aquaexcel at wageningen university metabolic research unit ... · - influent o 2 and/or co 2...

Use of control and monitoring systems for analysing nutritional and environmental effects on fish metabolism

Ep Eding (WU), Tuesday 20th May 2014, 08:30 –09:15 Sealab Auditorium, NTNU, Trondheim, Norway

Lecture (power point) in the AQUAEXCEL Short Intensive Training Course 1 #4:

Efficient Utilisation of New Monitoring and Control Systems in Fish Experiments

AquaExcel at Wageningen University

Metabolic Research Unit (WU-MRU)

Use of control and monitoring systems for analysis of

nutritional and environmental effects on

fish metabolism

Ep Eding

Aquaculture and Fisheries Group

Wageningen University

AQUAEXCEL 20 May, 2014, NTNU, Trondheim, Norway

Use of control and monitoring systems for analysis of

nutritional and environmental effects on

fish metabolism

Ep Eding

Aquaculture and Fisheries Group

Wageningen University

AQUAEXCEL 20 May, 2014, NTNU, Trondheim, Norway

Metabolic Research Unit (MRU)

� 12 metabolic chambers of 200 L each

� Linked to one recirculation system


� 12 metabolic chambers of 200 L each

� on-line measurement actual water flow;

� cumulative water flow;

� actual and cumulative airflow;

� on line O2 concentration in the air and water and CO2 in air;

� on line temperature, pH, salinity;

� TAN, Urea, NO2-N, NO3-N, dissolved protein, and PO4-P in

the rearing water (auto-analyzer measurements)


� Control over environmental factors:

� Water temperature 15-30 °C

� Salinity 0-35 ppt

� Gasses: 4 influent concentrations for O2 or CO2 can be chosen

� The unit can be connected to two identical RAS

Factors affecting feed intake Factors affecting feed intake

Fish factors

Feed quality

Water quality

Feed intake

� Size� Species� Behavior

� Dietary energy � Macro & micro nutrient availability

� Temperature� Dissolved oxygen� Carbondioxide� pH, ammonia

� Research questions in the MRU relate to how:

� Animal factors (genetics, phenotypic differences health status)

� Nutritional factors � Environmental factors (temperature, O2, CO2, .....)

affect responses of animals.



� Used in our research for:

� Nutrient and energy balance studies over a production cycle

� For measurement of within day variations (O2, CO2, TAN, …..)

� Adaptive physiology studies.

� Fish species:

� Sea bass, Turbot, Yellow tail

� Rainbow trout, Tilapia, European eel, African cafish

� Response parameters:

� Feed efficiency� Feeding behaviour � Apparent digestibility nutrient� Heat production� Behaviour (camera’s available)� Energy and Nitrogen balance� Within day variation in O2, CO2, TAN.

� Maesurements are combined with:

� Blood parameters� Anything what you can measure at slaughter


12 Metabolic Units for NUTRIENT & ENERGY BALANCE studies In: Fresh, Marine, Warm and Cold water fish species

Water flow meters

Front sideFish tank


12 Metabolic Chambers


Back side MRU

Air flow metersFeces collection

On line measurement in the air of:O2 and CO2

On line measurement in water of:TAN, Urea, NO2-N, NO3-N, PO4-P, CO2

12 Metabolic Units for NUTRIENT & ENERGY BALANCE studies In: Fresh, Marine, Warm and Cold water fish species


From Tran Duy et al., 2012

Metabolic Research Unit (MRU) Gas control

Membrane contractor

Liqui-cell ®, Membrane Contactor, Charlotte, North Carolina, USA. (http://www.liquicel.com/)


From Tran Duy et al., 2012


No Fish tank Records

Water flow In Data logging in PC 1

pH In and out Data logging in PC 1

Oxygen In and out Data logging in PC 1

Temperature In and out Data logging in PC 1

Salinity In and out Data logging in PC 1

TAN, urea In and out Data logging in PC 3 auto analyzer file

NO2-N In and out Data logging in PC 3 auto analyzer file

NO3-N In and out Data logging in PC 3 auto-analyzer file

HCO3 In and out Data logging in PC 3 auto-analyzer file

PO4-P In and out Data logging in PC 3 auto-analyzer file

Airflow Out Manual

CO2 and oxygen In and out Data logging in PC1 (n.a))

Metabolic Research Unit (MRU) Data logging

- Influent O2 and/or CO2 concentrations can be changed through a combination of degassing and controlled oxygen supply enabling studies on the response of fish to various O2/CO2 ratios in the environment. -The unit is further equipped with a mobile feeding registration system, and mobile faecal collection units. Mobile webcams (N=16) and an imaging analysis software are additionally available to record and analyse behavioural data. All data can be stored in a data acquisition system and made available in excel spreadsheets for later analysis.

