A functional gel feed for Anthocidaris crassispina and its preparation method

Abstract

The present invention discloses a functional gel feed for Anthocidaris crassispina and a preparation method thereof. Among them, the mass fraction of 5-HT in the functional gel feed is 0.05-0.5‰, and the energy of the functional gel feed is 1,800-2,000 kcal / kg; after feeding the Anthocidaris crassispina with the functional gel feed for 20-40 days, the content of vitellin in the Anthocidaris crassispina is 400-497 mg / g; and the gonadosomatic index is 9-13%. On the one hand, the functional gel feed of the present invention has a balanced nutrition, is rich in various nutrients required for the growth of Anthocidaris crassispina, can meet the growth and development needs of Anthocidaris crassispina, has a high feed digestibility, can reduce fecal emissions, and reduce the pollution of aquaculture to the environment. On the other hand, the feed is in a gel state, has good palatability, and is easy for Anthocidaris crassispina to ingest. In addition, the functional gel feed of the present invention has good stability in water, is not easy to disperse, and can improve the utilization rate of the feed.

Description

Technical Field

[0001] The present invention relates to the technical field of aquaculture feeds, and more particularly to a functional gel feed for Anthocidaris crassispina and a preparation method thereof. Background Art

[0002] Anthocidaris crassispina is an important marine economic organism and has high value in aquaculture. Currently, the cultivation of Anthocidaris crassispina mainly relies on natural bait. On the one hand, there are problems such as unstable supply and unbalanced nutritional components in natural bait. Moreover, one of the most commonly used methods in the prior art is to purchase natural kelp from Changdao and Daqin Island in Shandong. For areas far away, it is not only inconvenient to purchase, but also costly. On the other hand, the artificial feeds in the prior art also have disadvantages such as poor palatability and low digestibility, which are not conducive to the growth and development of Anthocidaris crassispina. Summary of the Invention

[0003] The purpose of the present invention is to provide a functional gel feed for Anthocidaris crassispina with balanced nutrition, good palatability and high digestibility to meet the needs of Anthocidaris crassispina cultivation.

[0004] The present invention provides an application of 5-HT in increasing the vitellin content and gonadosomatic index of Anthocidaris crassispina.

[0005] Further, the application is to prepare a functional gel feed for Anthocidaris crassispina by adding 5-HT. The mass fraction of 5-HT in the functional gel feed is 0.05 - 0.5‰, and the energy of the functional gel feed is 1800 - 2000 kcal / kg; the mass fraction of crude protein in the functional gel feed is 40 - 45%, the mass fraction of crude fat is 7 - 10%, the mass fraction of crude fiber is 4 - 6%, the mass fraction of water is 40 - 45%, and the balance is other components, and the other components include vitamins and minerals;

[0006] After feeding Anthocidaris crassispina with the functional gel feed for 20 - 40 days, the vitellin content of Anthocidaris crassispina is 400 - 497 mg / g; and the gonadosomatic index is 9 - 13%.

[0007] The 5-HT is 5-hydroxytryptamine, which is an indole derivative with the chemical formula C 10 H 12 N 2 O and a molecular weight of 176.22. The definition of crude protein is the nitrogen content in the feed sample multiplied by the coefficient 6.25. Most proteins generally contain 16% nitrogen, and this coefficient is derived from this. Therefore, multiplying the percentage of nitrogen in the feed by 100÷16, or multiplying by 6.25, can calculate the crude protein content.

[0008] Serotonin (5-HT), although an important neurotransmitter in the human body and also known as serotonin, can promote gonad maturation and increase the content of vitellin in sea urchins. Serotonin and its receptors can regulate follicle development, oocyte maturation, and early embryonic development. Serotonin participates in a wide range of physiological activities by activating multiple signaling pathways through its interaction with its receptor family. In sea urchins, serotonin may affect the synthesis and accumulation of vitellin by influencing the maturation process of oocytes. Moreover, during the cultivation of purple sea urchins, using the functional gel feed described in the present invention containing 0.05 - 0.5‰ of 5-HT and 40 - 45% of crude protein can ensure sufficient vitellin content and no excessive hormones in the purple sea urchins, thus ensuring that purple sea urchins are both nutritious and safe as food. If the 5-HT content in the functional gel feed is too low, the purpose of cultivating and fattening purple sea urchins cannot be achieved, while conversely, if the content is too high, it will have an adverse effect on the human body as food.

