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MYCOBACTERIA  OF  MEDICAL IMPORTANCE Prof M.I.N. Matee, PhD Department of Microbiology and Immunology, MUCHS
Mycobacteria: overview ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
MYCOBACTERIUM ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
TB: Epidemiology ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Burden of TB in Tanzania   ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Estimated Global TB incidence rates, 2000 The boundaries and names shown and the designations used on this map do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement. Global Tuberculosis Control. WHO Report 2002 . WHO/CDS/TB/2002.295 25 - 49 50 - 99 100 - 300 0 - 9 10 - 24 ,[object Object],No estimate Africa  > 300 /100,000
Prevalence of HIV among TB patients ,[object Object],[object Object],[object Object],[object Object],NTLP - MOH
 
Tanzania: TB Cases - trends, 1979 to 2001
Impact of HIV on the Epidemiology of Tuberculosis
Impact HIV in treatment outcomes High Mortality in HIV Co-infected patients van den Broek  & Mfinanga, 1998 NTLP reports 2003
THE GENUS  MYCOBACTERIUM  CAN BE DIVIDED  INTO FOUR BROAD GROUPS ,[object Object],[object Object],[object Object],[object Object]
THE TUBERCULOSIS COMPLEX (Organisms that resemble  M. tuberculosis; Causing a similar type of disease in humans) ,[object Object],[object Object],[object Object]
THE RUNYON GROUPING:  An older idea that  Remains useful for differentiating mott
RUNYON GROUPS I,II,III: Slow growing  mycobacterium Visible colonies on solid media After more than 5 days incubation GROUP I:  PHOTOCHROMOGENS  Produce pigment  When grown in the light but not the dark EXAMPLES:  M. kansasii ;  M. marinum ;  M. simiae GROUP II:  SCOTOCHROMOGENS   Pigment when grown in both light and dark EXAMPLES:  M. scrofulaceum ;  M. szulgai ;  M. xenopi GROUP III: NONCHROMOGENS No pigment when grown in both light and dark EXAMPLES:  M. avium-intracellulare ;  M. genevense M. ulcerans  and  M. leprae  do not fit in this scheme
RUNYON GROUP IV: Rapidly growing  mycobacterium Visible colonies on solid medium In less than 5 days incubation EXAMPLES: M. fortuitum ;  M. chelonae ;  M. abscessus
Robert Koch 1843-1910
MYCOBACTERIUM TUBERCULOSIS   Prototypic facultative intracellular pathogen Primary human pathogen Transmitted from human to human  Closely related to  M. bovis   It is an intracellular pathogen (typically inside alveolar macrophages).  This organism does not secrete an identifiable exotoxin.  Host immunity to  M tuberculosis  appears to be cell mediated (CMI).
CELL ASSOCIATED TOXINS MAY BE IMPORTANT VIRULENCE FACTORS FOR MANY MYCOBACTERIA The glycolipids, such as trehalose dimycolates, of  Mycobacterium  tuberculosis  and related organisms appear to be related to their virulence .
Bacterial cell wall structure Gram + Gram - Mycobacterium Lipid  Peptido-  Lipid +  Porins  Mycolate  Acyl  LAM  arabino- Bilayer  glycan  LPS  Lipids  Lipo-arabino  galactan mannin
 
Biochemical properties ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Reaction to physical and chemical agents ,[object Object],[object Object],[object Object]
Constituents of tubercle bacilli ,[object Object],[object Object],[object Object],[object Object]
Virulence factors ( M. tb .) ,[object Object],[object Object],[object Object],[object Object],[object Object]
Tuberculosis – primary infection route
Tuberculosis in humans ,[object Object],INTRACELLULAR  pathogen   (facultative extracellular) Exposed Infected  (2 billion, 8 million new cases per year) Primary TB Latent TB Reactivation 30% 80-90% 5-10% 5-10% Clearance 70% Death (2 million)
Pathogenesis   ,[object Object],[object Object],[object Object],[object Object]
T helper Cell Differentiation ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],IFN  TNF  IL4 IL13 IL10 Th0 Th2 Th1
 
 
 
 
Pathology   ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Large caseating tubercle Miliary tubercles HUMAN LUNG HUMAN LUNG TUBERCULOSIS
Spread of organisms in the host ,[object Object],[object Object],[object Object]
Tuberculosis – hematogenous extension Extrapulmonary  Lymph nodes Liver  Bones
Primary and reaction TB ,[object Object],[object Object],[object Object],[object Object],[object Object]
Reactivation TB ,[object Object],[object Object],[object Object],[object Object]
M. tuberculosis
Symptoms of TB  Disease ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Clinical Manifestations of TB ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],DIAGNOSIS
Chest Radiograph Abnormalities often seen in apical  or posterior segments of upper  lobe or superior segments of  lower lobe May have unusual appearance in  HIV-positive persons  Cannot confirm diagnosis of TB Arrow points to cavity in  patient's right upper lobe.
Diagnosis of tuberculosis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Administering Tuberculin Skin Test ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Reading the Tuberculin Skin Test ,[object Object],[object Object],[object Object],[object Object]
Factors that affect the PPD Reaction  Type of Reaction   Possible Cause False-positive   Nontuberculous mycobacteria BCG vaccination   Anergy False-negative   Recent TB infection Very young age (< 6 months old) Live-virus vaccination Overwhelming TB disease
 
