FEED COMPOSITION FOR FARMED FISH USING FERMENTED PORCINE BLOOD

Abstract

A feed composition for farmed fish using fermented porcine blood having an effect of promoting the growth of farmed fish is proposed.

Claims

1.-8. (canceled)

9. A method for preparing a feed composition for fish, the method comprising: mixing porcine blood with a proteolytic enzyme and a blood anticoagulant; inoculating the porcine blood with a mixed strain of Lactobacillus johnsonii and Saccharomyces cerevisiae; and culturing the inoculated porcine blood for 3 to 30 days at 30 to 50 C. to prepare the feed composition comprising fermented porcine blood.

10. The method of claim 9, wherein the porcine blood is mixed with 6% (v/v) of the mixed strain.

11. The method of claim 9, wherein the porcine blood and the mixed strain are mixed with 5 to 7 wt % of a carbon source.

12. The method of claim 11, wherein the carbon source is selected from the group consisting of lactose, glucose, fructose, sugar, molasses, dextrose, and combinations thereof.

13. The method of claim 12, wherein the culture is performed at 40 C. with stirring at 50-70 rpm.

14. The method of claim 9, further comprising mixing the fermented porcine blood with rice bran, soybean meal, or a combination thereof; and culturing the mixture to prepare a secondary fermentation product.

15. The method of claim 14, wherein the fermented porcine blood is mixed with 25 to 75 parts by weight of the rice bran and 25 to 75 parts by weight of the soybean meal based on 100 parts by weight of the fermented porcine blood.

16. The method of claim 15, wherein the composition has a solid content of 30 brix.

17. The method of claim 9, further comprising mixing the secondary fermentation product with one or more of a plant components, animal components, and/or mineral components.

18. The method of claim 17, wherein the one or more plant components are selected from the group consisting of grains, brans, oilseed meals, root and tuber crops, food-processing byproducts, algae, microalgae, dietary fibers, oils and fats, legumes, vegetables, mushrooms, and combinations thereof.

19. The method of claim 18, wherein the plant components comprise between about 3 and about 80 wt % of the composition.

20. The method of claim 17, wherein the one or more animal components are selected from the group consisting of dried poultry byproducts, dried fish, crab meal, egg powder, Neverita didyma powder, fish meal, bone meal, bone ash, fish bone meal, oyster shell powder, chicken oil, animal edible residual oil, pork fat, beef tallow, mealworm larva, dried crickets, dried grasshoppers, rotifers, mysis shrimp, daphnia, lactose, whole milk powder, cheese, skim milk powder, and combinations thereof.

21. The method of claim 20, wherein the animal components comprise between about 3 and about 80 wt % of the composition.

22. The method of claim 17, wherein the one or more mineral components are selected from the group consisting of limestone powder, calcium carbonate, calcium sulfate, iron citrate, sodium molybdate, ammonium molybdate, sodium selenate, sodium selenite, chromium chloride, potassium iodate, calcium iodate, potassium iodide, manganese carbonate, copper sulfate, manganese sulfate, zinc sulfate, and combinations thereof.

23. The method of claim 22, wherein the mineral components comprise between about 0.0001 to about 5 wt % of the composition.

24. The method of claim 9, wherein the porcine blood is mixed with 1% (w/v) proteolytic enzyme and 6% (w/v) sugar; the mixture is inoculated with 6% (v/v) of the mixed strain; cultured for 3-30 days at 40 C. with stirring at 50-70 rpm to prepare fermented porcine blood with a solid content of 20 brix and a pH of 4.0 to 4.3.

25. The method of claim 24, further comprising: mixing the fermented porcine blood at a 1:1 weight ratio with a mixture of rice bran powder and soybean meal powder to prepare a secondary fermentation product; and drying the secondary fermentation product, wherein the rice bran powder and the soybean meal powder are mixed at a 1:1 weight ratio.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG. 1 shows changes in body weight during the experimental period in a simple water-tank experiment;

[0044] FIG. 2 shows changes in total length during the experimental period in a simple water-tank experiment; and

[0045] FIGS. 3A and 3B show changes in total length and body weight during the experimental period in a field application experiment.

DETAILED DESCRIPTION

[0046] Hereinafter, a detailed description will be given of the present disclosure through the following examples. However, these examples are not to be construed as limiting the scope of the present disclosure.

