Development of genetic and biotechnological tools and services for the salmonid industry

Development of genetic and biotechnological tools and services for the salmonid industry Developments and projections María Eugenia Cabrejos M., PhD

RESEARCH, DEVELOPMENT AND INNOVATION STRATEGY Technology transfer biotechnology STRATEGIC OBJECTIVES 1. Promoting the creation and development of knowledge, with view on obtaining high level, internationally competitive, results 2. Increasing the competitiveness of the economy, by promoting the diffusion and transfer of knowledge and the innovation processes with strong economic impact 3. Increasing the quality of life, through the development of solutions with high benefits for society.

Who we are Company Government University it Basically, Chilean Company 5 year project (started on 2007) US$ 10 Millions (50% Innova CORFO y 50% E. Aquachile) Property of the company: 83% AquaChile, 15% Universidad of Chile and 2% Biomar 3

Our compromises To build a laboratory and an experimental hatchery to develop biotechnological tools to study, by an integrated point of view, nutrition health, genetic and environmental conditions of fishes To develop molecular biological tools which we will introduce into new approaches to manage genetic programs in aquaculture species To invert into qualified people who will work doing research and development activities iii with ihthe aim to produce technological ltransfer of products or services and to add knowledge into this field To develop techniques to select individuals which are more resistant to diseases To develop a software to store databases and to manage and analyze the dt data from genetic programs To search for others genetic markers of productive interest to select individuals

Our challenge: To work in an integrated Way FENOTIPO (F) (PERFORMANCE) Genetic Integrated Genetic Nutrition and Health Environment 5

Location of the Research Station in Lenca Three water concession: 1200 l/s River 500 l/s River 30 l/s Fresh water 30 l/s Salt water T up to 28 C 2,5 Ha beside to the Lenca River 6

7 Design of the Research Station in Lenca

Laboratory Unit 1) Diagnostic area and 2) R&D&i Diagnostic Analysis Reproduction Disease Challenge Genetic Markers Nutrition and Health Platforms to assist Breeding Programs Vaccine Evaluation Salmon Genome and Bioinformatics Salmonids and Tilapia

Experience of work: Standardization of analysis ISAv Comparison between OIE v/s Snow (2006) conditions 9

Validation of real time RT PCR assay Comparison between OIE v/s Snow (2006) conditions 10

Development of a real time RT PCR assay using Taqman probes to discriminate HPR 0 from others HPR C+ Muestras positivas C 500 pb 00 pb 300 pb 300 app 200 pb 100 pb 11

Results from some cases presented in the summer of 2010 Taqman HPR0 dsdna sequencing Taqman F+/F A590_9 Negativo HPR 2 F A590_ Negativo HPR 2 F A596_9 Positivo HPR 0 F A597_7 Positivo HPR 0 F A620_ Positivo HPR 0 F A626_3 Positivo HPR 0 F A626_ Positivo HPR 0 F A63_ Negativo SR SR A63_8 Negativo SR SR A636_1 Negativo HPR 7b F+ A636_3 Negativo HPR 7b F+ A636_9 Negativo HPR 7b F+ A6_3 Positivo SR F A666_1 Negativo HPR 7b F+ SR= Sin resultado 12

Laboratory Accreditations: Sernapesca: Broodstock Screening Program (PSGR) Sernapesca: Contingency Program for ISA Virus (PSEVA ISAV) Corfo: I+D Laboratory Accreditation Sernapesca: Active Vigilance il Program (PSEVA waiting i for the permission) i ISO 025: In process. 13

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Disease Challenge ISAv ISAv Challenge with intraperitoneal injection. After day 22 there was between 20 to 30% survivors. Virus Amoun nt illions M 12 10 8 6 Survivors (20 a 30%) 2 0 12 13 1 15 18 20 21 22 31 Days Pos Treatment 15