 
 
 
 
 
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Mycobacteria: auramine stain
Nucleic Acid Hybridization ,[object Object],[object Object]
DNA fingerprinting Steps: 1. Digest DNA into smaller fragments 2. Separate fragments by size 3. Visualize bands 4. Each band corresponds to 1 fragment size 5. Unique pattern of bands identifies donor of DNA
Mycobacteria colonies on LJ medium
MGIT System ,[object Object],[object Object],[object Object],[object Object],[object Object],http://labmed.ucsf.edu/CP/SFGH/Microbiology/images/MAI.jpeg
 
 
Tuberculosis and  the Immune Reconstitution Inflammatory Syndrome (IRIS)
Definition ,[object Object]
Names ,[object Object],[object Object],[object Object]
Pathogenesis ,[object Object],[object Object],[object Object]
Risk factors for TB/IRIS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Types of TB IRIS ,[object Object],[object Object]
Timing of IRIS ,[object Object],[object Object],[object Object]
Prevention and control of tuberculosis   1.  Prompt and effective treatment of patients with active TB 2. Careful follow-up of contacts – tuberculin test, X-rays etc 3. Prophylaxis of asymptomatics, tuberculin positives 4. Correct underlying cause of immune suppression 5. Immunization with live-attenuated tubercle bacilli e.g. BCG 6. Eradication of TB in cattle and pasteurization of milk
OTHER IMPORTANT MYCOBACTERIA ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Mycobacterium ulcerans   ,[object Object],[object Object],[object Object]
Mycobacterium marinum ,[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Mycobacterium scrofulaceum ,[object Object],[object Object],[object Object]
Mycobacterium fortuitum  complex ,[object Object],[object Object],[object Object],[object Object]
HANSEN’S DISEASE (Leprosy) caused by  M. leprae Hansen’s disease is a chronic, slowly progressive  Granulomatous Disease involve ectodermally derived tissues e.g. skin and peripheral nerves.  Usually limited to the cooler parts of the body e.g. skin, nose and upper respiratory tract.  It rarely affects internal organs such as the brain, liver, spleen, kidneys, and bones.  It has a specific predilection for peripheral nerves.
Mycobacterium leprae is an acid-fast rod that grows in macrophages, nerve cells, the foot pads of thymectimized mice, and the nine banded armadillo.  It has never been grown in artificial medium,  an obligate intracellular pathogen.  has a generation time of 12 to 14 days,  the incubation period for onset of disease is prolonged,  therapy must also be prolonged (perhaps the lifetime of the individual).
There are 3 forms of the disease: ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
~ 500,000 cases Worldwide Treatment  –  Dapsone (1940s-1970s)   Rifampicin (1970s-)   Multidrug Therapy- Dapsone, Rifampicin, Clofazamine

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Mycobacteria slides for lecture

