MICROBIAL TAXONOMY - MICROBIOLOGY

Taxonomy of the Enterobacteriaceae
 The taxanomy of the Enterobacteriaceae is complex and is rapidly changing as further DNA homology studies are performed. A comprehensive approach to the classification of this family is presented by Kelly, Brenner and farmer in manual of clinical microbiology.
        The family Enterobacteriaceae in characterized biochemically by the ability to reduce nitrates to nitrites and to ferment glucose with the production of acid or acid and gas. The Enterobacteriaceae do not require increased amounts of sodium chloride for growth and are oxidase-nagative.

        Different species of Enterobacteriaceae are classified or grouped using series of Biochemical tests.
        Table 1.1 shows Biochemical reaction pattern in primary tests for the common clinically significant Enterobacteriaceae.
        Typically organism of Enterobacteriaceae is seen in growth on solid media as short gram-nagative rods often capsulated in some species.

1.   Eschherichia:- E. coli typically produces positive tests for indole, lysine decarboxylase, and mannitol fermentation and produces gas from glucose. An isolate from urine can quickly be classified as E.coli by its heamolysis on blood agar with an iridescent sheen on differential media like EMB.  

2.  Klebsiella- Enterobacter –Serratia group:-
Klebsiella species exhibit mucoid growth, large polysaccharide capsules, lack motility, gives positive tests for lysine decarboxylase and citrate.
Enterobacter species give positive tests for motility, citrate, and produces gas from glucose.

3. Proteus- Morganella- providencia group:
Members of this group on potassium cyanide medium (KCM), and ferment xylose proteus spp. Move very actively by means of peritrichous flagella resulting in swarming on solid media. Proteus and morganella are urease positive while providencia are urease negative.

4. Citrobacter: They are citrate positive and differ from the salmonellae in that they do not decarboxylate lysine ferment lactose very slowly.

5. Samonellae: They are motile rods, ferments glucose and mannose without producing gas. Do not ferment lactose or sucrose. They produce H2S.

6. Shigellae: These are non-motile and usually ferment lactose (but do not ferment other carbohydrates and producing acid but not gas they do not produces H2S.
                                                                
Antigenic classification  
    A part from biochemical characteristics, Enterobacteriaceae are classified by more than 150 different heat stable somatic O (Oligopolysaccharide) antigen, more than 100 heat labile K (Capsular) antigen, and more than 50 H (flagella antigens. Antigenic classification of Enterobacteriaceae often indicate the presence of each specific antigen for organisms to share a common environment, they must have something in common (characteristic)
G + C content
        – haemolysis
Sterner
    The Enterobacteriaceae are a large, heterogeneous group of gram negetive rods whose natural habitat is the intestinal tract of humans and animals. Some enteric organisms, e.g E. coli are part of the normal flora and incidentally cause disease, while others, the salmonellae and shigellae, are regularly pathogenic for humans. The enterobacteriaeae are facultative anaerobes or aerobes, ferment a wide range of carbohydrates, posses a complex antigenic structure and produce a variety of toxics and other virulent factors. These enteric gram-negative rods or enteric bacteria may also be called coliforms.      

    Carbohydrate fermentation patterns and the activity of amino acid decarboxylase and other enzymes are commonly used in biochemical differentiation and classification. Culture on differential media that contain special dyes and carbohydrates e.g EMB, MacConkey, etc distinguishes lactose fermenters from non-lactose fermenters and may allow rapid presumptive identification/classification of enteric bacteria together with production of indole for tryptophan.
    One complex medium have been devised to differentiate between salmonella and shigella from other coliforms. The medium is called triple sugar iron agar.

Rapid presumptive identification/classification of gram-negative enteric bacteria into 3 groups
Lactose fermented rapidly
Lactose fermented slowly
Lactose not fermented
E. coli: metallic sheen on differential media; motile, flat; non-viscous colonies.
Edward siella, Serratia, citrobacter
Shigella Spp: Non Motile; No Gas From Dextrose
Salmonella Spp: Motile, acid and usually gas from dextrose.
Enterobacter aerogenes: Raised colonies, no metallic sheen, often motile, more viscous growth.
Arizona,
Providencia
Erwinia.
Proteus spp: ‘swarming’ on agar; urea rapidly hydrolyzed (smell of ammonia).
Pseudomonas spp:
Soluble pigments, blue-green and fluorescing; Sweetish smell
           
TABLE: BIOCHEMICAL REACTION PATTERN IN PRIMARY TEST FOR COMMON CLINICALLY SIGNIFICANT ENTEROBACTERIACEA

Tests
Indole
Lysine
decarboxylase
Mannitol
Glucose
Citrate
xylose
Urease
lactose
Mannose
1
E. coli
+ve
+ve
+ve
Pdtn of Gas
+ve




2
Klebsiella spp

+ve

Pdtn of Gas
-




3
Proteus
 morganella poovidencia
-
-
-
-
+ve
+ve
+ve
+ve
-ve
-
-
4
Citrobacter
-


Pdtn of Gas



slowly

5
Salmonellae
Pattern of H2S



+ve



+ve
+ve
6
NO H2O pattern
Shigellae







+ve


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