Compound exocarpium citri rubrum feed additive for preventing and controlling enterohepatic cytozoonosis of South America white shrimps

Abstract

The invention belongs to the field of aquaculture feed additives, and particularly relates to a compound preparation with a synergistic effect of an exocarpium citri grandis extract and probiotics and a preparation method of the compound preparation, and the compound preparation is used for preventing and treating enterohepatic cytozoonosis of penaeus vannamei. Comprising an exocarpium citri grandis extract preparation step, a bacillus subtilis and clostridium butyricum culture step and a compound exocarpium citri grandis feed additive preparation step. 0.1%-0.5% of the compound exocarpium citri rubrum feed additive is added into shrimp feed, and penaeus vannamei is fed; compared with a direct feeding feed, the compound exocarpium citri rubrum feed additive has the advantages that the morbidity of the enterocytozoonosis hepatopenaei of the shrimps is effectively reduced, and meanwhile, the occurrence of other diseases is reduced.

Description

Technical Field

[0001] The invention belongs to the field of aquaculture feed additives, and particularly relates to a compound preparation with synergistic effects of tangerine peel extract and probiotics and a preparation method thereof, which is used for preventing and treating hepatoenterocytosis of white shrimp Penaeus vannamei. Background Art

[0002] In recent years, my country's whiteleg shrimp aquaculture industry has faced significant challenges, with enterocolitica (EHP) becoming a key disease limiting its development. This pathogen infects the shrimp's digestive tract, destroying intestinal tissue structure and leading to nutrient absorption disorders, growth retardation, and even death. More seriously, EHP infection often triggers secondary bacterial infections, further exacerbating aquaculture losses. Current prevention and control measures primarily rely on chemical fungicides, but these have three major drawbacks: fungicides have difficulty penetrating the insect cell wall, resulting in an insect inhibition rate of less than 30%; residual fungicides disrupt the microecological balance of the water body, leading to secondary pollution; and long-term use accelerates the evolution of antibiotic resistance in pathogens.

[0003] There is an urgent need to develop a new prevention and control solution that is safe, efficient, green and environmentally friendly. Summary of the Invention

[0004] The present invention aims to provide a compound feed additive based on the synergistic effect of natural plant extracts and probiotics, achieving the following technological breakthroughs:

[0005] 1. Targeted insect inhibition: Using flavonoids from tangerine peel to specifically interfere with insect energy metabolism;

[0006] 2. Microecological regulation: Optimizing the intestinal environment through reconstruction of probiotic communities;

[0007] 3. Sustained release and enhanced efficacy: Encapsulation technology is used to improve the stability and bioavailability of active ingredients;

[0008] 4. Zero residue safety: avoid chemical residues and ensure the safety of aquatic products.

[0009] The technical solution of the present invention is:

[0010] A compound tangerine peel feed additive for preventing and controlling hepatoenterocytosis of white shrimp, comprising:

[0011] The invention relates to the preparation steps of tangerine peel extract, the cultivation steps of Bacillus subtilis and Clostridium butyricum and the preparation steps of compound tangerine peel feed additive.

[0012] The preparation steps of the Citrus aurantium extract include: crushing Citrus aurantium fruit, adding 60% ethanol at a ratio of 1:50, ultrasonically extracting for 1 hour, and centrifuging to obtain Citrus aurantium flavonoid primary extract.

[0013] The preparation step of the Citrus aurantium extract further comprises concentrating the Citrus aurantium flavonoid primary extract to obtain a distilled concentrated extract and recovering ethanol.

[0014] The Bacillus subtilis and Clostridium butyricum culturing steps include: preparing a culture medium according to 1L distilled water + 20g glucose + 15g peptone + 5g sodium chloride + 0.5g beef extract, followed by sterilization at 121 degrees for 15 minutes; adding Bacillus subtilis or Clostridium butyricum after cooling, placing the culture medium at 28 degrees for 36 hours with shaking, and using the culture medium to prepare the additive when the OD value of the culture medium is 1.0.

[0015] The preparation steps of the compound tangerine peel feed additive include: adding 200 ml of Bacillus subtilis and Clostridium butyricum to 1 L of tangerine peel extract concentrate, mixing them evenly, then adding sodium alginate and calcium chloride for embedding, and freeze-drying to obtain the compound tangerine peel feed additive.

