Issue 9 of the BRCGS Global Food Safety Standard highlights the importance of validation in regards to cleaning procedures and performance. Depending on the risks associated with each site, acceptable levels may be defined by visual appearance, microbiological testing, allergen testing, or chemical testing. Another method referenced in the Standard is: ATP bioluminescence techniques.

These bioluminescence techniques include conventional ATP tests, along with the more accurate and reliable A3 system. But which option is best suited to your site?

This article hopes to provide an answer, highlighting the importance of effective Hygiene Monitoring within the meat processing industry. It will also explain the recent evolution of the ATP test from standard ATP tests to the Total Adenylate Tests (AMP + ADP + ATP) carried out by the A3 system. There are valuable takeaways from each of the points below, but you can also use the clickable links to skip to the issue that is most relevant to your site.

Why is Hygiene Monitoring so important?

In the meat production industry, hygiene monitoring is most commonly used to:

  • Identify high-risk sites and zones
  • Validate cleaning methods
  • Monitor the cleanliness of a factory
  • Educate and train operatives
  • Support the implementation of HACCP and HARPC programs
  • Reduce the risk of allergen contamination (as one part of a wider process)
  • Support the control of microorganisms to prevent foodborne illnesses and low quality products

One of the most common methods for hygiene monitoring is ATP testing. This is because ATP (adenosine triphosphate) is the universal energy molecule found in animals, plants, and microorganisms. A positive ATP test indicates organic residue is still present, so a lot of sites use this testing method to quickly find out if a surface is in need of further cleaning. When combined with visual inspections, conventional ATP tests are widely regarded as an effective approach to hygiene monitoring.

So, why did Kikkoman Biochemifa Company develop the A3 system?

As an unstable molecule, ATP degrades to ADP and AMP when exposed to heat, acids, alkali, liquids, and enzymes. This is likely to happen in an active food production site, with common practices such as cleaning, heating, processing, blanching, and fermentation capable of triggering this chemical shift. The problem with this limitation is that because ADP and AMP aren’t the target molecules for ATP-only assessments, a conventional ATP test will not be able to detect them. Instead, if the ATP has already been degraded, a conventional test could generate a pass result even if the surface it is testing is still covered in organic residue.

Recognising the shortcomings of conventional ATP tests, Kikkoman Biochemifa Company released a new hygiene monitoring technology in April 2017. Termed the ‘Kikkoman A3’, this system consists of the A3 Lumitester Smart and Lucipac Swabs and is capable of detecting all three forms of the adenosine molecule: ATP, ADP, and AMP. As a consequence, it can identify organic residue on a surface even after the degradation of ATP has taken place.

Used in the same way as a conventional ATP test, the A3 system features a lightweight, app based, handheld, auto calibrated device that allows you to set your own limits for pass/fail results. By running your choice of Surface Swabs, Water Swabs, or Pre-moistened Surface Swabs across the relevant test point, before inserting it into the meter, you can generate a reading in around ten seconds. The key difference is that the swabs used in the A3 system introduce recycling enzymes that allow for conversion between all three adenosine molecules. This means the reading you receive will relate to total organic residue.

Both conventional ATP tests and the more sensitive A3 system display results in Relative Light Units (RLU). This is because the technology behind them is based on the firefly luciferase reaction. Luciferase is the enzyme that produces light in the presence of ATP, so the intensity of the luminescence emitted during a test is a direct indicator of the amount of ATP on a surface.

When ATP degrades, conventional ATP assays are unable to detect the remaining organic residue – generating a low RLU result. However, the recycled enzymes mentioned above correct this issue, with the PPDK Enzyme converting AMP to ATP and the PK Enzyme converting ADP to ATP. This process increases the amount of luminescence produced and allows the A3 system to carry out a simultaneous measurement of all three adenosine molecules.

The video below offers a demonstration of a conventional ATP test and the A3 system at work:

How different are the results produced by the A3 system and conventional ATP tests?

To demonstrate the difference between the results produced by conventional ATP tests and the more sensitive A3 system, Kikkoman Biochemifa Company used both methods to detect organic residue on the same five surfaces (Figure One). When testing for the residue of sausage and bacon, along with raw chicken, beef, and pork, the results of the two systems differed dramatically. For example, when testing an area that had been exposed to bacon, the A3 system generated a reading of 400,000 RLU whereas the ATP test generated a reading of close to 0 RLU.

Kikkoman continued these tests with a wider range of organic residue(s), incorporating products such as beef brisket, sausages, and pork loin (Figure Two). When testing the A3 system against two of the most popular ATP meters, the Hygiena and the 3M, the RLU readings continued to differ. For example, when all three systems were used to test a surface that had been exposed to sausage, the A3 system generated a reading of over 10,000 RLU, whereas the 3M meter generated a reading of just over 10 RLU and the Hygiena system generated a reading of 0.

These findings are supported by research first published in the 2018 Journal of Food Protection, 81 (Figure Three). Data gathered by Bakke and Suzuki, illustrated the impact of ATP degradation over time, highlighting the limitations of conventional ATP tests when detecting organic residue once ATP has degraded. Again, they found that surfaces contaminated with products such as sausage and raw chicken were clearly detected by the highly sensitive A3 test, but passed a conventional ATP test, with the latter methodology unable to provide an accurate reflection of the amount of organic residue.

Why is A3 technology especially suited to a meat processing site?