[0009] The functional gel feed is nutritionally balanced, and it is particularly necessary to ensure an appropriate amount of crude protein. If the mass fraction of crude protein is less than 40, the protein source for the growth of sea urchin gonads and the synthesis of vitellin cannot be guaranteed, which is not conducive to the purpose of cultivation and fattening; conversely, if the mass fraction of crude protein is higher than 45%, most of the excess protein will enter the environment in the form of feces, which does not meet the comprehensive goals of environmental benefits and economic benefits.

[0010] The present invention also provides a functional gel feed for purple sea urchins, and the functional gel feed is the above-mentioned functional gel feed;

[0011] The functional gel feed, by weight, comprises the following components: 40 - 60 parts of protein source, 30 - 40 parts of carbohydrate source, 20 - 30 parts of carrots, 3 - 4 parts of mineral mixture source, 0.05 - 0.1 part of 5-HT, and 6 - 10 parts of gel former;

[0012] Among them, the protein source is a combination of fish meal and soybeans; the carbohydrate source is corn starch; the gel former includes gelatin or sodium alginate.

[0013] Furthermore, the weight ratio of corn starch, soybeans, and carrots in the functional gel feed of the present invention is (1 - 1.5):(1 - 1.5):(1 - 1.5).

[0014] Furthermore, the crushing particle size of soybeans and carrots in the functional gel feed of the present invention is 20 - 200 microns.

[0015] Furthermore, in the gelling agent of the functional gel feed of the present invention, the weight ratio of gelatin to water is 1:(3 - 5); or the weight ratio of sodium alginate to water in the gelling agent is 1:(75 - 100).

[0016] Furthermore, the mineral mixture source in the functional gel feed of the present invention consists of calcium dihydrogen phosphate and multi-vitamin and multi-mineral. The weight ratio of calcium, phosphorus, magnesium, iron, and zinc in the mineral mixture source is (1 - 1.2):(0.8 - 1):(0.5 - 0.6):(0.02 - 0.05):(0.01 - 0.03), and the mineral mixture source includes vitamin A, vitamin C, vitamin D, vitamin E, and vitamin K.

[0017] Furthermore, in the protein source of the functional gel feed of the present invention, the weight ratio of fish meal to soybean is (1.5 - 2):(1 - 1.2).

[0018] The present invention also provides a preparation method of a functional gel feed for purple sea urchins based on any one of the above, and the specific steps are as follows:

[0019] S1: Selection of materials: Prepare the protein source, carbohydrate source, carrot, and mineral mixture source according to the ratio.

[0020] S2: Processing of the food: First, mix corn starch, crushed soybean, and carrot to obtain a first mixture, then mix the first mixture with fish meal in a mass ratio of 10:(1 - 4) to obtain a second mixture, then add 5-HT to the second mixture, and finally add a gelling agent and stir to obtain the functional gel feed.

[0021] S3: Shaping of the food: Pour the functional gel feed obtained in S2 into a mold, demold after solidification, and cut and roll the demolded functional gel feed into shape.

[0022] S4: Storage of the food: Place the shaped functional gel feed obtained in S3 in a storage container and store it in a freezer at -22~-18°C.

[0023] Furthermore, the stirring speed in step S2 of the preparation method of the present invention is 80 - 100 rpm / min.

[0024] Furthermore, the solidification environment in step S3 of the preparation method of the present invention is to solidify for 300 - 360 min at 2 - 5°C and 50 - 60 min at -18~-22°C.

