KR102006258B1 - Composition for preventing, improving or treating earyl mortality sydrome infection of shrimps and method for manufacturing the same - Google Patents

Abstract

The present invention relates to a composition for improving infectious diseases of shrimp by Vibrio germs, and more particularly to a composition for improving vibriosus infection comprising an Omija extract and / or a wicker extract as an effective ingredient.
The vibrio species of the present invention are vibrio parahaemolyticus, vibrio harveyi species, and compositions for improving early mortality syndrome and / or acute hepatic pancreatic necrosis caused by the infection of the bacterium can be obtained by extracting and / Or as an active ingredient, the antimicrobial activity of Vibrio sp.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for prevention, treatment, or amelioration of infection with shrimp pneumonia, and to a method for manufacturing the same,

The present invention relates to a composition for preventing, treating and improving infection using natural materials for antibiotic against Vibrio spp. Causing serious injury by causing shrimp premature death syndrome or acute liver pancreatic traumatic syndrome which occurs when shrimp are cultured. More particularly, the present invention relates to an antimicrobial feed composition or a composition for veterinary medicine for vibrio parahaemolyticus, vibrio harveyi species.

In recent years, pathogens causing large-scale deaths of aquaculture have become a big problem. It is known as EMS (Earyl Mortality Sydrome) or Acute Hepatopancreatic Necrosis Syndrome (AHPNS) and shows a mortality rate close to extinction at the early stage of shrimp culture. At the time of the first occurrence, the cause of the shrimp early mortality was not known, but the cause of death in early stage of shrimp farming was discovered by the bacteria called Vibrio parahaemolyticus which exists in seawater at the University of Arizona in 2013. In addition, researchers at the Vietnam National Fisheries Research and Development Institute (RIA) have found that vibrio harveyi as well as Vibrio parahaemolyticus is the causative agent of the shrimp early pandemic.

In Thailand in 2013, shrimp early pest syndrome caused a shrinkage of nearly 50% of annual shrimp production, and in China and Vietnam, the loss of such infectious diseases caused a great decrease in the production of aquaculture shrimp worldwide, Resulting in sales of shrimp rising more than 30%.

Currently, there are no cases of pneumoconiosis epidemic in Korea. However, the importation of shrimp pneumoconiosis syndrome from Southeast Asia and the country is expected to cause great damage to the domestic shrimp farming industry.

In the prior art, abuse of the peroxidized water tank, formalin or antibiotics to prevent the occurrence of such an epidemic of premature ejaculation epidemic and to prevent the spread of the shrimp in the early stage may adversely affect the aquaculture environment and the health of the end consumer.

Under these circumstances, the inventors of the present invention have made extensive efforts to develop a method for preventing or treating shrimp early pox syndrome, and as a result, they have found a material having an antibacterial effect against Vibrio spp. Respectively.

KR 1020030068748 A KR 1020130090988 A

VETERINARSKI ARHIV., 2016, 86 (1), pp. 125-134

It is an object of the present invention to provide a pharmaceutical and feed composition for the prevention, treatment, or improvement of shrimp premature death syndrome or acute liver pancreatic traumatic syndrome including plant extracts.

It is another object of the present invention to provide an animal pharmaceutical or feed composition comprising an omega and / or anchovy extract.

 Another object of the present invention is to provide a vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2 or vibrio harveyi KC 13.17.5, which is a pathogen produced in shrimp culture in Vietnam, Thailand, And to provide a composition having an antimicrobial effect on the skin.

The present invention provides a pharmaceutical composition and an animal feed composition for preventing or treating shrimp premature death syndrome or shrimp acute hepatic pancreatic necrosis comprising an extract of Omija or a mixture of Omija and anchovy extract as an active ingredient.

In the present invention, the shrimp premature death syndrome or pancreatic acute hepatic pancreatic necrosis is infected by Vibrio sp., And the Vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2 or vibrio harveyi KC 13.17.5 .

The composition comprising the extracts of the various plants of the present invention has an effect of inhibiting the activity against the causative infectious Vibrio fungus in the premature death of shrimp and acute liver pancreatic necrosis, It can be widely used for increasing stability.

Fig. 1 shows the results of the first step of the antimicrobial activity test for the Korean wild-type strain Vibrio parahaemolyticus ATCC 17802 for the detection of antimicrobial activity.
Figure 2 shows that there is an effect on Vibrio parahaemolyticus KC 12.020 and 13.14.2, Vibrio hebei KC13.17.5 as a result of the antimicrobial activity of Omiza extract.
FIG. 3 shows the results after 2 hours and 5 hours of the effect on Vibrio parahaemolyticus KC 12.020 and 13.14.2, Vibrio harvey KC13.17.5 as a result of the antimicrobial activity of the Schizandra chinensis extract.
FIG. 4 shows the results of administration of 2 mg of the effect on Vibrio parahaemolyticus KC 12.020 and 13.14.2, Vibrio hebei KC13.17.5 as a result of the antimicrobial activity of the Schizandra chinensis extract.
Figure 5 shows that there is an effect on Vibrio parahaemolyticus KC 12.020 and 13.14.2, Vibrio hebei KC13.17.5 as a result of the antimicrobial activity of the detached extract.
Figure 6 shows the 16S-rRNA base sequence result of Vibrio parahaemolyticus KC 12.020.
Figure 7 shows the results of the Vibrio parahaemolyticus KC 13.14.2 16S-rRNA base sequence.
Figure 8 shows the results of the Vibrio levei KC13.17.5 16S-rRNA base sequence.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Vibrio parahaemolyticus KC 12.020 and 13.14.2 used as pathogens in the present invention were collected in Vietnam seawater in the summer of 2012 and Vibrio harbei KC13.17.5 was collected from Vietnam Nghe An.