<Examples> Preparation of Fermented Porcine Blood and Experiment on Effect of Fermented Porcine Blood as Feed Additive for Farmed Fish

[0047] 1. Preparation of Primary Fermentation Product of Porcine Blood

[0048] Porcine blood was obtained from the Jeju Pig-Breeding Nonghyup Livestock Cooperative Processing Center on Jeju Island, and the blood sample was added with 10% (v/v) of a 4% sodium citrate solution, serving as a blood anticoagulant, and was stored frozen until the experiment.

[0049] The frozen blood sample was thawed at room temperature, added with 1% (w/v) of a proteolytic enzyme Alcalase (Novozyme, Co.) and with 6% (w/v) of sugar as a carbon source, mixed therewith, inoculated with 6% (v/v) of a mixed strain of a Lactobacillus johnsonii (this strain was isolated and identified by the inventors of the present invention) culture solution and a Saccharomyces cerevisiae KCTC 7083 culture solution, and cultured for 3 days to 30 days while performing hydrolysis with stirring at 50-70 rpm in the temperature range of 40 C.10 C., thus manufacturing fermented porcine blood.

[0050] The fermented porcine blood thus manufactured had a solid content of 20 brix, and the pH thereof was 4.3 after culture for 3 days and was maintained at 4.0 after culture for 4 days, and thus long-term storage thereof for about 12 months at room temperature was determined to be possible.

[0051] Based on the results of analysis of components thereof, heavy metals such as lead, mercury and the like were not detected, and antibiotics were not detected. These results were found to satisfy the Standards and Specifications of Feed and the like as set forth by the Ministry of Agriculture, Food and Rural Affairs.

[0052] 2. Preparation of Secondary Fermentation Product of Porcine Blood

[0053] The concentrate (solid content: 30 brix) of the fermented porcine blood (pH 4.0) obtained through culture for 30 days in Preparation Example 1 was mixed at a weight ratio of 1:1 with a combination of rice bran powder and soybean meal powder at the same weight ratio (1:1) (the fermented porcine blood and the combination were mixed at a weight ratio of 1:1), and the resulting mixture was cultured at a temperature of about 30 C. for 7 days and then dried in a vacuum oven, thus manufacturing a secondary fermentation product of porcine blood.

[0054] 3. Simple Water-Tank Experiment

[0055] 3.1 Experimental Method

[0056] 3.1.1 Breeding Method

[0057] The experimental fish that was used was flatfish, and the sample that was used was the primary fermentation product of porcine blood (liquid, 20 brix). Depending on the amount of the primary fermentation product of porcine blood that was added to the compound feed, experimental groups were divided into 0% (untreated group added with compound feed alone, control), and 1%, 2% and 3% (w/w) treatment groups, the flatfish were reared in a continuous flow-through cylindrical water tank (90 L), and the experiment was conducted.

[0058] At the start of the experiment, the total length and body weight of flatfish were 17.80.2 cm and 38.91.1 g, respectively. 30 flatfish were placed in each water tank, after which acclimation was carried out in a manner in which only feed was provided for a week before the start of the experiment, and the experiment was then conducted.

[0059] During the experimental period, the compound feed was provided twice a day (09:00 AM and 17:00 PM), and the primary fermentation product of porcine blood was provided in a mixture with the compound feed.

[0060] During the experimental period, the water temperature and dissolved oxygen content were measured every day using a dissolved oxygen meter (YSI-85).

[0061] 3.1.2 Measurement of Extent of Growth Promotion, Feed Efficiency, Etc.

[0062] The total length and body weight of the experimental fish were measured every 4 weeks after the start of the experiment, and the feed supply and the feed intake were measured. The total weight gain, feed coefficient, and food conversion rate were calculated using the following equations.

Total weight gain=Total body weight at starttotal body weight at end

Feed coefficient=Feed supply/total weight gain

Food conversion rate={(Average body weight at end of experiment)(average body weight at start of experiment)}/(feed intake)100

[0063] For the measured total length and body weight, a one-way variance analysis (ONE-WAY ANOVA) was performed to verify the significance at the 5% level, and whether the difference between the averages was significant was determined.

[0064] 3.2 Experimental Results

[0065] 3.2.1 Breeding Environment and Viability

[0066] During the experimental period, the breeding water temperature fell in the range of 16.0 to 24.7 C. (average water temperature: 21.10.1 C.), and the dissolved oxygen was 4.5 to 7.7 mg/L (average dissolved oxygen: 6.30.4 mg/L). During the breeding period, no individuals died in any of the experimental groups.