[0025] The functional gel feed for sea urchin of the present invention has a balanced nutrition. The energy of the functional gel feed is 1800 - 2000 kcal / kg; the mass fraction of crude protein in the functional gel feed is 40 - 45%, the mass fraction of crude fat is 7 - 10%, the mass fraction of crude fiber is 4 - 6%, the mass fraction of water is 40 - 45%, and the balance is others, where the others include vitamins and minerals. The functional gel feed is rich in various nutrients required for the growth of sea urchin, and can meet the growth and development needs of sea urchin. After feeding sea urchin with the functional gel feed of the present invention for 20 - 40 days, the net weight increase of sea urchin is greater than or equal to 2.58 g, and the diameter increase is greater than 20%. And after feeding sea urchin with the functional gel feed for 20 - 40 days, the vitellin content of sea urchin is 400 - 497 mg / g; and the gonadosomatic index is 9 - 13%. The functional gel feed is in gel form. The functional gel feed fed at a feeding amount of 1.5 - 2 g / sea urchin per day is completely eaten by sea urchin within 24 h, and the functional gel feed can maintain the gel form before being eaten up, has good palatability, is easy for sea urchin to ingest, and has good stability in water, is not easy to disperse, and can improve the utilization rate of feed. In addition, the digestibility of the functional gel feed is greater than 85%, which can reduce fecal discharge and reduce the pollution of aquaculture to the environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a sample diagram of the functional gel feed for sea urchin of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0027] In order to have a clearer understanding of the technical features, objectives and effects of the present invention, the specific embodiments of the present invention will be described in detail with reference to the accompanying drawings. The described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention. Those not specified in the specific embodiments are carried out according to the conventional conditions or the conditions provided by the manufacturer.

[0028] When the following description refers to the accompanying drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all the embodiments consistent with the present disclosure. The terms used in the present disclosure are only for the purpose of describing specific embodiments, and are not intended to limit the present disclosure.

[0029] The present invention will be further described in detail with specific embodiments below. DETAILED DESCRIPTION OF THE INVENTION I:

[0031] Application of 5-HT in increasing the vitellin content and gonad index of Strongylocentrotus nudus

[0032] In other embodiments, the application is to prepare a functional gel feed for Strongylocentrotus nudus by adding 5-HT. The mass fraction of 5-HT in the functional gel feed is 0.05 - 0.5‰, and the energy of the functional gel feed is 1800 - 2000 kcal / kg; the mass fraction of crude protein in the functional gel feed is 40 - 45%, the mass fraction of crude fat is 7 - 10%, the mass fraction of crude fiber is 4 - 6%, the mass fraction of water is 40 - 45%, and the balance is other substances, and the other substances include vitamins and minerals;

[0033] As shown in Table 1, after feeding Strongylocentrotus nudus with the functional gel feed for 20 - 40 days, the vitellin content of Strongylocentrotus nudus is 400 - 497 mg / g; and the gonad index is 9 - 13%. Specific Embodiment 2:

[0035] A functional gel feed for Strongylocentrotus nudus, and the functional gel feed is the functional gel feed in the above Specific Embodiment 1, as Figure 1 shown;

[0036] And by weight, the functional gel feed comprises the following components: 40 - 60 parts of protein source, 30 - 40 parts of carbohydrate source, 20 - 30 parts of carrot, 3 - 4 parts of mineral mixture source, 0.05 - 0.1 part of 5-HT, and 6 - 10 parts of gel former;

[0037] Wherein, the protein source is a combination of fish meal and soybean; the carbohydrate source is corn starch; the gel former includes gelatin or sodium alginate.

[0038] In other embodiments, the weight ratio of corn starch, soybean, and carrot in the functional gel feed is (1 - 1.5):(1 - 1.5):(1 - 1.5).

[0039] In other embodiments, the crushing particle size of soybean and carrot in the functional gel feed is 20 - 200 microns.

[0040] In other embodiments, the weight ratio of gelatin to water in the gel former is 1:(3 - 5); or the weight ratio of sodium alginate to water in the gel former is 1:(75 - 100).

[0041] In other embodiments, the mineral mixture source consists of calcium dihydrogen phosphate and multi-vitamin and multi-mineral, and the weight ratio of calcium, phosphorus, magnesium, iron, and zinc in the mineral mixture source is (1 - 1.2):(0.8 - 1):(0.5 - 0.6):(0.02 - 0.05):(0.01 - 0.03). The mineral mixture source includes vitamin A, vitamin C, vitamin D, vitamin E, and vitamin K.

[0042] In other embodiments, in the protein source, the weight ratio of fish meal to soybean is (1.5 - 2):(1 - 1.2).

[0043] In other embodiments, the functional gel feed further includes an antioxidant and a preservative, and the total mass fraction of the antioxidant and the preservative is 0.1% to extend the storage time of the food.

