EP3727018A1

EP3727018A1 - Use of dha biomass, ara residue and corn gluten meal in white shrimp feed

Info

Publication number: EP3727018A1
Application number: EP18833028.6A
Authority: EP
Inventors: Bernard Pora; Ling Lu; Chaolan TANG; Jia Li
Original assignee: Roquette Freres SA
Current assignee: Roquette Freres SA
Priority date: 2017-12-20
Filing date: 2018-12-19
Publication date: 2020-10-28
Also published as: WO2019121878A1; CN111386047A; CN109938193A

Abstract

Feed for aquaculture comprising fish meal, corn gluten meal, and a source of docosahexaenoic acid.

Description

Use of DHA biomass, ARA residue and corn gluten meal in White shrimp feed

The present invention relates to the field of aquaculture and feed for fish and crustaceans of breeding. Feeds used in the field of aquaculture are traditionally made from fish meal as a protein source.

Fish meal is an expensive ingredient which production is not ideal for the preservation of the environment.

As sustainable development arises, it is therefore necessary to find alternatives to animal meal and especially fish meal.

In order to spare the production lines already in place, these alternatives must be able to only partially replace fish meal, and are to be used in combination with fish meal as a protein source.

Indeed, as part of a farm, it is good not to change feeding of fish and/or crustaceans too drastically. In addition, it is good to avoid a sudden drying up the fish meal production lines.

It is known to use soybean meal, poultry by-product meal, wheat gluten, soy protein isolate, sorghum, rapeseed meal, and even peas as protein sources for feed for aquaculture. However, these ingredients are rather expensive and lend themselves poorly to a partial substitution of fish meal, due to their imbalance essential amino acids, limited amounts of highly unsaturated fatty acids, anti-nutritional factors and/or toxins.

The partial replacement of fish meal by com gluten meal is advantageous economically speaking. Com gluten meal is a by-product from the wet milling of com kernels to produce native starch. Com gluten meal normally contains 55-64% protein, 15-25% starch, 0-3% extractable lipid, and 0-3% crude fiber. Indeed, com gluten meal is an underexploited by-product of the food industry, which production has a low impact on the environment and that also happens to be cheap. However, partial replacement of fish meal by com gluten meal alone led to a sharp decline in the survival and growth rates of the fed fish and/or crustaceans.

An object of the invention is therefore to provide a feed for aquaculture based on fish meal comprising a cheap substitute to fish meal, which production is environment-friendly, and which has no significant impact on the rate of survival of the fed fish and/or crustaceans.

For this purpose, it is provided a feed for aquaculture comprising fish meal, com gluten meal, and a source of docosahexaenoic acid.

The combined use of com gluten meal and docosahexaenoic acid (DHA) has a synergistic effect which allows limiting the negative effects of com gluten meal taken alone on the survival and growth rates of fed fish and/or cmstaceans fed.

The docosahexaenoic acid used in the feed according to the present invention is preferably obtained through algal biomass. A preferred way of obtaining the docosahexaenoic acid is thought whole cell powder of Schizochytrium sp., which is obtained through fermentation, followed by centrifugation to remove excess water and a further drying technology, such as spray drying . An example of obtaining Schizochytrium sp. biomass powder is mentioned in CN104968779A. The Schizochytrium sp. biomass contains at least 12% DHA.

Preferably, the feed for aquaculture according to the invention further comprises a source of arachidonic acid (ARA).

The arachidonic acid used in the feed according to the present invention is preferably through the addition of the residue obtained after the cmde oil extraction of a dried and washed Mortierella alpine biomass cells. The Mortierella alpine biomass cells are obtained through fermentation followed by plate pressure filtration and a further drying technology, such as fluidized bed drying. The cmde oil was extracted from the dried biomass cells using solvent, such as hexanes, and the obtained oil high in ARA is further refined and used for ARA fortification in infant formula. The Mortierella alpine biomass residue after the cmde oil extraction normally contains 1-6% ARA and 30-40% protein, and hence it can also act as another protein source in the feed.

Mortierella alpine biomass residue (a source of arachidonic acid and protein) also has an effect synergistic with the combined use of com gluten meal and docosahexaenoic acid. Indeed, Mortierella alpine biomass residue improves greatly the survival and growth rate of the fish and/or crustaceans fed by the feed according to the invention.

Mortierella alpine biomass residue is a by-product of food industry and as such, it is very advantageous to use it and to value it. The ratio of Mortierella alpine biomass residue comprised in the present invention preferably ranges from 0 to 10% in weight.

The feed for aquaculture according to the present inventions preferably comprises from 1% to 15 % in weight of com gluten meal.

