Was 2011 shrimp sensorial test - krill oil

1

Otavio S. Castro¹, Alberto J. P. Nunes¹, Sigve Nordrum, Marcelo V. C. Sá¹

Marine Science Institute (LABOMAR)Federal University of Ceará

WAS 2011

Natal, Brazil

June 09, 2011

Feed Manipulation¹

• Screening feed ingredients to attain

target desired characteristics

• Product color, flavor, qualitative

composition etc...

Consumer Behaviour¹

• Organoleptic Characteristics:

Standards of Quality and Freshness

• Willingness to pay more

• Healthier diet and lifestyle

Omega-3 Fatty Acids³

• Human health benefits

• Enrichment of Poultry eggs; Milk,

Meat, Broiler, Pork etc..

• Aquaculture Products

¹(SYLVIA et al., 1996; LING et al., 1999; WOOD et al., 2005; STEINE, ALFNES & RØRÅ, 2005; ALFNES et al., 2006; SIDHU, 2003); ²(UAUY, 1996; SIMOPOULOS, 1999; LEE et al, 2006:

PHETTPLACE & WATKINS, 1989; OFFER et al., 1999; KOOK et al., 2002; CACHALDORA et al., 2008))

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Rationale

Farm-Raised X Wild-Caught *

Fresh Water X Marine Species*

Naturally Enriched X Artificially*

Adding Value and/or Marketing

Segmentation Opportunities to

Aqua-products

*(MEYERS, 1994; VISENTAINER, 2003; TURCHINI et al., 2007; SHAPIRA et al., 2009; ALFNES et al., 2006)

To evaluate if Krill oil (QRILL™, Aker

Biomarine ASA, Oslo, Norway)

inclusion in experimental diets can

improve consumer’s acceptance of

white shrimp Litopenaeus vannamei

cooked tails.

4

Experimental SiteLANOA – Laboratory of Aquatic Animals NutritionCEAC – Costal and Environmental Studies Center (LABOMAR/UFC)

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Figures 1 and 2 – Aerial view of the Laboratory of Aquatic Animal Nutrition Facility

(LANOA). Situated on the Pacoti River Estuary, Eusébio Municipal district (25 km far from

the Fortaleza City – Ceará State). Geographic coordinates 35º00’0,25’’S e 38º25’22,57’’W

.

Foto: Alberto J. P. Nunes

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Rearing System50 Indoor Clear Water Tanks (500 L total water volume with total bottom

area of 0.57 m²)10 cells with 5 tanks - 14-h water filtering and recirculation regime with continuous

water aeration.

Fig. 3 – Indoor tanks at LANOA/LABOMAR where the

krill oil evaluation was conducted with the white

shrimp L. vannamei. Fig. 4 – Internal view of stocked

experimental tank. 6

3

4

Photo: Otávio S. Castro

Photo: Otávio S. Castro

Experimental Design4 Isonutritive Diets except in regards to the fatty acid content

3 Lipid Sources: Marine fish oil (Engraulis ringers), Antartic

Krill oil (Euphausia superba) and Soybean oil (Glycine max)

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Table 1: Experimental Diets Design

TratamentFatty Acid Requirements* Main Lipid

Source4LOA and LNA¹ EPA and DHA²

T1 (FISH) Satisfied Satisfied Fish Oil

T2 (KRILL) Satisfied Satisfied Krill Oil³

T3 (SOY) Satisfied 50% Deficient Soybean Oil

T4 (KRILL+) Satisfied 25% Above Krill Oil³

*based on 80% of that required by Penaeus monodon - Glencross & Smith (1997, 1999, 2001)

and Glencross et. al (2002a,b)

¹LOA: Linoleic acid (C18:2n-6) and LNA: Linolenic acid (C18:3n-3)

²EPA: Eicosapentaenoic acid (C20:5n-3) and DHA: Docosahexaenoic acid (C22:6n-3)

³ QRILL™ oil, Aker Biomarine ASA (Oslo, Norway) 4Soybean oil was included on the diets FISH, KRILL and KRILL+ in order to meet the LOA and

