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Method for inducing spider to generate antibiotic activity substance

An antibacterial active substance, spider technology, applied in the field of biomedicine, to achieve the effect of simple and convenient operation and good antibacterial effect

Inactive Publication Date: 2008-08-06
HUBEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few research reports on spider antibacterial substances at home and abroad, only the isolation, purification and molecular cloning of antibacterial peptides from Acanthoscurria gomesiana (a kind of tarantula)

Method used

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  • Method for inducing spider to generate antibiotic activity substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Collect 90 horned fat spiders and divide them into two groups. The first group is 80 for induction, and the second group is 10 for control. The 80 spiders of the first group were placed in a petri dish, irradiated for 1 hour at a distance of 1 meter under a 20w ultraviolet lamp (50 spiders could be irradiated at a time), and then continued to be raised under normal conditions. The 10 spiders in the second group were not irradiated and were fed under normal conditions as a control.

[0034] 2. 80 spiders after ultraviolet irradiation and 10 spiders without induction were single-headed in glass finger tubes (length * diameter = 12cm * 4cm), and there was a sponge at the bottom of the tube to absorb enough water for the spiders to drink. Seal it with a cotton plug, and then place the glass vial containing the spider in a 25°C incubator with a light cycle of 12:12 (Light:Dark). Glass vials were cleaned every 2 days and fed with adult fruit flies (Drosophila sp.) and mid...

Embodiment 2

[0041] 1. Collect 40 spiders, put them in a petri dish, and irradiate them at a distance of 1 meter under a 20w ultraviolet lamp for 1 hour.

[0042] 2. Raise 40 spiders single-headed in a glass finger tube (length×diameter=12cm×4cm) after ultraviolet irradiation. There is a sponge at the bottom of the tube to absorb enough water for the spiders to drink. The glass vials with spiders were placed in a 25°C incubator with a light cycle of 12:12 (Light:Dark). Glass vials were cleaned every 2 days and fed with adult fruit flies (Drosophilasp.) and midges (Tendipes sp.).

[0043] 3. After feeding the spiders irradiated by ultraviolet rays for 60 hours, take the spiders to extract hemolymph. Disinfect the spider with 75% alcohol, then place it in a 1.5ml centrifuge tube in an ice bath, add pre-cooled sterile water homogenate (when the external temperature is high, you need to add some mercaptoethanol to prevent the blackening of antibacterial substances) , The homogenate was centr...

Embodiment 3

[0047] 1. Collect 40 horned fat spiders and divide them into four groups, 10 in each group. The first three groups were induced by ultraviolet irradiation, bacterial injection and starvation respectively, and the fourth group was not induced and served as the control group.

[0048] Ultraviolet radiation induction: put 10 test spiders in a petri dish, irradiate them at a distance of 1 meter under a 20w ultraviolet lamp for 1 hour, and then continue to raise them under normal conditions.

[0049] Bacterial injection method: After 10 test spiders were anesthetized on ice, they were injected on the abdomen with a micro-injector on the ultra-clean workbench, and each spider was injected with 1 μl of prepared Escherichia coli suspension (concentration of 1× 10 6 cfu / ml), and then continue to feed under normal conditions.

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Abstract

The invention provides a simple and high-efficiency method for inducing spiders to produce antibiotic active substances, which comprises the following steps: a. ultraviolet irradiation is used for inducement: larinioides cornuta is put into a culture vessel and irradiated for 0.5-3 hours under 20W ultraviolet light and in the distance of 1 meter; then the larinioides cornuta is raised continuously; b. hemolymph of the spiders 1-120 hours after induced is extracted and collected by applying a tissue homogenate method; c. B bronchiseptica is employed as indicator bacteria in a coated flat drilling method to test antibacterial activity of the hemolymph. The antibacterial activity of extracted spider hemolymph after irradiated and induced by ultraviolet is high, and the antibiotic active substances can be applied to development of antibiotics.

Description

Technical field: [0001] The invention relates to a method for inducing spiders to produce antibacterial active substances, belonging to the field of biomedicine. technical background: [0002] The extensive use of antibiotics has caused many pathogenic bacteria to develop drug resistance, which has brought difficulties to the treatment of some diseases. Antimicrobial peptides from arthropods such as insects, spiders, shrimps and crabs have high-efficiency and broad-spectrum killing effects on Gram-positive bacteria and Gram-negative bacteria, and can kill pathogenic mutants that have developed drug resistance without inducing resistance. The emergence of drug strains. Antimicrobial peptides can not only act on bacteria, viruses and other prokaryotes, but also play a certain role in tumor treatment. Antimicrobial peptides are very likely to become a new source of antibiotics, antivirals and antitumor drugs. [0003] The antibacterial substances in insects, spiders, shrimps...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033A61K35/64A61P31/04A61P35/00C12Q1/18A61K35/646
CPCY02A50/30
Inventor 彭宇汪淼徐慧君刘凤想陈建焦晓国
Owner HUBEI UNIV
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