[0067] 3.2.1 Extent of Growth

[0068] (1) Change in Body Weight

[0069] The average body weight at the start of the experiment was 39.20.9 g for the control, 38.81.1 g for the 1% treatment group, 38.21.3 g for the 2% treatment group, and 39.51.1 g for the 3% treatment group. Also, the average body weight on the 4th week was 70.23.8 g for the control, 76.23.6 g for the 1% treatment group, 80.44.8 g for the 2% treatment group, and 87.13.7 g for the 3% treatment group. The growth of flatfish was significantly high in the 3% treatment group on the 4th week. The average body weight on the 8th week was 114.37.0 g for the control, 127.94.8 g for the 1% treatment group, 134.98.0 g for the 2% treatment group, and 135.46.5 g for the 3% treatment group. In the 2% and 3% treatment groups on the 8.sup.th week, significantly high results appeared (P>0.05). The results thereof are shown in FIG. 1.

[0070] (2) Change in Total Length

[0071] The average total length at the start of the experiment was 17.80.2 cm for the control, 17.70.2 cm for the 1% treatment group, 17.80.2 cm for the 2% treatment group, and 17.70.2 cm for the 3% treatment group. The average total length on the 4th week was 19.70.3 cm for the control, 20.20.3 cm for the 1% treatment group, 20.60.3 cm for the 2% treatment group, and 20.90.3 cm for the 3% treatment group. The effect of increase in the total length on the 4th week was significantly high in the 2% and 3% treatment groups. The average total length on the 8th week was 22.40.5 cm for the control, 23.60.3 cm for the 1% treatment group, 23.70.4 cm for the 2% treatment group, and 23.80.3 cm for the 3% treatment group. All of the treatment groups on the 8th week were significantly longer than the control (P>0.05). The results thereof are shown in FIG. 2.

[0072] (3) Feed Efficiency

[0073] The average feed intake was 2108.8 g for the control, 2093.7 g for the 1% treatment group, 2272.5 g for the 2% treatment group, and 2279.4 g for the 3% treatment group. Here, in the 2% and 3% treatment groups, the feed intake tended to be somewhat higher.

[0074] The food conversion rate was 3.6% for the control, 4.3% for the 1% treatment group, 4.3% for the 2% treatment group, and 4.2% for the 3% treatment group. The food conversion rate appeared high in all of the treatment groups.

[0075] The results for feed efficiency are summarized in Table 1 below.

TABLE-US-00001 TABLE 1 Classification Control A-1% B-2% C-3% Feed intake (g) 2108.8 2093.7 2272.5 2279.4 Feed conversion rate 3.6 4.3 4.3 4.2

[0076] 4. Field Application Experiment

[0077] 4.1 Experimental Method

[0078] 4.1.1 Breeding Method and Water Quality Environment

[0079] The field application experiment was conducted at the Haeyeon Fish Farm (located in Gujwa-eup, Jeju-si, Jeju Island).

[0080] The sample that was used was the secondary fermentation product of porcine blood.

[0081] In a control, general compound feed not added with the secondary fermentation product of porcine blood was provided, and in a treatment group, compound feed added with 4% (w/w) of the secondary fermentation product of porcine blood was provided. Other feed additives, drugs and the like were equally applied to the control and the treatment group.

[0082] As for the experimental fish, 80,000 flatfish fry (8-10 cm) were equally divided into 40,000 fish for the control and 40,000 fish for the treatment group.

[0083] The experimental water tank was a 10 m10 m tank, and 5 water tanks for the treatment group and 5 water tanks for the control were used.

[0084] The experimental period was 3 months, and a total of 6 tons of compound feed, corresponding to 2 tons per month, was used (3 tons for the control and 3 tons for the treatment group).

[0085] The seawater that was supplied to the experimental farm was sub-surface seawater, in which water characteristics such as water temperature, salinity, pH and the like were constant. Based on the results of on-site water quality measurement during the experimental period, the water temperature was 16.51.0 C., the salinity was 321.0 psu, the dissolved oxygen (DO) was 9.231.25 mg/L, and the pH was 8.020.24.

[0086] Moreover, the results of measurement of water quality of the experimental farm during the experimental period are shown as average values in Table 2 below.

