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How To Take Bacteria Samples

Sampling Air For Bacteria
Bacteria air samples are taken by impacting air onto some suitable growth media. Two of the commonly used samplers are Andersen (N6 Single Stage) and Reuter Centrifugal sampler (RCS). The Andersen sampler uses petri-dishes while the RCS uses agar strips. For environmental bacteria, a general purpose media such as Tryptic (Trypticase) Soy Agar (TSA) can be used.

Sampling Surfaces For Bacteria
Sterile cotton swabs can be used to sample for bacteria contaminating surfaces. The swabs are then sent immediately to a laboratory for culturing and identification of the bacteria to genus or species.

Mold Sampling And Identification Methods

Mold Sampling

The mold sampling method one chooses should be determined by the objective of the investigation. One may sample air or surfaces for mold contamination. The standard method for sampling air is to use a volumetric sampler e.g. RCS or Andersen N6 for viable airborne spores and hyphal fragments and Air-O-Cell, VersaTrap, Allergenco and others such cassettes for total spore counts. Testing of surfaces may involve use of RODAC agar plates for smooth surfaces, and swabs and adhesive tape on all other surfaces. It is important to note that adhesive tapes may not work well on wet and porous surfaces. Bulk samples can also be taken and plated onto agar plates or analysed by direct microscopic examination. Dust samples can be collect from surfaces such as carpets, upholstered furniture and textiles.

Media For Mold Sampling

It’s important to select media for mold sampling wisely. If one decides to collect viable air samples, the choice of media to use is very
important. Generally, malt extract agar (MEA) is used. It is a “broad spectrum” medium that supports the growth of a wide range of fungal species. However, antibiotics may have to be incorporated to surpress bacteria growth. Its main disadvantage is that fast growing molds tend to overgrow slow growers making it difficult to count colonies. To overcome this problem, DG18 and Rose Bengal can be used. These media have compounds added to them to slow down fast growing fungi and inhibit bacterial growth. If one is sampling a relatively dry environment, MEA+40% sucrose would be recommended for detecting xerophilic (dry loving) fungi.

Mold Identification

Currently, the only reliable means for routine identification of mold species is to perform traditional mycological methods. This requires years of training and practice. Be sure to use a lab that has a qualified Mycologist on-board (preferably at PhD level). The lab should also be regularly participating in a recognised proficiency testing program such as the AIHA EMPAT program.

Performing Effective Mold Sampling

If you need to take mold samples, use properly trained personnel or to get yourself trained. If you decide to undergo training, select a mold training course that provides skills and background information to enable you recognize indoor mold, develop effective mold sampling strategies, and interpret laboratory results.

Sampling for Airborne Mould: When Should One Use Viable, Non-viable or Both Methods?

An air quality investigation may require determining airborne mould (spores and hyphal fragments) concentration. Air can either be sampled onto some growth media for culture analysis or on a sticky surface or a filter membrane for direct microscopic examination. It is sometimes debated as to whether one should take non-viable samples, viable samples or a combination of the two. Either method can be used without the other or both can be used together (at the same time) depending on the objectives of the investigation.

Due to lack of standardization some terminologies used in air sampling are technically incorrect or misleading. Let’s discuss these terms first.

Non-viable Air Samples
“Non-viable air samples” refer to samples that are taken on some sticky media or on a filter membrane or tape and subsequently examined directly under a microscope for enumeration and identification of mould spores and hyphal fragments without culturing. In other words, the samples are taken for analyses by direct microscopic examination (DME). Results are presented as a listing of various categories of moulds and the corresponding number of spores or hyphal fragments per cubic meter of air (Spores/m3). This term is technically inaccurate since viable and non-viable propagules are indistinguishable under the microscope and hence both are enumerated.

Viable Air Samples
“Viable air samples” refer to samples that are taken on some growth media and subsequently incubated for mould propagules (spores and/or hyphal fragments) to germinate and form colonies. The resulting colonies are then enumerated and/or transferred to other media for identification to genus or species. Results are presented as a listing of the recovered moulds and their corresponding number of colony forming units per cubic meter of air (CFU/m3). That is, the analysis of viable air samples involves culturing. The term is also technically inaccurate because some (sometimes most) of the propagules impacted on the growth media may not germinate not because they are not viable but because of the selectivity of the growth media used, competition from fast growing moulds or that some moulds can only grow on living hosts.

Spore traps
“Spore traps” is commonly used to refer to non-viable air samples. However, whether sampling is done for culture analysis with an RCS, Andersen or for DME with Air-O-Cell or other similar cassettes it involves spore trapping. “Spore traps” is therefore applicable to both viable and non-viable samples.

When should one use viable, non-viable or both sampling methods?

The easiest way to decide on this is first to define the objectives of air sampling, data required from sample analysis and the questions these data are meant to answer. The objective might be broad or very specific.

  • When to use non-viable sampling

If the objective of air sampling was to have an idea of how contaminated the air is, then the data required would be total counts. Non-viable samples would then be the best to take because counting includes both those propagules that can grow on laboratory media and those which cannot grow either because they are dead or would not grow on the selected media. Non-viable sampling may also be selected when the objective of air sampling is to determine the total counts for airborne spores prior to and after remediation to assess the effectiveness of remediation. In this case viable air samples would not be necessary.

  • When to use viable sampling

If the objective of air sampling was to find whether the air contains a specific species of mould e.g., Aspergillus fumigatus, then identification to species would be required. Since non-viable analysis would not distinguish A. fumigatus from other Aspergillus species and not even from Penicillium species and related genera, then sampling for viable analysis would be selected. For detecting a specific species, a selective media that would support the growth of the mould of interest would also be selected. If identification to species was required for a broad range of moulds, then media that support growth of a wide range of moulds would be selected.

  • When to use both non-viable and viable sampling

If the objective of air sampling was to determine the total airborne mould concentration and at the same time determine the proportion of viable propagules, then both sampling methods would be used. This would possibly be the case in hospitals where concern is not only the total concentration of airborne mould but also the viable species present.

Conclusion
These are not the only reasons why one may sample for non-viable, viable or both non-viable and viable analysis. It all depends on the objectives of air sampling, the data required and the questions these data are intended to answer. Read Interpreting Numerical Data of Viable Airborne Mould Samples and Guidelines for Interpreting Numerical Data of Non-viable (Spore Traps) and Viable Airborne Mould Samples.

To get hands-on experience on the application of these guidelines register for our Mould Training Seminars today!

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