system

World Aquaculture 2012
EVALUATION OF THE PHYSIOLOGICAL
RESPONSE OF THE CRAYFISH Cherax
quadricarinatus TO DIFFERENT GROWING
CONDITIONS
Diana Carreño (ITSON-CIBNOR) ([email protected])
Humberto Villarreal (CIBNOR)
Lucía Ocampo (CIBNOR)
Ramón Casillas (ITSON)
José Naranjo (CIBNOR)
Armando Monge (ITSON-CIBNOR)
Las Vegas, NV.
February 29, 2012
INTRODUCTION
Redclaw Cherax quadricarinartus
An omnivorous species, with favorable characteristics for cultivation:
 Has a short life cycle, with great reproductive potential.
 It is adaptable to environmental variations
 Has high growth rate, (reaches commercial size in 6 months )
 Has good commercial value and global market acceptance.
(Villarreal, 2000)
BACKGROUND
• Diets
NUTRITION
• Enzymes
(López et al., 2004; López-López et al., 2005)
(Cortés et al., 2003; Cortés et al., 2004;
Campaña et al., 2005; Cortés et al., 2005;
Rodríguez-González et al., 2006)
SYSTEM
• Engineering
• Production System
(Villarreal & Hutchings, 1994)
• Vitellogenesis
• Embryology
(García-Guerrero et al., 2003)
(Rodríguez et al., 2002; Serrano-Pinto et
al., 2004)
PATHOLOGY
• Immunology
• Bacteriology
GENETICS
• Population
• Quantitative
BIOECONOMY
• Financial Analysis
ENVIRONMENTAL
• Water quality
• Environmental impact
•Oxidative stress
• Temperature - Salinity
PHYSIOLOGY
(Macaranas, et al., 1995)
(Campaña et al., 2005; Campaña
et al., 2006; Campaña et al.,
2008;)
(Naranjo-Páramo et al., 2004)
REPRODUCTION
(Claydon, et al., 2004)
• Digestibility
• FecundityMaturation
(Zapata et al., 2003; Cahansky
et al., 2008 Rodríguez-González
et al., 2009)
(Edgerton et al., 2004)
(Jones, et al., 2000)
(Villarreal & Naranjo, AVANCE-CONACYT 2008)
(Villarreal & Hutchings, 1994)
(Zenteno-Savin et al., 2008; Cortes et al.,
2009)
(Horwitz, 1990)
(Meade, 2002, Prymaczok et al., 2008)
• Toxicity
(Meade & Watts, 1995)
PHYSIOLOGY
The species is highly tolerant to environmental variations and has shown to be physiologically
robust, but there are few scientific reports.
PARAMETER
EXTREME
OPTIMUM
5 – 40 °C
28 °C
6 ups
(Tolerates brackish water without affecting
its growth significantly )
Freshwater
150 mg/L de CaCO3
-
pH
-
7 – 8.5
Oxygen
-
~ 4 mg O2/L
Metabolic rate
-
-
Temperature
Salinity
Hardness
Meade (2002) reported a Q10 = 2.44
OBJECTIVE
To determine the physiological response, through
metabolism, in juvenile freshwater crayfish Cherax
quadricarinatus, to critical environmental variations
presented in intensive culture conditions.
METHODOLOGY

Experiment 1. Growth of juveniles under different conditions

Experiment 2. Determination of the physiological response
through the metabolic rate and critical oxygen level
METHODOLOGY
Experiment 1. Growth of juveniles under different conditions with
multitrophic system
TREATMENTS
 System with
 System with
addition of natural
productivity
added probiotics
 Control
All treatments received a commercial pelleted shrimp feed with 35% protein, supplied at 2% of
total biomass in 2 rations per day (Cortés et al., 2003)
METHODOLOGY
General conditions
4 replicates per
treatment
25 Juveniles
1.5 g/tank
Photoperiod (h)
14:10
(light: dark)
Registered daily:
T, DO, pH, molts
and mortality
Tanks
0.6 X 0.4 m
(Density:
92 animals/m2)
water
exchange
20% / day
Temperature
28°C
Biometry every
15 days / end of
60 days
METHODOLOGY
 Parameters
Growth:
Especific growth rate =
FCR:
Survival:
100 ∗ (ln final weight − ln Initial weight)
time (days)
Food fed (g)
FCR =
Weight gain (g)
% Survival = ( final # of organisms ) ×100
(Initial # of
organisms )
METHODOLOGY
Experiment 2. Physiological response to intensive cultivation system
a) BIOCHEMICAL ANALYSIS (Haemolymph)
 200 µl haemolymph, using
potassium oxalate (5%) as
anticoagulant.
