Development of BioProducts for Animal and Human Health J. J. Lin, Ph.D. CEO/Chief Scientist Imagilin Technology, LLC

Who Are We: Biotechnology company, Frederick, Maryland Technology platforms of probiotics as biomedicines Innovative probiotic products

Emerging of Antibiotics Resistance Bacteria-Superbugs January 16, 2006, The Washington Post

Definition of Probiotics The Food and Agricultural Organization of the United Nations (FAO): " live microorganisms administered in adequate amounts which confer a beneficial health effect on the host.“

Market Size of Animals Farm Animals - USA, US $79.9 Billions Meat production Pet Foods -USA, 15.7 Billions Pet Nutrition Supplements -US $1.00 billion, % growth rate

Novel strains selection systems: -In vitro and in vivo selection of strains with enhanced immune responses. Microencapsulation of probiotics: - preservation of probiotics under harsh environment Probiotics as biomedicine: -against infectious diseases related to digestive disorders Innovative Technology

Effects of different pH on growth of P. acidilactici and L. acidophilus in vitro Pediococcus acididlactici Lactobacillus acidophilus pH 6.2 pH 1.5 pH 2.0

E. acervulina E. maxima E. tenella Poultry Coccidiosis

No toxic effects of P. acidilactici on broilers infected by Eimeria acervulii

on broilers infected by Eimeria acervulii Effects of P. acidilactici on broilers infected by Eimeria acervulii *: 20 chickens per group; chickens were orally infected with 5,000 sporulated oocytes and fed with or without P. acidilactici (P.a.). Effects of Pediococcus acidilactici on broiler chickens* infected with Eimeria acervulii. USDA, ARS, Beltsville, MD (X 10 8 Oocysts) (gm)

on broilers inefected by Eimeria Spp. Effects of serological responses of P. acidilactici on broilers inefected by Eimeria Spp.

Product Lines Nutraceutical Products: - MITOMAX R : Premium Probiotics for Dogs, Cats, Small animals - MITOBIRD TM : Premium Probiotics for Birds - MITOFISH TM : Premium Probiotics for Fish - MITOBOOST TM : Natural Antioxidant and Immune Enhancers for Pets Farm Animal Feeds Additives: -MITOGROW TM : Premium probiotics for Poultry -MITOSWINE TM : Premium probiotics for Pigs

Summary Two scientific publications in peer review journals Lead to develop the probiotic product lines to pet and farm animal business Develop international business partnerships in Japan, China, Hong Kong, Taiwan, and Brazil

37C P. acidilactici 65C* Effects of different temperature on growth of P. acidilactici and E. faecium in vitro E. faecium * Treated bacteria at 65C for 3 hrs, plated onto MRS plate,and incubated at 37C overnight

Characteristics of P. acidilactici Natural existence in animals and humans Generally Recognized As Safe (GRAS) by FDA Enhanced immune responses Less sensitive to exposure of oxygen than Lactobacillus and Bifidobacterium. More resistant to heat than Lactobacillus and Bifidobacterium. More resistant to acidic environment

Trends Emerging Antibiotics resistant SuperBugs -Abandon Antibiotics -Alternative Medicines Demanding Nutracetutical products -Healthy food -Nutrition supplement -Therapeutical effects Requiring Green (organic) foods -Antibiotics free -Growth hormone free -Pesticide free

Animal feedLive weight (g) Feed conversion (g/g) DWG* (g) Feed consumption (g) PEI Mortality* (%) Probiotic full2.7811,59566, a7,14a No additive2.7521,61465, b13,21b With promoter2.7231,62164, b7,85a With enzyme2.7371,64162, b10,35ab Performance of chicks(Cobb) fed or not with probiotic or growth promoter during all the breeding period (1 to 42 days of age) Means followed by different letters in the column are different among themselves by the Tukey test (p<0,05), 40 birds per treatment PEI: Viability X Weight gain/ convertion rate X 10. Brazil, Sao Paulo

Effect of Levofloxacin the growth of Pediococcus acidilactici Control50 ug/ml 100 ug/ml 200 ug/ml 4 hours Mixed 300 mg P. acidilactici with 3 ml of MRS medium, added 50 ug/ml, 100 ug/ml, 200 ug/ml of levofloxacin, followed by incubating at 37 C for two nights, and plated 100 ul at 10,000 dilution onto MRS plate. The colony formation was obtained after incubated overnight at 37C

Poultry Coccidiosis