TABLE-US-00002 TABLE 2 Experimental results No. Test items Unit Raw water Control (B5) Treatment group (B3) 1 General bacteria CFU/mL 8.3 3.1 12.7 3.1 26.0 14.7 (Total Colony Counts)* 2 Hydrogen ion concentration (pH) 7.80 0.10 7.80 0.10 7.87 0.06 3 Nitrate nitrogen (NO.sub.3N) mg/L Not detected Not detected Not detected 4 Nitrite nitrogen (NO.sub.2N) mg/L Not detected Not detected Not detected 5 Ammonia nitrogen (NH.sub.3N) mg/L 0.07 0.00 0.87 0.03 0.08 0.01 *General bacteria include non-pathogenic bacteria, pathogenic bacteria, etc.

[0087] As is apparent from Table 2, for most of the items, it was determined that there was no change in water quality characteristics for the raw water, the control and the treatment group. Here, the total colony count in the treatment group was appropriately doubled compared to that of the control, but the total colony count in all of the experimental groups for 3 months was maintained at 30 CFU/mL or less.

[0088] 4.1.2 Measurement of Extent of Growth Promotion, Feed Efficiency, Etc.

[0089] Flatfish were divided into a treatment group and a control, and 40,000 flatfish in each group were reared. At the start of the experiment, flatfish were classified so that there was no significant difference in size between the groups. The flatfish were bred for a total of 12 weeks. In order to investigate changes in body growth, the total length and the body weight were measured a total of 8 times, including at the start of the experiment and on the 2.sup.nd, 4.sup.th, 6.sup.th, 8.sup.th, 10.sup.th and 12.sup.th weeks of the experiment. Measurement was performed using 100 flatfish randomly selected from each group.

[0090] The measured total length and body weight of flatfish were analyzed for significant differences (P<0.05) through independent sample t-test analysis using the statistical processing program SPSS 21.0.

[0091] In addition, the growth rate, feed coefficient, and feed efficiency of flatfish were calculated using the following equations.

Growth rate=(Fish body weight at endfish body weight at start)/fish body weight at start100(%)

Feed efficiency=Total supply/(fish body weight at endfish body weight at start)

Feed coefficient=10/(feed efficiency10)100(%)

[0092] 4.2 Experimental Results

[0093] 4.2.1 Change in Body Weight

[0094] At the start of the experiment, the body weight was 10.090.12 g in the treatment group and 10.060.17 g in the control, based on which there was determined to be no significant difference between the groups. From the 8th week of the experiment, there was a significant difference between 52.050.82 g in the treatment group and 46.850.93 g in the control, indicating that the treatment group exhibited 11.10% higher total weight gain than the control. On the 10th week, the treatment group and the control were 68.100.95 g and 60.950.92 g, respectively, indicating that the total weight gain of the treatment group was 11.73% higher. At the end of the experiment, the treatment group and the control were 78.020.91 g and g, respectively, indicating that the total weight gain of the treatment group was higher. The results thereof are shown in FIG. 3.

[0095] 4.2.2 Change in Total Length

[0096] At the start of the experiment, the total length of the treatment group was 9.360.15 cm, and the total length of the control was 9.390.12 cm, based on which there was determined to be no significant difference between the groups. At the end of the experiment, the total lengths of the treatment group and the control group were 19.490.36 cm and 18.640.36 cm, respectively. Although there was no significant difference between the groups, the total length of the treatment group was greater than that of the control. The results thereof are shown in FIG. 3.

[0097] 4.2.3 Growth Rate, Feed Coefficient and Feed Efficiency

[0098] Based on the results of measurement of the growth rate, feed coefficient, and feed efficiency of the treatment group and the control at the end of the experiment, the growth rate of the treatment group was 563.11%, which was higher than 499.40% for the control. The feed coefficient of the treatment group was 2.36, which was lower than 2.67 for the control. The feed efficiency of the treatment group was 42.29%, which was higher than 37.41% for the control. The results thereof are summarized in Table 3 below.

TABLE-US-00003 TABLE 3 Classification Growth rate (%) Feed coefficient Feed efficiency Treatment group 563.11 2.36 42.29 Control 499.40 2.67 37.41

[0099] Although preferred embodiments of the present disclosure have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications are possible without departing from the scope and spirit of the disclosure as disclosed in the accompanying claims, and such modifications should not be understood separately from the technical ideas or essential characteristics of the present disclosure.