The term aseptic, as it is used in regards to sampling, means to avoid contamination by one’s actions during sampling. An aseptic sample is collected in such a way to avoid contamination during the collection process itself.  The sample is then placed in a sterile container. Aseptic samples are collected primarily to support observations that are made during inspections concerning the sanitary conditions in a food facility.

PREPARATION FOR COLLECTING SAMPLES

 Preparation is essential to collecting aseptic samples. Identify the product to be sampled in order to understand the microbiological hazards associated with the product and to assure that the appropriate equipment is available for sampling.

·       Identify production schedules.  Discuss collection times for aseptic samples with the analyzing laboratory so sample analysis can be planned within their work schedule.

·       Review product flow and develop an aseptic sampling strategy.  If familiar with the product flow, or a flow diagram is available from a previous inspection, a preliminary plan for collecting samples is advised. If no information is available, sample locations will be determined at the facility during the inspection. Normally, two sets of samples are collected during an inspection, one during the first part of the production shift and a duplicate set later in the day. As a rule, two finished product samples are collected; one that represents product produced on the day of the sampling, and another that represents a previous day’s production.

·       Pack the aseptic sampling equipment.  Having the right equipment to collect aseptic samples of a particular product and/or process is critical. Unless proper collection equipment is used, sample integrity might be questioned, making the sample meaningless.  Create an aseptic equipment checklist and use it when gathering equipment for sampling.

·       Pre-identify containers.  Record the sample number, collection date, and collector’s initials prior to entering the process facility on the container.  This can make sample collection easier under adverse plant conditions.  Sample numbers are typically assigned during the sample collection and are therefore not pre-marked.  Always include enough sample containers and equipment to account for open and closed controls. These controls are used to demonstrate that any microbial contamination found during analysis did not originate in the sample containers or sampling equipment. Personal equipment, such as lab coats, hair nets or disposable or cleanable boots, help to insure that the collector does not contaminate the food product or samples.

·       Review regulatory and inspectional programs.  Read and understand compliance programs and guidance documents for the Food that will be aseptically sampled. Programs such as: 

      ·         Domestic and Imported Cheese and Cheese Food;

      ·         Domestic Food Safety Program;

        ·         Domestic Acidified and Low-Acid Canned Foods Program.

All of these give valuable information concerning sampling strategy and sample sizes. If no sample guide exists for the product being sampled, the sample size should be discussed with the analyzing laboratory.

SAMPLING SITES

Bracketing samples means that a sample is collected every time the product changes. This includes any step in processing that changes the product, such as adding breading to a fish fillet or any time an employee touches the product, such as an employee who removes cheese from a vat.  

In-line Samples

In-line samples are typically the raw materials -- product, processing water, packages, or anything else used “in-line” during production.  In-line samples are collected to determine if the source of bacterial contamination occurred with the raw materials or at some point in the process.  

Environmental Samples

Environmental samples are bacterial swabs.  (NOTE: Environmental swab sampling does not give quantitative results because the sample is very small and microorganisms of significance are often missed).  The swabs are taken from places like food-contact surfaces, floor drains, walls, overhead pipes, and other potential sources of contamination that are observed during the inspection. Documenting the possible link between the source of the environmental sample and contamination of the food product makes the sample meaningful and is advised.

Examples are:  

·         Floor drain -- Were employees walking through the area of the floor drain and back to the processing area?

·         Wall -- Did an insect land on the wall and then land on in-process product?

·         Ceiling -- Is condensate or flaking paint located over in-process product and was it observed to contact the product?  

Open and Closed Controls

An open control is a sterile container that has been briefly exposed to the environment that the sample has been collected and then closed. Closed controls are those pieces of sterile sampling equipment that were never opened. These control samples must be included in the sample to show the condition of sampling equipment and the sampling environment at the time of sampling.  

EQUIPMENT

The equipment needed for sampling varies for different food. Compliance programs, inspection manuals and guides, or other guidance documents, should be reviewed to determine sample equipment needs for selected food. Again, it is useful to prepare an equipment checklist to insure that all the required aseptic sampling supplies are available before departing for the firm.

Equipment needed include:  

·         Alcohol wipes -- These are good for wiping the stem on a thermometer before and after one takes a temperature.

·         Hand antiseptics -- If the facility does not have handwashing and sanitizing solutions, inspectors can use antimicrobial soap wash and sanitize their hands.

·         Blue ice or  gel packs -- A cooling medium is necessary if one needs to keep the sample cold during storage and transport. Check to see if the bags of blue ice are intact, a leaker can contaminate a sample. One can also use wet ice. Wet ice might be available at the firm, however, one should be aware of this before picking up samples. Dry ice must be obtained prior to the inspection if one needs to keep the sample frozen.

·         Boxes or coolers -- If the samples do not need to be refrigerated, then a clean box can be used.  However, if the sample needs to be kept cold, a standard cooler or insulated carton must be used.  Typically, the cooler is lined with a plastic bag. Samples are then put inside the bag and the cooling medium like blue ice or dry ice is placed outside the bag so contamination of the sample by the ice is prevented.

·         Chlorine, iodine and other sanitizer check strips – Used to check hand dip or equipment sanitizing solutions.

·         Sterile containers – Vary from plastic bags to sterilized gallon paint cans for food with sharp edges, such as crabs and shrimp.

·         Sampling tools --  Spoons, scoops, specific types of triers, forceps, tongs, dippers, and cups.  The type of tool used is determined by the product to be sampled.  

Check the sterilization date on all sampling equipment and containers. The date should be on the label or package of the equipment. Some equipment might be sterilized in one’s local laboratory or purchased sterile.  Equipment sterilized in a local laboratory is normally sterile for at least two months. After that time the equipment should be re-sterilized.  

·         Sterile gloves -- If a product has to be touched during sample collection have one of the facility workers, who are involved with handling the product, place the sample into the collection container. Because employees are normally involved in handling the product during production, we would not consider that they had added additional contamination to the product. When gloves are used they must be put on in a manner that does not contaminate the glove. Gloves must be of the right size to work properly.

·         Sterile swabs -- These are used to swab equipment and environmental areas of the plant. There is a correct procedure for using the swab. One unwrap the swab by peeling back the cover, then one have to be careful to hold on to the top of the tube and the top of the swab to make sure one don't contaminate the swab. Next one swab the area one want to sample ­like a table top or an overhead pipe.  Then one remove the top of the tube and carefully put the swab inside -- and push it all the way down until it is in the medium.

·         Sterile Whirlpack bags -- These bags are purchased sterile. To use the bag one just tear off the top, hold the bag by the provided  tabs to open it, put the sample inside, then roll down the top of the bag and secure it with the wire ends. The ends should be folded twice so that the wire doesn't poke through the bag and cause the sample to leak.  As the sample is collected, the conditions under which the sample was collected, such as product temperature, location etc., along with the sub sample number, are recorded in the inspector’s notes. Samples are identified with the sample number, date of collection, sample number, investigators initials, and other identifying information.  

One of the most important rules when collecting aseptic samples is to do nothing to contaminate the sample. This requires the sample collectors to collect all samples with caution, to make sure this rule is not violated. The results of aseptic sample analysis are questionable, and of no use, unless the integrity of the sample can be assured from collection, to shipment, and through analysis.

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