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Isolation and Characterization of Novel Oxalate Degrading Lactic Acid Bacteria for Possible Probiotic Management of Kidney Stone View project
All content following this page was uploaded by Dr. Santosh Kumar Mishra on 14 June 2021.
College of Dairy Science and Technology, Guru Angad Dev Veterinary and
Animal Sciences University (GADVASU), Ludhiana 141004, Punjab, India.
Email: manjugdsc@gmail.com
Received: 19th November 2020 | Accepted: 15th January 2021
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of surface to be tested, frequency of sampling depending upon the product being processed.
and scope of analysis. It should be based on FCSs are made of various materials with
risk analysis and consider the previous history varying surface roughness and irregularities.
of sampling points (Zacharski et al., 2018). The microorganisms are attached to the surface
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being tested and they have to be removed process. The collected rinse water is tested for
using specific or generic sampling techniques, microbial load. This method has the advantage
it is further detected using different methods. that sampling of a large surface area can be
Therefore sampling protocol essentially should done and the equipment dismantling is not
ensure maximum recovery of microorganisms necessary. Alternatively, surfaces such as plastic
from the surface of interest. Sometimes the or wooden cutting boards are kept soaked in
sampling protocols need to be validated a rinse solution for a few minutes before the
with the detection method for recovery solution is tested for microorganisms. However
and repeatability to describe the surface as this rinse water sampling is not suitable to
clean from microorganisms/ready to start collect biofilm formed as the shear force
processing. In practice, sampling method is generated during flow may not be sufficient
either qualitative to determine whether the for their removal. In practice, using either a
number of microorganisms is within threshold specific or generic sampling technique any
level or quantitative to determine number of residue or microorganisms from the surface
microorganisms on the surface. Nevertheless it is transferred to an extraction or swabbing
is also important to maintain necessary hygiene solution.
precautions to avoid contamination of samples.
Swab method
Rinsing, Immersion Sampling The swab method is a traditional way
It is an indirect type of sampling used of direct contact sampling of surfaces which
to sample areas or equipment which are is most commonly used for many decades.
TARs
inaccessible for direct sampling for example The traditional swabbing procedure involves
tanks, pipelines, fillers etc. A sterile water is an applicator stick with one end with sterile
added upstream of the process and samples are bud (e.g. cotton, rayon, calcium alginate etc.)
collected downstream at various points of the remaining dipped in an extraction solution,
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usually 0.1 % sterile peptone water. Sampling tested for a few seconds. The attached cells
is done using the bud; the microorganisms are counted after incubation at optimum
on a defined area are extracted by rubbing growth temperatures. This method is more
thoroughly. After sampling the swab is cut into suitable to obtain quantitative data on the flat
the extraction solution to release recovered surfaces which are cleaned and sanitized prior
microorganisms. Finally the extraction solution to sampling.
is enumerated for microorganisms using
method of choice. It is also important that the Commercially available sampling methods
solution used for swab should not harm the Commercial test kits are simple, easy
recovered microorganisms and be compatible and more practical to perform and they require
with detection methods. Once the swabbing is less material as well as labor gives results faster
done the left over solution is wiped or rinsed (Table 1).
off from the sampled surface.
Petrifilm™ plate: composed by a double film
Scrubbing Methods system, comes in sealed foil pouches. On film
is the basis and is coated with dehydrated
An alternative to conventional swabs nutrients, indicator dye and gelling agents are
are sterile sponges or fabrics (e.g. Cellulose to be moistened using sterile distilled water
or polyurethane or polyester rayon blend) before sampling. When the gel is solid, the top
which have relatively large surfaces that can film is lifted and the gel is allowed to contact
be used to sample larger areas (>100 cm2). the sample surface by gently pressing to ensure
The sponges are pre-moistened with a rinse better contact. The top film is overlaid back
solution by rubbing over the surface. Sterile on the gel and incubated to obtain the results.
dry sponges are used to sample the surface This system is suitable only for direct contact
in moisture free processing sections. After sampling of flat or curved surfaces which are
sampling the swab is placed in the diluent free from crevices.
or enrichment broth. Before microbiological
analysis vortexing is necessary to facilitate Dip slides: These are plastic paddles coated
release of microorganisms from the sponge. with microbiological media used to sample
surfaces packed in capped vials. The media
Wipe sampling it is another sampling used is specific for microorganisms to be tested
method for assessing surface contamination. if present on the surface. While sampling agar
Microorganisms on the surface are picked up slide is removed from the vial by holding
by the wipe. When using the wipe for sampling the stopper. The agar is pressed against the
the surface of interest sometimes is moistened surface to be sampled with gentle press to
with an appropriate solvent. The percent ensure contact. The other side of the slide was
recovery is influenced by characteristics of also used similarly. The dip slide is returned
surface, solvent used and wiping technique to its original vial and incubated for 24-48 h
during wiping of surface. for bacterial growth and 48-120 h for fungal
Printing or RODAC growth. The microbial level is assessed by
using the chart provided by the manufacturer.
Another common direct sampling
method is using microbiological media
Microbiological Assay
TARs
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Table 1. Few examples of surface sampling methods available in the market (Ismaïl et al., 2013).
Type of method Name and description Manufacturer
Swabs 3M™ Swab-Samplers, 3M™ Quick Swab 3M, Minnesota, USA
HiMedia Foam Tipped Swab HiMedia, India
Q-Swab™ Hygiena/Scigiene Corporation
Path-Chek Hygiene surface swabs for Microgen Bioproducts Ltd
Salmonella spp., Listeria spp. and coliform
Spatula 3M™ Sponge-Sticks 3M, Minnesota, USA
3M™ Hydrated-Sponges 3M, Minnesota, USA
Swab rinse kits, SRK® COPAN Diagnostics, USA
HiMedia Nasco Sponge HiMedia, India
Microbiological Aerobic microbes, Hygicult® TPC Orion Diagnostica, Finland
dipslide Enterobacteria / *β-glucuronidase-positive organisms
(e.g. E. coli), Hygicult® E–/β -Gur
Yeasts and molds, Hygicult® Y&F
Total bacteria / *enterobacteria
Petrifilm™ Aerobic Count Plates 3M, Minnesota, USA
3M™ Petrifilm™ E. coli/Coliform Count Plates
Yeasts & molds,
RIDA® Count Aerobic Count Plates, E. coli/Coliform Count, R-Biopharm AG
Yeasts and moulds, Salmonella
enterobacteria
HYCON contact Total bacterial Count, Coliform Merck KGaA, Darmstadt, Germany
slides Yeasts and moulds
numbers if the low counts are expected. This microorganisms as an alternative to pour
approach is still proven effective at targeting plates as described earlier is widely used
indicator organisms or specific pathogens and in food microbiology. It is a sample-ready
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in swab are then detected calorimetrically with testing cannot differentiate the ATP from non-
a secondary antibody conjugated to specific microbial sources, such as food residues are
colored nanobeads. The entire analysis was a source of energy for bacteria. Surface which
rapid, simple and of low cost. have less microbial number but improperly
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being manufactured are made under sanitary of the Food Processing Environment.
conditions. In: Compendium Methods for the
Microbiological Examination of Foods.
(Eds.: F.P. Downes and K. Ito). 4th Edn.,
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