PACKAGE INSERT

The organism suspensions are provided in a sterile screw cap container with glass beads (100 or 20 unit vials, each dose unit is 100 micro liters or 0.1 ml ).

Note: - All reagents must be kept at recommended temperatures ( generally 0 to 4 o C) for most time and vortex minimally as this causes oxidation and germination, hence affects shelf life . Remove ascetically in the presence of a flame/hood, the required quantity of suspension for use. Close the container by flaming the opening, cap it, and return it to the appropriate storage temp as recommended (Other configuration and single use configurations are also available).

Pour plate ; typically 100 micro liters of microorganism suspension is placed in a Petri dish or inoculated for testing. Growth medium at temperate liquid form (37 to 40 o C) is poured on it enough to cover the surface, and the Petri dish is gently swirled in circles for TWO minute (IT IS IMPERATIVE THAT PROPER TEMPERATURE (37 TO 40 o C) AND MIXING IS PERFORMED IN CLOCK WISE AND COUNTER CLOCKWISE, experience show the figure 8 rotations is much better, GENTLE yet THOROUGH MOTION OR RESULTS WILL BE INCONSISTANT), to make sure the microorganisms are uniformly mixed and distributed, not clumped together, improper or incomplete mixing or premixing solidification of agar will result in clumps of organism in one area resulting in false counts. Allow the agar to solidify and then incubate as detailed below : If you are using a surface spread method note some airophilic organisms will show a higher count . ( Environmental enrichment is critical for full CFU development). Please note the CFU counts are based on a pour plate method; please contact ACD Inc. if you use other methods.

Aspergillus niger (16404) Incubate at 30 to 35 o C in a zip lock bag or in a container for 24-48 hours and continue incubation for a minimum of 5 to 7 days hrs to observe extent of fuzzy grey to black large spreading colonies. Early reading helps distinguishing the separate CFU.

Bacillus subtilis (6633) Incubate at 30 o to 35 o C for 24-48 hours and then continue incubation for a minimum of 5 to 7 days to observe extent of irregular white to creamy colored dry colonies. Inadequate mixing results in aggregation or cluster of colonies.

Bacillus/Geobacillus stearothermophilus (7953) Incubate at 55 to 60 o C in a zip lock bag with moisture pads or in a container with wet paper towels for a minimum of 5 to 7 days to observe extent of small irregular to circular creamy white colonies.

Candida albicans (10231) Incubate at 35 to 37 o C in a zip lock bag or in a container for 48-72 hours (preferred but not required CO2 incubator or Jar), and then continue incubation for a minimum of 5 to 7 days to observe extent of smooth and creamy white colonies. Prolonged incubation may cause diffuse growth on agar surface. Do not leave vials at room temp.

Clostridium sporogenes (11437) Incubate at 35 to 37 o C in an anaerobic environment place plates in a zip lock bag or anaerobic container for 48-72 hours to observe round circular to irregular fuzzy looking colonies (typically embedded in the agar). A minimum of 5 to 7 days is required to rule out negative cultures. For Fluid Thioglycollate Broth it is imperative that the broth be freshly prepared or reheated for 10 min. to drive of dissolved oxygen.

Escherichia coli (8739) Incubate at 35 to 37 o C in a zip lock bag (preferred but not required CO2 incubator or Jar) for 48 hours and then continue incubation for a minimum of 5 to 7 days to observe extent of moist slimy or shinny colonies (usually light colored dependant on dye used in medium). Inadequate mixing results in aggregation or cluster of colonies. Prolonged incubation may cause diffuse growth on agar surface . DO NOT STORE BELOW 0 o C, ice formation will affect viability . Higher temperatures (above 4-5 o C may cause germination.

Micrococcus luteus/ Kocuria rhizophila (9341). Incubate at 35 to 37 o C in a zip lock bag or in a container for 48 hours and then continue incubation for a minimum of 5 to 7 days to observe extent of bright yellow colonies. Inadequate mixing results in aggregation or cluster of colonies. DO NOT STORE BELOW 0 o C, ice formation will affect viability .

Salmonella typhimurium (14028) Incubate at 35 to 37 o C in a zip lock bag or in a container (preferred but not required CO2 incubator or Jar) for minimum of 5 to 7 days to observe extent of colonies. DO NOT STORE BELOW 0 o C, ice formation will affect viability .

Staphylococcus aureus (6538). Incubate at 35 to 37 o C in a zip lock bag or in a container (preferred but not required CO2 incubator or Jar) for 24-48 hours and then continue incubation for a minimum of 5 to 7 days to observe extent of moist white to dull white colonies (some times a dull white to light straw color dependant on medium). Do not leave vials at room temp Higher temperatures (above 4-5 o C may cause germination.

Pseudomonas aeruginosa (9027) Incubate at 35 to 37 o C in a zip lock bag or in a container (preferred but not required CO2 incubator or Jar) for 24-48 hours and then continue incubation for a minimum of 5 to 7 days to observe extent of moist slimy or shinny colonies (usually light green color is observed in medium ). DO NOT STORE BELOW 0 o C, ice formation will affect viability .

NOTE: - IN GROWTH PROMOTION TESTS THE COLONIES EMBADED IN AGAR DO NOT SHOW THE ABOVE CHARACTERISTS MORPHOLOGY AS THE COLONIES GROWING ON THE SURFACE OF AGAR.

Please follow incubation instructions as some organisms require a longer period of time to show visible CFU. Others are sensitive to temperature of poured agar or incubation temperatures.

IT IS IMPORTANT THAT PROPER GROWTH MEDIUM BE USED AS CFU NUMBERS, MORPHOLOGY AND EFFICIENCY ARE OPTIMUM WHEN RECOMEMDED GROWTH MEDIUM IS USED.

SHELF LIFE IS DIRECTLY AFFECTED BY THE EXTENT OF EXPOSURE OF MICRO ORGANISM SUSPENSIONS TO HIGHER TEMPERATURES ( above 0 to 4 o C ). HENCE IT IS IMPERATIVE THAT THE VIALS BE KEPT REFRIGERATED AT ALL TIMES . “DO NOT FREEZE”.