[0044] In other embodiments, the functional gel feed further includes a feeding attractant. Specific Embodiment Three:

[0046] A preparation method of the functional gel feed for purple sea urchins according to any one of the above embodiments, the specific steps are as follows:

[0047] S1: Selection of materials: Prepare the protein source, carbohydrate source, carrot, and mineral mixture source in proportion;

[0048] S2: Processing of the food: First, mix corn starch, crushed soybean, and carrot to obtain a first mixture, then mix the first mixture with fish meal in a mass ratio of 10:(1 - 4) to obtain a second mixture, then add 5-HT to the second mixture, and finally add a gel former and stir to obtain the functional gel feed;

[0049] S3: Shaping of the food: Pour the functional gel feed obtained in S2 into a mold, demold after solidification, and cut and roll the demolded functional gel feed into shape;

[0050] S4: Storage of the food: Place the shaped functional gel feed obtained in S3 in a storage container and store it in a freezer at -22~-18 °C.

[0051] In other embodiments, the stirring speed in step S2 of the preparation method is 80 - 100 rpm / min.

[0052] In other embodiments, the solidification environment in step S3 of the preparation method is to solidify at 2 - 5 °C for 300 - 360 min and at -22~-18 °C for 50 - 60 min.

[0053] Example 1:

[0054] The preparation method of a functional gel feed for Anthocidaris crassispina according to the present invention is as follows:

[0055] S1: Selection of materials: Prepare a protein source, a carbohydrate source, carrots, and a mineral mixture source in proportion; by weight, it includes the following components: 40 parts of the protein source, 30 parts of the carbohydrate source, 20 parts of carrots, 3 parts of the mineral mixture source, 0.05 part of 5-HT, and 6.9 parts of a gelling agent; the mineral mixture source consists of calcium dihydrogen phosphate and multi-vitamin and multi-mineral, and the weight ratio of calcium, phosphorus, magnesium, iron, and zinc is 1:1:0.5:0.02:0.01; the mineral mixture source includes vitamin A, vitamin C, vitamin D, vitamin E, and vitamin K.

[0056] In the protein source, the weight ratio of fish meal to soybeans is 1.5 - 2:1 - 1.2; the carbohydrate source is corn starch; in Example 1, the gelling agent includes gelatin, and the weight ratio of gelatin to water in the gelling agent is 1:3; the weight ratio of corn starch, soybeans, and carrots is 1.5:1:1.

[0057] S2: Processing of the feed: First, mix corn starch, crushed soybeans, and carrots to obtain a first mixture, then mix the first mixture with fish meal in a mass ratio of 10:(1 - 4) to obtain a second mixture, then add 5-HT to the second mixture, and finally add the gelling agent and stir to obtain the functional gel feed; in Example 1, the crushing particle size of the soybeans and carrots is 200 μm, and the stirring speed is 90 rpm / min.

[0058] S3: Shaping of the food: Pour the functional gel feed obtained in S2 into a mold, demold after solidification, and cut and roll the demolded functional gel feed into shape. In Example 1, the solidification environment is to solidify for 360 min at 4°C and 50 min at -20°C, and finally form into 1×1 cm 2 square small pieces with a thickness of 0.1 cm.

[0059] S4: Storage of the food: Place the shaped functional gel feed obtained in S3 in a storage container and store it at -20°C in a freezer.

[0060] In Example 1, the functional gel feed is Sample No. 1, the mass fraction of 5-HT in the Sample No. 1 is 0.5‰, and the mass fraction of crude protein in the functional gel feed is 40%.

[0061] Example 2:

[0062] The difference between Example 2 and Example 1 is only that: the functional gel feed is Sample 2, and the mass fraction of 5-HT in Sample 2 is 0.05‰.

[0063] Example 3:

[0064] The difference between Example 3 and Example 1 is only that: the functional gel feed is Sample 3, and the mass fraction of crude protein in Sample 3 is 42%.

[0065] Example 4:

[0066] The difference between Example 4 and Example 1 is only that: the functional gel feed is Sample 4, and the mass fraction of crude protein in Sample 4 is 45%.

