Non sporing anaerobes Naresh

Non Sporing anaerobes

1. Non Sporing Anaerobes Mr. Naresh Pokhrel M.Sc Clinical Microbiology Department of Microbiology Father Muller Medical College, Mangalore

2. OVERVIEW :- INTRODUCTION CLASSIFICATION VIRULENCE FACTORS PATHOGENESIS PREDISPOSING FACTO R LABORATORY DIAGNOSIS TREATMENT REFERENCES

3. INTRODUCTION:- Anaerobes :- Microorganisms that grow only in complete or nearly complete absence of molecular oxygen. Non- sporing Anaerobes :- These do not form spores and are those which usually form the Normal flora Of Human beings and usually present in Skin, Oral cavity, GIT, Genitourinary tract and are opportunistic in nature.

4. CLASSIFICATION :- Gram negative bacilli :-   Bacteroides Prevotella Porphyromonas Fusobacterium Leptotrichia Gram positive bacilli :- Bifidobacterium Actinomyces  Propionibacterium Lactobacillus Mobiluncus  

5. Gram negative cocci:- Veillonela Gram positive cocci:- Peptostreptococcus Peptococcus Sporing forming anaerobes(Clostridia) Gram Positive Bacilli - C.tetani - C.perfringens - C.botulinum - C.difficle etc.

6. Virulence Factors :- Capsular polysaccharide :- Acts as antiphagocytic and chemo tactic . Adherence factors:- pili and fimbriae It is found in B.fragilis & Porphyromonas gingivalis.

7. Enzymes:- collagenase , neuraminidase, heparinase & fibrinolysins are produced by B.fragilis . Phospholipase A Super oxide dismutase Lipopolysacharide Endotoxin: - It is produced by Fusobacterium spp & Bacteroides . Succinic acid: - Helps to inhibit phagocytosis. Ex- B.fragilis .

8. Pathogenesis: 1. Brain Abscess:- 2. Oral & dental infection :- Root canal infections : Dental abscess : Periodontitis : 3. ENT , Head & neck infections :- Chronic suppurative otitis media : Acute bacterial Pharyngitis & tonsillitis : Cervicofacial Actinomycosis :

9. 4. Skin & soft tissue infection :- Acne vulgaris 5. Infections of lung & pleural space:- Aspiration pneumonia Lung abscess Empyema 6. Infections of female genital tract :- Pelvic inflammatory disease: Bacterial vaginosis : 7. Intradermal infections :- Intradermal sepsis : Pyogenic abscess :

10. PREDISPOSING FACTORS • Trauma, Tissue necrosis, Impaired circulation, hematoma formation or the presence of foreign bodies • Diabetes, Malnutrition, Malignancy or prolonged treatment with antibiotics.

11. Lab Diagnosis :- All the clinical specimen must be handled in very anaerobic condition. Long exposure to oxygen may kill obligate anaerobes and results in failure to isolate them in the Clinical Lab.

12. Accepted Specimen:- tissue bits, necrotic material, aspirated body fluids or pus in syringes. Unaccepted Specimen :- all the swabs, sputum or voided urine. Specimen should be immediately put into RCM broth or other anaerobic transport media and brought to the Laboratory as soon as possible.

13. Microscopy :- All the clinical specimen from suspected anaerobic infection should be done Gram stain and examined for characteristic morphology. Cul t ur e medi a Bruce l l a b l oo d ag ar (BR B A) P h eny l ethy l a l coho l ag ar (PE A ) K a namyci n -vanc o my c in- l a k e d b l oo d ag ar ( K V L B A) B a cteroi d e s b i l e esc u l i n ag a r ( B E A) Ne o myci n b l oo d ag ar ( N B A)

14. Cul t ur e met h od s :- A n aerobi c ja r Gaspak Candle Jar Bio Bag ( Anaerobic Sachets) Anaerobic Chamber

15. Anae r ob ic Jar Tightly sealed container in which oxygen is completely eliminated by hydrogen and a catalyst. palladium coated aluminum pellets acts as a catalyst . Methylene blue anaerobic indicator .

16. Bio Bag ( Anaerobic Sachets)

17. Anaerobic Chamber

18. Identification of Anaerobes: Biochemical tests Susceptibility to antibiotic Disks Gas Liquid Chromatography Ide n t i f i c a t i o n b y means o f sp e cial- po t ency An t i mic r ob ial a g e n t d i sk

19. Treatment:- Choice of antibiotics depends on the site of infection , type of anaerobe involved and susceptibility to antibiotics. Antimicrobial resistance in anaerobic bacteria is an increasing problem. Penicillin & Metronidazole. Clindamycin, Chloramphenicol, Cefoxitin, Cefotetan Bacteroides fragilis group Combination of Penicillin with β- Lactamase inhibitor (Co- Amoxyclav ) and to Carbapenems.

20. Reference: Anananthnarayan and Paniker’s text book of microbiology. Koneman’s color atlas and text book of diagnostic microbiology.