Romer Labs on the future of mycotoxin testing

Sampling methods to detect mycotoxins have evolved a great deal over the years. Yet, the quest is to make them even savvier and quicker, for a food/feed safety and economical reason. The aim is to get results closer to the point of purchase. On-site testing in the form of test strips is a very practical way to test for mycotoxins in the feed mill or storage facility. Romer Labs, based in Austria, is an expert in mycotoxin testing and focuses on a constant increase of accuracy and usability through continuous improvements of the applied technologies.

Biography:

Kurt Brunner studied Technical Chemistry in Vienna, Austria and worked during his PhD at the Vienna University of Technology and at the University of Naples, Italy. After several PostDoc years at different Universities in Austria he lead the “Molecular Diagnostics Workgroup” at Institute of Agrobiotechnology (IFA-Tulln), Austria and since 2015 he is Head of R&D at Romer Labs. Kurt Brunner has more than ten years of experience in the field of mycotoxin research.

How has mycotoxin research evolved over the last years?

“Mycotoxin research already started in the late 60s. During these years, the only method used and available was thin layer chromatography, where thin layers of silica were interpreted under UV light. This way you could detect the mycotoxins. However, this method was not sensitive enough so, soon after new detection tools were developed in the 80s. One of them was high-performance liquid chromatography (HPLC). The latter is still used today. Also in the 80s, ELISA tests were developed. These are sensitive and allow high throughput analysis, but can only be used in laboratories. A great development was seen in the mid-2000s, when lateral flow devices hit the market. These test strips first started as pure qualitative tests. This means that the test would give a simple yes or no if the mycotoxin was present at a certain cut off level. Year by year, these tests have been improved (such as the use of water as a solvent, instead of toxic solvents). Now these strips are considered as fully quantitative tests, which means you get a number (ppb level) as a result. A great improvement over the years has been the speed of testing. It would takes hours, but now we are talking just minutes. Tests have also been made much simpler. Everyone can use these, after a some basic training.”

Romer Labs has environmentally friendly tests. What does this mean?

“A lot of on-site testing is done with lateral flow devices (strips). To do the testing, an organic solvent is used for sample extraction. These have to be disposed. If you are working in a laboratory, you have disposal organic waste collection but this is not the case in a feed mill or other non-laboratory environments. Organic solvents are practical and efficient, easy to use. But on the other hand, they are toxic to both the operator and the environment. Romer Labs developed test strips that use water for extraction instead of an organic solvent. So the diluted samples do not have to be disposed with organic waste. Another advantage is that the extracted sample can be used to test for multiple mycotoxins. It is not only the environment that we have in mind, as we also aim to make these tests easier and quicker to use. We are trying to cut down the whole work flow to just a few steps, as each extra step can add analysis time and cause additional errors.”

The lateral flow device strips are put in the incubator and very quickly the operator can see how much ppb is present in the sample. The upper line on the strips tells that the test worked correctly. The intensity of the lower line is indirectly proportional to the mycotoxin concentration. The right sample is heavily contaminated.

How do you guarantee that your tests produce a reliable result?

“We can never guarantee perfect results, as we simply cannot control our customers. But next to our products, we offer services. These include proper training where we educate people on how to use the tests and how to take a representative sample. Next to this, we have reliable tests because we invest a lot of effort in validations. You cannot sell one test for every commodity or environment. This is often a misconception with customers. They buy a test and they think they can use it to test corn, wheat or finished feed. But every feed formulation has different ingredients and each ingredient is influencing the test in its own way. This can lead to higher results (overestimating) or underestimation. Especially changing the content of soy (protein and fat) significantly influences the test results. For this reason, we strongly work on validations. This means that we start validating every test for the simple raw materials such as corn, wheat, barley, etc. After that, we do the same for the more complex raw materials (such as DDGS and corn gluten meal). The next (and most complicated step) is finished feed. This is a complex issue, as you never know what it contains! But it is very important to tailor each test for our clients to make sure that the feed matrix is not influencing the test results. The effect of ambient temperature (think of tropical conditions, compared to northern European conditions) is avoided as the test strips are measured in an incubator at a constant temperature of 45° Celsius. We also keep on challenging our own tests to make them better.”

What can be improved in general mycotoxin testing / sampling?