Metabolic Research Unit -Sensors-Webcams - Auto analyzer

Host PC (PC1)- Data base- Back-up

www.LogMeIn.com

LogMeIn Database

LogMeIn Gateway

FirewallInternetLogMeIn

Internet

Firewall

Remote user (Guest)

Remote WU-Staf

MRU-Lab PC (PC2) (PC3)- Data base- WU staff

(Controller data acquisition)

Administrator (remote access authorization)

LogMeIn Architecture for remote access

Remote access Metabolic Research Unit (MRU)

Bioenergetic approach for growth analysisBioenergetic approach for growth analysis

Utilization of dietary energy by fishUtilization of dietary energy by fish

E(in) = E (out) + E (p)

Bioenergetic approach Bioenergetic approach

� 1 g Protein = 5.65 kcal - 23.64 kJ� 1 g Carbohydrate = 4.2 kcal - 17.15 kJ � 1 g Fat = 9.4 kcal - 39.54 kJ

� 1 kJ = 0.24 kcal� 1 kcal = 4.186 kJ

� Gross energy (GE) measured in food and body tissues with bomb calorimeter

Energy metabolism

Biological process of utilization and transformation of absorbednutrients for energy, for own body synthesis,

maintenance and growth


Gross Energy (GE)

Digestible Energy (DE)

Metabolizable Energy (ME)

Retained Energy (RE)

Fecal losses (FE)

Branchial and Urinary Energy (BUE)

Heat production (H)

GE (kJ/g) = RE + H + BUE + FE


Feed, Fish Feces (DM) = Protein + Fat + Carbohydrates + Ash

Methiod 1 . E-liberation by combustion

Method 2. E-calculation from chemical composition E (kJ/g) = 0.2364*Protein (%) + 0.3954*Fat (%) + 0.1715*Carbohydrates (%)

Method 3. Chemical Oxygen Demand (see Henken et al., 1985)

Determination:

� Gross energy feed (GE) � Retained Energy (RE)� Fecall losse (FE)

Bioenergetic approach App. digestibility

Settled Feces (stored on ice during collection

Stripped feces Feces collector

Bioenergetic approach DEBioenergetic approach DE

Apparent digestible Energy & Nitrogen determination

Methiod 1 . ADC(%) = (1-feces/feed consumed) *100 (Quantitative method)

Method 2. ADC (%) = 100-100 * (IFeed/IFeces)*(E Feces/EFeed) (Indicator method)

Method 3. DE (%) = (RE + H + BUE)/GE (Energy budget)

Determination:

� Digestible Energy (DE)

Bioenergetic approach BUEBioenergetic approach BUE


Methiod 1 . BUN = Flow rate tank * (CN,Out – CN,In ) (Direct measurement)

Method 2. BUN= DN - RN (Nitrogen budget)

BUE = BUN *24.85 (kJ/gN) (Energy budget)

Determination:

� Branchial and Urinary Energy (BUE)

Bioenergetic approach MEBioenergetic approach ME


Methiod 1 . ME = GE - FE - BUE = DE - BUE (From Geintake , FE and BUE)

Method 2. ME = RE + H (From RE and H (heat prod.))

Determination:

� Metabolizable Energy (ME)

Bioenergetic approach HBioenergetic approach H

Determination Heat production (kJ/g)

Methiod 1 . H = ME - RE (Energy budget)

Method 2. H = measured heat production (Direct calorimetry, difficult)

Method 3. H = 11.18 O2 + 2.61 CO2 - 9.55 NH3 (Indirect calory metry, MRU)H = 11.18O2 + 2.61CO2-7.86 BUNH = Qox * O2

Determination:

� Heat production (H)

Bioenergetic approach for growth REBioenergetic approach for growth RE

Determination Energy retention (kJ/g)

Methiod 1 . RE = ME - H (Energy budget)

Method 2. RE = Wt * E t – W0 * E0 (From body composition)

Exp. Period should be enough to create sign. E-gain during exeprimental period.

Determination:

� Retained energy (RE)


Gross Nitrogen (GE)

Digestible Nitrogen (DN)

Retained Nitrogen(`RE)

Fecal losses (FN)

Branchial and Urinary Nitrogen (BUN)

Nitrogen balanceNitrogen balance

Nitrogen balance (kJ/g) GNIntake = Feed intake * N Feed

BUN = Flow rate tank * (CN,Out – CN,In ) (CN in or out by autoanlyzer MRU)BUN = DN – RN (From DN & BUN)

RN = DN - BUN (From DN & BUN)RN = Wt * N t – W0 * N0 (From body composition, more accurate

� N content feed, fish and feces determined by Kjeldalh-N Protein content = 6.25* N

Oxygen consumption constrains food intake in fishOxygen consumption constrains food intake in fish

Saravanan et al., 2013. PLOSONE (8)8.

Rainbow trout (Oncorhynchus mykiss).

Saravanan et al., 2013. PLOSONE (8)8.

Oxygen consumption constrains food intake in fishOxygen consumption constrains food intake in fish

Rainbow trout (Oncorhynchus mykiss).

Santos et al., 2013. Aquaculture Research 44

Effect CO2 on energy metabolism and stress responseEffect CO2 on energy metabolism and stress response

Seabass (Dicentrachus labrax).

iDO = 3 mg/L

Determination incipient DO concentration Determination incipient DO concentration

Nile tilapia (Oreochromis niloticus).

After Tran-Duy et al., 2012 Aquaculture Research,43, 730–744

Feeding and diurnal variation in waste production Feeding and diurnal variation in waste production

Euroepean eel (Anguilla anguilla).

Heinsbroek et al., 2008. Aquacult. Int. 16:93–108

19 kg N

61 g N

(S.I. = 32%)

12 kg P

2 g P

5 g P Feaces

Excretion

(S.I. = 43%) 377 g COD

1192 g COD

Excretion

512 g COD

179 g COD

124 g COD

Faeces

Rest

Respiration

(S.I. = 32%)

Feed composition (g/kg feed)

5 g P

35 kg N

6 g N Feaces

BUN

Fish composition (retention) (g/kg feed)Source: http://www.sustainaqua.org/images/handbook/EN.pdf

N, P and COD mass balance & RAS design

aquaexcel at wageningen university metabolic research unit ... · - influent o 2 and/or co 2...

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