of ARMADILLO  Under the microscope it forms the so-called Cigar Packet arrangement  Diagnosis is often made based on clinical symptoms SCREENING TEST: ACID FAST STAINING 1) Ziehl Neelsen _____________________________ 2) Kinyoun’s Method _________________________ DO CULTURE NON-SELECTIVE AGAR BASED SELECTIVE LIQUID MEDIA egg based media with malachite green, inhibits contaminating organism; clear media, easy with these are growth in 6-10 weeks examination of antibiotics non- colonies growth in conventional 10-12 days media Common Disadvantage: Lowenstein Jensen Media: ____________________________________________ Middlebrook 7H10 Gruft Bactec 12B, Petragnani- more inhibitory: ___________________________________________ with dextrose Modified Bactec 13A American Thoracic Society /ATS ______________________________________ Middlebrook 7H11 Lowenstein Septi-chek Dorset Egg Media with casein Jensen Middlebrook Wallenstein medium: _________________________________________________ hydrolysate Selective 7H9 Mitchison 7H11 Middlebrook 7H11 Mitchison Selective 7H11 IF THERE IS (+) GROWTH IN ANY OF THESE MEDIA, NEXT STEP IS TO DO SERIES OF BIOCHEMICAL TESTS TO IDENTIFY SPECIES BIOCHEMICAL TEST REQUIRED REAGENT (+) RESULT (-) RESULT POSITIVE NEGATIVE MEDIA CONTROL CONTROL pg. 1 1ST SEMESTER 2023-2024  MIDTERM - BACTE LECTURE NOTES # 3 - prepared by: MAM LIGHT NIACIN TEST Lowenstein Strip impregnated with YELLOW No color M. M. Jensen Media CYANOGEN BROMIDE change tuberculosis intracellulare NITRATE Sodium Nitrate After incubation we Initial (+) RED M. M. REDUCTION Broth add: (-) no color change tuberculosis intracellulare n-n -dimethyl-l- naphthylamine After adding zinc powder: sulfanilic acid (+) no color change (-) RED to confirm a (-) result we add: Use of Strip (+) Blue zinc dust/zinc powder IRON UPTAKE TEST Lowenstein No color x Jensen Media change ARYLSULFATASE Phenolphthalein 2N Sodium carbonate. M. fortuitum M. TEST media intracellulare Detection of rapid growers PYRAZINAMIDASE Pyrazinamidas 1% ferrous ammonium (+) red TEST e agar sulfate HEAT STABLE Tween 80 Heating of colonies at QUALITATIVE M. fortuitum M. CATALASE TEST Media 68 degC prior to the (+) ____________________ M. gordonae tuberculosis To detect heat labile addition of reagent is (-) ______________________ catalase required Mycobacterium like M. QUANTITATIVE tuberculosis (+) height of bubbling more than 45 mm (-) less than 45 mm TWEEN 80 Useful in the identification of M. kansasii ____________________ M. kansasii M. HYDROLYSIS TEST M. tuberculosis (+) in 10-20 days intracellulare (+) result: _____________________________ (-) result: no color change Indicator: _____________________________ TELLURITE For the identification of M. avium M. avium M. REDUCTION TEST (+) result: smooth, fine black precipitate (smoke like action) tuiberculosis (-) result:gray clumps (no smoke like action) SALT TOLERANCE For identification of rapid growers & M. triviale M. fortuitum M. gordonae TEST 5% NaCl (+) growth while (-) no growth T2H/ TCH May be done to differentiate M. tuberculosis and M. bovis M. bovis M. susceptibility (+) growth inhibition (no growth) tuberculosis Thiopene-2- (-) growth carboxylic hydrazide Growth in  To identify M. fortuitum -chelonae complex Mac Conkey agar without crystal violet NIACIN TEST NITRATE REDUCTION PYRAZINAMIDASE TWEEN 80 HYDROLYSIS HEAT STABLE CATALASE Mycobacterium tuberculosis  a.k.a Koch’s Bacillus or Tubercle Bacillus; obligate aerobe, gram (+) or gram neutral pg. 2 1ST SEMESTER 2023-2024  MIDTERM - BACTE LECTURE NOTES # 3 - prepared by: MAM LIGHT  Virulence Factors: (1) CORD FACTOR and (2) SULFATIDES _____________________________________________  With MUCH GRANULES, length is 1-4 u while diameter is 0.2 – 0.6  Produces “cauli flower like growth at 35 – 37 degC;  Causes Pulmonary TB; BCG vaccine / Bacillus Calmette Guerin __________________________________ Types of TB: (Primary, Secondary, MDR-TB – shows resistance to PRIMARY TB DRUGS), XDR-TB- shows resistance not only to primary but also to some 2nd line TB drugs POTT’s disease – TB of spine while MILIARY – disseminated TB, extrapulmonary PRIMARY TB DRUGS – RIPES – Rifampin-Isoniazid- Pyrazinamide-Ethambutol- Streptomycin 2ND LINE TB DRUGS – ofloxacin, kanamycin, ciprofloxacin DIAGNOSIS 1. Skin Testing – MANTOUX TEST – involves intradermal injection of PPD on the skin PPD (purified Protein Derivative) – protein extracted and purified from M. tuberculosis cell wall OT (old tuberculin) Von Pirquet _____________ (old tuberculin mixed with lanolin to produce ointment w/c is scratch onto the skin) 2. Chest X-Ray 3. Sputum exam – collect _______________in a day (1st ______________________, 2nd ______________________) 4. Culture – maintained for 8 wks / 2 months  CAULI FLOWER LIKE GROWTH – tan to buff, dry, rough, warty, granular 5. QUANTIFERON TB (ELISA) rapid