LNA requirements

Feed ManufacturingSmall scale laboratory process,

Laboratory of Aquatic Animals Nutrition of LABOMAR, Brazil

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INGREDIENTS MILL

400 µmCOOKING – 90ºC, 25

min.EXTRUSION – 2mm

OVEN DRYING –

70ºC, 3 h

PELLET GRINDING

5mm

SIEVING STORAGING-22ºC

3 MIXTURES

Diets Allotment6 replicate tanks were assigned for each treatment

Random block design

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Ingredients (g kg-1 diet)

Diet

FISH SOY KRILL KRILL+

Soybean meal 350.0 350.0 350.0 350.0

Wheat flour 298.7 300.0 299.8 291.5

Poultry by-product meal 100.0 105.0 100.0 67.5

Fishmeal, Anchovy 60.2 70.6 71.6 68.9

Rice broken 40.0 40.0 40.0 40.0

Soybean protein concentrate 32.5 16.6 19.1 20.0

Fish oil 26.6 0.0 0.0 0.0

Krill oil 0.0 0.0 48.3 55.0

Soybean oil 10.0 34.5 4.4 3.8

Soybean lecithin 15.0 15.0 0.0 0.0

Cholesterol 0.0 1.3 0.0 0.4

Squid meal, whole 10.0 10.0 10.0 10.0

Phosphate monodicalcium 13.0 13.0 13.0 13.0

Potassium chloride (KCl) 10.0 10.0 10.0 10.0

Salt common 10.0 10.0 10.0 10.0

Vitamin-mineral premix 10.0 10.0 10.0 10.0

Corn gluten meal, 65% CP 0.0 0.0 0.0 40.0

DL-methionine, crystalline,

99%8.0 8.0 8.0

4.6

Synthetic binder 4.0 4.0 4.0 4.0

Ascorbic acid polyphosphate 2.0 2.0 1.8 1.3

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Table 2: Ingredient composition of the experimental diets

Experimental Diets

• Minimum Cost Formulation

• Software Feedsoft

Professional 3.1

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Table 3: Nutritional levels of experimental diets

Composition(g kg-1 diet)Diet

FISH SOY KRILL KRILL+

Crude Protein1,2 351.8 353.5 354.4 353.1

Crude Fat1 88.8 80.8 90.0 91.3

Fiber1,2 12.0 13.3 9.0 12.0

Ash1 93.7 91.0 89.5 82.3

Calcium1,5 10.1 10.4 10.6 9.0

Total Phosporus1,5 8.8 8.6 9.0 7.9

Phospholipidis1,3 14.25 14.00 14.25 15.95

Cholesterol1,3 1.70 1.70 2.97 1.50

Gross Energy (kcal/kg) 1,4 4,184 4,273 4,209 4,3601 AOAC,1990² Amino acid requirements - Akiyama, Dominy & Lawrence (1992) e Fox, Lawrence & Li-Chanb

(1995)

³Phospholipids and Cholesterol: Gong et al. (2000)4Gross Energy: Cousin et al., (1993) 5Calcium and Phosphorus: Davis, Lawrence & Gatlin (1993)

Starch: Cuzon et al. (2000)

Experimental Diets

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Table 4. Fatty acid profile² and astaxanthin content³ of the experimental feeds