A3 technology and Stainless Steel

Bakke and Suzuki’s research also examined the levels of residual ATP on stainless steel surfaces that had been exposed to raw meat. Six stainless steel plates were exposed to samples of chicken, beef, and pork, before being rinsed with 20°C tap water, 55°C hot tap water, then finally 20°C tap water, a sponge, and dishwashing detergent. This process was carried out twice and after each stage the surfaces were tested by an ATP test and an A3 system. The results of these tests are plotted in the graph below, with a standard Benchmark value of 200 RLU represented by the blue dotted line:

  • Blue: Raw Chicken
  • Orange: ATP levels
  • Pink: Raw Pork
  • Yellow: ATP and AMP levels
  • Purple: Raw Beef
  • Green: A3 (ATP+AMP+ADP)

As you can see, the RLU values of the surfaces exposed to raw chicken and beef fall below the benchmark value after the cold water wash, with the RLU levels for raw pork sitting just above this point. At this stage, a conventional ATP test would pass the surfaces exposed to the chicken and beef. By the time the surface has been cleaned by cold and hot water, the ATP test would pass all three surfaces, with a system that detects both ATP and AMP passing all but the surface exposed to raw pork. In contrast, the A3 system, only passes the surfaces on the final round of testing, following the clean with a sponge and dishwashing detergent.

If a meat processing site relied on conventional ATP tests to ascertain the best approach to cleaning stainless steel equipment, it would have every reason to assume a cold wash was sufficient for the removal of organic material. However, the findings of Bakke and Suzuki clearly illustrates that only the A3 system is able to accurately detect organic residue, sensitive enough to detect all three adenosine molecules even after the degradation of ATP.

A3 technology and Processed Meat

The same research paper determined the ratios of ATP, ADP, and AMP in different food samples. It established that large amounts of ADP and AMP were found in food products such as meat, seafood, dairy, and fruits.

The research paper also confirmed that raw meat contains mainly ADP and processed meat is abundant in AMP, with the levels of ATP fluctuating independently of the actual soil level. Published in the Journal of Food Protection, the Madison Study from the University of Wisconsin examined the ATP levels in different types of raw meat. As you can see the levels of ATP degrade rapidly over time, with the levels of ADP and AMP remaining fairly constant. This suggests that the best approach to detecting raw meat residue is to use a test that not only detects ATP, but also the degradation products of ATP.

A3 technology and Handwashing

A Case Study carried out in 2013-2015 at the Shiga Meat Centre, explored the on-site applicability of the A3 system in meat processing plants – examining the impact of A3 technology on the hygiene standards of operatives working on a cow slaughter line. The Shiga operatives were required to wash their hands thoroughly after processing each carcass, but damage, contamination, fungi, and wetness was still being transferred from container to container.

To understand more about what was causing this issue, A3 tests were conducted on the hands and fingers of each operative. With results available in around ten seconds, this process motivated workers to wash their hands more effectively. It also allowed the site to set realistic cleanliness benchmarks in different zones and establish the most effective hand washing technique. At the end of this Case Study, A3 and bacteria tests were conducted again. It was found that the detection of E.coli was reduced by approximately 30% and all RLU values, general bacteria counts, and coliform bacteria counts were reduced.

What are the benefits of introducing the A3 system to your site?

Ongoing Support

Introducing new technology to your site can be an intimidating prospect, but you will not have to manage this alone. Here at Klipspringer, we provide ongoing support during every step of the process. Working closely with a team of micro-biologists, we will help you to establish and validate the referential benchmarks for your A3 testing. We can also help you to identify suitable test points and select the right swab design for your site. Finally, we can provide training for your operatives – sharing resources and being on-call to answer key questions and encourage your team to engage with the new equipment.

Eliminate False Negative Readings

A conventional ATP test will often display a reading of 0 after swabbing. Sites will understandably process this as a pass result. However, a reading of 0 could also mean that a swab has been exposed to a high concentration of chemical detergent, is faulty in some way, or has been used incorrectly. In contrast, the A3 meter eliminates the possibility of false negative readings, as it always produces a reading (normally between 1-8) even if it is extremely low. For the A3 meter, a reading of 0 will only ever indicate a potential fault that requires investigation – reducing the risk levels surrounding your hygiene monitoring.

Access your data via the Lumitester App

The data gathered from your Hygiene Monitoring can be stored on the Lumitester App. This data analysis tool will automatically turn your inspection pass rates into graphs that can be downloaded onto Microsoft Excel or stored on the cloud to be accessed any time, anywhere, and from any device. The Lumitester App automatically records the results of every test alongside the date it took place and the operative responsible. This feature should help you to drive accountability across your site. The system also allows you to set test points and benchmark values remotely.


Why are undetected residues so dangerous in meat production?   

In the context of a food production site, residues are thought of as ‘undesirable substances’ with the potential to undermine standards of food safety. Chemical or biological in nature, residues can arise for several reasons, including modern processing practices and the incorrect storage of food. If residue levels sit above the permissible limits, there is the risk of a serious health hazard.  

In the case of the meat processing industry, biological residues are a major concern. This is because animals naturally carry bacterial species such as Salmonella and E.coli in their intestines, and during the slaughter process, the meat can become contaminated. What’s more, the pH of meat combined with its high water content, can promote microbial growth. This is a real concern as the consumption of pathogen-contaminated meat can cause gastrointestinal ailments and damage to the digestive system. 

So that brings us to the end of our guide to Hygiene Monitoring within the meat processing industry. We hope this article has highlighted the key differences between conventional ATP tests and the more sensitive A3 system, and helped you to understand the importance of accurate hygiene monitoring when it comes to driving standards across your site.

If you would like any further guidance, the Klipspringer team would be happy to help with your enquiries. Our in-house A3 expert Radek Tameczka will also be available to provide support and relevant resources. You can contact us on 01473 461800 or Alternatively, you can use the form below to arrange a consultation.

If you would like further guidance relating to Hygiene Monitoring, the Klipspringer team would be happy to help. Share your details below to arrange a free consultation.