How does Mesa report the spore population on their Certificate of Analysis?
The spore population on the Certificate of Analysis is reported as the heat shocked population per unit (biological indicator) with the exception of spore suspensions which are reported per 0.1 mL. Each product has a specific recommended population assay method. Follow the link below to access the population assay procedures for each Mesa Labs product.
I accidentally left my ampoules out of the refrigerator overnight. Are they ok or should I throw them out?
Geobacillus stearothermophilus, which is contained in Mesa Labs liquid ampoule biological indicators (MagnaAmp, SterilAmp II, ProSpore and ProAmp), has a growth range of approximately 40 – 70ᵒC and therefore cannot grow at room temperature. If the ampoules are still purple and have not turned yellow, they should be ok. If the ampoules have experienced temperatures higher than 40ᵒC, the spores would begin to germinate and lose their heat resistance.
I have a heat-sensitive, liquid pharmaceutical that must be sterilized. I want to use BIs with Bacillus subtilis “5230” to monitor my low-temperature steam cycle. Will regulatory agencies consider this an acceptable choice of BI?
The typical spore forming organism used to monitor steam sterilization cycles is Geobacillus stearothermophilus. This bacillus, however, may be too resistant to use in low temperature sterilization. BIs manufactured with Bacillus subtilis “5230” are acceptable but you will want to have data available to demonstrate that the reduced resistance capabilities of the Bacillus subtilis “5230” still represents an adequate challenge to your sterilization process when compared to the resistance capabilities of the product bioburden. For example, if your product bioburden is capable of “out-surviving” the spores in the BI, the particular BI in question is not appropriate. In such cases, one will need to perform bioburden testing to identify and document the resistance capabilities of the product bioburden and compare the results to the BI resistance. For additional information, please read Spore News Volume 1, Number 2 and Spore News Volume 1, Number 3 at the following links.
I have a product that must be sterilized by steam but is not heat-stable. BIs with Geobacillus stearothermophilus are too resistant. Is there another BI option to monitor my low temperature steam exposure?
Bacillus subtilis “5230” is a spore forming bacillus that has a lower resistance to steam than Geobacillus stearothermophilus. Use of this organism is recognized in ISO 11138-3:2006. Mesa manufactures Strips, SterilAmp and Suspensions using this organism for customers that have heat sensitive products.
BIs manufactured with Bacillus subtilis “5230” are acceptable but you will want to have data available to demonstrate that the reduced resistance capabilities of the Bacillus subtilis “5230” still represents an adequate challenge to your sterilization process when compared to the resistance capabilities of the product bioburden. For example, if your product bioburden is capable of “out-surviving” the spores in the BI, the particular BI in question is not appropriate. In such cases, one will need to perform bioburden testing to identify and document the resistance capabilities of the product bioburden and compare the results to the BI resistance. For additional information, please read Spore News, Volume 1, Number 2 and Spore News Volume 1, Number 3 at the following links.
I need to validate sterilization of a steam piping system. What type of biological indicator should I use and how do I secure the BIs at various locations?
The EZTest Steam biological indicator is perfectly suited for applications such as these. We also have an array of specially manufactured gaskets which will securely hold the BI in place. For additional information about Sterilization in Place (SIP) validation efforts, please read Spore News Volume 3, Number 4 at the following link. http://biologicalindicators.mesalabs.com/wp-content/uploads/sites/31/2014/03/V3-N4-Rick-SIP-problems-and-solution.pdf
I need to verify spore population label claim as part of our incoming inspection. What aspects of this test do I need to know to provide best chances for successful verification?
When trying to verify or duplicate someone else’s data, rule number 1 is to eliminate as many potential sources for variation as possible. Essentially, you want to execute the test in as close to an identical manner as is was performed to obtain the original data. Follow the link below to access the population assay procedures for each Mesa Labs product.
For additional information, please read Spore News Volume 4, Number 6 and Spore News Volume 10, Number 2 at the following links.