The 16S rRNA sequence of the obtained Vibrio species strain showed the closest homology to each vibrio species as compared with the microorganisms registered in GenBank, and the 16S rRNA sequence of the strain obtained as a result of the 16S rDNA sequence analysis was 6 to 8.

Example  1. Plant extract preparation

The plant and medicinal materials used in the present invention can be easily collected or purchased from nature, and the extracts are pulverized and used in the form of flakes to obtain the extract efficiently.

The natural materials selected for use in the present invention are Schisandra chinensis and Cinnamomum cassia, which are each subcultured in 2 Kg, immersed in 2 L of organic solvent, alcohol or methanol, filtered, concentrated under reduced pressure, .

Example  2. Shrimp Premature Mortality Syndrome  Causative bacteria On the Vibrio species  Agar diffusion method for paper disk

The vibrio strains used in the present invention were vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2, and vibrio harveyi KC 13.17.5, three species of Vibrio spp.

In order to complete Example 2 of the present invention, the pathogenic strain (4.3 × 10 ⁸ CFU / mL) previously cultured in sterilized maring agar for 24 hours was inoculated 0.5% and solidified in a petri dish uniformly mixed in the medium. Then, 100 νL of the extract (10 mg / mL) prepared in Example 1 was dispensed on an 8 mm paper disk, dried without moisture and placed on a marine agar, and incubated in an incubator at 25 ° C. for 24 hours. After that, a clear zone was formed by inhibiting the growth of pathogens around the paper disk, and its range was measured to evaluate the antimicrobial activity. The results of the antimicrobial activity of these antimicrobials are shown in Table 1. At this time, sulfamethoxazole 23.75 g + trimethoprim 1.25 g was used as a control antibiotic.

Example  3. Using liquid medium dilution method MIC  Measurement method

The vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2, and vibrio harveyi KC 13.17.5 species of the present invention were inoculated into 3 mL of the liquid medium and cultured overnight at 37 ° C. 1000 and mixed with each extract in a 96-well plate for each concentration. The cells were incubated overnight at 37 ° C and assayed at 600 nm in an ultraviolet light absorber. The results are shown in Table 2.

Experimental results 1. Natural plant extract In Vibrio  Antibacterial effect

As shown in FIG. 1, the antimicrobial activity of various natural extracts of vibrio parahaemolyticus ATTCC 17802, which is a known strain, was examined in order to select natural products having antimicrobial activity in Vibrio sp. The activity was confirmed in the gill, but it was not found in the omija extract.

However, as shown in Table 1 below, the omija extract and the guinea gum extract were found in vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2, and vibrio harveyi KC 13.17.5 species, which are causative agents of shrimp infectious diseases in Southeast Asia including Vietnam, Thailand and China Was found to have antimicrobial activity. From these results, it was confirmed that each extract composition had higher antimicrobial activity than the control group.

<Antimicrobial Activity Evaluation of Vibrio Species of Omiza and Artichoke Extracts (mm)>

Strain Schisandra Anchorage Control v. parahaemolyticus KC 12.020 17.0 20.9 18.6 v. parahaemolyticus KC 13.14.2 10.2 13.7 - v. harveyi KC 13.17.5 8.9 14.3 -

<Measurement of MIC of Schizandra chinensis>

Strain Schisandra
(vg / mL) Anchorage
(vg / mL) v. parahaemolyticus KC 12.020 330.45 220.14 v. parahaemolyticus KC 13.14.2 563.82 410.93 v. harveyi KC 13.17.5 800.44 663.86

The MIC concentration in the treatment of each extract against vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2, and vibrio parveemolyticus KC 13.17.5, and vibrio harveyi KC 13.17.5 as shown in Table 2 above was found to be 330.45, 563.82, 800.44 νg / Extract, 220.14, 410.93, 663.86 νg / mL, respectively.

In addition, in FIGS. 2 to 5, it was confirmed that the antibacterial activity against the Vibrio sp.

In addition, as shown in the following Table 3, it can be seen that the mixture of omiza and guacamole has an excellent antimicrobial effect against vibrio hemolyticus or harbei bacteria when omiza or anchovy extract is used alone, It was confirmed that the antimicrobial effect was still not observed in Hemolacticus.

Strain Schisandra
(mg / mL) Anchorage
(mg / mL) Omija + interlocking mixture v. parahaemolyticus KC 12.020 330.45 220.14 201.35 v. parahaemolyticus KC 13.14.2 563.82 410.93 396.22 v. harveyi KC 13.17.5 800.44 663.86 482.25 v. parahaemolyticus ATTCC 17802 > 1000 855.78 868.54

Therefore, the composition of the present invention, which is effective as an ingredient, is effective for prevention, treatment and improvement of shrimp preterm delivery syndrome or acute liver pancreatic traumatic syndrome occurring in shrimp culture in Vietnam, Thailand and China &Lt; / RTI &gt;

Claims (6)

A mixture of omija and anchovy extract as an active ingredient,
A pharmaceutical composition for preventing or treating shrimp acute pancreatic necrosis or shrimp infestation by Vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2 or vibrio harveyi KC 13.17.5.
delete delete A mixture of omija and anchovy extract as an active ingredient,
Animal feed composition for improving shrimp premature death syndrome or shrimp acute hepatic pancreatic necrosis infected by Vibrio parahaemolyticus KC 12.020, vibrio parahaemolyticus KC 13.14.2 or vibrio harveyi KC 13.17.5


delete delete

Images