CN111066781A - Carvacoline disinfectant suitable for low-temperature use - Google Patents
Carvacoline disinfectant suitable for low-temperature use Download PDFInfo
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- CN111066781A CN111066781A CN201911413917.3A CN201911413917A CN111066781A CN 111066781 A CN111066781 A CN 111066781A CN 201911413917 A CN201911413917 A CN 201911413917A CN 111066781 A CN111066781 A CN 111066781A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/08—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having one or more single bonds to nitrogen atoms
- A01N47/10—Carbamic acid derivatives, i.e. containing the group —O—CO—N<; Thio analogues thereof
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Abstract
The invention discloses a disinfectant for a Caravan suitable for low-temperature use, which is suitable for use in a low-temperature environment (40 ℃ below zero to 0 ℃) and comprises the components of the Caravan, a deicing agent and a solvent which are mixed in proportion, is suitable for use in the low-temperature environment (40 ℃ below zero to 0 ℃), has good sterilization stability, has good sterilization effect on reproduction bacteria, viruses or spore bacteria by different formula components, and is suitable for sterilization of farms, public places, equipment, eggs and the like; in addition, the sterilization rate of the disinfectant for the delivery vehicles exceeds 99.9 percent, the disinfectant meets the regulation of No. 101-veterinary disinfectant identification technical specification (trial) of the Ministry of agriculture, veterinary drug review center 1992, and the formula has low corrosivity on metals and can meet the disinfection requirement of metal compartments of the delivery vehicles.
Description
The technical field is as follows:
the invention relates to a disinfectant, in particular to a methomyl disinfectant suitable for low-temperature use.
Background art:
with the global and deep development of the present day, the communication between countries and regions and between regions is more closely and frequently. The increase of the movement of the entry and exit personnel and materials brings great pressure to disease prevention and control, and the epidemic diseases can not only threaten the lives and properties of the personnel, but also influence the health safety situation, the import and export trade and the economic culture communication activities. Particularly, in recent years, the frequent occurrence of serious animal epidemic at home and abroad not only causes the death of a large amount of livestock and poultry, seriously influences the quality safety of animal products, hinders the development of animal husbandry and the income of farmers, but also directly harms human health and causes huge loss to animal husbandry and public health service. The transportation process of port animals and animal products is one of the main ways for spreading epidemic situations at home and abroad.
The disinfection of transportation vehicles has been taken as an important part in the prevention and control of animal epidemic diseases at home and abroad, and is also the most effective measure for controlling the spread of the epidemic diseases. The disinfection of transportation means mainly comprises a physical disinfection method and a chemical disinfection method, wherein the physical disinfection method mainly uses ultraviolet rays, microwaves, pressure, mechanical removal and other methods to kill or remove pathogenic microorganisms. Because the vehicles are continuously accessed and accessed, methods such as ultraviolet irradiation and the like are difficult to implement, the vehicle surface is mainly cleaned mechanically on the disinfection port site, the purpose of reducing the quantity of pathogenic microorganisms can be achieved, but all pathogenic microorganisms are difficult to kill; the chemical disinfection method mainly plays a role in killing pathogenic microorganisms through chemical disinfectants such as quaternary ammonium salts, aldehydes, chlorines and the like, and the disinfectants commonly used at home and abroad mainly comprise benzalkonium bromide (benzalkonium bromide), benzalkonium chloride (benzalkonium chloride), decamethylammonium bromide and the like, have low toxicity and irritation, good water solubility, convenient use and stable property, and are widely applied to the indoor environment, drinking water and animal body surface spray disinfection of various farms and pet clinics, and the disinfection of various equipment, instruments and vehicles. However, the disinfectant has poor effect of killing partial microorganisms, is greatly influenced by organic matters, has relatively high price, and has low temperature and long time in winter and spring in northern areas, and the disinfection effect of almost all the disinfectants is obviously reduced or even ineffective along with the reduction of the environmental temperature.
The invention content is as follows:
the invention aims to improve the existing formula and develop an environment-friendly disinfectant for the methomyl forest, which has the characteristic of good killing effect at low temperature (-40-0 ℃) and no corrosion aiming at reproduction bacteria, viruses or spore bacteria.
The invention is implemented by the following technical scheme: the disinfectant for the methomyl forest suitable for low-temperature use is used at the temperature of minus 40-0 ℃, and comprises the components of methomyl forest, a deicing agent and a solvent which are mixed in proportion.
Further, when the disinfectant is used for disinfecting breeding bacteria and/or viruses, the concentration of the methomyl is 0.1-0.5% (v/v), and the solvent is water.
Furthermore, the using temperature is-20-0 ℃, the deicing agent is ethanol, and the content of the deicing agent is 5-41% (v/v).
Furthermore, the using temperature is-20-0 ℃, and the deicing agent is glycol, and the content of the deicing agent is 10-50% (v/v).
Furthermore, the using temperature is-20-0 ℃, the deicing agent is propylene glycol, and the content of the deicing agent is 5-50% (v/v).
Furthermore, the using temperature is-20-0 ℃, and the deicing agent is sodium chloride, and the content of the deicing agent is 0.3-23 wt%.
Further, when the using temperature is-40 to-20 ℃, the deicing agent is ethanol, and the content of the deicing agent is 35-64% (v/v).
Furthermore, when the using temperature is-40 to-20 ℃, the deicing agent is ethylene glycol or propylene glycol, and the content of the deicing agent is 35-55% (v/v).
