**5.3.8 Full automatic bacteria identification device**

Some amount of material is taken and transferred into selenite F bouillon (bio-Merieux SA-France). It is incubated at 37°C for 16 to 24 hours in the drying oven. Single colony planting is performed to *Salmonella* -Shigella agar (bio-Merieux SA-France) after the period has passed. It is incubated at 37°C for 16 to 24 hours in the drying oven. Bacteria is made suspension to provide 0,40-0,60 McFarlantd turbidity from reproduced suspicious colonies to Phoenix ID broth.

1 drop of Phoenix AST indicator (Phonex AST Indicator solution, BD Sparks, Benex Limited, Shannon, Ireland) to Phoenix AST broth (Phonex AST borth, BD Sparks, Benex Limited, Shannon, Ireland). 25 µl ID broth is taken and pipetted into AST Broth (BD Phoenix NMIC/ID-82 Sparks- USA). ID and AST broths are transferred to ID and AST pplate. Identification and antibiogram process are performed as full automatically in Phoenix-100 (BD Sparks-USA).

Isolation and Identification of Salmonellas from Different Samples 153

4. If formalin culture includes granular particles or thin membrane or sediment, control is performed by adding formalin salty water instead of antiserum (salty water control). For this, 0,02 ml Formalin salty water is put into the tube with the same length and 1 ml

5. Antigen-serum mixture and antigen-salty water mixture are incubated at 50°C warm water bath for one hour. It is controlled by 15 minutes of interval first and the final

a. If agglutination is present in culture+formalin salty water + serum mixture and agglutination is absent in culture- formalin salty water mixture, reaction is

b. If there is no agglutination in culture+ formalin salty water + serum mixture,

7. Immotile *Salmonella* cultures or *Salmonella* polyvalent H (flagella) negative cultures are

1. Various dilutions of antiserums are performed and it is processed with a known culture

2. The test is applied as told in section II. But, "0" group antiserums are used in here

3. Intense suspensions of cultures which give positive reactions with Vi antiserums in 1 ml physiological saline and it is heated in boiling water for 20-30 minutes and left for cooling. The test is repeated by using D, C and Vi antiserums of 0 group with these heated cultures. Vi positive cultures which react with Soamtic D group antiserums are likely *Salmonella* typhi. Vi positive cultures which react with Somatic C1 group antiserum are probably *Salmonella* paratyphi C. If Vi positive cultures heated which does not react with any of = group antiserum continues to give positive reaction after heating, they are not probably *Salmonella*. They belong

4. The culture is accepted as belonging to the group that the culture reacted positively with which of "0" group antiserums. Cultures that do not react positively with any of 0

These tests may be used instead of polyvalent H test which was specified in 4.4.2. It is used

1. Various dilutions of antiserums are prepared and it is processed with a known culture

2. Every seven Spiecer-Edwards H antiserum is processed with each of them. This examination is as told in section III. Spiecer-Edwards H antiserums are used instead of

3. Positive agglutination shows presence of H antigen. Antigen is detected according to Spiecer-Edwards antiserum agglutinins which was shown in the sollowing table 3.

c. If there is agglutination in both mixtures, reaction is non-specific.

This test is performed to determine the "0" group that the culture belongs to.

of formalin bouillon culture is added on it.

assessed according to Edwards and Ewing.

and reliability of antiserum is controlled.

instead of 0 polyvalent. ( Including Vi).

group antiserums are accepted as negative.

and reliability of antiserum is detected.

result is read after one hour. 6. Polyvalent H test is assessed as follows.

reaction is negative.

**5.4.3 "0" antiserum groups test** 

to Citrobacter group.

in determination of H antigens.

polyvalent H antiserum.

**5.4.4 Spicer-Edwards H (flagella) test** 

positive.