The feed for aquaculture according to the present inventions preferably comprises from 1% to 5% in weight of Schizochytrium sp. biomass.

The feed for aquaculture according to the present inventions preferably comprises fish meal, such as Menhanden, Pern fish meal, and white fish meal.

The feed for aquaculture according to the present inventions is preferably devoid of additional fish oil, such as Menhaden or fish liver oil, including cod liver oil.

Another object of the present invention is a use of a feed for aquaculture according to the present invention to feed shrimps, preferably shrimps chosen amongst Penaeus monodon and Litopenaeus vannamei.

Another object of the present invention is a use of a feed for aquaculture according to the present invention to feed fish, preferably fish chosen among Japanese seabass, gilthead seabream, Solea senegalensis, turbot, common carp, Paralichthys olivaceus, and puffer. Another object of the present invention is a use of a combination of com gluten meal and a source of docosahexaenoic acid as a substitute for fish meal, preferably for aquaculture. Advantageously, such combination can also comprise a source of arachidonic acid. The invention can be better understood at the reading of the detailed examples below, which constitute non-limitative embodiments of the present invention and which illustrate the properties of the food according to the present invention.

Seven different samples of feeds were prepared in order to test the properties of the feeds according to the present invention, among which were two control compositions with low and high contents of fish meal. The protein content of the low fish meal control was compensated by adding com gluten meal. In order to prepare the samples, the major components of the ingredients were smashed and passed through an 80-mesh sieve (180 pm openings), and gradually mixed with the micro components (such as vitamins and minerals). The mixture was granulated using an electric meat grinder, dried at 80°C in an oven , smashed and passed through a 40-mesh sieve (425 pm openings) for feed at early growth stage and passed through a 20-mesh sieve (850 pm openings) for feed at latter growth stage. The feed samples were kept at 4°C until they were used, where they were mixed with water to obtain a dough consistency for feeding.

The five other feeds, which are also referred to as trial samples, comprised com gluten meal and different amounts of Schizochytrium mangrovei biomass as a source of DHA, which was obtained through fermentation, followed by centrifugation to remove excess water and spray drying as mentioned in CN104968779A. The Schizochytrium mangrovei biomass contained 12% DHA. The other ingredients were supplied by Hangzhou Haihuang Feed Development Co., Ltd., Hangzhou, China. Two groups further comprised Mortierella alpine biomass residue as a source of ARA, as per illustrated in Table 1. The Mortierella alpine biomass residue was obtained through, in sequence, fermentation, plate pressure filtration, fluidized bed drying, and finally solvent extraction. The Mortierella alpine biomass residue after the crude oil extraction contained 4% ARA and 36% protein.

Chromic oxide (0.5%) was mixed with a 1 diets to determine apparent digestibility coefficients.

Table 1

In Table 1, one kilogram of vitamin mix contained vitamin A 100,000 IU, vitamin D 200,000 IU, tocopherol acetate 3 g, menadione 1 g, thiamine 0.5 g, riboflavin 1.5 g, nicotinic acid 4 g, Ca-pantothenate2.5 g, pyridoxine 0.8 g, vitamin B12 2 mg, folic acid 0.25 g, biotin 8 mg, and inositol 15 g. One kilogram of mineral mix contained Ca(H₂P0₄) 2,600 g, KCr(S0₄) 20.55 g, CuC0₃ 0.3 g, FeC₆H₅0₇ 10 g, MgO 30 g, MnS0₄ 3.5 g, C₆H₅K₃0rH₂0 220 g, KI 0.02 g, K₂S0₄ 52 g, NaCl 74 g, Na₂SeO 30.02 g, and ZnC0₃ 3 g.

The properties of the seven trial samples were analyzed and are reported in Tables 2 and 3.

Table 2 shows the contents of crude proteins (CP), crude fat (EE or ether extract), crude ash, water and gross energy (GE) of the seven trial samples. GE was a calculated value and others were measured values. GE (MJ/kg dry matter) = 23.4 x CP (%) + 39.2 x EE (%) + 17.2 x CARB (%), where CARB (%) = 1 - (CP (%) + EE (%) + Moisture (%)).

The water content (moisture) was determined by drying method following GB /T 6435-2014. The crude ash content was determined at high temperature (550°C) following GB /T 6438-2007.

