Natural compound anti-aeromonas-hydrophila agent, natural feed and preparation method thereof

[0057] Example 1: Preparation of natural product anti-Fish Aeromonas hydrophila infection feed

[0058] 1) Add the 6 extracts to the oil at a concentration of 1g/kg (referring to adding one extract per kg of basic feed) according to the following table 1, mix them evenly and add to the basic feed to make the corresponding 8 types Pellets are labeled as A0, A1, A2, A3, A4, A5, A6, A7.

[0059] Table 1

[0060] Additives A0 A1 A2 A3 A4 A5 A6 A7 Pomegranate peel ethanol extract × × Turmeric ethanol extract × × Eucommia Ethanol Extract × × Myrobalan Ethanol Extract × × Ebony Ethanol Extract × × Scutellaria baicalensis ethanol extract × ×

[0061] in FIG. 1:

[0062] A0 refers to the basic feed without adding any extracts. In the following examples, A0 takes two copies, which are labeled A0(1) and A0(2).

[0063] A1 refers to the addition of 1 g pomegranate peel ethanol extract, 1 g turmeric ethanol extract, 1 g Eucommia ulmoides ethanol extract, 1 g myrobalan ethanol extract, 1 g ebony ethanol extract, and 1 g scutellaria baicalensis ethanol extract per kg of basic feed.

[0064] A2 means that 1g turmeric ethanol extract, 1g eucommia ulmoides ethanol extract, 1g myrobalan ethanol extract, 1g ebony ethanol extract and 1g scutellaria baicalensis ethanol extract are added per kg of basic feed.

[0065] A3 means that 1 g of pomegranate peel ethanol extract, 1 g of Eucommia ulmoides ethanol extract, 1 g of myrobalan ethanol extract, 1 g of ebony ethanol extract, and 1 g of scutellaria baicalensis ethanol extract are added to each kg of basic feed.

[0066] A4 refers to the addition of 1g pomegranate peel ethanol extract, 1g turmeric ethanol extract, 1g myrobalan ethanol extract, 1g ebony ethanol extract, and 1g scutellaria baicalensis ethanol extract per kg of basic feed.

[0067] A5 refers to the addition of 1g pomegranate peel ethanol extract, 1g turmeric ethanol extract, 1g Eucommia ulmoides ethanol extract, 1g ebony ethanol extract and 1g scutellaria baicalensis ethanol extract per kg of basic feed.

[0068] A6 refers to the addition of 1 g pomegranate peel ethanol extract, 1 g turmeric ethanol extract, 1 g eucommia ulmoides ethanol extract, 1 g myrobalan ethanol extract and 1 g scutellaria baicalensis ethanol extract per kg of basic feed.

[0069] A7 means that 1g of pomegranate peel ethanol extract, 1g of turmeric ethanol extract, 1g of Eucommia ulmoides ethanol extract, 1g of myrobalan ethanol extract, and 1g of black plum ethanol extract are added to each kg of basic feed.

[0070] 2) Add the 6 extracts to the water at a concentration of 2g/kg (referring to adding one extract per kg of basic feed) as shown in Table 2, and add them to the basic feed after mixing to make the corresponding 8 types of pellets. The feed is marked as B0, B1, B2, B3, B4, B5, B6, B7.

[0071] Table 2

[0072] Additives B0 B1 B2 B3 B4 B5 B6 B7 Pomegranate peel water extract × × Turmeric water extract × × Eucommia Water Extract × × Myrobalan Water Extract × × Ebony water extract × × Scutellaria baicalensis water extract × ×

[0073] In Table 2:

[0074] B0 refers to the basic feed without adding any extracts. In the following examples, two copies of B0 are taken, and they are labeled B0(1) and B0(2).

[0075] B1 refers to the addition of 2g pomegranate peel water extract, 2g turmeric water extract, 2g Eucommia ulmoides water extract, 2g myrobalan water extract, 2g black plum water extract, and 2g scutellaria baicalensis water extract per kg of basic feed.

[0076] B2 means that 2g turmeric water extract, 2g eucommia ulmoides water extract, 2g myrobalan water extract, 2g ebony water extract, and 2g scutellaria baicalensis water extract are added per kg of basic feed.

[0077] B3 refers to the addition of 2g pomegranate peel water extract, 2g eucommia ulmoides water extract, 2g myrobalan water extract, 2g ebony water extract and 2g scutellaria baicalensis water extract per kg of basic feed.

[0078] B4 refers to the addition of 2g pomegranate peel water extract, 2g turmeric water extract, 2g myrobalan water extract, 2g ebony water extract and 2g scutellaria water extract per kg of basic feed.

[0079] B5 refers to the addition of 2g pomegranate peel water extract, 2g turmeric water extract, 2g eucommia water extract, 2g ebony water extract, and 2g scutellaria baicalensis water extract per kg of basic feed.

[0080] B6 refers to the addition of 2g pomegranate peel water extract, 2g turmeric water extract, 2g Eucommia ulmoides water extract, 2g myrobalan water extract and 2g scutellaria baicalensis water extract per kg of basic feed.

[0081] B7 means that 2g pomegranate peel water extract, 2g turmeric water extract, 2g eucommia water extract, 2g myrobalan water extract and 2g ebony water extract are added to each kg of basic feed.

[0082] 3) According to the following table 3, 12 kinds of extracts were added to the concentration of 1 or 2g/kg (referring to adding one extract per kg of basic feed), in which the water extract was added to the water, the ethanol extract was added to the oil, and mixed After being evenly added to the basic feed, 6 kinds of pellets are made into corresponding pellet feeds and labeled as C0, C1, C2, C3, C4, C5, and the addition amount of C5 is 0.5g/kg.