 After 60 days: 20 animals were
selected / treatment.
 Plasma: was centrifuged at 3600
rpm
 Preserved at -80°C.
ANALYSIS
METHOD
Glucose
GOD-PAP enzymatic method
Lactate
PAP enzymatic method
Lipids
Phospho-vanillin (Barnes and
Blackstock, 1973)
Proteins
Bradford (1976)
Hemocyanin
Direct absorbance (Chen et al.,
1994)
METHODOLOGY
Experiment 2. Physiological response to intensive cultivation system
b) METABOLIC RATE AND CRITICAL POINT
Selection and conditioning
 Selected animals
(experiment 1)
 20 juveniles (replicate)
 Starvation of 24 hours
(Scelzo andZúñiga, 1987; Villarreal and
Ocampo, 1991; Rivera, 1992).
Experimental system
 Closed respirometry
 28 ± 0.5°C.
 One animal /
experimental unit
 2 controls / 10
respirometers
O2 Measuring system
PreSens Precision with
optical fiber oxygen
microsensor
METHODOLOGY
Experiment 2. Physiological response to intensive cultivation system
b) METABOLIC RATE AND CRITICAL POINT
Haemolymph
sampling
Initial DO
Acclimation
DO/15 min for
1 hr
Juvenile
Weight
RESULTS
Experiment 1. Growth of juveniles under different conditions
Culture condition
Control: Filtered
freshwater
System with addition of
natural productivity
System with added
probiotics
Final
Weight (g)
Specific
growth rate
(g/week)
FCR
n
5.57 ± 1.91
2.2 ± 0.2
0.5 ± 0.10
47
5.26 ± 1.89
2.1 ± 0.1
0.4 ± 0.03
50
5.45 ± 1.75
2.1 ± 0.1
0.4 ± 0.02
50
Mean values ​± standard deviation, ANOVA p>0.05
n = number of juveniles / treatment
ANOVA p>0.05
85.0
73.5
δ
20
SURVIVAL
(%)
63.7
REPORTED BY
Naranjo et al., 2004 (pond)
20
61.0
Thompson et al., 2004 (pond)
20
85.0
Cortés et al., 2003 (aquarium)
SURVIVAL (%)
80.0
75.0
73.5
69.1
70.0
65.0
60.0
55.0
50.0
Filtered water
Primary productivity
TREATMENTS
Probiotics
(δ: 92/m2)
RESULTS
Experiment 2. Physiological response to intensive cultivation system
Juvenile
condition
Without
metabolic
evaluation
Culture condition
Culture ponds
Filtered Water
System with primary
productivity
System with probiotics.
Filtered Water
Post metabolic System with primary
evaluation
productivity
System with probiotics.
Total Proteins Total Lipids Hemocyanin
Ratio
n
(mg/ml)
(mmol/L) Hemocyanin/
(mg/ml)
Protein (%)
443.46 ± 85.4 2.30 ± 0.60 2.71 ± 0.61
39.0 ± 3.15 20
483.27 ± 84.2 2.46 ± 0.86 3.14 ± 0.82
40.1 ± 5.39 20
498.42 ± 79.8
2.60 ± 1.46
3.18 ± 0.82
38.9 ± 6.38
20
488.53 ± 73.5
525.50 ± 44.1
3.48 ± 0.87
3.05 ± 1.36
3.55 ± 0.77
2.96 ± 0.34
43.4 ± 4.91
34.9 ± 8.90
20
20
494.58 ± 44.6
3.10 ± 1.50
4.00 ± 0.30
54.3 ± 10.4
20
481.78 ± 37.0
1.90 ± 0.62
2.64 ± 0.82
37.1 ± 10.6
20
Mean values ​± standard deviation, ANOVA p>0.05. n = number of juveniles / treatment
Lower than that reported for shrimp (60-90%) (Rose et al., 2004, Pascual et al., 2006).
 The ratio of 40% HC / PT, shows an efficient species for:
• Oxygen transport
• Utilization of protein for growth.