and requires heparinized whole blood 6. GeneXpert – PCR Based NAAT, rapid and sensitive Mycobacterium bovis  _____________________________________________________________________________________________  ______________________________________________________________________________________________  Nitrate and Niacin Test __________________ Mycobacterium leprae a.k.a __________________________  Forms the so-called ____________________________________________________________  Non culturable on artificial media, but can be maintained in the lab using ________________________________________________________  Causes Leprosy MOT – prolong skin to skin contact with infected person, droplets from nose & mouth (inhalation)  Clin manifestation LEONINE FACE  Tuberculoid type /paucibacillary_____________________________________________________________________  Lepromatous type /multibacillary ____________________________________________________________________  Basis of diagnosis: _____________________________________________________Lepromin Test____________________________________  Specimen: TISSUE JUICE (earlobes, nasal scrapings) NON-TUBERCULOUS MYCOBACTERIA PHOTOCHROMOGENS  Mycobacterium kansasii – regarded as the # 2 NTM to cause TB in those with AIDS; a.k.a Cross Barred Bacillus  Mycobacterium marinum- can cause Fish Tank granuloma or _________________________________  Mycobacterium asiaticum, M. simiae , M. intermedium & M. novocastrense SCOTOCHROMOGENS  Mycobacterium szulgai - pulmonary disease, predominantly in middle-aged men; cervical adenitis; bursitis  Mycobacterium scrofulaceum - Cervical adenitis in children, bacteremia, pulmonary disease, skin infections  Mycobacterium gordonae / M. aquae __________________________________  M. xenopi - Primarily pulmonary infections in adults. ______________________colonies on CORN MEAL AGAR  M. flavescens , M. thermoresistible  M. interjectum - Unknown Potentially pathogenic Chronic lymphadenitis, pulmonary disease  Others: M. heckeshornense, M. tusciae, M. kubicae, M. cookie, M. hiberniae NON-PHOTOCHROMOGENS  M. avium complex - pulmonary infections in patients with preexisting pulmonary disease; cervical lymphadenitis; and disseminated disease (M. avium pathogenic – LADY WINDERMERE SYNDROME while M. intracellulare = Battey Bacillus)  M. terrae _____________________________ M. triviale - ___________________________  M. gastri , M. ulcerans - mycobacteriosis, also referred to as Buruli ulcer, biochemically inert  M. malmoense M. genovense -Disseminated disease in patients with AIDS  , M. haemophilum , Others: M. heidelbergense; M. simiae; M. shimoidei RAPID GROWERS  M. abscessus subsp. Abscessus - Disseminated disease, primarily in immunocompromised individuals; skin and soft tissue infections; pulmonary infections; postoperative infections  M. fortuitum - Postoperative infections in breast augmentation and median sternotomy; skin and soft tissue infections; pulmonary infections, usually single. localized lesions. Central nervous system (CNS) disease is rare but has high morbidity and mortality  M. chelonei - Skin and soft tissue infections, postoperative wound infections, keratitis  M. phlei – breadcrumbs growth on LJ medium  Other: M. smegmatis – Urine AFB + Biochemical tests to speciate Mycobacterium pg. 3 1ST SEMESTER 2023-2024  MIDTERM - BACTE LECTURE NOTES # 3 - prepared by: MAM LIGHT Tests Mycobacterium tuberculosis Mycobacterium bovis Niacin test + - Nitrate Reduction Test + - Heat Stable Catalase test - - Tween 80 Hydrolysis Test V - Arylsulfatase Test (3days) - - Iron Uptake Test - - Pyrazinamidase Test + - Tests M. kansasii M. marinum M. simiae M. asiaticum Niacin test - V + - Nitrate Reduction Test + - - - Heat Stable Catalase test + - + + Tween 80 Hydrolysis Test + + Slow + + Arylsulfatase Test (3days) - V - - Iron Uptake Test - - - - Pyrazinamidase Test - + + - Tests M. scrofulaceum M. szulgai M. gordonae M. thermoresistible M. flavescens Niacin test - - - - - Nitrate Reduction Test - + - - + Heat Stable Catalase test + + + + + Tween 80 Hydrolysis Test - Slow + + + + Arylsulfatase Test (3days) - - - - - Iron Uptake Test - - - - - Pyrazinamidase Test V - - NA + Tests M. Avium – Intracellulare M. terrae – triviale M. malmoense M. haemophilum M. gastri Complex complex Niacin test - - - - - Nitrate Reduction Test - + - - - Heat Stable Catalase test + + + - - Tween 80 Hydrolysis Test - + + - + Arylsulfatase Test (3days) - - - - - Iron Uptake Test - - - - - Pyrazinamidase Test + V + + - Tests M. fortuitum M. chelonei M. phlei M. smegmatis Niacin test - - - - Nitrate Reduction Test + - + + Heat Stable Catalase test + + + + Tween 80 Hydrolysis Test V - + + Arylsulfatase Test (3days) + + - - Iron Uptake Test + - + + Pyrazinamidase Test + + - NA DO NOT ANYMORE READ THE MYCOBACTERIUM CHAPTER in the BOOK BY MAHON pg. 4 1ST SEMESTER 2023-2024