[0016] The beneficial effects of the present invention are as follows: the compound tangerine peel feed additive is added to shrimp feed at a rate of 0.1%-0.5% and then fed to white shrimp. Compared with direct feeding, the use of the compound tangerine peel feed additive effectively reduces the incidence of shrimp hepatoenteric cytozoonosis and the occurrence of other diseases. Tangerine peel flavonoids can inhibit the growth of shrimp hepatoenteric cytozoonosis and improve shrimp immunity. Bacillus subtilis and Clostridium butyricum can adjust the shrimp intestinal microbial flora, build a beneficial bacterial community, inhibit the growth of harmful bacteria, and improve shrimp immunity. The use of sodium alginate and calcium chloride to embed tangerine peel extract and probiotics can improve the bioavailability of the product, promote the absorption of active ingredients by shrimp, and enhance the effect of the active ingredients. DETAILED DESCRIPTION

[0017] The following is further explained in conjunction with specific implementation methods:

[0018] Example 1

[0019] 1. Preparation of the Citrus aurantium extract of the present invention: crush the Citrus aurantium fruit, add 60% ethanol at a ratio of 1:50, perform ultrasonic extraction for 1 hour, and centrifuge to obtain the Citrus aurantium flavonoid primary extract. Distill and concentrate the extract and recover the ethanol.

[0020] 2. Culture of Bacillus subtilis and Clostridium butyricum: Prepare a culture medium containing 1 L distilled water, 20 g glucose, 15 g peptone, 5 g sodium chloride, and 0.5 g beef extract. Sterilize at 121°C for 15 min. After cooling, add Bacillus subtilis and incubate at 28°C with shaking for 36 h. When the culture reaches an OD value of 1.0, use it to prepare the additive.

[0021] 3. Preparation of compound tangerine peel feed additive: 200 ml of Bacillus subtilis and Clostridium butyricum were added to 1 L of tangerine peel extract concentrate, mixed evenly, and then sodium alginate and calcium chloride were added for embedding, and the compound tangerine peel feed additive was prepared by freeze drying.

[0022] Verification experiment

[0023] 600 adult whiteleg shrimp were divided into 6 groups (100 shrimp in each group). The feed types of each group are shown in Table 1. The morbidity rate was statistically analyzed after 30 days of feeding. The statistical results are shown in Table 2.

[0024] Table 1 Record of feed types used in each control group or additive group

[0025]

[0026] Note: The placebo in Table 1 is dextrin

[0027] Table 2 Statistics of the incidence rates of the control group or additive group

[0028]

[0029] Result analysis: The additive group (especially the 0.3% dosage group) significantly inhibited the proliferation of the worms (Ct value difference > 3.5).

[0030] Experiments have verified that the product obtained by the present invention can be used to prevent and treat hepatoenteric cytosis of white shrimp.

[0031] The above embodiments and descriptions are only for explaining the principles and best embodiments of the present invention. Without departing from the spirit and scope of the present invention, the present invention may be subject to various changes and improvements, which shall fall within the scope of the invention to be protected.

Claims

1. A compound tangerine peel feed additive for preventing and controlling hepatoenterocytosis of white shrimp, characterized in that: include: The invention relates to the preparation steps of tangerine peel extract, the cultivation steps of Bacillus subtilis and Clostridium butyricum and the preparation steps of compound tangerine peel feed additive.

2. The compound tangerine peel feed additive for preventing and controlling hepatoenterocytosis of white shrimp according to claim 1, characterized in that: The preparation steps of the Citrus aurantium extract include: crushing Citrus aurantium fruit, adding 60% ethanol at a ratio of 1:50, ultrasonically extracting for 1 hour, and centrifuging and filtering to obtain the Citrus aurantium flavonoid primary extract.

3. The compound tangerine peel feed additive for preventing and controlling hepatoenterocytosis of white shrimp according to claim 2, characterized in that: The preparation step of the Citrus aurantium extract further includes concentrating the Citrus aurantium flavonoid primary extract to obtain a distilled concentrated extract and recovering ethanol.

4. The compound tangerine peel feed additive for preventing and controlling hepatoenterocytosis of white shrimp according to claim 3, characterized in that: The Bacillus subtilis and Clostridium butyricum culturing steps include: preparing a culture medium according to 1L distilled water + 20g glucose + 15g peptone + 5g sodium chloride + 0.5g beef extract, followed by sterilization at 121 degrees for 15 minutes; adding Bacillus subtilis or Clostridium butyricum after cooling, placing the culture medium at 28 degrees for 36 hours with shaking, and using the culture medium for preparing the additive when the OD value of the culture medium is 1.

0.

5. The compound tangerine peel feed additive for preventing and controlling hepatoenterocytosis of white shrimp according to claim 4, characterized in that: The preparation steps of the compound tangerine peel feed additive include: adding 200 ml of Bacillus subtilis and Clostridium butyricum to 1 L of tangerine peel extract concentrate, mixing evenly, adding sodium alginate and calcium chloride for embedding, and freeze-drying to obtain the compound tangerine peel feed additive.

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