Crude protein (N = 6.25) was determined using an automatic Kjeltec 2300 (Foss Tecator AB, Hoganas, Sweden). Crude lipid was determined by Folch method. Sample was extracted using Folch solution at a concentration of 20 mL/g sample, where the Folch solution contained 2 parts by volume of chloroform and 1 part by volume of methanol. After standing for 24 h, 10 mL of the Folch solution was filtered into a preweighed glass tube, and was mixed with 2 mL 0.4% CaCl₂. After standing for 30 minutes, additional 2 mL of chloroform-methanol- water solution (8:4:3 by volume) was used to rinse the tube wall three times. The solutions were combined and then dried in a vacuum drying oven (DZF-6050; Boxun Industry & 5 Commerce Co., Ltd., Shanghai, China). The crude fat content was determined gravimetrically, and the lipids were resolubilized in 10 mL chloroform and stored at -20°C until further analysis.

The different superscript letters behind the numbers within the same line represent significant different at p < 0.05.

10 Table 2

Table 3 shows the essential amino acid contents of the seven trial samples. The proteins in the feed samples were completely hydrolyzed using hydrochloric acid and the amino acid was analyzed using an automatic amino acid analyzer S- 433D (Sykam Co., Germany).

The different superscript etters behind the numbers within the same line represent significant different at p < 0.05.

Table 3

The seven trial samples were tested on White shrimp

( Litopenaeusvannamei ), which has been selected because of its remarkably stable weight distribution (2.45±0.l0g).

Three replicates of each feeding were performed. For each replicate, white shrimps were kept at a density of 40 shrimps per cage, which had a volume of one cubic meter. Every morning, the uneaten feed and feces were be removed, and the

dead shrimps were counted and removed. The water quality was monitored at room

temperature and pH was maintained at 8.0+0.5. The dissolved oxygen concentration

was maintained above 6.0 mg/L, and total ammonia nitrogen was kept under 0.03

mg/L.

The measurements lasted for 55 days. The shrimps were fed thrice a day, at

fixed hours: 7:00, 12:00 and 17:00. The shrimps were weighed every 2 weeks, and

the feeding rate was adapted based on the weight of the shrimps (5-10%). The

uneaten feed was collected at one hour after each feeding, and the dry weight was

obtained after drying. Feces collection was performed two hours after feeding. The

amounts of protein, amino acid, and chromic oxide (a marker for digestibility) of

dried fecal and feed samples were analyzed. At the end of the experiment, the

shrimps were weighed.

Different parameters are reported in Table 4, as follows.

• Initial body weight per shrimp = (Body weight of total shrimps at the

beginning of the trial)/ Number of total shrimps at the beginning trial

• Final body weight per shrimp = (Body weight of total shrimps at the end

of the trial)/Number of total shrimps per cage

• Body weight gain per shrimp = Final body weight gain - Initial body

weight

• Specific growth rate = 100*(1h average (Final body weight) - ln average

(Initial body weight))/Total trial dates

• Feed conversion ratio = Feed intake per shrimp/Body weight gain per

shrimp, wherein the feed intake per shrimp is the sum of daily feed intake over the final number of total shrimp per cage/trial dates

• Survival rate = (Number of total shrimps at the beginning trial - Number

of total shrimps at the end of trial)/Number of total shrimp at the

beginning of the trial per cage

Group Initial body Final body Final body Specific Feed conversion Survival rate weight weight weight gain growth rate ratio (%)

(g/shrimp) (g/shrimp) (g/shrimp) Group 1 (low 2.45±0.10 9.05±0.25^b 6.60±0.32^b 2.21±0.10^b 1.49±0.06^! 53.89±2.55' fish meal)

Group 2 (high 2.44±0.07 10.09±0.17^a 7.65±0.18^a 2.41±0.05^a 1.14±0.07^d 68.89±3.47^abc fish meal)

Group 3 (1% 2.42±0.06 9.19±0.34^b 6.77±0.37^b 2.26±0.09^b 1.35±0.03^b 65.55±2.54^bc biomass)

Group 4 (2% 2.39±0.05 9.78±0.11^a 7.39±0.10^a 2.39±0.02^a 1.33±0.04^bc 66.66±1.67^bc biomass)

Group 5 (5% 2.41±0.06 10.01±0.21^a 7.60±0.22^a 2.41±0.06^a 1.25±0.05^c 70.55±2.54^ab biomass)

Group 6 2.41±0.09 9.85±0.22^a 7.44±0.13^a 2.38±0.03^a 1.29±0.02 73.33±4.41^a Group 7 2.42±0.04 9.12±0.09 6.69±0.09 2.24±0.03 1.35±0.04 63.33±4.41

The different superscript letters behind the numbers within the same column

represent significant different at p < 0.05.

Table 4

All statistical analysis were performed using SPSS 11.5 software. The data were compared by ANOVA. Where ANOVA revealed significant differences, the

Duncan’s multiple tests were applied so as to determine the significant difference between treatments. Statistically significant were determined at p < 0.05. The results of palatability were tested by /-test.