[0067] Example 5:

[0068] The difference between Example 5 and Example 1 is only that: the functional gel feed is Sample 5, the gelling agent in Sample 5 includes sodium alginate, and the weight ratio of sodium alginate to water in the gelling agent is 1:100.

[0069] Comparative Example 1:

[0070] The difference between Comparative Example 1 and Example 1 is only that: the functional gel feed is Sample 6, and Sample 6 does not contain 5-HT.

[0071] Comparative Example 2:

[0072] The difference between Comparative Example 2 and Example 1 is only that: the functional gel feed is Sample 7, and the mass fraction of 5-HT in Sample 7 is 0.01‰.

[0073] Comparative Example 3:

[0074] The difference between Comparative Example 3 and Example 1 is only that: the functional gel feed is Sample 8, and the mass fraction of 5-HT in Sample 8 is 1‰.

[0075] Comparative Example 4:

[0076] The difference between Comparative Example 4 and Example 1 is only that: the functional gel feed is Sample 9, and the mass fraction of 5-HT in Sample 9 is 2‰.

[0077] Comparative Example 5:

[0078] The difference between Comparative Example 5 and Example 1 is only that: the functional gel feed is Sample 10, and the mass fraction of crude protein in Sample 10 is 30%.

[0079] Comparative Example 6:

[0080] The difference between Comparative Example 6 and Example 1 is only that: the functional gel feed is Sample No. 11, and the crude protein mass fraction in Sample No. 11 is 65%.

[0081] Example 6:

[0082] Use the functional gel feeds in the above Examples 1-5 and Comparative Examples 1-6 to culture purple sea urchins respectively. The specific culture environment is: natural seawater, dissolved oxygen is 5.45-5.6 g / L; temperature is 22-25 °C; salinity is 28-33 ppt, and pH is 8.3-8.4.

[0083] Select purple sea urchin samples with a body weight of 60-80 g, a diameter of 2.2-2.3 cm, and a gonad index of 3-5%. Before the culture process of this Example 6, the selected purple sea urchin samples have not been fed with functional gel feed and have only eaten natural kelp.

[0084] First, starve the selected purple sea urchin samples for 7 days, and then feed them once a day according to the feeding amount of 1.5-2 g / sea urchin for 20-40 days. The specific results are shown in Table 1.

[0085] Table 1 Culturing purple sea urchins with the functional gel feeds in Examples 1-5 and Comparative Examples 1-6

[0086]

[0087] The specific measurement method of the vitellin content is as follows:

[0088] Sample preparation: Take a certain amount of vitellin sample and add concentrated sulfuric acid to digest it at high temperature to decompose it into ammonia (NH 3 )

[0089] Distillation of ammonia: Through distillation, absorb the generated ammonia gas into an excessive standard acid solution;

[0090] Titration: Determine the ammonia content in the absorption solution by titration, and thus calculate the protein content.

[0091] The calculation method of the gonad index GSI is: GSI = gonad / body weight.

[0092] Feed digestibility = (food - feces) / food.

[0093] Observing the feeding of purple sea urchins with the functional gel feed during the feeding period, it can be seen that when using the functional gel feeds in Examples 1-5 and feeding them at a feeding rate of 1.5-2 g / sea urchin per day, the purple sea urchins completely consumed them within 24 hours, and the functional gel feeds remained in a gel state before being completely eaten, indicating that the functional gel feeds have good palatability, are easy for purple sea urchins to ingest, and have good stability in water and are not easily dispersed.

[0094] Observing the growth of purple sea urchins during and after the feeding period, it was found that the body weight, diameter, swing amplitude of the spines, activity level of the tube feet, etc. of the purple sea urchins were all in good condition.

[0095] Example 7:

[0096] Purple sea urchins are farmed on a large scale in a farm. The farm includes 4 pools, each pool includes 12-15 nets, the volume of each net is 1-1.2 m³, and the number of purple sea urchins on each net is 150-200. Using the same breeding environment and feeding method as in the examples, it was found that the functional gel feed of the present invention can fully meet the growth and development needs of purple sea urchins, has good palatability, good stability, and high digestibility of purple sea urchins to the functional gel feed.

