6/7/2024

Integrated Environmental
Monitoring Program
Vanessa Agustine | Professional Service | Neogen Corporation

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01 Integrated Environmental Monitoring Overview

02 Sampling Handling

Hygiene Monitoring with ATP

Agenda
03

04 Allergens

05 Indicator and Spoilage Organisms

05 Pathogens

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Integrated Portfolio Solution

Neogen® Petrifilm Plates Soleris Automated Testing Culture Media Sample Collection
and Petrifilm Plate
Reader Advanced

Hygiene Monitoring Allergen Testing Natural Toxin Testing MLS UHT/ESL Testing

Pathogen Detection Genomics Nutritional Analysis Data Analytics

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The importance of Environmental Monitoring

Just because it looks clean,
doesn’t mean it is clean

Microbiological
Visual Inspection testing ATP
(swab & plate) bioluminesence

Cleaning and sanitation to remove or prevent All have their place in a robust environmental monitoring program
pathogens, spoilage microorganisms and
allergens.
Environmental monitoring generates data to determine if a
Environmental monitoring to assess the surface has been cleaned sufficiently and enables the high-risk
effectiveness of your cleaning and sanitation decision to start food production
programs.

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The goals of Environmental Monitoring

• to find pathogens in the environment before they contaminate product
• to find spoilage microorganisms in the environment before they affect product
• to assess effectiveness of cleaning, sanitation, and hygiene practices

Investigate growth Selection of the

niches: a hidden sample collection
home for device and sampling
microorganisms methods

Determine the
Selection of the
frequency of A negative result doesn’t
sampling neutralizers
sampling always mean absence –
maybe it wasn’t found.

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Integrated Environmental Monitoring

Not just ATP…

Not just Allergens…
Not just Indicator Organisms… ATP Allergen

Not just Pathogens… Sample

Collection

Indicator/
A good EMP should be an Pathogen
Spoilage
integrated approach!

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Integrated Environmental Monitoring

Based on zoning

Zone 1
Food contact surfaces
Slicers, peelers, fillers, hoppers, screens, conveyor belts, air
blowers, employee hands, knives, racks, work tables

Zone 2
Non-food contact surfaces in close proximity to food and food Allergens
ATP Indicators
contact surfaces
Processing equipment exterior and framework, refrigeration units,
equipment control panels, switches

Spoilage
Zone 3
Pathogens
More remote non-food contact surfaces located in or near the
processing areas
Forklifts, hand trucks, carts, wheels, air return covers, hoses, walls,
floors, drains

Zone 4
Non-food contact surfaces outside of the processing areas
Locker rooms, cafeterias, entry/access ways, loading bays, finished
product storage areas, maintenance areas

Higher focus/frequency Lower focus/frequency

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Integrated Environmental Monitoring

“Every Test Starts with Sample Collection”

ATP Allergen

Sample
Collection

Indicator/
Pathogen
Spoilage

Choosing the right sample collection

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Choosing the right collecting device

Sponges Swabs
Larger sampling devices and are available Smaller sampling devices consisting of a tip or bud for collecting the
in a variety of formats sample attached to a long flexible stem

Preferred choice when the area being Due to their smaller size and ease-of-use for sampling a defined area,
sampled is large and readily accessible they are preferred for small crevices and penetrations (areas of 100
(greater than 100 cm2) cm2 or less)

Preferred if qualitative pathogen testing is Can be particularly useful for quantitative environmental testing (e.g.,
to be conducted indicator organisms)

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Environmental sampling; sample collection device selection

Important
Considerations

Easy accessibility to sampling

Efficacy of locations and surfaces
neutralization
Sampling surface material
Sanitization Neutralization
Disruption of bacteria
immobilized in build up or
biofilm
Compatibility to
downstream tests Absence of biocide in sample
collection device

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Environmental monitoring and surface sampling

Sampling surfaces for microorganisms is a main

component of microbiological safety.

Sampling means removing the organisms from the surface.

Microbes are not inert inanimate particles and can quickly

respond to changing environmental conditions and attaching
and colonizing surfaces.

Current directions of research aimed at improvement of quality

and efficiency of microbiological surface sampling.

Adapted from: Digel I., Akimbekov N.S., Kistaubayeva A., Zhubanova A.A. (2018)
Microbial Sampling from Dry Surfaces: Current Challenges and Solutions. In: Artmann
G., Artmann A., Zhubanova A., Digel I. (eds) Biological, Physical and Technical Basics of
Cell Engineering. Springer, Singapore.

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Environmental sampling: Importance of neutralization

240,000

–
200,000

160,000
RLU

120,000

80,000

40,000

0
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75
Time (minutes)

Sample collection device WITHOUT Bacteria may die-off due to

ineffective neutralization
Potential false negative result
appropriate neutralizer

240,000

200,000

160,000
+
RLU

120,000

80,000

40,000

0
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75

Time (minutes)

Sample collection device WITH Effective bacterial recovery

appropriate neutralizer Accurate result
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Methods and Tools for Environmental Monitoring

ATP Method

ATP
Allergen

Sample
Collection

Indicator/
Pathogen
Spoilage

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ATP-based method review

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Sampling zone recommendations:

ATP Testing

Forklift Cart

Packaging Warehouse
RM
Warehouse

Finished Goods

Production Line

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Collection devices and simple testing steps