“We are constantly improving. When we have launched a new product, we are already busy with the next generation of that product. Improvements are made also when you get feedback from the market. In addition, we are always looking for better performing antibodies, which are needed to make the tests more sensitive. In practice, this means that you can dilute the sample even more. And by doing this, you reduce the effect that different ingredients in the sample can have on the test result. Some people think that this will create more problems as we find more, but this is not the case. We don’t find more mycotoxins, but get more reliable results. And even if the user now measures a concentration of 300 or 400 ppb DON in a sample, they know that this is way below the regulatory limits. Regarding sampling, there are many helpful tools and guidelines (for example from the EU and FAO). These will guide the feed professional from taking a sample out of a big batch to turn it into an analytical sample (which is a different thing). These EU guidelines were published in 2006 and are very good so I don’t expect these to change in the coming years.”

How do you develop a test for new mycotoxins (or masked mycotoxins/ metabolites)?

“Coming up with a novel test for a new mycotoxin always starts with antibody development. And this is usually the longest phase. At Romer Labs we try to develop these antibodies in-house. But it can also be that we use antibodies from external parties. You can’t buy them from a catalogue. It is a very complex and time consuming business. Antibodies are nowadays produced by cell cultures. This enables us to have a constant quality of the antibodies. Once you have the antibodies that you want for that specific (new) mycotoxin, you also have to develop the different reagents. These are used to create the test and control line on the test strips, which are based on modified gold nano-particles. These have to be adjusted to the specific antibody used. Once this is done we have a prototype strip, and then the real work starts. We have to test this prototype strip for all the major commodities, the complex ingredients and finished feed. But we also test for different concentrations, different mycotoxins (to avoid undesired cross-reactivity) and different sources of the raw materials. You can imagine that we need extra hands during this phase.”

What are the current research topics Romer Labs is working on?

“First of all, we have a broad focus on research topics. Our general strategy is to improve our existing products. But most of our time is dedicated to developing new products, which also entails some fundamental research. Often market and regulation changes decide which direction our research efforts are steered towards. At the moment, the upcoming regulation for ergot alkaloids (expected this summer) is a hot topic. There are almost no simple test methods for ergots at the moment, so as soon as the EFSA came up with their opinion about this topic, we started to develop products serving these demands. This is the clear ‘pull strategy’ within our company. We don’t want to push products onto the market. But want to pull information from our clients and develop according to their needs. This is why our people keep a close eye on these EFSA opinions for example, as often these opinions will be translated into new regulations.”

Romer Labs has its own collection of fungal strains. This is very important as these are used for mycotoxin standard production for fundamental research, validations and for developing new tests. Photo: Emmy Koeleman

What will mycotoxin testing look like in 5 years?

“Frankly, I don’t expect very big changes in mycotoxin testing in the next few years. Of course, products will be improved, and hopefully we will be able to make tests that can measure multiple mycotoxins at the same time. But this is a challenge. For major breakthroughs we might have to look at a ten year span. This is because new technologies become available in the clinical diagnostics sector first. However, the money that can be spent on testing in the food and feed sector is less than what people are willing to spend in the medical area. There is often a tenfold difference. But I expect that novel technologies will become cheaper in the future and can be used for feed and food testing as well. Think of biosensors and chip technology for example. The possibilities to use these tools for mycotoxin testing in the future is exciting.”

Romer Labs, part of the Erber Group, is a global supplier of diagnostic solutions for mycotoxins, food pathogens, food allergens, gluten, GMO, veterinary drug residues, and other food contaminants. It has four fully accredited service labs in USA, Austria, UK and Singapore. Romer Labs was founded in 1982 in Washington, USA as a testing service for mycotoxins. In 1999, the company was acquired by Erber AG. In 2002, Romer Labs Diagnostic GmbH was founded in Austria. The move to the campus grounds close to the world’s leading mycotoxin research institute IFA Tulln in 2005 has been an important and strategic step. The IFA-Tulln was founded in 1994 as a joint research institution of three major universities in Vienna, the University of Veterinary Medicine Vienna (VetMed), the Vienna University of Technology (TUW) and the University of Natural Resources and Life Sciences Vienna (BOKU). The idea has been to enable the collaboration of scientists with complementary backgrounds in the interdisciplinary area of agro-biotechnology under one roof. “The proximity to the research institutes at the Campus Tulln and arising joint development projects have been beneficial for both parties, academia and Romer Labs. Campus Tulln is often seen as the worldwide hotspot for mycotoxin research and we are very proud of being part of it” explains Cristian Ilea, Head of Marketing at Romer Labs.