[0097] When feeding the functional gel feed at a feeding rate of 1.5-2 g / sea urchin per day, the purple sea urchins completely consumed it within 24 hours, and the functional gel feed remained in a gel state before being completely eaten, had good palatability, was easy for purple sea urchins to ingest, had good stability in water and was not easily dispersed, and could improve the utilization rate of the feed. After feeding purple sea urchins with the functional gel feed of the present invention for 20-40 days, the net increase in body weight of the purple sea urchins was greater than or equal to 2.58 g, the diameter increase was greater than 20%, and the digestibility of the functional gel feed was greater than 82%.

[0098] The present invention is illustrated by the above specific embodiments. Those skilled in the art should understand that various transformations and equivalent substitutions can be made to the present invention without departing from the scope of the present invention. The parts not detailed in the specification of the present invention are well-known technologies to those skilled in the art. In addition, various modifications can be made to the present invention for specific situations or circumstances without departing from the scope of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed, but should include all embodiments falling within the scope of the present invention.

Claims

1. A gel feed for purple sea urchin, characterized in that: The gel feed comprises the following components by weight: 40-60 parts of a protein source, 30-40 parts of a carbohydrate source, 20-30 parts of carrots, 3-4 parts of a mineral mixture source, 0.05-0.1 parts of 5-HT, and 6-10 parts of a gel forming agent; Wherein, the protein source is a combination of fish meal and soybean; the carbohydrate source is corn starch; the gel forming agent includes gelatin or sodium alginate; The mass fraction of 5-HT in the gel feed is 0.05-0.5‰, and the energy of the gel feed is 1800-2000 kcal / kg; the mass fraction of crude protein in the gel feed is 40-45%, the mass fraction of crude fat is 7-10%, the mass fraction of crude fiber is 4-6%, the mass fraction of water is 40-45%, and the rest is others, and the others include vitamins and minerals; After the purple sea urchin is fed with the gel feed for 20-40 days, the yolk protein content of the purple sea urchin is 400-497 mg / g, and the gonad index is 9-13%.

2. The gel feed of purple sea urchin according to claim 1, characterized in that: The weight ratio of the corn starch, soybeans and carrots is (1-1.5): (1-1.5): (1-1.5).

3. The gel feed of purple sea urchin according to claim 2, characterized in that: The soybean and carrot have a crushed particle size of 20-200 μm.

4. The gel feed of purple sea urchin according to claim 3, characterized in that: The weight ratio of gelatin to water in the gel forming agent is 1:(3-5); or the weight ratio of sodium alginate to water in the gel forming agent is 1:(75-100).

5. The gel feed of purple sea urchin according to claim 4, characterized in that: The mineral mixture source is composed of monocalcium phosphate and multivitamin. The weight ratio of calcium, phosphorus, magnesium, iron and zinc in the mineral mixture source is (1-1.2): (0.8-1): (0.5-0.6): (0.02-0.05): (0.01-0.03). The mineral mixture source includes vitamin A, vitamin C, vitamin D, vitamin E and vitamin K.

6. The gel feed of purple sea urchin according to claim 5, characterized in that: In the protein source, the weight ratio of fish meal to soybean is (1.5-2): (1-1.2).

7. A method for preparing a gel feed of purple sea urchin based on any one of claims 1 to 6, characterized in that: The preparation steps are as follows: S1: Material selection: prepare protein source, carbohydrate source, carrot and mineral mixture source in proportion; S2: Processing of feed: firstly, corn starch is mixed with crushed soybeans and carrots to obtain a first mixture, then the first mixture is mixed with fish meal in a mass ratio of 10:(1-4) to obtain a second mixture, then 5-HT is added to the second mixture, and finally a gel forming agent is added and stirred to obtain the gel feed; S3: shaping the food: pouring the gel feed obtained in S2 into a mold, re-molding after solidification, and cutting the re-molded gel feed into pieces and rolling them into shape; S4: Food storage: Place the formed gel feed obtained in S3 in a storage container and store it in a freezer at -18~-22 ℃.

8. The method for preparing the gel feed of purple sea urchin according to claim 7, characterized in that: The stirring speed in step S2 is 80-100 rpm / min; the solidification environment in step S3 is 2-5°C for solidification for 300-360 min, and -18~-22°C for solidification for 50-60 min.

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