Health Through Nutrition

Health Through Nutrition

Aquaculture:The nutrient conflict Limited Aqua nutrient supply from raw materials Limited raw materials from marine origin High prices for protein and lipid sources Variable quality of raw materials Increased nutrient requirements Increasing total aqua production and lower margins Genetically improved faster growing strains Better farming/higher yield expectations To reduce costs, nutritionists are increasingly using alternatives for marine proteins by effectively supplementing essential amino acids 2

Feed cost reduction Feed cost Economical return for farm: ROI Formulation Raw material cost Operational efficiencies Finance Survival: health FCR & growth Fish/shrimp quality Role of Nutritional Additives? 3

Fish meal replacement research Number of journal articles found in ASFA search for keywords fish and meal and replacement 18 16 14 Number of articles 12 10 8 6 4 2 0 1981 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 Increasing information on strategies and alternatives for marine protein replacement but many gaps remain 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 4

Cost reduction in commercial formulations Carp, Tilapia, Catfish Little or no fish meal Diets without micronutrients Salmonids, Marine fish Significant reductions in marine ingredients The laws of unintended consequences Shrimp Lower fish meal levels Less attention to micronutrients 5

What are the unintended consequences? Basing formulations on minimum nutrient requirements is complicated by a number of key issues: Much research is conducted over a short time (eg 8 weeks) on small fish Research may be outdated, using poor protocols with limited practical relevance Functional levels of nutrients are higher than minimum requirements Information gaps for key alternative protein sources Consequences may be seen in: Production Efficiency Animal Health End product quality 6

Type two errors Conclusion: The data suggest that up to 140 g/kg of peanut meal could be included in practical diets 7

Type two errors Conclusion: The data suggest that up to 140 g/kg of peanut meal could be included in practical diets 8

Type two errors Conclusion: The data suggest that up to 140 g/kg of peanut meal could be included in practical diets 9

Diets did not affect growth or blood chemistry When challenged, fish got sicker faster higher prevalence and intensity of infection higher percentage with parasite in entire intestinal tract faster parasite establishment Nutrition predisposing factors Reduced feed intake Increased intestinal damage Osmoregulatory failure Immune status 10

Feed cost reduction and health Feed cost reduction is not only about supplying and balancing nutrients. Assuring nutrient availability and improving animal health is equally important More bioavailable micronutrients Attractants and pallatants Gut health modifiers Immunostimulants, nucleotides Enzymes 11

Minerals in fishmeal replacement Mineral Fish Meal Anchovy 65% CP Soy Bean Meal 43-44%CP Corn gluten Mn 1 17 40 8 Zn 1 82 48 26 Cu 1 7 15 12 Se 2 1.36 0.2 0.83 P soluble in H 2 O 3 0.55 0.2-0.4 0.12 1 CVB 2011, 2 NRC 2011, 3 lab analyses (2005-2012) 12

Mineral immobilization and antagonisms 3-4 times inorganic copper for equivalent performance 10.20 10.00 Chelated Inorganic Final Weight (g) 9.80 9.60 9.40 9.20 9.00 8.80 8.60 1.2% Phytic Acid in diets CuSO4 Mintrex Copper 0 50 100 150 200 250 300 350 400 13

Copper deficiency in crustaceans Poor growth Reduced tissue copper levels Exoskeleton about 18% Hepatopancreas about 25% Hemolymph about 40% Enlargement of the heart Hemocyanin deficiency? Enzyme deficiencies? lysyl oxidase cytochrome c oxidase tyrosinase superoxide dismutase Cu tissue distribution Palaemon spp. Hemolymph Hepatopancreas Exoskeleton Other 14

Shrimp digestive system The anatomy of the shrimp digestive system includes barriers to protect against pathogen infection Gastric Sieve Bell and Lightner 1988 Digestive fluid circulation Ceccaldi 1997 Peritrophic membrane in dissected midgut Wang et al. 2012 Stomach cuticular lining Bell and Lightner 1988 15

The stomach, intestine and hepatopancreas all contained diverse Vibrio spp. Presence in hepatopancreas is not necessarily indicative of disease. This differs from previous findings from diseased shrimp, where one or two species predominate. stomach intestine HP hemolymph N 28 of 28 25 of 25 26 of 26 4 of 28 Mean 1.29x10 6 2.1x10 6 4.30x10 4 Maximum 4.40x10 6 1.03x10 7 2.67x10 5 3x10 3 Minimum 3.33x10 4 1.01x10 4 1.11x10 2 2x10 2 16

Vibriosis Vibriosis continues to be a major problem as a primary or as a secondary infection Nyan Taw 2010 Bacterial colonization in posterior stomach Lightner 1996 Lightner AHPNS presentation NACA 2012 17

Gut environment modifiers Antimicrobials Prebiotics Antibiotic growth promoters Essential Oil blends Organic acid blends Oligosacharides Gut Health and Microbial Community Equilibrium Probiotics Bacillus Pediococcus 18

Gut health is critical for overall performance Enhanced beneficial bacterial populations in the gut Reduction in enteric pathogen load Improved integrity of gut epithelium Control of vibriosis Improved digestibility Direct and indirect effects on immunocompetence Improved growth (reduced fishmeal formulations) Improved survivability Return on investment 19

