non sporing anaerobes

1. NON SPORINGNON SPORING
ANAEROBESANAEROBES
DR.P.HEMA SUGANYA M.D.DR.P.HEMA SUGANYA M.D.
Clinical microbiologistClinical microbiologist

2. CONTENTCONTENT
 DEFINITIONDEFINITION
 CLASSIFICATIONCLASSIFICATION
 NON SPORING ANAEROBE ASNON SPORING ANAEROBE AS
COMMENSALCOMMENSAL
 NON SPORING ANAEROBE ASNON SPORING ANAEROBE AS
PATHOGENPATHOGEN
 LAB DIAGNOSISLAB DIAGNOSIS
 TREATMENTTREATMENT

3. ANAEROBIC BACTERIAANAEROBIC BACTERIA
There is no universally accepted definitionThere is no universally accepted definition
of anaerobic bacteria-these require aof anaerobic bacteria-these require a
reduced oxygen tension for growth and failreduced oxygen tension for growth and fail
to grow on surface media when incubated into grow on surface media when incubated in
air or 5-10% CO2.air or 5-10% CO2.
►►Obligate anaerobesObligate anaerobes
►►Aerotolerant anaerobeAerotolerant anaerobe
►►Facultative anaerobes.Facultative anaerobes.
►►Capnophilic bacteriaCapnophilic bacteria
►►Microaerophilic bacteriaMicroaerophilic bacteria

5. Why anaerobes are killed byWhy anaerobes are killed by
oxygen?oxygen?

6. Why anaerobes are killed byWhy anaerobes are killed by
oxygenoxygen
 Aerobic organisms use oxygen as terminal electron acceptorAerobic organisms use oxygen as terminal electron acceptor
andand generate toxic oxygen reduction productsgenerate toxic oxygen reduction products includingincluding
HH22OO22,hydroxy radicals, singlet oxygen and superoxide anions,,hydroxy radicals, singlet oxygen and superoxide anions,
such reactive metabolites destroy the lipid components of cellssuch reactive metabolites destroy the lipid components of cells
and damage the DNA.and damage the DNA.
 However, aerobes and facultative anaerobes are protectedHowever, aerobes and facultative anaerobes are protected
from these reactive by-products due to certain enzymes likefrom these reactive by-products due to certain enzymes like
peroxidases and superoxide dismutasesperoxidases and superoxide dismutases

7. CLASSIFICATIONOF NONSPORING ANAEROBESCLASSIFICATIONOF NONSPORING ANAEROBES
Gram Positive Cocci:Gram Positive Cocci:
PEPTOCOCCUSPEPTOCOCCUS
PEPTOSTREPTOCOCCUSPEPTOSTREPTOCOCCUS
GramPositiveBacilli:GramPositiveBacilli:
BIFIDOBACTERIUMBIFIDOBACTERIUM
PROPIONIBACTERIUMPROPIONIBACTERIUM
MOBILUNCUSMOBILUNCUS
ACTINOMYCESACTINOMYCES
LACTOBACILLUSLACTOBACILLUS
EUBACTERIUMEUBACTERIUM
GramNegativeCocci:GramNegativeCocci:
VEILLONELLAVEILLONELLA
GramNegativeBacilli:GramNegativeBacilli:
FUSOBACTERIUMFUSOBACTERIUM
BACTERIODESBACTERIODES
PREVOTELLAPREVOTELLA
PORPHYROMONASPORPHYROMONAS
LEPTOTRICHIALEPTOTRICHIA

8. NON SPORINGANAEROBES AS NORMAL FLORANON SPORINGANAEROBES AS NORMAL FLORA
►►Most muco-cutaneous surfaces ofMost muco-cutaneous surfaces of
humans harbor a rich flora ofhumans harbor a rich flora of
anaerobic bacteria which varies atanaerobic bacteria which varies at
different anatomic sites in terms ofdifferent anatomic sites in terms of
concentrations and microbialspecies.concentrations and microbialspecies.
►►Upper air waysUpper air ways
►►Saliva (approximately 10Saliva (approximately 1088
/ml )/ml )
►►Gingival crevices (almost 10Gingival crevices (almost 101212
/ml)/ml)