Experimental Diets

Composition(g kg-1 diet)¹Diet

FISH SOY KRILL KRILL+

Crude Fat (g kg-1 diet) 88.80 94.00 80.80 91.30

Fatty Acid Profile (% of Crude Fat)²

C12:0 (ácido láurico) 0,0 0,0 0,59 0,67

C14:0 (ác. mirístico) 3,56 0.98 12,29 13,99

C16:0 (ácido palmítico) 20,39 17,84 23,49 26,75

C16:1 cis (ácido palmitoléico) 5,28 2,07 10,73 12,22

C18:0 (ácido esteárico) 4,70 4,45 3,76 3,24

C18:1 cis (ácido oléico) 15,59 16,03 14,07 12,20

C18:2 cis (ácido linoléico) – LOA 28,32 44,71 16,21 13,96

C18:3 cis (ácido linolênico) – LNA 3,40 4,91 1,52 1,31

C20:4n6 (ácido araquidônico) – ARA 0,38 0,0 0,0 0,0

C20:5n3 (ác. eicosapentaenóico) – EPA 5,09 0,65 5,34 6,80

C22:6n3 (ác. docosahexaenóico) – DHA 2,54 0,28 1,59 1,81

Sum PUFA 8,01 0,93 6,93 8,61

Sum LOA+LNA 31,72 49,62 17,73 15,72

PUFA/LOA+LNA Ratio 0,25 0,02 0,39 0,55

Sum EFA 39,72 50,55 24,66 24,42

Esterified Astaxanthin (ug/100 ul)³ 14,70 13,80 21,40 24,70

¹ Analyzed according to AOAC (1990) by Animal Nutrition Laboratory from Federal University of Ceará State, Fortaleza, CE,

Brazil. ²Analyzed by gas chromatography method in the Laboratory of Biochemistry and Instrumental Analysis from São Paulo

University (USP), Piracicaba, SP, Brazil. ³ Analyzed by high performance liquid chromatography (HPLC) by Plant

Morphogenesis and Biochemistry Laboratory from Federal University of Santa Catarina (UFSC), Florianópolis, SC, Brazil.

Experimental Period64 days of rearing on experimental diets

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Fig. 8 – Feed tray used on the

experimental feeding period.

Feeding Management

• Feed offer: daily at 7:30 and 16:00 h

• 1 Feed trail per tank - 14.3 x 3.5 cm

(diameter x height)

Water Quality

• Salinity: 44,0 ± 0,2 ‰

• pH: 7,35 ± 0,31

• Temperature: 27,5 ± 0,46 ºC

Shrimp Weight

• Initial: 2,79 ± 0,6 g

• Final: 11,4 ± 2,0 g (At harvest)

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Photo: Otávio S. Castro

Sensorial TrialBest-worse scaling methodology with 4 dietary treatments (JAEGER

et. al, 2008)

20 Non-trained panelists (10 men and 10 women) – 4 Taste Sets with 3

shrimps

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SHRIMP # TASTE SET SAMPLE TASTE PLATE NUMBER DIET

1

1

B 1 KRIL+

2 C 2 KRILL

3 A 3 FISH

4

2

D 1 SOY

5 A 2 FISH

6 B 3 KRILL+

7

3

A 1 FISH

8 D 2 SOY

9 C 3 KRILL

10

4

C 1 KRILL

11 B 2 KRILL+

12 D 3 SOY

TABLE 5. Experimental set-up of the best-worse scaling sensory test

to evaluate the dietary effect of oil source an inclusion

level on shrimp tail color, texture and flavor.

Sensorial TrialTest procedures:

• 240 shrimps: 60 for each treatment (11,4 ± 2,0 g, n = 48)

• Defrosting (After 1-week period: -22ºC), De-heading and Mineral

Water Rinsing

• 5-min Boiling in Water (3,3 mg/ml NaCl)

• Panelists Instruction – Test procedures

• Panelists Tasting and Evaluation

Fig. 1 – Example of a referee form used in the best-worse scaling sensory test to evaluate

shrimp tail sensory characteristics. Colored “X’s” exemplify how forms were filled..15

SET ONE

COLOR TEXTURE FLAVOR

LEAST

LIKEDSHRIMP

MOST

LIKED

LEAST

LIKEDSHRIMP

MOST

LIKED

LEAST

LIKEDSHRIMP

MOST

LIKED

X 1 1 X X 1

2 X 2 2

3 X 3 3 X

Note: Note: Note:

1 2 3

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COLOR TEXTURE FLAVOR

PANELIST FISH KRILL SOY KRILL+ FISH KRILL SOY KRILL+ FISH KRILL SOY KRILL+

1 -1 2 -3 2 0 1 -1 0 2 0 -3 1

2 1 2 -2 -1 0 1 -1 0 -3 0 0 3

3 0 1 -3 2 0 0 2 -2 -1 1 0 0

4 -1 1 -1 1 1 0 0 -1 1 -2 1 0

5 -2 1 -2 3 2 2 -2 -2 0 2 -3 1

6 -1 2 -3 2 1 -2 3 -2 2 -1 0 -1

7 -1 3 -3 1 0 -1 1 0 -1 0 2 -1

8 0 2 -3 1 1 -1 -1 1 1 -2 0 1

9 -3 1 -1 3 0 1 -1 0 0 2 -1 -1

10 -2 0 -1 3 0 -1 -1 2 -2 2 -2 2

11 -3 1 -1 3 0 -2 1 1 0 -1 0 1

12 -2 1 -2 3 2 1 0 -3 -1 2 -1 0

13 -1 1 -2 2 2 -1 -2 1 -1 -1 -1 3

14 1 0 -3 2 0 2 -1 -1 0 0 1 -1

15 1 0 0 -1 2 -1 -2 1 1 2 -3 0

16 -2 2 0 0 -2 1 1 0 -2 1 1 0

17 -1 1 -3 3 0 0 -2 2 -2 2 0 0

18 0 0 -2 2 -1 -2 3 0 1 -1 0 0

19 -2 3 -2 1 0 3 -1 -2 1 0 -3 2

20 -2 -1 0 3 -1 0 0 1 1 -1 -1 1

TOTAL

SCORE-21 23 -37 35 7 1 -4 -4 -3 5 -13 11

Table 6 - B-W scores, each value represent the sum of three evaluated

tastes for each sample. Positive choices valued +1, neuter choice valued 0

and negative choice valued -1, B-W did vary between -3 and +3.

Results and Discussion

¹ Aggregate values of Best-worse scores for Shrimp Color

Preference. Values represents the sum of B-W scores of 20

judgers for each sample, different letters in the bar denote

significant difference at the = 0.05 level by Mann-Whitney U test

for frequency of choices.17

ab

-40

-30

-20

-10

0

10

20

30

40

- 21

23

- 37

35

BE

ST-W

OR

SE

SC

AL

E

CONSUMERS COLOR PREFERENCE ¹

KRILL+

SOY

KRILL

FISH

a

c

c

b

18

ab

-10

-8

-6

-4

-2

0

2

4

6

8

10

7

1

- 4 - 4BE

ST-W

OR

SE

SC

AL

E

CONSUMERS TEXTURE PREFERENCE ¹

KRILL+SOY

KRILLFISH

¹ Aggregate values of Best-worse scores for Shrimp Texture

Preference. Values represents the sum of B-W scores of 20

judgers for each sample, different letters in the bar denote

significant difference at the = 0.05 level by Mann-Whitney U test

for frequency of choices.

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MOLT STAGEFISH

(%)

SOY

(%)

KRILL

(%)

KRILL+

(%)

A - (early postmolt) - 8.33 8.33 16.67

B - (late postmolt) 16.67 16.67 8.33 25.00

C - (intermolt) 8.33 25.00 8.33 -

D0 - (onset of premolt) 16.67 - 33.33 16.67

D1 - (early premolt) 16.67 33.33 41.67 8.33

D2 - (intermediate

premolt)8.33 8.33 - 16.67

D3 - (late premolt) 33.33 8.33 - 16.67

E - (molt) - - - -

TOTAL 100.00 100.00 100.00 100.00

TABLE 6. Shrimp Molt Stages observed in experimental shrimp

samples according to Oliveira Cesar et al. (2006); the

setogenesis of the endopodite of the shrimp uropod

were analyzed (n = 12 for each treatment).