Atlantic salmon family challenge Tests Year Class Year Pathogen N Families Data 2007 2008 P.salmonis 83 1150 2008 2009 P.salmonis 0 800 2008 2009 C.rogercresseyi 75 1500 SRS Caligus (grs) Mortlidad (%) - Peso ( 200,0 180,0 160,0 10,0 120,0 100,0 80,0 60,0 0,0 20,0 0,0 0,3000 0,2000 0,1000 0,0000-0,1000-0,2000-0,3000 0 Number of parasites 20 0 YC 2007YC 2008 Family mean Abundance in adult stage 1 7 10 13 16 19 22 25 28 31 3 37 0 3 6 9 52 55 58 61 6 67 70 73 76 79 82 100% 1 710131619222528313370369525558616677073 Familias Families BV_Probit (0-1) SRS by Family BV Chalimus III IV abundance in fish PBV CH3 3 abundance 20 15 10 5 6181 6121 6129 6086 6061 610 6083 6180 6036 621 60 6231 6213 615 6087 6161 6060 6098 60 6035 6192 0 5 10 15 80% 60% 0% 20% 1,5000 1,0000 0,5000 0,0000 0% -0,5000-1,0000 1 5 9 13 21 25 29 33 37 1 5 9 53 57 61 65 69 73 Families 7183 YC 2008 Mortalidad SRS Prueba Desafío 3 - Febrero 2010 7181 722 BV_Probit (0-1) SRS by Family 79 7107 7112 7061 7060 7058 716 7211 75 7075 707 7101 7210 7135 7129 7120 7123 16

Overview of the form by which the laboratory works today Diagnostics Health Genetics Products & Services R & D & i FONDEF Project Ch$ 60 MM/ 3 y Collaborations (a) Companies (b) TT Expert (c)eu Laboratories: NTI, Aarhus, Denmark AFBI, Belfast, Irland (d) Authorities CORFO INNOVA Project Ch$ 577 MM/ 3 y Collaborations (a) Companies (b) Universities (c) Nutrition Expert Ronald Hardy, Idaho University, USA Master Thesis bioinformatic field Incorporation PhD CORFO INNOVA Project into preparation Salmon Genome Sequence Project Collaborations: (a) SoniaConsuegra, Aberystwyth y University, Wales, UK (SSR multiplex, MH)

FONDEF de CONICYT entrega más de 13 mil millones de pesos para la Investigación aplicada en Chile 18

Gene Expression in different Tissues Evaluation of the immune system and oxidative stress condition OBJECTIVES 1. Evaluation of normalparameters Tissue Selected Genes 1. Behaviour during a challenge 2. Normalization of the parameters for the administration of various additives in the diet 19

Complete Genome Sequence of Atlantic salmon: 2011 2012 20

Increase of the density of the Markers for Selection ALOZIMAS RFLP Microsatélites (*) SNPs (*) 00 2.000 5.000 Marcadores Marcadores Marcadores 10 cm 0,5 cm 0,2 cm Saturación del mapa genético humano, Adaptado de S. Primrose & R. Twyman.200. Genomics, Aplications in Human Biology. Cristian Araneda, U de Chile Fac. Cs. Agronómicas 21

PhD Thesis: omic s era Evaluación de resistencia genética específica y conjunta a patógenos, Caligus rogercresseyiy Piscirickettsiasalmonisen Salmón del Atlántico (Salmo salar) Jean Paul L horente Lhorente Identificación de Biomarcadores moleculares y análisis del efecto del Selenio sobre la resistenciaa Piscirickettsiasalmonis en Salmón del Atlántico (Salmo salar) Rodrigo Pulgar Estudio de expresión génica y validación de polimorfismos SNPs en genes candidatos asociados al crecimiento en el pez cebra (Danio rerio) en respuesta a una dieta con proteína de origen vegetal Pilar Ulloa 22

Selections by Integrated Genetics Genomics Transcriptomics Proteomics Production cycle Performance Objective Questions Work Plan Projects Business: Costs, Feasibility, Economic evaluation 80 20 23

Impact of improvement of some productive traits of importance Características Efecto en costo 1% de mayor crecimiento (sgr) 6 /kg 1% menos mortalidad 2 /kg 5% mejor retención de pigmento 1 /kg 2

The people Projects & Adm R&D&i Lab Vet Technical Assistant 25

Muchas Gracias Thank you 26