Rethink in ovo protection

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1 Discover the potential of innate immunity.

2 The battle against E. coli begins at incubation Innate vs. adaptive immunity Egg transmission of Escherichia coli (E. coli) is common and can lead to omphalitis, colibacillosis, colisepticemia or even death in chickens. Mortality may range from 5-20%, with more severe infections occurring in young chicks. The risk for early transmission of E. coli presents unique challenges to poultry operations that current vaccines and antibiotics can t solve alone. The diversity in serotypes of E. coli bacteria complicates the development of vaccines, and early use of antibiotics can increase the potential for antibiotic resistance. In an effort to meet the needs of the poultry industry, Bayer set out to explore other viable options in the fight against E. coli. The result: Victrio DNA Immunostimulant, a non-antibiotic innovation from Bayer. Victrio is the first and only DNA immunostimulant approved for in ovo administration in 18-day-old embryonated eggs (or during transfer stage at hatchery) as an aid in the reduction of mortality associated with E. coli in embryonated eggs and newborn chicks. Victrio jumpstarts the innate immune system in poultry, which has been shown to provide a rapid, potent and broad protective response to infectious agents. Victrio works fast, within 24 hours, and is proven to significantly reduce mortality under E. coli challenge. 1 The immune response is commonly thought of as one process, but there are actually two types of immunity: innate and adaptive. The innate immune system is known as the first line of defense, because it develops earlier (at birth) and has a faster onset of action than the adaptive immune system in fact, adaptive immunity is dependent upon an innate immune system response. Immune response Linked Innate immunity Broad s spread Directs adaptive response Adaptive immunity Specific Clears infection Elicits a memory response hours-days Onset of action* *Adapted from Ian Tizard: Veterinary Immunology days-weeks While vaccines work with the adaptive immune response to target specific pathogens, immunostimulants trigger the innate immune system in poultry for a rapid and broad response to infection. Innate immunity is another approach in infectious disease management. Read on to discover how Victrio offers new possibilities for non-antibiotic, in ovo protection.

3 The next generation of immunostimulants Victrio DNA Immunostimulant is taking immunostimulatory technology to another level with its revolutionary DNA-liposome complex. Plasmid DNA Victrio is made up of a special type of plasmid DNA surrounded by a lipid carrier, or liposome. This DNA has CpG-motifs that mimic infection and jumpstart the innate immune system in birds. The liposome protects this DNA as it moves through the bird. The liposome is also believed to enhance the uptake of DNA by immune cells and to mediate DNA delivery to the cytosol. Liposome How Victrio works Once administered, Victrio is engulfed by immune cells, where it begins the process of jumpstarting the innate immune system. Immune cell Within the cell, the The plasmid DNA PAMP The activated immune cell The bird s Victrio liposome breaks (mixture of CpG motifs and releases cytokines, which immune system down, exposing the DNA other immuno-stimulatory help activate other immune is now ready to to endosomal toll-like DNA), is recognized cells and tell them to be fight infection. receptors (TLRs), and through endosomal TLR21 prepared for the encounter delivering the plasmid to as well as cytoplasmic DNA of pathogens. the cytosol. The Victrio recognition pathways. This DNA has a pathogen- results in activation of the associated molecular immune cells. pattern, or PAMP, that the animal s immune cell recognizes as foreign.

4 E. coli challenge studies Pre-challenge (prophylactic) administration The objective of this study was to determine the efficacy of in ovo administration of Victrio DNA Immunostimulant one day prior to E. coli challenge, as measured by reduction in mortality seven days post-hatch. Pre-challenge administration The objective of this study was to determine the efficacy of pre-challenge in ovo administration of Victrio against E. coli challenge, as measured by reduction in mortality seven days post-hatch. 7-day post-hatch mortality 2 7-day post-hatch mortality 3 7-day post-hatch mortality 20% 18% 16% 1 12% 10% 8% 6% 2% 0% 0.7% Unchallenged 17.7% Challenged 7.8% 1X Victrio 8.2% 1.5X Victrio Victrio treatments were administered in 18-day-old embryonated eggs. E. coli challenge occurred one day later, in 19-day-old embryonated eggs. 7-day post-hatch mortality 20% 18% 16% 1 12% 10% 8% 6% 2% 0% 1.6% Unchallenged 17.8% Challenged 1.3% Administered Victrio in ovo pre-challenge In ovo treatments were administered in 18-day-old embryonated eggs, at which time treatment groups were also administered Merial Inc. MAREK S DISEASE VACCINE (HVT + SB1). E. coli challenge occurred one day later, in 19-day-old embryonated eggs. Results: Victrio treated groups had significantly (P P ) lower mortality than the challenged group. Results: Groups treated with Victrio pre-challenge were significantly protected (P P ) from E. coli.

