High density liver improvement

For Aquaculture

LiverCall GCG TM

- product description

▣ Culture environment of aquaculture fish

In order to culture fish within limited period, the cultured fish usually are fed feeds containing more nutrients than they required and fish are cultured in high density in limited space, therefore arise the unbalanced metabolism and give fish many artificial stress.

Because of these problems in aquaculture, liver is the most affected intestines and most of the parts in liver are wound or the metabolism can't be proceeded smoothly, then arise nutritional diseases or are easy to be infected by bacteria or virus because their immune resistance are depressed.

▣ Function of liver

(1) The center of metabolism Liver is the place for metabolism of carbohydrate, protein and lipid

Liver has the important metabolism function of protein synthesis.

If the liver is wound or its function is depressed

- the balance of body materials metabolism will be destroyed

- Arise nutritional diseases, the normal metabolism could not be maintained.

(2) Maintaining the balance of body hormones Hormones are secreted by the endocrine organs.

If the disease of liver is serious, the secretion of hormones will be

unbalanced and arise many problems of body functions.

If the liver function is depressed, such secretion of hormones will

become unbalanced and decrease the growth of fish.

(3) Antidotal function Liver is the organ that all medicines absorbed in fish body (antibiotic, formalin etc.) are transferred to other forms and the organ that could secrete ammonia produced from protein metabolism.

Specially in summer when water temperature is high, or fish are cultured in high density or fish were over fed, the concentration of ammonia in blood is easy to become high to be toxic to fish.

If the concentration of ammonia continuously accumulate, fish could become comatose or even die.

(4) Immune function Liver has the function of storing antibody to against abnormal materials.

However, if the function of liver is depressed, the immune resistance ability is also depressed, and the fish is infected by bacteria and virus.

In order to increase immune resistance ability, use the additives to improve its own immune resistance ability through improvement of liver function.

(1) LiverCall GCG could completely replace the existed liver improvement - glutathione. LiverCall GCG contains several kinds of amino acids, minerals and immune stimulants.

(2) LiverCall GCG could improve the liver function, release stress and increase disease resistance through increase of immune resistance.

(3) LiverCall GCG could greatly improve feed intake as feed additive to improve liver function, almost has no adverse effect and has high absorption.

▣ Specific character of LiverCall GCG

▣ Merits of LiverCall GCG

(1) Improve feed intake, growth and feed efficiency

(2) Improve metabolism of protein and amino acids

(3) Improve the synthesis of glutathione and glutathione peroxidase.

(4) Prevent fat liver and improve antioxidation ability

(protect body against oxygen free radical and peroxide)

(5) Release stress

(6) Strengthen disease resistance through improve immune resistance

(7) Improve secretion of liver ammonia, heavy metal and antibiotic

(8) Colorless through reducing of melanin color

▣ The composition of LiverCall GCG

(1) GCG could completely replace glutathione, could improve

synthesis of glutathione, strengthen liver function and cou

nteracting poison function, could improve body metabolis

m as a strong reducing agent (As the improvement of liver

function, LiverCall GCG is in the application processing o

f patent)

(2) Several kinds of essential nutrients : containing 18 kinds o

f essential and non-essential free amino acids, could impro

ve growth and feed efficiency through increasing of feed i

ntake, could increase disease resistance through increasin

g of non-specific immune resistance.

(1) Improve growth

1) Experimental method Test animal : juvenile flounder flatfish (average weight 22.5 ± 0.1 g) Test condition : test period 6 weeks Experimental design The basal diet was used as control (Cont), GCG (liver improvement material of Liver Call) and GSH (glutathione) were supplemented in diets as 10 and 20 mg/kg fish/day

(Cont, GCG10, GCG20, GSH10 and GSH20).

▣ Function of LiverCall GCG proved through experiment

Figure 1. Weight gain of fish fed

diets containing GCG and

GSH for 6 weeksb

a a a a

80

105

130

155

180

(%)

Control GCG10 GCG20 GSH10 GSH20

WG

(1) Improve liver function 1) Experimental method Test animal : juvenile flounder flatfish (average weight 22.5 ± 0.1 g) Test condition : test period 10 days Experimental design The basal diet was used as control (Cont), GCG (liver function improvement material of Liver Call) and GSH (glutathione) were supplemented in diets as 10 and 20 mg/kg fish/day (Cont, GCG10, GCG20, GSH10 and GSH20).

