Ppt Natal Fenucci & Pereira

8/3/2019 Ppt Natal Fenucci & Pereira

1/37

Facultad de Ciencias Exactas y Naturales

ALKALINE PHOSPHATASE INSHRIMPArtemesia longinaris:RESPONSE TO VITAMIN D IN

DIET.Fenucci, J.L.*; Pereira,N.A.;

FernndezGimenez, A.V. and Harn,N.S.


2/37

The techniques utilized in the production of peneidos

shrimps have evolved due to the economic pressureand the environmental protectionThe development of methods for the formulation andelaboration of comercial food has reduced 30% to40% the operating costs. Nevertheless, nutrition isstill a limited economic factor.

It is escential to know the nutritional needs ofthe cultived species in order to select the dietand the ingredients for its preparation.


3/37

Pleoticus muelleri and Artemesia longinaris are the shrimpspecies exploited within the Argentinian sea.

Both species are part of the benthic fauna and play animportant role in the trophic chain as primary andsecondary consummers.


4/37

PLEOTICUS

MUELLERI

Distribution: from 20 S to 50S

Temperature: between 6 and 24 C Salinity: between 31.5 and 35.5

ups. Feeding: Benthonic invertebrates,detritus, and organism particles.P.muelleri is omnivore-carnivoreand can be cannibal under captivityconditions.


5/37

ARTEMESIALONGINARIS

Distribution: from 23 S to 43S

Temperature: between 6 and 24 C Salinity: between 31.5 and 35.5

ups. Feeding: Benthonic invertebrates,

detritus, crustaceans remains,algae, and organism particles.A.longinaris is omnivore-carnivoreand can be cannibal under captivity

conditions.


6/37

Between 1992 and 2007, the anual catchespresented a varied range, from 8,000 to80,000 tons of P. muelleri and 50 to 400tons ofA. longinaris.

The exportation of these species producean income of 60 to 100 million dollarsper year!

In Argentina are more than 90,000 acres toculture shrimps; Baha Blanca and San Blas(Provincia de Buenos Aires) and the coastal


7/37

The researching team called Acuicultura (Facultad de

Ciencias Exactas y Naturales de la Universidad Nacionalde Mar del Plata), has developed:

Maduration and reproductionMassive culture of larvaeNutritionPond growing


8/37

The vitamins are one of the mostimportant ingredients of the diet, and,

in some way, they are essential.


9/37

15 vitamins liposolubles: A, D, E y K;

hydrosolubles, B1,B2, B6, B12, Pantothenic acid, nicotinicacid, biotine, folic acid, C, coline andinositol

Within nutritional studies of shrimps, therequirement of vitamins is considered aless-studied area.


10/37

Vertebrates: Vitamin D takes part in the calcium andphosphorus metabolism by promoting its intestinalabsortion and stimulating bones mineralization. It alsocontrols calcium levels in blood.

Crustaceans: Vitamin D can also be involved inmineralization processes of exoskeleton.

The formation of 1,25-OH-D3, catalized by specificenzymes present in vertebrates, has not been observedin crustaceans yet.


11/37

Vitamin D is important for the survival ofMarsupenaeus japonicus larvaes.

Kanazawa (1985)

The lack of vitamin D in the diet ofLitopenaeus vannameiproduced a lossin weight and appetite. He et al. (1992)

Levels that are higher and lower than 0.1mg/kg diet of vitamin D3 forPeneausmonodon, decreases the alkalinephosphatase activity. This enzyme wouldbe involved in the calcium and phosphorus

absorption and carrying. Shiau & Hwang,1994


12/37

The development of biochemical andhistological tools allows to evaluate in themidgut gland the nutritional state and thehealth of the organisms under cultivityconditions.


13/37

MATERIALS &

MTHODSEXPERIMENTAL CONDITIONS6weeks108 prawns(1.5g intitialmeanweight);

108 shrimps(4.20ginitialmean weight)150 l glass aquariaDensity: 8 animals/mEnvironmentalconditions:33salinity,18C,

pH7,11:13hs photoperiod.Feedingregime:5%biomass.Eachdiet was tested in three replicategroups


14/37

COLLECTION OF SPECIMENS

Shrimps were obtained from a commercialfisherman and placed in 150 l glass aquaria(33 salinity, 20C, pH 7, 13:11 hphotoperiod).


15/37

DIET ELABORATION

Five semi purified diets (vitamin free casein, maniocstarch, squid protein concentrate and vitamin freefish oil) were designed containing 0 (D0); 8,000 (D8),15,000 (D15); 30,000 (D30) and 40,000 (D40) UI

vitamin D3/kg diet.

Diet PL4 was applied as a control treatmentsince it is considered nutritrionalappropriated for the studied species.(Daz

& Fenucci, 2002)


16/37

COMPOSITION OF SEMIPURIFIED DIETS


17/37

COMPOSITION OF DIET PL4

Ingredient Porcentual %Harina de pescado 48

Harina de soja 17

Trigo 8,5

Fcula de mandioca 20

Protena de calamara 1

Aceite de pescado 2

Soluble de pescado 2

Lecithin 0,5

CHolesterol 0,5

Vitamin 0,5


18/37

HISTOLOGICALANALYSIS

At the end of trials,animals wereanesthetized on ice and the midgutgland and haemolymphof allindividuals, in intermolt, were

aseptically removed.Tissues were fixed in Davidson fluidfor 24h ,dehydrated in a progressiveseries of ethanol and embebbed inparaffin.Sections of 5 m were stained with

hematoxylin-eosin.