  • 1. MYCOBACTERIA OF MEDICAL IMPORTANCE Prof M.I.N. Matee, PhD Department of Microbiology and Immunology, MUCHS
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  • 9. Tanzania: TB Cases - trends, 1979 to 2001
  • 10. Impact of HIV on the Epidemiology of Tuberculosis
  • 11. Impact HIV in treatment outcomes High Mortality in HIV Co-infected patients van den Broek & Mfinanga, 1998 NTLP reports 2003
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  • 14. THE RUNYON GROUPING: An older idea that Remains useful for differentiating mott
  • 15. RUNYON GROUPS I,II,III: Slow growing mycobacterium Visible colonies on solid media After more than 5 days incubation GROUP I: PHOTOCHROMOGENS Produce pigment When grown in the light but not the dark EXAMPLES: M. kansasii ; M. marinum ; M. simiae GROUP II: SCOTOCHROMOGENS Pigment when grown in both light and dark EXAMPLES: M. scrofulaceum ; M. szulgai ; M. xenopi GROUP III: NONCHROMOGENS No pigment when grown in both light and dark EXAMPLES: M. avium-intracellulare ; M. genevense M. ulcerans and M. leprae do not fit in this scheme
  • 16. RUNYON GROUP IV: Rapidly growing mycobacterium Visible colonies on solid medium In less than 5 days incubation EXAMPLES: M. fortuitum ; M. chelonae ; M. abscessus
  • 18. MYCOBACTERIUM TUBERCULOSIS Prototypic facultative intracellular pathogen Primary human pathogen Transmitted from human to human Closely related to M. bovis It is an intracellular pathogen (typically inside alveolar macrophages). This organism does not secrete an identifiable exotoxin. Host immunity to M tuberculosis appears to be cell mediated (CMI).
  • 19. CELL ASSOCIATED TOXINS MAY BE IMPORTANT VIRULENCE FACTORS FOR MANY MYCOBACTERIA The glycolipids, such as trehalose dimycolates, of Mycobacterium tuberculosis and related organisms appear to be related to their virulence .
  • 20. Bacterial cell wall structure Gram + Gram - Mycobacterium Lipid Peptido- Lipid + Porins Mycolate Acyl LAM arabino- Bilayer glycan LPS Lipids Lipo-arabino galactan mannin
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  • 26. Tuberculosis – primary infection route
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  • 36. Large caseating tubercle Miliary tubercles HUMAN LUNG HUMAN LUNG TUBERCULOSIS
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  • 38. Tuberculosis – hematogenous extension Extrapulmonary Lymph nodes Liver Bones
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  • 45. Chest Radiograph Abnormalities often seen in apical or posterior segments of upper lobe or superior segments of lower lobe May have unusual appearance in HIV-positive persons Cannot confirm diagnosis of TB Arrow points to cavity in patient's right upper lobe.
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  • 49. Factors that affect the PPD Reaction Type of Reaction Possible Cause False-positive Nontuberculous mycobacteria BCG vaccination Anergy False-negative Recent TB infection Very young age (< 6 months old) Live-virus vaccination Overwhelming TB disease
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  • 59. DNA fingerprinting Steps: 1. Digest DNA into smaller fragments 2. Separate fragments by size 3. Visualize bands 4. Each band corresponds to 1 fragment size 5. Unique pattern of bands identifies donor of DNA
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  • 64. Tuberculosis and the Immune Reconstitution Inflammatory Syndrome (IRIS)
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  • 71. Prevention and control of tuberculosis 1. Prompt and effective treatment of patients with active TB 2. Careful follow-up of contacts – tuberculin test, X-rays etc 3. Prophylaxis of asymptomatics, tuberculin positives 4. Correct underlying cause of immune suppression 5. Immunization with live-attenuated tubercle bacilli e.g. BCG 6. Eradication of TB in cattle and pasteurization of milk
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  • 78. HANSEN’S DISEASE (Leprosy) caused by M. leprae Hansen’s disease is a chronic, slowly progressive Granulomatous Disease involve ectodermally derived tissues e.g. skin and peripheral nerves. Usually limited to the cooler parts of the body e.g. skin, nose and upper respiratory tract. It rarely affects internal organs such as the brain, liver, spleen, kidneys, and bones. It has a specific predilection for peripheral nerves.
  • 79. Mycobacterium leprae is an acid-fast rod that grows in macrophages, nerve cells, the foot pads of thymectimized mice, and the nine banded armadillo. It has never been grown in artificial medium, an obligate intracellular pathogen. has a generation time of 12 to 14 days, the incubation period for onset of disease is prolonged, therapy must also be prolonged (perhaps the lifetime of the individual).
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  • 82. ~ 500,000 cases Worldwide Treatment – Dapsone (1940s-1970s) Rifampicin (1970s-) Multidrug Therapy- Dapsone, Rifampicin, Clofazamine

Notas do Editor

  1. 3 The global burden of tuberculosis is truly enormous with some 1.9 million deaths per year according to the most detailed country-based analysis yet carried out and reported last year by the surveillance and monitoring unit of WHO&apos;s Communicable Disease Operational and Epidemiological Research Unit. Some 380,000 of these deaths occur in HIV infected individuals largely, but not solely, in Africa. And 98% of these deaths occur in the developing world. Staggeringly, for a disease many of us believed would disappear in our lifetime, TB is estimated to be the commonest cause of death of young women worldwide. Worse still, but at the same time providing cause for hope, these deaths are preventable. However, persistent poor practice in many countries is raising the spectre of multi-drug resistant (MDR) TB. This form of TB, if it became widely established, would make the control of the disease beyond the financial capacity of many countries.
  2. 3 The global burden of tuberculosis is truly enormous with some 1.9 million deaths per year according to the most detailed country-based analysis yet carried out and reported last year by the surveillance and monitoring unit of WHO&apos;s Communicable Disease Operational and Epidemiological Research Unit. Some 380,000 of these deaths occur in HIV infected individuals largely, but not solely, in Africa. And 98% of these deaths occur in the developing world. Staggeringly, for a disease many of us believed would disappear in our lifetime, TB is estimated to be the commonest cause of death of young women worldwide. Worse still, but at the same time providing cause for hope, these deaths are preventable. However, persistent poor practice in many countries is raising the spectre of multi-drug resistant (MDR) TB. This form of TB, if it became widely established, would make the control of the disease beyond the financial capacity of many countries.
  3. With decreasing CD4 T cells, the clinical manifestation changes from pulmonary to more severe forms of TB Like disseminated tuberculosis With increasing immunosuppression not only the risk of tuberculosis increases, but also the clinical manifestation of the disease changes as this compilation of several studies demonstrates. Early in the course of immunosuppression, pulmonary tuberculosis is the most frequent manifestation.  At the other extreme with very low CD4 cell counts, disseminated forms of tuberculosis are frequent.
  4. Period 1984 –2000 Death rate between 5% and 10% failure rate between 1% and 5% Study - HIV positive patients had higher mortality rate. 35% in HIV positive, and 13% in HIV negative patients. Mortality was strongly related to HIV infection, during treatment and three years after treatment.