When Feedinfo chatted to Germany’s Provita Supplements last year about its functional fermentation product, MAXFERM/Pro, our discussion centred around its technology boosting the availability of protein in feed ingredients to improve pig performance. We also addressed how it could impact feed costs by reducing the total protein content needed in formulations.
At the time the company also mentioned that it was starting to investigate the potential of functional fermentation products to improve gut integrity in monogastrics, with initial studies showing a positive influence on gene expression of tight junction proteins.
Now, a little more than 11 months later, Provita Supplements is sharing the results of these investigations with Feedinfo. In today’s Industry Perspectives, the company’s Head of Sales and Marketing, Dr. Karoline Reckmann talks us through its gut health studies involving MAXFERM/Pro, touching not only on its impact on tight junction proteins, but also its ability to breakdown complex sugar-protein chains to provide easily fermentable carbohydrates and how it can prevent the adhesion of certain enteric pathogens in the pig gut.
[Feedinfo] When we last spoke last year, there was mention of further investigating how MAXFERM/Pro can potentially promote intestinal health. Why did Provita Supplements see this as vital for the company to explore?
[Karoline Reckmann] Together with sustainability and raw material availabilities, gut health is one of the major areas of interest for the global livestock industry. We as Provita Supplements wanted to contribute to its understanding while aiming for antibiotic-free weaning and, therefore, more sustainable pig production. This is why we developed a holistic nutritional approach which made gut health the centre of attention. And we have employed our years of experience gained in Germany and Europe to develop recommendations that can successfully be applied all over the world.
Thus, when developing and evaluating products, we do not only assess their effects on performance but also on environmental parameters and the vitality of animals.
[Feedinfo] So, what exactly has this investigation entailed? What has Provita Supplements been doing to further its understanding/research on MAXFERM/Pro’s potential gut health benefits since we last spoke? And what were some of the key findings?
[Karoline Reckmann] The main effect of MAXFERM/Pro is the improvement of protein availability in monogastrics. By using select strains of Aspergillus fungi with carbohydrase activity and a wide range of fermentable substrates, our solid-state fermentation (SSF) product can break up the protein-rich cell structures and cell residues in the feed very efficiently. Through this process protein from the colloidal system of the cytoplasm and from the cell wall is mobilised.
Outside of this main mode of action, we wanted to investigate the potential increased expression of tight junction proteins leading to a lower overall risk of leaky gut and a strengthened intestinal barrier. Our trials also highlighted impacts on the degradation of carbohydrates promoting desirable microbiota and the suppression of pathogens. We also discovered positive effects on the wound healing process.
[Feedinfo] Let’s unpack things a bit more. What did you find out on the impact that MAXFERM/Pro can have on the expression of tight junction proteins? How does this contribute to improved gut health?
[Karoline Reckmann] The maintenance of the intestinal barrier against pathogenic invasion is an important milestone in promoting gut health. This barrier is preserved by the formation of protein complexes between epithelial cells. Oxidative stress and toxic substances are known to inhibit the expression of tight junction proteins and, thus, increase the permeability of the intestinal epithelium, resulting in the occurrence of a leaky gut.
In a study, using a porcine IPEC-J2 cell culture from jejunal epithelial cells of weanling piglets, we observed the effects of our SSF product on the expression of the tight junction proteins occludin, claudin-12 and zonula occludens-1. The addition of the mycotoxin deoxynivalenol (DON) to the cell culture had a detrimental effect on the expression of these tight junction proteins, leading to an 8 to 19% decrease compared to the untreated control. Through the simultaneous addition of the SSF product, the negative effect of DON could be more than compensated for, resulting in an upregulation of gene expression of up to 5% compared to the control. The same positive effect was also observed in the absence of DON.
[Feedinfo] What about MAXFERM/Pro’s ability to provide easily fermentable carbohydrates and how this can benefit the microbiome? Please tell us a bit more.
[Karoline Reckmann] The main advantage of SSF products over single enzymes is their targeted spectrum of different enzyme activities. MAXFERM/Pro can very efficiently break up complex sugar-protein chains, thereby releasing protein from the plant cells and cell walls as well as easily fermentable carbohydrates. The breakdown of these carbohydrates promotes desirable bacterial populations that have a positive effect on animal health. At the same time, the proportion of undesirable populations decreased because less undigested crude protein reaches the hindgut.
Investigating the microbial composition of the faeces of broiler chickens using 16s rRNA analysis, we discovered a threefold increase in the number of lactobacilli and as much as a tenfold increase in the number of bifidobacteria in the animals whose diets were supplemented with the SSF product. The E. coli bacteria were reduced by half.