• Similar to other species of crabs and crayfish (Hagerman and Uglow, 1985, Spicer and Baden, 2000, Paschke et
al., 2010).
 It is considered an adaptive physiological condition of the species.
RESULTS
Experiment 2. Physiological response to intensive cultivation system
(a)
Metabolic Rate (mg O2/g/h)
0.12
ANOVA p>0.05, n=20
a
0.1
0.08
METABOLIC RATE
a
a
Filtered water
Primary productivity
0.06
0.04
0.02
0
Probiotics
(This work)
(b)
Critical oxygen (mg O2/L)
0.6
0.5
CRITICAL OXYGEN LEVEL
ANOVA p>0.05, n=20
a
a
Filtered water
Primary productivity
a
0.4
0.3
0.2
0.1
0
Probiotics
(This work)
RESULTS
Experiment 2. Physiological response to intensive cultivation system
 Cherax obtain energy from
0.50
gluconeogenic pathway:
Without metabolic evaluation
ANOVA p<0.05, n=20
b
1. Have greater availability of
Glucose (mg/ml)
0.40
0.30
nutrients in the medium, which
were used to generate a reserve
of glycogen
2. Glycogen is converted to lactate
by anaerobic pathway for energy
(glucose)
a
a
0.20
a
0.10
0.00
Lactate (mg/ml)
2.50
ANOVA p<0.05, n=20
2.00
c
1.50
lactate is not oxidized mainly by
anaerobic metabolism, possible
conversion to glucose or glycogen
1.00
b
b
0.50
a
0.00
Culture ponds
Filtered Water
Primary productivity
 Possible recycling of products:
Probiotics
(Gade et al., 1986)
RESULTS
Experiment 2. Physiological response to intensive cultivation system
 There are reports that diets with high protein availability increased capacity for
gluconeogenesis (Pellegrino et al., 2008)
ENVIRONMENT
(multi-trophic)
Total Proteins
(mg/g)
Primary productivity
46.21
Probiotics
121.36
AUTHOR
SPECIES
 There are reports of gluconeogenic capacity and
glucogenogenesis in some crustaceans using as
substrate: lactate, pyruvate or alanine.
COMMENTS
Stetten, 1982
L. polyphemus
Gluconeogenesis by lactate or pyruvate
Gade et al., 1986;
Hervan et al., 1999
M. mercenaria, L.
polyphemus, N. virei
Glucogenogenesis and gluconeogenesis using as
substrate lactate.
Oliveira y Da Silva,
1997
C. granulata.
Synthesis of glucose via alanine or lactate in
hepatopancreas (gluconeogenesis)
Schein et al., 2005
C. granulata.
Effect of seasonal and environmental changes on
capacity for gluconeogenesis
Pellegrino,
et al., 2008
C. granulata.
Effect of diets high in carbohydrate or protein, on
gluconeogenic capacity and glucogenoneogenesis.
RESULTS
Experiment 2. Physiological response to intensive cultivation system
Post metabolic evaluation
0.50
b
ANOVA p<0.05
b
Glucose (mg/ml)
0.40
0.30
a
a
0.20
0.10
0.00
2.50
Anaerobic
metabolism
Lactate (mg/ml)
2.00
1.50
b
1.00
c
ANOVA p<0.05
Aerobic
metabolism
0.50
a
a
0.00
Culture ponds
Filtered Water
Primary productivity
Probiotics
O2 Condition 
CONCLUSIONS
The results showed that the species:
 Tolerates intensive culture conditions without affecting its growth and survival.
 Uses nutritional environmental sources such as multi-trophic sources
 Keeps a low metabolic rate, thus explaining why redclaw is a species with high
energy efficiency
 Is highly tolerant to limited oxygen conditions.
 Has the physiological ability for aerobic and anaerobic metabolism.
 Shows a gluconeogenic capacity as source of energy production.
 The metabolic rate (0.07 mg O2/g/h) and the critical level of oxygen (0.483 mg
O2 / L) of juvenile C. quadricarinatus are very low compared with those
reported for other crustaceans.
World Aquaculture 2012
Thank you
Diana Carreño (ITSON-CIBNOR) ([email protected])
Humberto Villarreal (CIBNOR)
Lucía Ocampo (CIBNOR)
Ramón Casillas (ITSON)
José Naranjo (CIBNOR)
Armando Monge (ITSON-CIBNOR)
Las Vegas, NV.
February 29, 2012