Date post: | 07-Apr-2018 |
Category: |
Documents |
Upload: | nair-de-los-angeles |
View: | 219 times |
Download: | 0 times |
of 37
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
8/37
The vitamins are one of the mostimportant ingredients of the diet, and,
in some way, they are essential.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
12/37
The development of biochemical andhistological tools allows to evaluate in themidgut gland the nutritional state and thehealth of the organisms under cultivityconditions.
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
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).
8/3/2019 Ppt Natal Fenucci & Pereira
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)
8/3/2019 Ppt Natal Fenucci & Pereira
16/37
COMPOSITION OF SEMIPURIFIED DIETS
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
21/37
Histology of midgut gland of
Artemesia longinaris
8/3/2019 Ppt Natal Fenucci & Pereira
22/37
ARTEMESIA LONGINARIS15000UI of vitamin D3/ kg diet
8/3/2019 Ppt Natal Fenucci & Pereira
23/37
ARTEMESIA LONGINARIS 40000UI of vitamin D3/ kg diet
Foamy cells!!
8/3/2019 Ppt Natal Fenucci & Pereira
24/37
THE BEST.
Wild Prawn
A longinaris 15000 UI ofvitamin D3/ kg diet
8/3/2019 Ppt Natal Fenucci & Pereira
25/37
THE WORST
Foamy cells!!
Artemesia longinaris D40
8/3/2019 Ppt Natal Fenucci & Pereira
26/37
ALKALINE PHOSPHATASE ACTIVITY IN
Artemesia longinaris.
Determinacin del pHptimo
SUSTRATUM CONCENTRATION
8/3/2019 Ppt Natal Fenucci & Pereira
27/37
SUSTRATUM CONCENTRATION
EFFECT ON ENZYMATIC
ACTIVITY.
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
30/37
ALKALINE PHOSPHATASE ACTIVITY INMIDGUT GLAND OF ANDARTEMESIA
LONGINARIS FEED ON DIFFERENTDIETS.
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
32/37
THE DIETARY REQUIREMENT OF JUVENILE
GRASS SHRIMP
(PENAEUS MONODON) FOR VITAMIN D.SHIAU & HWANG
8/3/2019 Ppt Natal Fenucci & Pereira
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
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
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.
8/3/2019 Ppt Natal Fenucci & Pereira
37/37
THANK YOU FOR
YOUR ATTENTION!