Click & Shake 5 Measure

seconds

Surface Swab

3 steps; <30 seconds

CIP Water
Dip & collect

2
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ATP system components
ATP swabs
• Surface and water options
• Sensitivity ≈3 femtomoles
• Optimized chemistry for use with
PMT technology = low %CV
17
Reliability of the AT P - based Method?
18
ATP reader
• Ergonomic design, one-handed
use
• Photomultiplier technology
• Storage of test points, sampling
plans and results
6/7/2024
Software
• Powerful enterprise level
software
• Data trending and reports
• Create and manage test points,
sample plans, users etc.
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Methods and Tools for Environmental Monitoring

Food Allergen Method

protein protein

Allergenic
protein A
Allergen protein
ATP Food product

Sample
Collection

Indicator/
Pathogen
Spoilage

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Sampling zone recommendations:

Allergen Testing

Forklift Cart

Packaging Warehouse
RM
Warehouse

Finished Goods

Production Line

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Cross-contact contamination with allergens

Almond

Almond
beverage
Rice

Plant-based beverage
production Rice beverage

Coconut

Coconut
beverage
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Allergen detection methods

Non-specific test Specific protein test

Quantification and detection of
ATP test swabs specific protein

Non-specific protein swabs Qualitative detection of specific protein

• Non-specific. • Highly specific and selective.

• Indirect measurement. • Direct measurement of specific proteins.
• Application: Cleaning procedure verification of • Application: Validation and verification of
complex (ingredient) production lines. cleaning procedures.

Preventing cross-contact contamination with food allergens is part of an environmental monitoring program

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Non-specific protein swabs

Clean-Trace surface protein (Altech60) AccuClean Advance

Green Clean
Grey Low level of contamination
Light Purple Moderate level of contamination
Dark Purple High level of contamination

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Allergen Protein Rapid Kits

Kit Format Lateral Flow Device, 3-D Technology, no QUALITATIVE

equipment

Limit of Low ppm levels of food residue

Detection

Suitable for Environmental swabs, CIP rinses

Testing

Samples per 10, 20, 25 samples or swabs

Kit (*depending on the type of kit)

Total Test 5 – 11 minutes

Time (*depending on the type of kit)

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Allergen Protein Rapid Kits

Reveal 3-D Allergen Rapid

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Allergen Protein ELISA Kits

QUANTITATIVE

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Methods and Tools for Environmental Monitoring

Swab Microbiology Method

ATP Allergen

Sample
Collection

Pathogen Indicator/
Spoilage

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Sampling zone recommendations:

Indicator / Spoilage Testing

Forklift Cart

Packaging Warehouse
RM
Warehouse

Finished Goods

Production Line

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Environmental Monitoring with microbiological methods

Air sampling

Swab sampling

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Indicator and spoilage test targets

Indicator Spoilage

Total Plate Count Total Plate Count

• Total population of bacteria capable of growing in the
presence of oxygen at mesophilic temperature.
• General indicator of sanitation.
Coliforms
• Indication of improper sanitation, insanitary conditions
or post process contamination.
Enterobacteriaceae
• Defined taxonomic group which includes more genera
than coliforms.
• Indication of improper sanitation, insanitary conditions
or post process contamination.
• Total microbial load in the processing environment.
• Relevant for highly perishable products.
Lactic Acid Bacteria
• Represent a diverse, functional collection of bacteria that
cause spoilage.
• Spoilage is categorized by off-flavor metabolites, slime and
carbon dioxide production.
Yeast and Mold
• Persist and propagate even under extremely harsh
environmental conditions.

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Methods and Tools for Environmental Monitoring

Swab Microbiology Method

ATP Allergen

Sample
Collection

Indicator/
Pathogen Spoilage

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Sampling zone recommendations:

Pathogen Testing

Forklift Cart

Packaging Warehouse
RM
Warehouse

Finished Goods

Production Line

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Pathogen testing

By focusing on the microbiological, physical and chemical safety of

the environment where food is being manufactured

Environment Key targets:

• L. monocytogenes (testing typically
targeting all Listeria spp.).
• Salmonella.
Raw End
Food product • Cronobacter spp. for powdered infant
material production formula (PIF) manufacturing.

Minimizes the risk of food contamination

in final product
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Swabs process

Pour Plate Process Petrifilm® Plate Process

Prepare agar Open plate package

Weigh out powder Prepare sample

Add water
Dilution(s)
Autoclave
Incubate
Temper

Prepare sample

Dilution(s)
Petrifilm Plates are a
convenient and reliable way
Pour agar to detect environmental
microbial contamination.
Wait for agar to solidify

Incubate

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Environmental Monitoring
Solutions
Allergen
ATP
Allergen Protein kits
Clean-Trace® System Reveal 3D, Veratox
Accupoint® NG
Sample
Collection

Pathogen Indicator/
Molecular Detection
Spoilage
System Petrifilm® Plates

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Contact Us
Thank you for listening and please contact me
with any needs or questions. Vanessa Agustine
Professional Service

Number Address Email Website

+62 812-9260-4356 The Manhattan Square, 12st VAgustine@neogen.com www.neogen.com
floor, Jln. TB Simatupang Kav 1s,
Cilandak Timur – Jakarta
Selatan 12560. LinkedIn : Vanessa Agustine

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