Phagocytosis Phenoloxidase activity Respiratory burst 20

Current research Antimicrobial Essential Oil blend Organic acid blends 21

Essential oil blend NEXT Enhance 150 is based on a unique formula combining thymol and carvacrol (active substances found naturally in Oreganum spp.) Microencapsulation to improve handling, protection of active ingredients and release in the gut Antimicrobial activity as hydrophobic molecules interact with cell membrane causing ion leakage. 1 mm 40 X 22

Essential oil blend laboratory growth and challenge trial a a a b Challenge mortality HP microbial load Kasetsart University, 2008 23

Organic acids Bacteriostatic and bacteriocidal activities Physiological status of the organism Physiochemical characteristics of the environment Undissociated acids penetrate lipid membrane of bacterial cell Dissociate in cytoplasm Disrupts homeostasis ph drop Depletion of cellular energy Cell death Interference with membrane structure and function 24

Shrimp Organic Acid Research CENIACUA Shrimp MERA Cid 0, 0.1, 0.3, 0.5 1.3 g 57d TSA, TCBS Gavage V. harveii Kasetsart Shrimp MERA Cid 0, 0.5, 1.0 PL12 60d Marine, TCBS Waterborne V. harveii Treatment ph Hepatopancreas* ph Gut* Control 6.156 ± 0.005 a 7.156 ± 0.033 a MeraCid 0.1% 6.250 ± 0.016 a 7.066 ± 0.069 b MeraCid 0.3% 5.872 ± 0.013 b 7.028 ± 0.030 b MeraCid 0.5% 5.872 ± 0.015 b 6.966 ± 0.005 b 25

Apparent Nutrient Digestibility Nutrient Experimental diets Control MeraCid 0.1% MeraCid 0.3% MeraCid 0.5% ADC DM 72.6±1.20 ab 71.2±1.89 b 74.5±0.75 a 74.8±1.6 a ADC protein 86.0±2.3 c 87.5±1.8 abc 89.1±0.8 ab 90.0±2.3 a ADC energy 84.9±0.9 a 83.2±3.2 ab 83.8±1.8 ab 81.2±2.1 bc Linear regression of inclusion level vs. protein digestibility Slope = 0.81 ± 0.14, r2=0.891 Dotted lines represent 95% confidence intervals Submitted for publication -CENIACUA, Colombia 26

Kaplan Meier Survival Estimates Mera Cid 0.5% Mera Cid 0.3% Control Mortality rates 6g L. vannamei 0% MeraCid - 37.5% 0.3% MeraCid - 22.5% 0.5% MeraCid - 11.4%. Significant differences (p<0.001) between control and 0.5% MERA Cid inclusion diets. Submitted for publication -CENIACUA, Colombia 27

Growth of L. vannamei juveniles from PL9 Weight (g) 4 3 2 1 0 * * Control 1% Mera Cid Day 40 Day 50 Day 60 Treatment Average Body Weight (g) Percentage Survival (%) Control 2.77 + 0.72 b 92.8 + 2.28 a MeraCid 1% 3.14 + 0.77 a 93.6 + 2.60 a Yankomut et al. Asian Pacific Aquaculture 09 Kasetsart University 28

Effects of MeraCid on shrimp gut flora Intestine homogenized and plated on PCA and TCBS 30000000 25000000 25000000 20000000 20000000 15000000 15000000 10000000 10000000 5000000 5000000 0 Control MERA Cid 1% 0 Control MERA Cid 1% Treatment Number of total bacteria (CFU/g) Treatment Number of total Vibrio (CFU/g) Control 1.52 + 1.21 x 10 7b Control 1.15 + 1.10 x 10 7b MeraCid 1% 3.06 + 1.95 x 10 6a MeraCid 1% 7.74 + 4.16 x 10 5a Yankomut et al. Asian Pacific Aquaculture 09 Kasetsart University 29

Survival of L. vannamei post challenge 120 100 Survival (%) 80 60 40 20 1,000 CFU/ml control Meracid 0.5% 120 100 100,000 CFU/ml 0 120 100 0 10 20 30 40 50 60 Days Survival (%) 80 60 40 control Meracid 0.5% Survival (%) 80 60 40 20 10,000 CFU/ml control Meracid 0.5% 20 0 0 10 20 30 40 50 60 Days 0 0 10 20 30 40 50 60 Days Limsuwan et al. Kasetsart University 30

Nutritional additives and Health: Farm trial- Ecuador Trial set up: Cage (1x1x1 m) trials in pond. 6 cages per treatment 20 shrimp per cage (20/m²), Initial weight: 10g; 71 days, final weight 23-24 g Treatments: Control Antibiotic treatment (Enroflox) Next Enhance 150 (30 ppm thymol+carvacrol) Meracid ( 5 kg/ton) 31

32 Cage trial Ecuador Growth 1.25 1.30 1.35 1.40 1.45 Growth (g/wk) a ab b ab 40% 45% 50% 55% 60% 65% 70% 75% 80% 85% 90% Survival (%) a b ab a 1.00 1.20 1.40 1.60 1.80 FCT a b ab a 2000.00 2500.00 3000.00 3500.00 4000.00 4500.00 Production (kg/ha) a b ab a

Focus on the shrimp! In vivo models Digestibility Challenges Acute Chronic Immunology Oxidative status Structure In vitro models Gut community Pathogens Ex vivo assays Product quality Omics Genomics Metabolomics Feed - pond interactions Functional feeds 33