Today we started by putting on scrubs and helped load 250 lbs milo grain, 750 cc of oyster shell dust and 25 gallons of water in each of 10 large stainless steel containers. After that we talk mushrooms with the director of mycology lab, then on to a 2 hour session with a 25 years teacher with a lot of enthusiasm who taught us about designing simple fruiting rooms and what the basic necessities were to maintain moisture, temperature and CO2 levels below 1000 for fruiting as well as about insulation and design. We had some hands on with low tek techniques. We made 3 trash barrels of 6 lbs each of straw hay (for low tek inoculation) one treated with one cup hand soap, one with 2 cups of bleach and one with 750 cc of lime. We submerged the hay to tomorrow’s inoculation of oyster mushrooms. I’ve read about this and have done some of this including cold formation (not attempted here) but again here it is hands on and the little nuances in 3 D really hit home the concepts. We had a 3 hour lecture and interchange regarding Sterilization and Clean room techniques with specific details on avoiding contamination in commercial product. The instructor the plant manager ,a stickler for sterility discussed the definition of a clean room, the meaning of sterile, the 7 different ways we can unknowing introduce contamination, the details of a HEPA filters (also trouble shooting them) and their importance, how air quality is measured with a Lazer particle counter and petri dish samples of the air, how much contamination is allowable, what involves work space attention (avoiding clutter, foot decontamination (90% of contamination), air filtration functioning, personal hygiene, hand washing clean clothes, minimum movement, dedicated equipment, nothing in the hood, and proper cleaning of the tools and work area every 10 min. We discussed the 7 sources for contamination ( Air, tools, inoculums, personnel, media or substrate, surface and mobile contamination units-MCUs) I had read and tried to practice take notes of all this at home but it is different again to see it and have excellence required. They did a hand washing test with with black light sensitive material that mimicked the adherence of bacteria. This to test our hand washing techniques. Before and after finger prints on agar were done for culture.
We discussed the proper cleaning techniques for clean rooms including wall and ceiling and wall cleaning in one direction with an autoclavable “swiftee” ($250 dollars each).
We are basically employees in training and must do all the chores and techniques of the other employees. No special priviledges will be afforded us and we will assist in all activities.
We ended the day discussing the effects of various medicinal mushrooms and blends.
We have a written test required for clean room certification before we can enter the clean rooms.
Sterilization and Clean Room Techniques and Concepts
Aloha medicinal Inc. runs a number of clean rooms and it is important that anyone working in these cleanroom areas understand the concepts of sterility and the procedures to be followed.
- What is a cleanroom?
- What is the meaning of Sterile?
Sterile means that all the microorganisms present are killed or removed.
Once something has been sterilized, it cannot get any more sterile.It can only pick up organisms as it is being processed further.This is why it is so crucial for us to maintain cleanrooms, so that the number of organism that get back into the product once it has been sterilized remains at a minimum
- What are the Different Ways we can unknowingly introduce contaminants into our products?
3.Inoculum (packaging for finished products)
5.Media or Substrate
7. Mobile Contamination Units (MCU’s) such as mites (in the personal cultivators space cats, dogs, and insects)
- What is a HEPA filter and why is a HEPA air filtration important?
Our HEPA filters clean the air down to the size of .3 micros.How big is that ?One micron is one millionth of a meter or 1,000,000 meters.Another way to look at this is there are 25,500 microns in an inch.A piece of paper is more than 100 micros thick.As you can see, a micron is very, very tiny.A typical bacterium is 5-20 microns in size.A spore from mold or mushrooms or other airborne life forms are also around 5-20 microns in size.Even the particles in cigarette smoke are larger than .3 microns and are too large to pass through a HEPA filter. So by passing the air through such a small , it effectively removes all the particles and microorganism such as bacteria and spores from the air. Essentially everything is removed from the air, leaving just pure air containing no living spores or bacteria that could contaminate our products.This clean filtered air is crucial to the sterility of the cleanroom.
- How is air quality measure?
The second method for determine air quality is to expose sterile Petri dishes to the ari to see what grows in them.