5 Clinical benefit studies Victrio vs. Garasol (gentamicin sulfate) Injection The objective of this study was to determine the efficacy of in ovo administration of Victrio DNA Immunostimulant vs. Garasol one day prior to E. coli challenge, as measured by reduction in mortality seven days post-hatch. Performance of progeny from old hen flocks The objective of this study was to evaluate the 39-day live performance of progeny from old breeder hen eggs. Progeny from Ross 308 broiler hens (56 weeks of age) and Ross 308 Parent Stock (PS, 54 weeks of age) were administered Victrio in ovo or kept as groups. 7-day post-hatch mortality 4 Cumulative post-hatch mortality 5 20% 12% 7-day post-hatch mortality 18% 16% 1 12% 10% 8% 6% 2% 0% 5. Unchallenged 17.2% Challenged 8.0% Administered Garasol in ovo 8.20% 6.6% Administered Victrio in ovo In ovo treatments were administered in 18-day-old embryonated eggs, at which time all treatment groups were also administered Merial Inc. MAREK S DISEASE VACCINE (HVT + SB1). E. coli challenge occurred one day later, in 19-day-old embryonated eggs. Cumulative post-hatch mortality 10% 8% 6% 2% 0% 7-day mortality 12-day mortality 39-day mortality Ross % 8.5% 10.1% Ross 308 Victrio 4.75% PS 7.85% % 8.20% 308 PS Victrio 3.12% 4.29% 6.45% Results: Victrio treated groups had significantly lower mortality compared to the E. coli challenge group. Victrio treated groups were almost significantly (P P < 0.059) different than the Garasol treated group. Results: Victrio groups had lower overall mortality, which was maintained throughout the grow period, compared to groups.

6 Victrio DNA Immunostimulant features and benefits The first and only DNA immunostimulant licensed for in ovo administration in 18-day-old embryonated eggs as an aid in the reduction of mortality associated with Escherichia coli in embryonated eggs and newborn chicks. Stimulates the innate immune system in poultry via a unique DNA liposome complex. Works fast, within 24 hours, and is proven to significantly reduce mortality under E. coli challenge. 1 Can be used in antibiotic-free hatcheries, because it does not contain antibiotics or preservatives. May be combined with Merial Inc. MAREK S DISEASE VACCINE (HVT + SB1) for in ovo administration. Victrio requires a multi-step mixing process that must be completed in the appropriate order. Refer to the complete application instructions on the label or product insert. Available in 4,000 and 16,000 dose package sizes. 1 Data on file with USDA. 2-5 Data on file. Bayer, Shawnee Mission, KS. Visit to learn more

7 For In Ovo Administration to 18-Day-Old Embryonated Eggs FOR VETERINA READ IN FULL RY USE ONLY DESCRIPTION VICTRIO is a bacterial-produced plasmid DNA with a liposome carrier to stimulate the innate immune system in poultry. The stimulation of the innate immune OR the reconstituted cake may be combined with Marek s Disease Vaccine, as described below. - INDICATION VICTRIO is indicated for use in 18-day-old embryonated eggs as an aid in the reduction of mortality associated with Escherichia coli in embryonated eggs and newborn n chicks. IMPORTANT STORAGE CONDITIONS Store Ref rigerated 2 C to 8 C (35 F to 46 F) DO NOT FREEZE. Diluent B can be stored at room temperature. SAFETY PRECAUTIONS For in ovo administration, this product may be combined with Merial Inc. MAREK S DISEASE VA CCINE (HVT + SB1). PREPAPA RATION OF THE PRODUCT This product requires a two-step mixing process that must be completed in the appropriate order. Step 1: Step 2: Step 3: Step 4: OR Step 4: Draw entire contents of Diluent A vial (silver cap) into a syringe. Reconstitute the lyophilized cake (gold cap) with entire contents of Diluent A vial (silver cap). Swirl gently to complete resuspension of the cake. Transfer entire contents of gold cap vial into Diluent B bag. Mix the contents of the diluent bag by swirling and inverting. Transfer entire contents ts of gold cap vial into Marek s Disease Vaccine cine diluent. Volume of diluent should correspond to 200 ml diluent per each 4,000 doses of DNA Immunostimulant. METHOD OF ADMINISTRATIONAT PRECAUTION Do not administer within 21 days of slaughter. OTHER INFORMATION Contains no antibiotics and no preservatives. HOW SUPPLIED Vials of 4,000 and 16,000 doses. MANUFA CTURED BY: Diamond Animal Health, Inc., Des Moines, IA U.S. Veterinary License No. 213 Made in U.S.A. DISTRIBUTED BY: Bayer HealthCare LLC, Animal Health Division, P. PO. Box 390 Shawnee Mission, KS U.S.A This product is based on technology developed by Juvaris BioT herapeutics and is patent protected. Animal health applications are being exclusively developed by Bayer HealthCare Animal Health and are protected by Bayer patent applications Bayer HealthCare LLC, Animal Health Division, Shawnee Mission, Kansas U.S.A. Bayer (reg d), the Bayer Cross (reg d) and VICTRIO are trademarks of Bayer , R , R B

8 2016 Bayer, Shawnee Mission, Kansas Bayer (reg d), the Bayer Cross (reg d), Victrio and are trademarks of Bayer. Garasol is a registered trademark of Intervet. VCT151070