[Feedinfo] There is also MAXFERM/Pro’s potentially inhibitory effect on pathogens in the gut. Can you tell us how you were able to prove this and what this potentially means for pig producers?
[Karoline Reckmann] A pathogen adherence test was carried out in an ex vivo mucus model using authentic intestinal mucus from the small intestine of pigs. The results showed a dose-depending effect of preventing potentially pathogenic bacteria from attaching to the intestinal epithelium. The inhibition of the adherence of E. coli was up to 77% and for S. enterica up to 60%, suggesting that MAXFERM/Pro effectively prevented these pathogens from binding their adhesins to epithelial cells, thereby preventing pathogen entry into the cells and subsequent cell damage. Instead of binding to the epithelium, an efficient co-flocculation of the pathogenic bacteria with carbohydrate moieties present in the SSF product occurs, leading to agglomerates which will finally be excreted via faeces. This protects the epithelial layer and preserves the intestinal barrier. As a result, less nutrients are needed for tissue renewal and immune response, and/or are lost through a leaky gut. More nutrients are thus available for retention and the occurrence of diarrhoea is reduced, which is especially important during the challenging post-weaning period.
[Feedinfo] You also explored MAXFERM/Pro’s impact on the healing process of intestinal lesions. What did you find and how were you able to prove this?
[Karoline Reckmann] Besides protective mucus, antimicrobial peptide secretion, and tight junction protein formation, epithelial wound healing is another important mechanism to maintain the intestinal barrier in repairing damages caused by foreign agents. Mycotoxins, such as DON, are known to affect epithelial cell migration, as the initial step in gastrointestinal wound healing.
To evaluate the effect of MAXFERM/Pro in affecting the homeostasis of intestinal epithelial cells, we performed a wound-healing assay using IPEC-J2 monolayers to mimic the porcine intestinal epithelium. Just after provoking the wound at the centre of the monolayer by gently scraping with a pipette tip, the cells were incubated with solutions containing either DON, MAXFERM/Pro, or a combination of both, in comparison to an untreated control. At recurring intervals, the extent of wound healing was determined in comparison to the size of the initial wound using video microscopy.
Results of the wound healing assay indicated that the treatment with DON clearly inhibited the wound healing processes. In comparison, treating the cell monolayer with a suspension of our SSF product was able to partially prevent or even completely salvage the detrimental effects caused by DON. This means using MAXFERM/Pro could be an effective strategy to positively affect gastrointestinal wound healing capacity and potentially prevent enteritis. However, this requires further research.
[Feedinfo] We also know that you conducted tests to evaluate the potential symbiotic effect of SSF products with other feed additives. What additives were considered in these tests and what were the main findings?
[Karoline Reckmann] We are still in the process of digging deeper into potential symbiotic effects of our SSF products with other feed additives from our portfolio. First tests have been conducted to evaluate potential synergistic effects of our SSF product with our spore-forming probiotic Bacillus subtilis. With the very first results it could be assumed that a combination of B. subtilis with our SSF products could improve the digestibility of cell wall components even further.
B.subtilis releases supporting proteins, such as expansins, which do not break down plant cell walls but lead to a loosening of the cell wall structure. This pre-damage enables the enzymes present in the SSF product to target and cleave the cell wall carbohydrates even more effectively. Initial tests showed an improved degradation of cell wall components present in the NDF fraction of the diet by up to 10 percentage points in vitro.
[Feedinfo] Am I correct in saying that most of the finding discussed today pertain to swine? How reliably can they be applied to poultry? Is this something that you are currently investigating? If so, what initial findings can you share with us?
[Karoline Reckmann] You are right that. So far we have mainly focused on the effects of MAXFERM/Pro in swine. However, as mentioned earlier, we have already completed some trials with broiler chickens. In general, both species are monogastric animals but there are differences in their diet composition.
From a sustainability standpoint and the efficient utilisation of limited resources in poultry diets, we would suggest using more local raw materials in their feeding. However, their application can often be limited due to the level of antinutritional factors, especially non-starch polysaccharides which cannot be hydrolysed by birds’ endogenous enzymes, leading to a negative effect on nutrient digestibility, digesta viscosity and litter quality. Using MAXFERM/Pro we were able to show that even in stress-inducing diets using high rye inclusion levels of 15% in the starter phase and 30% in the grower phase this did not negatively affect the growth performance of the broilers. In addition, we were able to reduce the crude protein content in the treatment group by 1.5 percentage points, which corresponded to a reduction of 13% soybean meal over the entire fattening period without compromising broilers growth and feed conversion.
Published in association with Provita Supplements