▣ Function of LiverCall GCG proved through experiment

2) Experimental result

Diets

Cont GCG10 GCG20 GSH10 GSH20GOT (IU/L) 56a 45b 42b 43b 47b

GPT (IU/L) 13a 8b 9b 13a 12a

Liver glutathione(micromole/g)

2.8b 4.2a 3.9a 3.8a 4.1a

Liver glutathione peroxidase (micromole/(minute×mg protein)) 20b 26a 29a 27a 30a

Liver antioxidation rate (%) 25b 43a 41a 42a 44a

b

aa a

a

0

2

4

6

Con. GCG10 GCG20 GSH10 GSH20

(micromoI/g)

Glutathione

Figure 2. Values of liver glutathione and glutathione peroxidase fed diets containing GCG

and GSH for 10 days.

b

aa

aa

0

10

20

30

40

Con. GCG10 GCG20GSH10 GSH20

(micromole/ (min. x mg protein))

Glutathione peroxidase

(2) Non-specific immune response

1) Experimental method Test animal : juvenile flounder flatfish (average weight 22.5 ± 0.1 g) Test condition : test period 10 days Experimental design The basal diet was used as control (Cont), GCG (liver function improvement material of Liver Call) and GSH (glutathione) were supplemented in diets as 10 and 20 mg/kg fish/day (Cont, GCG10, GCG20, GSH10 and GSH20).

2) Experimental result

Diets

Cont GCG10 GCG20 GSH10 GSH20

Peak value of CL (mV) 446b 631a 617a 472b 469b

Lysozyme activity (U/ml) 323b 425a 417a 341b 352b

(3) Stress response 1) Experimental method Test animal : juvenile flounder flatfish (average weight 22.5 ± 0.1 g) Experimental design After formalin bath, blood cortisol returned to normal level, and blood cortisol level of fish fed GCG and GSH returned to normal level after 6 hours of formalin bath.

▣ Function of LiverCall GCG proved through experiment

Blood cortisol

0

30

60

90

120

0 1 2 3 4 6 12 24 48

(ng/ml )

Hours

Con.GCG10GCG20GSH10GSH20

2) Experimental result

Figure 3. Stress responses during

48 hours after formalin

bath

Figure 4. Non-specific immune response of fish fed diets containing GCG and GSH

for 10 days.

b

a a

b b

300

400

500

600

700

Con. GCG10 GCG20 GSH10 GSH20

(mV)

Peak of CL

b

a a

b b

100

200

300

400

500

Con. GCG10 GCG20 GSH10 GSH20

(U/ml)Lysozyme activity

▣ Discussion

(1) Improve liver function

When liver cells are mysterious death or are infected - GOT and GPT could flow out to blood in high concentration - GOT and GPT will become high in blood quickly

GCG (improve liver function material in Liver Call) could help the metabolism of protein and amino acids in liver more smoothly than glutathione (GSH), and improve the function of liver.

Glutathione (GSH) - Existed in all animals in nature and yeast - The important function in liver to prevent the lipid accumulation and the formation of fat liver and peroxides. - GSH are absorbed through the effect of digestible enzymes from each amino acids, it can't be absorbed as the GSH form. - After releasing from each amino acids, GSH becomes part of active, and SH radical of GSH has the effect to improve liver function.

From the above mentioned experimental results, it is obvious that the supplementation of GCG and GSH could improve the synthesis of glutathione.

The supplementation of GCG and GSH - Improve the synthesis the glutathione peroxidase - Could improve the total antioxidation activity through glutathione and glutathione peroxidase

With the oxidation of accumulated lipid in liver - The toxic of produced peroxides become serious and destroy the liver of cultured fish - The glutathione peroxidase could prevent the formation of such peroxides and could be considered as an improtant enzyme in liver function

(2) Non-specific immune response

The function of liver is depressed - Immune resistance ability is also depress - The fish is easy to be infected by bacteria, parasite and virus

Non-specific immune resistance with the supplementation of GCG - The activities of Chemiluminescence (CL) and plasma lysozyme increased - Thus the disease resistance is also increased from the above mentioned experiments

(3) Stress response

The blood cortisol concentration - To be stressed, the cortisol level will be increased - Stress are released, the cortisol will recover to normal level again After the formalin bath, the plasma cortisol levels returned to normal levels within 6∼12 hours in fish supplemented with GCG and GSH, and within 24∼48 hours in fish from control group. GCG and GSH have strong effects on releasing stress.

There is no difference in GCG (improve liver function material in

Liver Call) and glutathione in liver function improvement.

Compared with the existed glutathione product (containing 2% of

glutathione), the concentration of GCG in LiverCall GCG is high

(containing 6% of GCG), similar results could be obtained even

using 1/3 concentration of Liver Call.

It is a big burden for fish farmers to use glutathione due to its high

price, however, only 1/3 of expense is possible if using Liver Call

that could get the similar results in improving liver function and it is

possible to use for long term.

▣ Economical merit of LiverCall GCG

▣ How to use LiverCall GCG effectively

Efficacy Medication amount

(g/ton fish weight)Medication period Remarks

Improve liver function 150-300g 1-2 weeks

Possible to extend or shorten

medication period according

to the status of fish

Counteracting medicine poison and medicine bath

150-300g 1-2 weeks

When culture fish in high water temperature and high density 150-300g

Medication biweekly

Prevent liver disease 50-100gContinuously

medication