- What is contamination in regards to our product?
- How much Contamination is Allowable?
- How is contamination measured?
At the DRYING STAGE, there are two ways to detect contamination- SIGHT and SMELL.
The second way contamination is measure is by analytical lab methods. This is the Stand Plate Count (SPC) test, usually called just PLATE COUNT.In this test, a one gram sample of material is put on a Petri dish and grown for 24 hrs.. The number of organisms that grow is then counted.That number is described as the number of Colony Forming Units per gram, written cfu/g. This method is used to evaluation the completeness of our sterilization process.A SPC number of <100 cfu/gram is considered Sterile.Each customer and each country will specify how many cfu/g is allowable, but in any case, a plate count of no more than 10,000 cfu/g is ever allowable. Even if we get 5000 cfu/g it indicateswe have a real problem in our process.
Assume that when we sterilize our dried material we get a plate count of <100 cfu/g then everything we do to that material from then on can only INCREASE the plate count number.There is no way the number can go down, but will always icreas as we handle it. That is why it is so important that we use our very best cleanroom techniques when we work with dry powder in the canning and packaging stage.
- What are the factors involved in keeping the contamination down?
- Clean and Sterile suiting including gloves, face masks and foot covers.
- De-contaminate your feet before entering the cleanroom area. 90% of contaminates come from the floor.
- Be CERTAIN the air system is working properly.
- Personal hygiene. Hand Washing, Clean clothes, etc.
- Minimum movement- and keep your hands above your waist.
- Attention to work space.
- Dedicated equipment. Do not take any cleanroom equipment out of the cleanroom. Do not take non-cleanroom equipment into the clean room.
- What does attention to work space involve?
- Nothing in the room or work area other than the day’s work EVER
- Nothing in the hood except your hands and what you are working on. The hoods are not storages place.
- Proper cleaning of the room before the work starts and throughout the day.
- Not leaving the work area and re-entering without full re-suiting and de-contamination.
- Proper cleaning and handling of all implements such as pens, scissors,tape,etc.
- Be ABSOLUTELY CERTAIN YOU USE THE FOOT WASH TRAYS. No exceptions to this rule.
- Do not leave any work uncovered, ever. If you do not have your hands in the container it should be covered. This applies to all materials and supplies used in the clean rooms. It does not matter whether it is cans or bottles or powder or cotton or caps, or anything else. Keep them covered.
- What are the proper cleaning techniques for cleanrooms?
After the non-essential are removed, the room needs to be 100% cleaned with the designated cleaning sterilants. The particular cleaner will be specified by the supervisor in charge of that area. By 100% cleaned, that means floor, walls, ceiling and all surfaces such as countertops, canning machine, on top of the hoods, everywhere. If you cannot eat off the surface it is not clean enough.
It is extremely important that different sterilants be cycled through on each subsequent cleaning. The sterilants Aloha Medicinals typically uses are Bleach, Nolvasan, Iodine,Quaternary Ammonia, and Pine Sol. These MUST be measured and mixed according to instructions and not just dumped into the water bucket for cleaning. By properly mixing and cycling different sterilants, the maximum cleanliness is maintained.
Mops need to be clean and of the proper type. The mops need to be rinsed thoroughly every few minutes and not just wiped around spreading the dirt from here to there. The idea is to remove everything that is not a part of the room and not jot just to make the surface look wet. Only the broom, dust pans, mops, and mop buckets assign to that particular cleanroom are to be used. DO NOT tak cleaning supplies from one room to another. If some supplies are missing form your work are, INFORM you superviso so he or she can take care of the issue.
Alcohol an /or Nolvasan solution should be available in spray bottles at all times and all surfaces regularly wiped or sprayed with these sterilants every 10-15 minutes. This should be done more often when working under the hoods. If you are out of spray bottle sterilants stops and contact your supervisor to get you some more. This is one of the things you should check when your first set up for the job to ensure you not run out of sterilants or other supplies during the working day.
- Check lists
Edited by coorsmikey, 26 May 2016 - 08:16 PM.