9. NON SPORINGANAEROBES AS NORMAL FLORANON SPORINGANAEROBES AS NORMAL FLORA
►►Small intestine (10Small intestine (1044
- 10- 1055
/ml)/ml)
►►Terminal ileum and colon (10Terminal ileum and colon (101111
/gm)/gm)
(Peptostreptococcus species, Bacteroides species,(Peptostreptococcus species, Bacteroides species,
Fusobacterium species, Clostridia and a variety ofFusobacterium species, Clostridia and a variety of
non spore forming gram positive rods likenon spore forming gram positive rods like
Bifidobacterium, Eubacterium etc)Bifidobacterium, Eubacterium etc)

10. NON SPORINGANAEROBES AS NORMAL FLORANON SPORINGANAEROBES AS NORMAL FLORA
►►Female genital tract (10Female genital tract (1055
-10-101111
/ml) shows shifts during various/ml) shows shifts during various
stages of the menstrual cycle that may be hormonallystages of the menstrual cycle that may be hormonally
influenced. Thedominantinfluenced. Thedominant
anaerobesareanaerobesareLactobacillus(Doderlein’sLactobacillus(Doderlein’s
bacillus)bacillus),Peptostreptococcus and Bacteroides species.,Peptostreptococcus and Bacteroides species.
►►Urethra in both sexes contains skin flora as well as BacteroidesUrethra in both sexes contains skin flora as well as Bacteroides
and Fusobacterium species.and Fusobacterium species.

11. ROLE AS COMMENSALROLE AS COMMENSAL
 Compete With Pathogens For Its NutritionCompete With Pathogens For Its Nutrition
 Modulate Host Innate Immune ResponseModulate Host Innate Immune Response
 Produce Vit K& Bile Acids In IntestineProduce Vit K& Bile Acids In Intestine
 lactobacilluslactobacillus maintains vaginal Phmaintains vaginal Ph
 Prevent IBS.Prevent IBS.

12. ANAEROBIC INFECTIONSANAEROBIC INFECTIONS
COMMENSAL AS PATHOGENSCOMMENSAL AS PATHOGENS
►►Anaerobic infections are usually endogenous that is theyAnaerobic infections are usually endogenous that is they
originate from the host’s own flora when there is a breech inoriginate from the host’s own flora when there is a breech in
the muco cutaneous barrier resulting in the displacement ofthe muco cutaneous barrier resulting in the displacement of
normal flora.normal flora.
►►Spillage of fecal contents into the peritoneum followingSpillage of fecal contents into the peritoneum following
appendicitis, penetrating trauma or cancer can lead to intraappendicitis, penetrating trauma or cancer can lead to intra
abdominal infections involving anaerobesabdominal infections involving anaerobes

13. Anaerobic infectionsAnaerobic infections

14. CLINICAL DISEASES- Anaerobic CocciCLINICAL DISEASES- Anaerobic Cocci
 PEPTOSTREPTOCOCCUS
P.anaerobius responsible for puerperal sepsis.
P.magnus causes abscesses- breast/ axillary
Infections of sebaceous cyst also can occur due to these
organisms.

15. CLINICAL DISEASES- Anaerobic GPBCLINICAL DISEASES- Anaerobic GPB
 EUBACTERIUM
Common cause of Periodontitis
 PROPIONIBACTERIUM
Implicated in Acne
 LACTOBACILLUS
L.catenaforme is associated with Broncho-pulmonary
infections.
 MOBILUNCUS
M.curtisii & M.mulieris isolated in Bacterial vaginosis

16. Bacterial vaginosis
 Polymicrobial infection characterised by rotten fish,Polymicrobial infection characterised by rotten fish,
vaginal dischargevaginal discharge
 A direct wet mount may reveal many characteristic “A direct wet mount may reveal many characteristic “clueclue
cellscells”= desquamated epithelial cells with attached”= desquamated epithelial cells with attached
organismsorganisms
 Gram variable organisms seen on stainingGram variable organisms seen on staining