I read in ISO 14161 that the resistance for the spore suspension I purchased may be altered when I use it to inoculate my products. Can you explain why the spores change their resistance capability?
The spores haven’t changed their resistance capability; rather the test conditions have been altered. To measure D-value, the spores must be placed onto a carrier material. Together, the spores and carrier form a system. If one changes the carrier material used, the system has changed and the measured resistance will also be impacted. For a detailed discussion, please review Spore News Volume 5, Number 1 at the following link.
Am I going to contaminate my environment by transporting biological indicators into clean areas?
The bacterial spores used to inoculate Mesa Labs biological indicators are nonpathogenic, naturally occurring soil bacteria that are probably already present in a facility due to being carried in by people via soiled shoes, clothing and general every day human use of the facility. The bacterial spores on the biological indicators are probably in a higher concentration than what would be introduced by people but will not be a problem in a facility that employs Good Lab Practices (GLP) and aseptic handling. For additional information, please read Spore News Volume 4, Number 5 at the following link.
Are all of Mesa's products registered with the FDA's 510(k) notification process?
No. FDA has divided the biological indicators into two distinct segments for regulatory purposes. Biological indicators that are intended for sale to healthcare providers are Class II medical devices. ALL of Mesa’s products sold to healthcare providers have been granted the right to market through the FDA 510(k) notification process.
Biological indicators sold to manufacturers of sterile medical devices or pharmaceuticals are classified as manufacturing components not medical devices. Therefore, products sold exclusively to manufacturers of FDA regulated sterile products are NOT required to be approved by the FDA through the 510(k) process. As manufacturing components it is the responsibility of our industrial customers to assure that our biological indicators meet GMP’s and that Mesa has an acceptable quality system in place. Mesa has been audited by dozens of pharmaceutical and medical device manufacturers and found to have an acceptable quality systems program.
At what temperatures does Mesa perform D-value studies on EZTest steam?
Mesa Labs performs D-value testing on EZTest steam at 121ᵒC, 124ᵒC, 127ᵒC, 132ᵒC, 134ᵒC and 135ᵒC. The Certificate of Analysis lists the D-values at 121ᵒC, 132ᵒC, 134ᵒC and 135ᵒC. The Z-value listed on the Certificate of Analysis is calculated using the D-values at 121ᵒC, 124ᵒC and 127ᵒC per the ISO 11138 requirement of using three D-values between 110ᵒC and 130ᵒC to calculate the Z-value.
At what temperatures does Mesa perform D-value studies on spore strips for steam?
Mesa Labs performs D-value testing on Mesa Strips for steam, manufactured at the Bozeman Manufacturing Facility, at 121ᵒC, 124ᵒC, 127ᵒC, and then extrapolates the D-value at 134ᵒC. The Certificate of Analysis lists the D-value at 121ᵒC and the extrapolated D-value at 134ᵒC. The Z-value listed on the Certificate of Analysis is calculated using the D-values at 121ᵒC, 124ᵒC and 127ᵒC per the ISO 11138 requirement of using three D-values between 110ᵒC and 130ᵒC to calculate the Z-value.
Mesa Labs performs D-value testing on Spore Strips for steam, manufactured at the Omaha Manufacturing Facility, at 118ᵒC, 121ᵒC and 124ᵒC and then extrapolates the D-value at 132ᵒC. The Certificate of Analysis lists the D-value at 121ᵒC and the extrapolated D-value at 132ᵒC. The Z-value listed on the Certificate of Analysis is calculated using the D-values at 118ᵒC, 121ᵒC and 124ᵒC per the ISO 11138 requirement of using three D-values between 110ᵒC and 130ᵒC to calculate the Z-value.
Can I break the tops off the MagnaAmp and pour the liquid out to perform the viable spore count?
