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Determination of Milbemectin Activity

Milbemectin is a new type of broad-spectrum, non-cross-resistant microbial natural product pesticide with broad-spectrum activity, high efficiency, and low toxicity against agricultural and forestry pests, not easily polluting the environment, and not easy to produce resistance.

Milbemectin is a hexadecyl macrolide mixture, compared with ivermectin in the C-13 position less disaccharide group, so its activity in the inheritance of ivermectin stability and long-lasting at the same time, but also has a higher fat solubility than ivermectin. It is a golden yellow semi-transparent viscous substance at room temperature, easily soluble in hexane, benzene, acetone, ethanol, methanol, and chloroform, but almost insoluble in water, and the mixture of its A3 and A4 components has high acaricidal activity.

Fig.1 Structural diagram of milbemectin α, milbemectin β, milbemectin A3, and milbemectin A4.Fig.1 Schematic structural diagram of milbemectin: (a) α series, (b) β series, (c) A3, (d) A4.

The mechanism of action of Milbemectin is different from that of γ-aminobutyric acid agonist ivermectin, which can cause the opening of a glutamate-gated chloride channel (glutamate-gated chloride), thus increasing the inward flow of chloride ions and preventing the release of normal action potentials, leading to the death of insect paralysis.

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CAS NO.Product NameInquiry
51596-10-2Milbemectin a3Inquiry
51596-11-3Milbemectin a4Inquiry

The experimental approach to the activity of milbemectin pesticide is evaluated by determining the virulence of milbemectin against the vermilion leaf mite at different developmental stages, as well as the ovicidal and oviposition inhibition of the leaf mite. The experiment used ivermectin as a control agent.

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Reagents, Materials, and Apparatus

Tetranychus cinnabarinus is selected as the test organism. 95% milbemectin (A3:A4=3:7) and 96% ivermectin are used as the agents and are configured in different mass concentrations. All other reagents are analytically pure.

Experimental equipment such as Potter spray tower, Olympus dissecting microscope, air compressor, pipette, brush, and punch are prepared.

Experimental Procedures

Determination of virulence of milbemectin against female adult of Tetranychus cinnabarinus.

  • Fava bean leaves of uniform growth are selected and leaf disks of 2 cm diameter are made using a hole punch.
  • The leaf discs are placed on skimmed cotton wool in the center of a plastic dish and three leaf discs are placed in each dish.
  • Female adult mites are inoculated onto the leaf disks with a small brush, 45 mites per leaf disk, and the appropriate amount of water is added.
  • The leaf discs are placed in an incubation room at (26±1) ℃ with a light intensity of 3000-4500 lx and a photoperiod of 14 hours.
  • The number of female adult mites is checked and recorded 2 h after inoculation to ensure that the number of mites on each leaf dish is not less than 30.
  • Spray treatments are carried out using a Potter spray tower at different mass concentrations of milbemectin and abamectin solutions.
  • The treated leaf disks are continued to be incubated for 24 hours and then examined microscopically and the number of dead and surviving mites is recorded.

Fig.2 Tetranychus cinnabarinus.

Determination of the virulence of milbemectin to female lavar of Tetranychus cinnabarinus.

  • Leaf disks containing eggs lay by female lavar of Tetranychus cinnabarinus are prepared according to the above procedure.
  • The young mites are collected 96 hours after the female adult mites lay eggs.
  • The young mites are treated with the same methods and agents, and the number of dead and surviving mites is recorded.

Determination of the virulence of milbemectin to egg of Tetranychus cinnabarinus.

  • Leaf disks containing eggs laid by female adult mites are prepared according to the above procedure.
  • 24 hours after the female adult mites lay eggs, the leaf disks are transferred to new plastic dishes.
  • The leaf dishes are treated using the same methods and agents and the hatching rate and the number of surviving juvenile mites are recorded.

Based on the results of the experiment, the activity of milbemectin on leaf mites at different developmental stages could be evaluated. By recording the number of dead and surviving mites, it is possible to determine the virulence of milbemectin as well as its ovicidal and oviposition inhibitory effects on leaf mites.

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