17. CLUE CELLSCLUE CELLS

18. CLINICAL DISEASES- Anaerobic GNBCLINICAL DISEASES- Anaerobic GNB
 BACTEROIDES
B.fragilis is the most common anaerobe isolated from various
clinical diseases viz., peritonitis, meningitis, brain abscess etc.
 PORPHYROMONAS
P.gingivalis- periodontal disease
P.endodontalis- root canal infections
 LEPTOTRICHIA
L.buccalis causes Vincent’s angina along with fusobacterium

19. CLINICAL DISEASES-CLINICAL DISEASES-
ANAEROBIC GNBANAEROBIC GNB
 FUSOBACTERIUM
F.necrophorum- produces a wide range of exotoxins: causes
Lemierre’s Disease ( necrosis of the lateral pharyngeal wall
with septic jugular vein thrombosis)
 PREVOTELLA
P.melaninogenica- Wound infections, mastoiditis, abscesses.
-Produces a black colour pigment (hemin derivative) & gives
red fluorescence when exposed to UV light

20. CLUES TO ANAEROBIC INFECTIONCLUES TO ANAEROBIC INFECTION
►►Infection adjacent to surfaces that harbour anaerobes as normal flora.Infection adjacent to surfaces that harbour anaerobes as normal flora.
►►Infections characterized byInfections characterized by abscess formation or tissue necrosis.abscess formation or tissue necrosis.
►►Gas formationGas formation
►►Grams stain of exudates showingGrams stain of exudates showing poly microbial florapoly microbial flora or organisms withor organisms with
morphologic features of anaerobes.morphologic features of anaerobes.
►►Foul odorFoul odor

21. Lab diagnosis ofLab diagnosis of
NonSporingAnaerobeNonSporingAnaerobe

22. Specimen collectionSpecimen collection
►► Liquid or tissue specimenLiquid or tissue specimen
is preferred .is preferred .
Swabs should be avoidedSwabs should be avoided (if used- must be(if used- must be transported intransported in
Stuart’s mediaStuart’s media))
►► Successful recovery requires rapid delivery to the laboratorySuccessful recovery requires rapid delivery to the laboratory
in a transport medium that maintains a moist, anaerobicin a transport medium that maintains a moist, anaerobic
atmosphere.atmosphere.

23. ANAEROBIC MEDIAANAEROBIC MEDIA
►► Non selective media having peptones, yeast extract, vitamin K, hemin, 5%Non selective media having peptones, yeast extract, vitamin K, hemin, 5%
sheep blood agar and reducing agent can be used for culturing anaerobes.sheep blood agar and reducing agent can be used for culturing anaerobes.
Variety of broth and solid media are commerciallyVariety of broth and solid media are commercially
available for the isolation of anaerobic bacteriaavailable for the isolation of anaerobic bacteria
►► Anaerobic blood agarAnaerobic blood agar
►► Kanamycin vancomycin laked blood agarKanamycin vancomycin laked blood agar
►► Phenylethyl alcohol agarPhenylethyl alcohol agar
►► Columbia colistin nalidixic blood agar mediumColumbia colistin nalidixic blood agar medium
►► Thioglycollate brothThioglycollate broth
►► Chopped meat brothChopped meat broth (Robertson’s Media)(Robertson’s Media)

24. GaspakGaspak

25. Anaerobic incubationAnaerobic incubation
systemssystems
A variety of systems are available forA variety of systems are available for
this purposethis purpose
►►Anaerobic jarsAnaerobic jars
►►Disposable bags & ChambersDisposable bags & Chambers
►►Roll tube methods etc.Roll tube methods etc.

27. TreatmentTreatment
 METRONIDAZOLEMETRONIDAZOLE
 CARBAPENEMSCARBAPENEMS
 AMPICILLIN /SULBACTAMAMPICILLIN /SULBACTAM

28. THANK YOU