[0083] table 3

[0084] Additives C0 C1 C2 C3 C4 C5 Pomegranate peel ethanol extract × × × Turmeric ethanol extract × × × Eucommia Ethanol Extract × × × × × × Myrobalan Ethanol Extract × × × Ebony Ethanol Extract × × × × × Scutellaria baicalensis ethanol extract × × × Pomegranate peel water extract × × × × × × Turmeric water extract × × × × × Eucommia Water Extract × × × × × Myrobalan Water Extract × × × Ebony water extract × × × Scutellaria baicalensis water extract × × ×

[0085] In Table 3:

[0086] C0 refers to the basic feed without adding any extracts. In the following examples, two copies of C0 are taken, and they are labeled C0(1) and C0(2).

[0087] C1 refers to the addition of 1 g pomegranate peel ethanol extract, 1 g turmeric ethanol extract, 1 g myrobalan ethanol extract, 1 g ebony ethanol extract, and 1 g scutellaria ethanol extract per kg of basic feed.

[0088] C2 refers to the addition of 1g pomegranate peel ethanol extract, 1g turmeric ethanol extract, 1g myrobalan ethanol extract and 1g scutellaria baicalensis ethanol extract per kg of basic feed.

[0089] C3 refers to the addition of 2g turmeric water extract, 2g eucommia ulmoides water extract, 2g myrobalan water extract, 2g ebony water extract and 2g scutellaria baicalensis water extract per kg of basic feed.

[0090] C4 means that 2g myrobalan water extract, 2g black plum water extract, and 2g scutellaria baicalensis water extract are added per kg of basic feed.

[0091] C5 refers to the addition of 0.5g pomegranate peel ethanol extract, 0.5g turmeric ethanol extract, 0.5g myrobalan ethanol extract, 0.5g scutellaria ethanolic extract, 1g myrobalan water extract, and 1g black plum water extract per kg of basic feed. And 1g Scutellaria baicalensis Georgi water extract.

[0092] Example 2

[0093] The purchased crucian carp fry were raised in a breeding room for one week, and 405 crucian fry of the same weight were selected and randomly divided into 9 treatment groups, each with 3 duplicate fish tanks. The shape, size and color of each fish tank are the same, the size is 30×30×40cm, and a built-in aerator with 800L/h flow. The water in the fish tank uses potable tap water as the water source, and the volume of water in the tank is 30L. The temperature of the breeding room is controlled to 22±1℃, and the aerator provides continuous oxygen. Nine treatment groups were fed separately labeled "A0(1)", "A0(2)", "A1", "A2", "A3", "A4", "A5", "A6" in Example 1. "And "A7" feed, fed 4 times a day with full food. After feeding for 7 days, the feeding was stopped for 1 day and the challenge experiment was conducted.

[0094] The challenge experiment uses intraperitoneal injection. The number of crucian carp per fish tank is 15. Fish fed "A0(1)" feed were injected with 0.1 mL of distilled water, and fish from other fish tanks were injected with 1.0×10 8 A/mL concentration of Aeromonas hydrophila solution 0.1mL. After injection and disinfection, return the fish to the original aquarium, and observe and record the number of fish deaths in each aquarium in the next week.

[0095] The test results are shown in Table 4 and figure 1.

[0096] Table 4 The effect of compound alcohol extract feed on the number of crucian carp deaths after challenge

[0097]

[0098]

[0099] In Table 4, the data are represented by the mean ± standard deviation of 3 replicates; in the same column of data, those with different superscript letters indicate significant differences (P <0.05).

[0100] From Table 4 and figure 1 It can be seen that the order of mortality: A2>A5=A7>A3>A1>A6>A4, 68.89%, 64.44%, 62.22%, 60.00%, 53.33%, 48.89%, 44.44%, respectively.

[0101] Example 3

[0102] The purchased crucian carp fry were raised in a breeding room for one week, and 405 crucian fry of the same weight were selected and randomly divided into 9 treatment groups, each with 3 duplicate fish tanks. The shape, size and color of each fish tank are the same, the size is 30×30×40cm, and a built-in aerator with 800L/h flow. The water in the fish tank uses potable tap water as the water source, and the volume of water in the tank is 30L. The temperature of the breeding room is controlled to 22±1℃, and the aerator provides continuous oxygen. Nine treatment groups were fed separately labeled "B0(1)", "B0(2)", "B1", "B2", "B3", "B4", "B5", "B6" in Example 1. "And "B7" feed, fed 4 times a day with full food. After feeding for 7 days, the feeding was stopped for 1 day and the challenge experiment was conducted.

[0103] The challenge experiment uses intraperitoneal injection. The number of crucian carp per fish tank is 15. Fish fed with "B0(1)" feed were injected with 0.1 mL of distilled water, and fish from other aquariums were injected with 1.0×10 8 A/mL concentration of Aeromonas hydrophila solution 0.1mL. After injection and disinfection, return the fish to the original aquarium, and observe and record the number of fish deaths in each aquarium in the next week.

[0104] The test results are shown in Table 5 and figure 2.

[0105] Table 5 The effect of feeding compound water extract on the number of crucian carp deaths after challenge

[0106]

[0107]

[0108] In Table 5, the data are represented by the mean ± standard deviation of 3 replicates; in the same column of data, those with different superscript letters indicate significant differences (P <0.05).

[0109] From Table 5 and figure 2 It can be seen that, compared with B0(2), the compound extract reduced the mortality of fish after challenge to varying degrees. The order of mortality: B6>B5>B7>B1>B3>B2=B4, respectively They are 68.89%, 62.22%, 60.00%, 51.11%, 48.89%, 42.22%, 42.22%.