It is very difficult to pour or pipette all the liquid out of the MagnaAmp ampoule. It is also very difficult to rinse all the spores off the inside glass surface of the ampoule. It is best to place the ampoules in a 250 mL Wheaton jar or other Pyrex jar. Crush the ampoule with a sterile stainless steel rod. This allows the capture of all of the media and allows the washing of the inside of the glass surfaces. Follow the link below to the MagnaAmp population assay instructions.
and for additional information, please read Spore News Volume 10, Number 2 at the following link. http://biologicalindicators.mesalabs.com/wp-content/uploads/sites/31/2014/03/Spore-News-Vol-10-No-2.pdf
Can I use SterilAmp and MagnaAmp to monitor steam sterilization processes other than sterilization liquids?
NO. There are three critical variables in a steam sterilization cycle, time, temperature and saturated steam. A liquid BI placed in the chamber will only be monitoring time and temperature and not saturated steam.
As the temperature inside the chamber rises, so will the temperature inside of the liquid BI which in turn will create a steam environment inside of the BI and possibly kill the spores. The spores inside of the BI may be killed but the bioburden on the dry goods may not be killed if there was not saturated steam inside of the sterilizer chamber. Since the liquid BI does not monitor saturated steam, the dead spores may simply create a false sense of security because in reality the dry goods may not be sterile. Saturated steam is about 10 times more lethal than dry heat so the time and temperature may appear sufficient but in reality the cycle is not sufficient if there is not saturated steam.
For example, steam sterilization of porous loads of fabrics or hard goods loads of glass or metal instruments or machine parts are extremely sensitive to the presence of residual air in the autoclave. SterilAmp and MagnaAmp are not sensitive to the presence of residual air in autoclaves. Air will accumulate in the coolest location in the chamber and that is usually in lumens of devices or in the center of porous loads. Sterilization is MUCH slower in a trapped air pocket versus saturated steam. The SterilAmp and MagnaAmp will only respond to temperature. Since they are sealed liquid ampoules they generate their own saturated steam inside to kill the spores. These sealed ampoules placed in air pockets could be killed when spores on paper carriers such as EZTest® or spore strips demonstrate survivors.
Do I need to use a biological indicator (BI) in EVERY sterilization load or is less frequent use acceptable; for example is once per day or once per week an acceptable approach?
Mesa Labs cannot know the requirements of every regulatory agency around the world, so it may be worth checking with your regulating body to see if they have any specifications or recommendations concerning the frequency of BI use. However, if a BI is not used to evaluate cycle performance, the load is being released based only on the physical data available about the cycle, i.e. parametric release. In a properly validated sterilization process the expectation is that the BIs will be killed but if a BI is not used in every load, an assumption is being made that biological kill was achieved. Relying on assumptions puts one at risk of releasing a load that would have been quarantined had the BI been used and it detected conditions that probes can miss. At Mesa Labs, we tell customers that BIs are your insurance to sterility assurance.
How are PCDs used?
Process Challenge Devices (PCDs) are used in routine production sterilization cycles as an external test pack. They are designed to help medical device manufacturers and contract sterilization providers efficiently and accurately confirm EO sterility assurance. PCD patented products nominally consist of any commercial Biological Indicator strip, disc or Self-Contained BI (SCBI) in a proprietary EO barrier pouch. Directions for use can be found here.
How do I Validate PCDs?
What is the EO resistance (Deo) of PCDs?
Laboratory testing of our various PCD ‘types’ (particular polymeric film/pouch specifications) is done using the Stumbo-Murphy-Cochran (SMC) fractional method in a Biological Indicator Evaluator Resistometer (BIER) vessel. The values listed are nominal and should be used for reference and comparison only. Actual D values vary dependent upon the type of BI and other inherent process variabilities. Please review our PCD Selection Guide or contact us for further information.
What is the shelf life of a PCD?
Based on real-time studies conducted, the shelf life of BIs in our PCD packaging is equal to, or greater than, the BI manufacturer’s published expiration date.
Is Mesa ISO Certified?
Yes, Mesa is ISO 9001:2000 certified. Please contact us if you have specific Quality System inquiries.