Environmental Swabbing of Production Areas

• What is the purpose of the sampling
• Think like a bacteria – consider the basic principles discussed
• Where is the product more at risk from the environment
• Where are the more likely niche locations
• Where are cleaning and sanitation activities more likely to be compromised
• What practices increase the chances of growth niche creation, cross contamination, transport of microorganisms from on area to another, etc
• Take into account the zone concept and assignment of the designated zones

• There is no set standard for establishing frequency
• What is the purpose of the sampling
• What is the risk to the product
• Initially established based on results obtained during initial risk assessments and surveys
• Can be adjusted as the data is accumulated and trended
• Rotation of sites
• Common practice to identify a list of sites and rotate complete sampling of list over a period of time
• Allow for out of rotation or additional sites based on observations of production environment by sampling personnel
• Adverse results should always result in adjustment of frequency until resolution identified

Each group of microorganisms (gram positives, gram negatives, spore-formers, biofilm-formers) will have general characteristics relative to how they adapt and survive in the environment.

Microbial growth niches, a.k.a harborage site

• Areas within the facility or equipment that allow ongoing propagation of microorganisms
• Allows dispersion of the microbial population into the environment or equipment

Microbial biofilms

• Bacteria attached to a solid surface that produce extracellular polymeric substances that enclose and anchor the bacteria to surfaces
• The cells are protected from general cleaning and sanitizing efforts by the extracellular materials and are very difficult to remove once formed
• Biofilms are considered a microbial growth niche

• Training of sampling personnel
• Sampling tools
• Sampling techniques
• Sample site selection
• Sampling frequency and numbers
• Organisms of interest
• Test methodology
• Criteria or specifications
• Data management

Educated in the concepts of aseptic sampling using aseptic techniques

• Collection of samples using sterile materials and utensils
• Practices and techniques that avoid/prevent contamination of sampling materials or samples
• Gloves are for single use only – their purpose is to protect the sample from contamination, not to protect the samplers hands

Sampling protocols

• General guidance often provided by manufacturer of supplies
• FDA/ORA has training guidelines for investigations
• ISO 18593:2004 provides a general platform for critical steps in the sampling procedure

Organized and prepared

• Identify sampling sites prior for routine monitoring
• Fill out as much information on records or labels as much as possible
• Single sample for each location – do not use same sponge/swab for multiple locations
• Apply pressure to remove organisms
• Use both sides of the sponge to optimize recovery and surface area sampled
• Begin sampling in the “cleanest” production areas and move backward through the higher risk areas

Maintain integrity of the samples prior to testing

• Close and seal containers with samples
• Maintain at ambient temperature for minimal time
• <1 hour if quantitative testing involved
• 1 – 2 hours for pathogens
• Hold in refrigeration (2 – 7° C) until testing
• If shipping to test site
• Refrigerated, not frozen
• Use appropriate coolers and ice packs (no dry ice or loose ice)
• Testing initiated within 48 hours after sampling