As it is shown in Table 4, the final body weight gains and the specific growth rates of Groups 4, 5, and 6 were significantly higher than those of the Group

1 (low fish meal control). The feed conversion ratios of Groups 3, 4, 5, 6, and 7were significantly lower than that of the Group 1 (low fish meal control), but were significantly higher than of the Group 2 (high fish meal control). Most importantly, the survival rates of Group 5 and 6 were significantly higher than that of the Group 1

(low fish meal control).

The result showed that Schizochytrium mangrovei biomass and Mortierella alpine biomass residue significantly improved the survival rate of the shrimps.

Apparent digestibility coefficients (ADC) of dry matter, lipid, protein, and amino acids were analyzed and results are reported in Table 5. Chromic oxide (0.5%) was mixed with all diets to determine apparent digestibility coefficients. Fecal samples were collected via a siphonage approach every day during the final 3 weeks. Fresh excrements with intact envelops were selected, centrifuged (3,200 rpm at 6°C for 15 minutes), pooled, dried, and ground. Proximate (AO AC 1990) and chromic oxide analyses (Bolin et al. 1952) were performed on the feed and fecal samples. The samples were analyzed for total crude protein (micro -Kjeldahl method, nitrogen analyzer, Fison instrument, N = 6.25), crude fat (dichloromethane extraction by Soxhlet method). ADC (%) = [1 - (feed Cr level /fecal Cr level)* (fecal nutrient/feed nutrient)] *100. Groups 3, 4, 5, and 6 show an improvement in the digestibility of crude proteins, crude fat, and dry matter compared to Group 1 (low fish meal control). There were no significant differences in the digestibility of the dry matter and crude proteins between Group 6 and the Group 2 (high fish meal control), but the digestibility of the crude fat of the Group 6 was significantly higher than that of the Group 2 (high fish meal control).

Digestibility of essential amino acids was significantly higher in Groups 3, 4, 5, and 6 than that of Group 1 (low fish meal control). There were no significant differences of the essential amino acids between Group 6 and the Group 2 (high fish meal control), except for ARG. The digestibility of Group 4 was lower than that of Group 6, but both of them were higher than that of Group 7.

Schizochytrium sp. biomass and Mortierella alpine biomass residue used in the feed according to the present invention therefore improve the digestibility of crude fat, crude protein and essential amino acids, so as to make up for the shortcomings of com gluten meal in low fish meal feed.

The different superscript letters numbers within the same line represent significant different at p < 0.05.

Table 5 It is understood that the described embodiments are not restrictive and that it is possible to make improvements to the invention without leaving the framework thereof. Thus, we can for example provide additional additives or modify the conditioning of the feed without departing from the scope of the present invention. Although exemplary absolute percentages of com gluten meal, Schizochytrium sp. biomass, and Mortierella alpine biomass residue are given all along the present description, the synergetic effects of these components also depend on their relative proportions: e.g. it is understood that a feed for aquaculture comprising less than twice as much Mortierella alpine biomass residue as Schizochytrium sp. biomass whilst containing com gluten meal would not depart from the scope of the invention.

Unless otherwise specified, the word “or” is equivalent to “and/or”. Similarly, the word 'one' is equivalent to 'at least one' unless the contrary is specified. Unless otherwise specified, all percentages are weight percentages.

Claims

1. Feed for aquaculture comprising fish meal, com gluten meal, and a source of docosahexaenoic acid.

2. Feed for aquaculture according to claim 1 further comprising a source of arachidonic acid.

3. Feed for aquaculture according to claim 1 or 2 comprising 0 to 10 % in weight of a source of arachidonic acid.

4. Feed for aquaculture according to any preceding claim comprising 1% to 15 % in weight of com gluten meal.

5. Feed for aquaculture according to any preceding claim including 1% to 5 %

Schizochytrium sp. biomass.

6. Feed for aquaculture according to any preceding claim, wherein the fish meal is chosen among Mehanden, Pern fish meal, and white fish meal

7. Feed for aquaculture according to any preceding claim being devoid of additional fish oil or fish liver oil.

8. Use of a feed for aquaculture according to any preceding claim to feed shrimp, preferably shrimp chosen amongst Penaeus monodon and Litopenaeus vannamei.

9. Use of a feed for aquaculture according to any of claims 1 to 7 to feed fish, preferably fish chosen among Japanese seabass, gilthead seabream, Solea senegalensis , turbot, common carp, Paralichthys olivaceus, and puffer.

10. Use of a combination of com gluten meal and Schizochytrium sp. biomass as a substitute for fish meal and fish oil, preferably for aquaculture.