This document discusses the identification of bacteria using the Biolog system. It begins by explaining the importance of bacterial identification and limitations of traditional methods. It then describes how the Biolog system works, using a panel of 95 carbon sources to generate a metabolic fingerprint to identify bacteria. The procedure for using the Biolog system is outlined, including inoculating the microplates, incubating them, and obtaining readings to compare to the Biolog database for species-level identification. Key features of the Biolog system are its ease of use, ability to identify both gram-positive and gram-negative bacteria from a small number of cells, and generation of an identification within 24 hours.

1. RANI LAKSHMI BAI CENTRAL AGRICULTURAL UNIVERSITY
COLLEGE OF AGRICULTURE, JHANSI UP
COURSE: Plant Bacteriology APP 503 (2+1)
TOPIC: Identification of bacteria using biolog

2. INTRODUCTION
Identification of bacteria is essential for
nomenclature, to know about its effectiveness,
pathogenicity and it's management.
 A large number of automated, semi-automated and
manual systems are practiced for identification of
pathogenic bacteria.

3.  Most of these systems relay on bacterial
growth and subsequent pH, directed colour
change to estimate and identification.
 And they are not good at identifying some
important genera like Acinetobacter,
moraxella, haemophilus etc

4. BIOLOG FOR MICROBIAL IDENTIFICATION

5. BIOLOG IDENTIFICATION SYSTEM
It is a new instrument that establishes an
identification of bacteria based on the exchange of
electron produced during an organisms respiration,
leading to a subsequent tetrazolium based colour change.
This system tests the ability of a microorganisms to
oxidize a panel of 95 different carbon source

7.  Biolog panel contain 95 tests and the manufacturer has
been abel to compile a data base that includes 434
species or group of bacteria.
 Using biolog database,. over 2650 species of bacteria,
yeast and filamentous fungi can be identified by just
preparing a cell suspension and inoculate the
appropriate microplate.

8. PRINCIPLE OF WORKING:
 Biolog's microplate use redox chemistry to
colorimetrically indicate respiration of live cell
suspension.
 All wells are initially colourless, when a chemical in a
well is oxidised there is a burst of respiration and the
reduce a tetrazolium dye, forming a purple colour
 And a reference well contain no carbon source
 The test yield a pattern of purple well which consists
a metabolic fingerprint of each organisms

9. BIOLOG MICRO PLATE

10. PROCEDURE:
 The biolog system consists of a microplate containing
95 different carbon source, a contol well, a
turbidimeter, a computer driven automatic plate
reader.
 Culture to be tested were removed from storage,
subculture twice on trupticase soy agar containing 5
% sheep blood, and incubate overnight at 35°c.

12.  The inoculum to be used for testing was
prepared from second subculture.
 The inoculum was prepared by rolling a cotton
swab over the agar plate and preparing
suspension in 18-20 ml of 0.85% saline to
establish the appropriate inoculum density
relative to that of the milk of magnesia, standard
specified by manufacturer.

13.  The resulting suspension was poured into a
multi-channel pipette reservoir, precisely 150
micro liter of the suspension was dispensed in
to each well of microplate.he microplate was
incubated at 35°c for 4 hours.
 Then the plate was placed in the reader and
read automatically.
 The plates then returned to the incubator for an
additional 14-20 hours of incubation, and then
the second and final reading was made.

14.  The plates then returned to the incubator for an
additional 14-20 hours of incubation, and then the
second and final reading was made.
 The phenotypic fingerprint of purple well is
compared to bilog's extensives species library. If a
match is found, a species level identification of the
isolate is made.

15. PHENOTYPIC MICRO ARRAYS FOR MICROBIAL CELL

17. 1. The simplest and easiest bacterial ID system.
2. No Gram stain needed.
3. No pretest or follow on tests needed.
4.
4. Requires very few cells OD=0.01.
5.
5. One minute setup.
6. ID both GN and GP bacteria is a single,
universal panel.
7. 96 tests selected after evaluating 1,920 tests
FEATURES:

18. APPLICATIONS OF BIOLOG

19. Thank you
Submitted to,
Dr. Prashant. P. Jambhulkar,
Department of plant pathology,
RLBCAU, Jhansi.
Thank you
Submitted to,
Dr. Prashant.P. Jambhulkar,
Associate professor,
Department of plant pathology,
RLBCAU, Jhansi
Submitted by,
Sanjay H B- Ag/Pg/0017/19
M SC Ag GPB
RLBCAU, Jhansi