Microgen Bioproducts LtdMicrogen Bioproducts LtdMicrogen Bioproducts LtdMicrogen Bioproducts Ltd

Bacillus ID MIDBacillus ID MIDBacillus ID MIDBacillus ID MID----66666666

An identification system for An identification system for An identification system for An identification system for MesophilicMesophilicMesophilicMesophilic Bacillus species and Bacillus species and Bacillus species and Bacillus species and related genera associated with related genera associated with related genera associated with related genera associated with food poisoning and food spoilagefood poisoning and food spoilagefood poisoning and food spoilagefood poisoning and food spoilage

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Bacillus ID MIDBacillus ID MIDBacillus ID MIDBacillus ID MID----66666666

�General information

�Procedure

�Competitors

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Bacillus ID MIDBacillus ID MIDBacillus ID MIDBacillus ID MID----66666666

�Biochemical identification system that consists of 24 substrates

�Identification of mesophilic Bacillus and related Genera

�Substrates based on conventional methods: direct correlation with standard methods

�The last well of the second strip is a control which facilitates differentiation between positive and negative reactions

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List of organisms List of organisms List of organisms List of organisms in databasein databasein databasein database

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ReactionsReactionsReactionsReactions

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Reactions cont.Reactions cont.Reactions cont.Reactions cont.

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Procedure

� Confirmation of Bacillus species � Gram-positive Bacillus

� Endospores

� Catalyse positive reaction

� Preparation of suspension

�Some colonies can be very dry and hard so advised

to use a moistened swab – use of a vortex mixer may

be necessary

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Procedure

�Distribute colonies in 5mL of the supplied suspension

�Add 3-4 drops of the suspension to each well.

�Overlay Arginine well with mineral oil, easy to recognise as it is marked with black outline (3-4 drops)

�Incubate for 18-24 hrs at 30˚C

�First interpretation

�Second incubation for 18-24 hrs at 30˚C

�Add reagents (Indole, Nitrate A & B, VP I & II)

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Procedure

�Interpretation by use of colour chart

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Procedure

�Record the results on the form and create the octal

code

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Procedure

� Analyse the results on the Microgen-ID

software

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Summary Summary Summary Summary - An identification system that consists of 24 substrates

- Packaging: 20 tests (BAC1 & BAC2-supplied separately)

20 bottles of suspension broth

Instructions for Use

Results form

� Code: MID-66

� Additional Reagents:

� Reagent: Indole Kovacs MID61f

� Reagent: VPI, VPII MID61c/d

� Reagent: Nitrates A,B MID61a/b

� Mineral oil: MID61h

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SummarySummarySummarySummary

�System of 24 wells, easy-to-inoculate

�Results in 48 hours

�Database employs current taxonomy

�The only system developed for the identification of mesophilic

Bacillus and related genera of food poisoning & food spoilage

importance.

�Presence of control well acts as a aid in identification

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Competitors Competitors Competitors Competitors

�Biomerieux (API 20E + API50CHB )

�BD (Crystal Gram positive)

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bioMerieuxbioMerieuxbioMerieuxbioMerieux, API, API, API, API

�System that consists of 70 substrates

�Not all substrates are relevant to result: extra cost

and wastage

�Adaptation of two separate systems (API 20E &

50CHB), not a system devoted to Bacillus

�Requires the purchase of two separate systems

�Taxonomy employed is out-of-date

�Expensive per test

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BD,Crystal Gram Positive kit

� System is incubated for 18 hours and consists of 30 substrates for the identification of 121 Gram-positive organisms

� Not a dedicated system for the identification of Bacillus

� Self-inoculating device which remains closed after inoculation

�Wells are 2mm in diameter and very difficult to read

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Microgen Bacillus-ID vs

API50CHB+API20E

� A total of 49 strains of “Bacillus and related genera” comprising strains from recognised culture collections and isolates from various food and beverage sources were identified using the Microgen®Bacillus ID (MID - 66) and the API System comprising the API 50CHB and the API 20E

Total

Tested

MID-66 API

B. firmius 2 2 2

B. cereus 11 11 11

B. licheniformis 11 11 11

P. macerans 1 1 1

B. megaterium 1 1 1

B. pumilus 4 4 4

B. sphaericus 3 3 3

B. subtilis(1) 11 11 11

B. thuringiensis(2) 2 2 0

P. alvei 1 1 1

B. circulans 2 2 2

Total 49 49 47

Percent Correct 100.0% 95.9%

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Microgen Bacillus-ID vsAPI50CHB+API20E

� The Microgen® Bacillus ID successfully identified all of the isolates tested. The API system combination of the 50CHB and 20E was also able to identify 100% of the isolates however some taxonomyemployed was found to be out of date (see below):

2 isolates identified a B.amyloliquefaciens - a member of the B.subtilis

group

2 B.thuringiensis not included in API database - isolates identified a B.cereus. B.thuringiensis is a member of the B.cereus group. If the ID

of B.cereus considered correct, API = 100%