Prepared for: Prepared by: Sustainable Nutrition Contact: Website

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1 Prepared for: Prepared by: Sustainable Nutrition Contact: Website

2 Content Introduction Sustainable Nutrition Glycerides of fatty acids Gut IMMUNITY and Mode of Action of MG ZnO reduction A pplications Introduction SN // Glycerides of fatty acids // Mode of action // Applications // Further developments

3 Introduction Sustainable Nutrition State of the art manufacturing plant Basic producer of mono-, di- and triglycerides of fatty acids Flexible and reliable partner MT output per year Fully equipped in-house laboratory Continuous focus on further product development All possibilities to esterify other fatty acids into mono-, di- and/or triglycerides

4 Overview of Inflammation and Immunity Monoglycerides? Chemical Advantages in their use Inflammation of the gut? Cause Results The gut Immunity Immunometabolism AGP : Antibiotic or Anti Inflammatory Activity AGP VERSUS MONOGLYCERIDES Conclusion

5 Monoglycerides Propionic Acid (C 3 H 6 O 2 ) Butyric Acid (C 4 H 8 O 2 ) Short chain FA Glycerol Caprylic Acid (C 8 H 16 O 2 ) α Capric Acid (C 10 H 20 O 2 ) Medium Chain FA Lauric Acid (C 12 H 24 O 2 )

6 Chemical This molecule is compatible with both HYDROPHILIC AND LIPOPHILIC membranes or cell walls and can penetrate into different types of bacteria Covalent bond LIPOPHILIC α Glycerol HYDROPHILIC H Covalent bond: attraction force between 2 atoms that share a couple of electrons The chemical bond between the fatty acid and glycerol is not an ionic bond(ph dependent), but a covalent bond (ph independent)

7 Advantages of Monoglycerides use Non-volatile Non smelling Non corrosive Non-ADR: ease of handling, transport and storage Heat stable during feed processing of either mash or pellets Neutral taste and odor FFA < 2% Water < 2% Liquid & dry Highest purity Very stable: up-to 230 C and in ph range from 1 to 7

8 What can cause an inflammation in the gut? Toxin Peroxides Mycotoxins Endotoxins Bacteria Clostridium Salmonella Lawsonia Endotoxins Coli Virus Rotavirus Coronavirus Feed Particel Size Saturated Fat Omega6 Starches Parasites Ascaris Coccidea Strongiloids Immune Cells Macrophages Neutrophils T Lymphocytes CYTOKINES Inflammatory Reaction Mucosa Destruction Appetite Reduction Nutrient Demanda Muscle Loss Desquamation Diarrhea

9 The Inflammation in the gut

10 And this is the gut inflammation result!

11 The Gut Immunity 70% 30%

12 The Immunometabolism

13 The Immunometabolism Effect of immunecell on organs that regulate the whole body metabolism Role of methabolic pathway within immunecell and how this regulate immune response outcome Decreasing immunologic stress in the intestinal mucosa would reduce the catabolic cost to the host increasing the energy available for muscle development and improving growth performance BASAL IMMUNE RESPONSE : NOT AFFECT TOO MUCH NUTRIENT BODY DEMAND

14 AGP : Antibiotic or Anti-Inflammatory Activity Antibiotics as growth promoters (AGP) in the feed of different animal species have been used for many years to improve feed efficiency and growth but the intensive use have contributed to the emergence of antimicrobial resistance in zoonotic pathogens, resulting in a ban by the European Union with suggestive restrictions by other countries. It has been demonstrated that antibiotics are able to inhibit one or more functions of inflammatory cells and pro inflammatory cytokine production. AGP prevent immunologic stress that is associated with metabolic changes, allowing more energy to be available for muscle development thus, improving growth. This seems especially likely when one considers the fact that AGP are administered at levels below the minimum inhibitory concentration for bacteria, thus suggesting their mode of action is not one that directly affects bacteria.

15 AGP : Antibiotic or Anti-Inflammatory Activity According to Costa et al. (2011), AGP could reduce immunologic stress in the host, since the intestinal mucosa is in continuous interaction with the microflora, and consequently in a state of physiologic inflammation. GUT Inhibiting pro inflammatory cytokine production would cause a decrease in acute phase response, which is linked to a high uptake of catabolic energy (Niewold, 2007). It has been demonstrated that antibiotics are able to inhibit one or more functions of inflammatory cells such as chemotaxis, reactive oxygen species production, and pro inflammatory cytokine production.