Results and Discussion

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¹ Aggregate values of Best-worse scores for Shrimp Flavor

Preference. Values represents the sum of B-W scores of 20

judgers for each sample, different letters in the bar denote

significant difference at the = 0.05 level by Mann-Whitney U test

for frequency of choices.

ab

KRILL+SOJA

KRILLPEIXE

-15

-10

-5

0

5

10

15

- 3

5

- 13

11

BE

ST

-WO

RS

E S

CA

LE

CONSUMERS FLAVOR PREFERENCE ¹

KRILL+

SOY

KRILL

FISH

a

ab

c

bc

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¹ Aggregate values of Best-worse scores for Shrimp Color

Preference. Values represents the sum of B-W scores of 20

judgers for each sample, different letters in the bar denote

significant difference at the = 0.05 level by Mann-Whitney U test

for frequency of choices.

ab

KRILL+SOJA

KRILLPEIXE

-15

-10

-5

0

5

10

15

- 3

5

- 13

11

BE

ST

-WO

RS

E S

CA

LE

CONSUMERS FLAVOR PREFERENCE¹

KRILL+

SOY

KRILL

FISH

a

ab

c

bc

Spearman’s

Correlation Test:

Flavor Responses vs.

Color Responses =

No significance (0.10)

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Fatty Acid Profile of Shrimp Tails

Fatty Acid Profile (% of total

Crude Fat)

Diets

PXE SJA KRL KRL+

C12:0 (ácido láurico) - - - 0.88

C14:0 (ác. mirístico) 29.03 24.63 27.16 27.18

C16:0 (ácido palmítico) 1.46 0.48 3.02 2.90

C16:1 cis (ácido palmitoléico) 0.99 0.60 1.21 1.04

C17:0 (ác. heptadecanóico) 12.04 11.59 10.98 10.25

C18:0 (ácido esteárico) 11.62 11.05 12.94 11.79

C18:1 cis (ácido oléico) 14.93 26.94 11.10 11.53

C18:2 cis (ácido linoléico) 0.63 1.30 0.54 -

C18:3 cis (ácido linolênico) - - - -

C20:1 cis (ác.cis-11-eicosanóico) - 1.80 0.76 0.74

C20:4n6 (ácido araquidônico) 1.74 1.33 0.94 1.20

C20:5n3 (ác. eicosapentaenóico) 10.10 6.60 11.50 13.92

C22:6n3 (ác. docosahexaenóico) 5.34 3.53 4.22 5.66

SUM PUFA4 15.44 10.13 15.72 19.58

SUM LOA+LNA5 15.56 28.24 11.64 11.53

PUFA/LOA+LNA Ratio 0.99 0.36 1.35 1.70

SUM EFA6 31.00 38.37 27.36 31.11

• Results Agree with:

Guary et al. (1976)

- M. japonicus

Catacutan (1991);

Kumaraguru Vasagam,

Ramesh &

Balasubramanian (2005)

- P. monodon

González-Félix et al.

(2002c, 2003); Hurtado

et al. (2006) - L.

vannamei

• FLAVOR:

Waagbø et al. (1993)

Positive Associations with

organoleptics characteriscs

in salmon filetswith

enhanced Omega-3 fatty

acids levels.

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ab

KRILL+SOJA

KRILLPEIXE

• Krill oil utilization in L. vannamei feeds

enhances consumer acceptance for the

color and flavor attributes in shell-on

shrimp cooked tails.

24

ab

KRILL+SOJA

KRILLPEIXE

FURTHER SUGGESTIONS

• Dose-response Test

• Lower effective dose/response to promote

changes in shrimp tails

• Minimum feeding period necessary

• Optimization of formulae costs

• Marketing study:

• Wich class of products have the greatest

potential of valorization?

• Who will be the consumers and how much

more they will pay for the new differentiated

product?

• Production Viability: Logistics; Additional

costs vs. Additional profits

THANK YOU!!!Gracias!!! Obrigado!!!

Merci Beaucoup!!!

[email protected][email protected]

+55 65 8159-4303 / 8112-0109

ACKNOWLEDGEMENTS......AKER BIOMARINE ASA, CNPq, LANOA staffs,

LABOMAR/Federal University of Ceará; EVIALIS