19/37

ALKALINE PHOSPATASE

Alkaline phosphatase activitywas determined inthe protein extract of midgut gland andhaemocytes (HLS) using p-nitrophenylphosphate

(p-NPP) (Sigma N4645) as substrate.


20/37

RESULTS AND DISCUSSION

Crecimiento y supervivencia enArtemesia longinaris

Treatmen

t

Mean weight Weight

gain

(gr.)

Weight

gain

(%)

Survival

(%)Initial

(gS)

final

(gS)

D0 2,88

0,564

3,50

0,688

0,62a 21,5 52a

D8000 1,50

0,238

1,60

0,206

0,10 b 6,7 71a

D15000 2,85

0,570

3,70

0,745

0,85 29,8 67a

D30000 1,49 0,266

1,71 0,353

0,21 c 14,4 75a

D40000 2,92

0,647

3,96

0,829

1,04 35,6 70a

PL4 2,52 0,325 2,90 0,431 0,38d

15,1 60a


21/37

Histology of midgut gland of

Artemesia longinaris


22/37

ARTEMESIA LONGINARIS15000UI of vitamin D3/ kg diet


23/37

ARTEMESIA LONGINARIS 40000UI of vitamin D3/ kg diet

Foamy cells!!


24/37

THE BEST.

Wild Prawn

A longinaris 15000 UI ofvitamin D3/ kg diet


25/37

THE WORST

Foamy cells!!

Artemesia longinaris D40


26/37

ALKALINE PHOSPHATASE ACTIVITY IN

Artemesia longinaris.

Determinacin del pHptimo

SUSTRATUM CONCENTRATION


27/37

SUSTRATUM CONCENTRATION

EFFECT ON ENZYMATIC

ACTIVITY.


28/37

EFFECTS OF DIFFERENT LEVELS OF VITAMIN

D3 IN THE DIET OF Artemesia longinaris ON

ALKALINE PHOSPHATASE ACTIVITY.

A. longinaris The lower activity levels wereobtained in wild animals, meanwhile the

highest activity levels were registered inorganisms feed on D40000 UI/kg diet

SOLUBLE PROTEINS AND ALKALINE PHOSPHATASE ACTIVITY IN


29/37

SOLUBLE PROTEINS AND ALKALINE PHOSPHATASE ACTIVITY INMIDGUT GLAND OF Artemesia longinaris FEED DIFFERENTDIETS. Artemesia longinaris

Soluble protein

(mg/ml)

Alkaline phosphatase

(Abs/min/mg prot.)

Wild 1,62+0,171a 0,05+ 0,003e

PL4 9,99+1,697c 0,250,009

Initial 2,77+1,345a 0,140,007b

D0 8,17+0,318b 0,240,002

D80000 7,34+0,974b 0,140,004b

D15000 8,62+0,415b 0,180,009c

D30000 8,58 +0,184c 0,140,005b

D40000 6,27+0,015e 0,300,016d

ALKALINE PHOSPHATASE ACTIVITY IN


30/37

ALKALINE PHOSPHATASE ACTIVITY INMIDGUT GLAND OF ANDARTEMESIA

LONGINARIS FEED ON DIFFERENTDIETS.


31/37

Alkaline phosphatase activity in the

haemocytes ofArtemesia longinaris

The present study shows the occurrence ofdifferent alkaline phosphatase activities inthe midgut gland and haemocytes of

Artemesia longinaris in response to varyinglevels of Vitamin D in diet.

THE DIETARY REQUIREMENT OF JUVENILE


32/37

THE DIETARY REQUIREMENT OF JUVENILE

GRASS SHRIMP

(PENAEUS MONODON) FOR VITAMIN D.SHIAU & HWANG


33/37

D 40000 UI de vitamina D3/ kg dieta

TREATMENT IN Artemesia longinaris

Foamycells!!

A longinaris 40000 UI of

vitamin D3/ kg diet

In human beings : cellular irreversible damages

alkaline phosphatase activity.

HE WORST!!!!

Artemesia longinaris

Soluble

protein

(mg/ml)

Alkaline

phosphatase

(Abs/min/mg

prot.)

Wild 1,62+0,171a 0,05+

0,003e

PL4 9,99+1,697c

0,250,009

Initial 2,77+1,345a 0,140,007b

D0 8,17+0,318b 0,240,002

D80000 7,34+0,974b 0,140,004b


34/37

CONCLUSIONS

Vitamin D3 is not a assential

nutrient for the survival of

Artemesia longinaris in culture.The inclusion of 8000 and

30000 UI of vitamin D3/ kg diet

in Artemesia longinaris acts ina negative way over thegrowing process.


35/37

CONCLUSIONS

In A. longinaris exists a relationshipbetween vitamin D3 and the alkalinephosphatase activity, however, it is

neccesary to do some extra researchesin order to stablish its role in thecalcium metabolism.

The supply of 40000 UI of vitamin D3

/kg diet may be considered ashypervitaminosis in Artemesialonginaris.


36/37

CONCLUSIONS

15000 UI of vitamin D3 /kg diet,

improve the sanitary conditions ofthe culturedA. longinaris.

The occurrence of different alkalinephosphatase activities in the midgutgland and haemocytes ofArtemesialonginaris in response to varyinglevels of Vitamin D in diet.


37/37

THANK YOU FOR

YOUR ATTENTION!

Documents

Ppt Natal Fenucci & Pereira