16 AGP : Antibiotic or Anti-Inflammatory Activity Niewold (2007) stated that AGP are able to accumulate in phagocytic inflammatory cells improving the intracellular killing of bacteria and partially inhibiting the innate immune response. As a result, treated animals would have a decrease in proinflammatory cytokines (Buret, 2010), which in turn would save catabolic energy facilitating its growth. As elucidated by Buret (2010),another contribution of the AGP in reducing inflammation could be explained by an induction of neutrophil apoptosis by some types of antibiotics such as macrolides, which has also been described to reduce pro inflammatory cytokines of both innate and acquired immune systems (Steel et al., 2012). T reg COMMENSAL ANIMAL HEALTH Th cell

17 AGP VERSUS MONOGLYCERIDES PAST Antibiotic Resistance FUTURE Natural Chemical Way with NO ANTIBIOTIC RESISTANCE

18 Where Monoglyceride are active in the gut?

19 Antimicrobial Acitvity Effects of MONOGLYCERIDE on the structure of B Strepto-coccus (GBS) Electron microscopy of sections of bacterial cells Control sample Bacteria cell after exposure to Monoglyceride MG Picture C shows cell from the control samples with intact plasma membrane and intact cytoplasmic granules (G). Picture D shows a cell from samples treated with 10 mm Monoglyceride (MG) for 30 min, demonstrating disappearance of plasma membranes and cytoplasmic granules. Some changes can be seen in the cell wall as well.

20 Antimicrobial Acitvity E. coli Staphylo coccus xylosus Zygosac- charomyces bailii Action of MONOGLYCERIDE on the cell surface MG Hyldgaard et al., 2012 Exposure of bacteria to monocaprylin produce pores in the membranes that lead to instability of the cell and death.

21 Control of proinflammatory citokine with MG The effect of MONOGLYCERIDES on levels of the cytokines on experimental animal in Entamoeba histolytica and Giardia lamblia infection

22 Biological Activity of IL-6

23 Control IL-6 increasing muscle production A simplified mechanism of myostatin-induced muscle atrophy MG Inflammation increases levels of circulating or local proinflammatory cytokines, such as IL 6. These cytokines stimulate phosphorylation of Stat3, which in turn stimulates SOCS 3 expression. The result is suppression of insulin/igf 1 signalling, leading to muscle atrophy. IL 6 induced activation of pstat3 also stimulates C/EBPδ, which activates myostatin and, therefore, results in muscle loss

24 EU Regulation for zinc oxid application Regardless of whether the use of zinc oxide really causes environmental problems and resistance to antibiotics, the truth is that the sector has to plan its withdrawal from feeds, because this opinion was ratified by the CVMP in its meeting of March A new era is coming and we need to devevelope and change our mentality in this new scenario. Apply more and better vaccination protocols (sows and piglets). Better farm biosecurity. Improve the cleaning, disinfection and drying protocols, and follow a strictly All In All Out policy in the weaner and finishing rooms. Restrict the mixing of animals from different origins. Improve the planning and sizing of the nursery and finishing units, so as to avoid mixing the animals. Increase the wean to finish systems, to avoid problems related to entrance to the finishing units. Improve management; give more attention to temperature and ventilation requirements. Lower protein levels as much as possible, using the available synthetic amino acids. Formulate feeds based on net energy, possibly with levels slightly lower than the current ones.

25 Weaning Transition in the gut COMMENSAL ANIMAL HEALTH T reg Th cell

26 Innate defense in the gastrointestinal tract

27 ZnO : Inflammatory Activity? In recent studies, mrna levels of TNF α, IL 6 and IFN γ decrease with increasing concentrantion of dietary Zinc Oxide in the jejunum of pigs (Hu et al., 2012; Hu et al., 2013b). TGF β a L 10 are anti inflammatory cytokines and can thereby help to maintain intestinal barrier function. The mrna levels of TGF β a L 10 were increased with high zinc oxide included in zeolite after weaning (Hu et al., 2013b). Zinc regulates the signaling pathways of TLRs, which may induce or inhibit the activation of NF κb (Foster and Samman, 2012)

28 Conclusion In the past three decades in our animal farms we move from Growth Promoter with anti inflammatory and low antibacterial activities to Antibiotics just with antibacterial activity. Unfortunately through these 2 steps we created one of the biggest antibiotics resistance for several bacteria. In our portfolio, we develop product base on several researches, conducted worldwide, that has reveal the anti inflammatory activity at different level in the gut immune system that is able to reduce the fat deposition and increase the muscle production. The base of our research is to modulate the inflammation, that is inversely related to growth, keep the gut barrier function active and control the growth of the pathogen in the Microbiome and we can obtained these results with the use of Monoglycerides that are able to balance both gastrointestinal microflora and immune stimulation, particularly by maintaining the equilibrium of proinflammatory and antiinflammatory cytokines.

29 Conclusion The multi tools activity of SN-MONOMIX NO inflammation Feed Efficiency Less antibiotics MG

30 Available products at Panadditív SN MONOMIX DRY SN MONOMIX LIQUID SN MONOLAURIN DRY