N13A-T013 TITLE: Probiotics for Maintaining Dolphin (Tursiops truncatus) Health and the Readiness of the U.S. Navy’s Marine Mammal Systems

TECHNOLOGY AREAS: Biomedical

ACQUISITION PROGRAM: Explosive Ordnance Disposal Underwater Programs (SEA00 EOD/CREW-2)

OBJECTIVE: To develop probiotic pharmaceuticals to treat and prevent gastrointestinal disease in dolphins and improve their health through the utilization of indigenous commensal microbes of these marine mammals.

DESCRIPTION: The U.S. Navy uses Atlantic bottlenose dolphins (Tursiops truncatus) in the Fleet’s operational Marine Mammal Systems to protect harbours and Navy assets and to detect and/or mark underwater mines. To contribute to the maintenance of the fitness of these marine mammals for duty and the readiness of the U.S. Navy Marine Mammal Systems, the U.S. Navy is interested in developing probiotics for these animals. Both general protective effects and efficacy on gastrointestinal diseases are desired in this probotic to help address the potential negative impacts of an unbalanced gastrointestinal microbiome, inflammatory disease, or potential pathogens (e.g., Clostridium difficile and Helicobacter spp.) on dolphin health (1).

Probiotics are defined by the Food and Agriculture Organization of the United Nations/World Health Organization as live microorganisms which can confer a health benefit for the host when administered in adequate quantities (2). Additionally, probiotics have been subcategorized into probiotic drugs (intended to cure, treat, or prevent disease), probiotic foods (which include foods, food ingredients, dietary supplements), direct –fed microbials (probiotics for animal use), and designer probiotics (genetically modified probiotics) (3). Both bacteria and yeast have been used as probiotics in humans and animals (4, 5). Once ingested, probiotic microorganisms can modulate the balance and activities of the intestinal microflora, inhibit the growth of harmful bacteria, promote good digestion, boost immune function, and increase resistance to infection (3, 4). Medical conditions for which evidence has been obtained that probiotics may help in the prevention and/or treatment include various diarrheal illnesses, especially antibiotic-induced diarrhea; urinary tract infections; irritable bowel syndrome; atopic eczema; respiratory infections; inflammatory bowel disease; and gastroenteritis relapses caused by Clostridium difficile bacteria after antibiotic therapy (6). Regulatory requirements differ for probiotics depending on their intended use (3, 7).

PHASE I: Provide an initial development effort that isolates and identifies potential probiotic candidates for Tusiops truncatus derived from indigenous commensal microbes of these marine animals. Tursiops truncatus indigenous bacterial species with in vitro immunomodulatory and dolphin gastric and enteric pathogen (e.g. Helicobacter spp, Campylobacter spp, enterohemorrhagic Escherichia coli, enterotoxigenic Escherichia coli, and Salmonella spp) growth inhibitory activities are desired. Microencapsulate the candidate probiotic microorganisms for Tursiops truncatus to enable selective delivery to, and release in, the dolphin intestine; enhance viability of the probiotics during delivery into the body; and enhance the viability of the organisms during storage.

PHASE II: Demonstrate the safety of the commensal-derived probiotic, and assess the efficacy of the probiotic for the treatment of gastrointestinal diseases, in Tursiops truncatus. Demonstrate that the microencapsulated probiotic mircroorganisms colonize the dolphin intestine better than corresponding uncoated probiotic microorganisms. Assess shelf life of product and general feasibility for long-term production of, and access to, a dolphin probiotic product.

PHASE III: Continue the program forward and address any Food and Drug Administration regulatory matters to reach full probiotic development. Commercialize the product.

PRIVATE SECTOR COMMERCIAL POTENTIAL/DUAL-USE APPLICATIONS: A probiotic for Tursiops truncatus would have application in the care of captive and managed populations of these animals world-wide (e.g. aquariums, marine parks).

REFERENCES:
1. S. Venn-Watson, et al, "Primary bacterial pathogens in bottlenose dolphins Tursiops truncates: needles in haystacks of commensal and environmental microbes" Dis. Aquat Organ., Vol. 79, No. 2, pp. 87-93 (2008).

2. Joint Food and Agriculture Organization of the United Nations/World Health Organization Working Group report on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria, Cordoba, Argentina, October 1-4, 2001. Accessed at ftp://ftp.fao.org/docrep/fao/009/a0512e/a0512e00.pdf on August 14, 2012.

3. M. E. Sanders, "How do we Know When Something Called "Probiotic: is Really a Probiotic? A Guideline for Consumes and Health Care Professional" Functional Food Reviews, Vol. 1, No. 1, pp. 3-12 (2009).

4. S. Parvez, et al, "Probiotics and their Fermented Food Products are Beneficial for Health", J. Appl. Microbio., Vol. 100, pp. 1171-1185 (2006).

5. F. Chaucheyras-Durand and H. Durand, "Probiotics in Animal Nutrition and Health" Beneficial Microbes, Vol. 1, No. 1, pp. 3-9 (2010).

6. B. R. Goldin and S. L. Gorbach, "Clinical Indications for Probiotics: An Overview", Clin. Infect. Dis., Vol. 46 Suppl2:S96-100; discussion S144-51 (2008).

7. V. Venugopalan, et al, "Regulatory oversight and safety of probiotic use. Emerg. Infect. Dis. (2010). Accessed at http://wwwnc.cdc.gov/edi/article/16/11/10-0574.htm.

8. M. Diaz, et al, Isolation, Isolation, Culture and Characterization of Lactobacillus Salivarius as Probiotic Candidates for the Bottlenose Dolphin (Tursiops truncatus), Abstract, 2011, 1 page (uploaded in SITIS 2/1/13).