Neogen project to evaluate sanitation validation

Neogen and the Alliance for Advanced Sanitation are to use Next Generation Sequencing (NGS) and metagenomics to evaluate sanitation validation.

The technologies can be used to understand the relationship between microbes, shelf stability, safety and organoleptic properties of food.

The study will look at how frequently sanitizers should be rotated, what is the microbiological impact of changing them and determining frequency of periodic infrastructure cleaning (PIC) and periodic environmental cleaning (PEC).

It is planned that work will start as samples are received for microbiome analysis from next month.

Potential to understand and improve food safety through sanitation

Joe Heinzelmann said Neogen submitted a research proposal on using metagenomics to understand some key elements for sanitation practices.

“The proposal we put together is monitoring a food production site over the course of around 10 weeks and we are taking samples around sanitation events, so pre and post sanitation. There is a couple of key things we are looking for and trying to understand, because we think it will add value to the industry in the long term,” he told FoodQualityNews.

“There is no set deadline, it is basically to generate this type of information that the alliance and its member companies can use to better understand their sanitation practices.

“Such an important part of food safety and quality is tied to sanitation practices, for us to better understand the impact of our sanitation practices on the bacteria in a facility is going to eventually improve food safety and quality.”

The work includes periodic infrastructure cleaning (PIC) and periodic environmental cleaning (PEC).

“These two practices occur at a lot of ready to eat facilities over here and it is a deep cleaning on both equipment and physical elements of a production facility,” said Heinzelmann.

“The idea is as production continues different types of bacteria communities evolve and change and affect production and if we can monitor that in real time over the course of a number of weeks we can understand when these PICs should occur as they are more laborious and time intensive and what the impact of carrying those out are.”

Changing bacterial profiles

Another critical element is watching bacterial profiles change over the course of 10 weeks. 

“We are trying to time it around the change of season from Spring to Summer; temperature and humidity can play a role in the bacterial communities that exist in a food production facility and we can watch those change and grow and understand how they are affected by different sanitation practices,” said Heinzelmann.  

“We are planning three to five different companies depending on how many people we can get signed up, samples on the order of 10-20 per week around that sanitation event for the course of 10 weeks so we are talking 700-1,100 samples.”

Other members of the alliance include 3M, Cargill, Ecolab, Hershey’s, Kellogg’s, Nestle and the study is looking for additional companies that represent different commodities.

“If you look at the members in the alliance we don’t have a great representation from a fruit and vegetable production company but it would be great to understand, especially in that type of environment, the role of microbiomes and sanitation - how they interact.

“We would hopefully be targeting zone 1 and zone 2 close to the production line but that will depend on a company’s willingness to send in these types of samples,” said Heinzelmann.

“One of the things we could pull out if we are looking at zone 2 and zone 3 are, for example, better indicators of pathogen outgrowth than what we are using today or a pre-cursor to Listeria becoming a resident strain within a production facility.”

Sanitizer rotation

Heinzelmann said one of the things that is less clear is how often should sanitizers be rotated.

“Over the course of these 10 weeks what type of communities are building up, how are they changing, when we do rotate what is the impact, do different microorganisms come in and play a bigger role with new sanitizers and how does that impact their ability to do cleaning and sanitation and ultimately provide safe food,” he said.

“We are doing some of this work as a fee for service and from what we’ve seen in some of the datasets I can say we are going to learn an awful lot just because of how these profiles are changing even within a processing line from beginning to end. Some people had an idea but what is coming through in the data is actually surprising a lot of the experts we have been talking to as well.”

Technology and media bias

Neogen uses a metagenomic approach with the V4 16S region with Illumina technology and bioinformatics from Metagenome Analytics.

“We take an environmental sponge, we extract all the bacteria from it and we use that to amplify this gene that we can use to identify the bacteria in the sample,” said Heinzelmann.

“One of the things we have shown some of the invited people is within our technology stack of sequencing services we can turn off Salmonella and Listeria at the genus level. Which allows them to rely on their current food safety plans and also gain an understanding of the impact of microbiome and sanitation and how they come together.

“What we are seeing with a lot of these techniques is the introduction of any type of growth is only a partial picture of the different types of bacteria present in the environment. These media, either Tryptic Soy Agar or any type of total aerobic count, is only a portion of that because of the bias that it is only growing mesophiles and aerobic bacteria which is only a small portion of the picture.”

Heinzelmann said a lot of awareness is being pushed by adoption of the technology through the regulatory agencies and an application of NGS called Whole Genome Sequencing.

“This is a different application of NGS and that is metagenomics,” he said.

“So, instead of building the genetic code of one microorganism to compare it against a thousand other genomes to understand how it is behaving and where it is coming from we’re looking at a thousand genes in a community to see how they are changing and what the impact of those microorganisms are and how they behave in different types of environments and contexts.

“Instead of going a mile deep on one organism we are going a mile wide on a lot of them.”