Further, when the disinfectant is used for disinfecting spores, the concentration of the methomyl is 15-30% (v/v), and the solvent is water.
Further, the deicing agent is ethanol, and the content of the deicing agent is 5-64% (v/v).
Further, the deicing agent is ethylene glycol, and the content of the ethylene glycol is 10-55% (v/v).
The invention has the advantages that: the disinfectant for the methomyl forest disclosed by the invention is suitable for being used in a low-temperature environment (minus 40-0 ℃), has good sterilization stability, has good sterilization effects on reproduction bacteria, viruses or spore bacteria by different formula components, and is suitable for sterilizing breeding farms, public places, equipment and instruments, hatching eggs and the like; in addition, the sterilization rate of the disinfectant on delivery vehicles exceeds 99.9 percent, and the disinfectant meets the regulation of No. 101 of the national veterinary drug evaluation center 1992 of agriculture (animal husbandry) in the technical specification (trial) for identifying the disinfectant for animals; and the formula has low metal corrosivity, and can meet the disinfection requirement of a metal carriage of a carrying vehicle.
The specific implementation mode is as follows:
description of the drawings:
FIG. 1: statistical plot of corrosion of the methomyl disinfectant on metals.
Example 1:
the disinfectant for the methomyl is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and comprises the components of 0.1% (v/v) of methomyl, 5% (v/v) of deicer ethanol and water as a solvent.
Example 2:
the disinfectant for the methomyl is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and comprises the components of 0.3% (v/v) of methomyl, 23% (v/v) of deicer ethanol and water as a solvent.
Example 3:
the disinfectant for the methomyl is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and comprises the components of 0.5% (v/v) of methomyl, 41% (v/v) of deicer ethanol and water as a solvent.
Example 4:
the disinfectant for the methomyl is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and comprises the components of 0.05% (v/v) of methomyl, 3% (v/v) of deicer ethanol and water as a solvent.
Example 5:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is propylene glycol, and the content of the propylene glycol is 5% (v/v).
Example 6:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is propylene glycol, and the content of the propylene glycol is 25% (v/v).
Example 7:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is propylene glycol, and the content of the propylene glycol is 50% (v/v).
Example 8:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is propylene glycol, and the content of the propylene glycol is 2% (v/v).
Example 9:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 10% (v/v).
Example 10:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 20% (v/v).
Example 11:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 50% (v/v).
Example 12:
the disinfectant for the Carvaclin aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 5% (v/v).
Example 13:
the disinfectant for the methomyl forest aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is sodium chloride, and the content of the deicer is 0.3 wt%.
Example 14:
the disinfectant for the methomyl disinfectant aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is sodium chloride, and the content of the deicer is 12 wt%.
Example 15:
the disinfectant for the methomyl forest aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is sodium chloride, and the content of the deicer is 23 wt%.
Example 16:
the disinfectant for the methomyl forest aims at the disinfection of escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-20-0 ℃, and compared with the disinfectant in the embodiment 2, the deicer is sodium chloride, and the content of the deicer is 0.1 wt%.
Example 17:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, and compared with the disinfectant in the embodiment 2, the ethanol content of the deicer is 35% (v/v).
Example 18:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, and compared with the disinfectant in the embodiment 2, the ethanol content of the deicer is 50% (v/v).
Example 19:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, and compared with the disinfectant in the embodiment 2, the ethanol content of the deicer is 64% (v/v).
Example 20:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, and compared with the disinfectant in the embodiment 2, the ethanol content of the deicer is 30% (v/v).
Example 21:
the disinfectant for the Carvaclin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicing agent is propylene glycol, and the content of the deicing agent is 35% (v/v).
Example 22:
the disinfectant for the Carvaclin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicing agent is propylene glycol, and the content of the deicing agent is 45% (v/v).
Example 23:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicer is propylene glycol, and the content of the deicer is 55% (v/v).
Example 24:
the disinfectant for the Carvaclin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicing agent is propylene glycol, and the content of the deicing agent is 30% (v/v).
Example 25:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 35% (v/v).
Example 26:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 45% (v/v).
Example 27:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicer is ethylene glycol, and the content of the deicer is 55% (v/v).
Example 28:
the disinfectant for the Weiweilin in the embodiment is used for disinfecting escherichia coli, staphylococcus aureus and Newcastle Disease Virus (NDV) at the temperature of-40 ℃ to-20 ℃, compared with the disinfectant in the embodiment 2, the deicer is glycol, and the content of the deicer is 30% (v/v).
Example 29:
the disinfectant for the Carmyl forest aims at the disinfection of the Bacillus cereus at the temperature of-40-0 ℃, wherein the Carmyl forest is 15% (v/v), the deicer ethanol is 5% (v/v), and the solvent is water.
Example 30:
the disinfectant for the Carmyl forest aims at the disinfection of the Bacillus cereus at the temperature of-40-0 ℃, wherein the Carmyl forest is 23% (v/v), the deicer ethanol is 30% (v/v), and the solvent is water.
Example 31:
the disinfectant for the Carmyl forest aims at the disinfection of the Bacillus cereus at the temperature of-40-0 ℃, wherein the Carmyl forest is 30% (v/v), the deicer ethanol is 64% (v/v), and the solvent is water.
Example 32:
the disinfectant for the Carmyl forest aims at the disinfection of the Bacillus cereus at the temperature of-40-0 ℃, wherein the Carmyl forest is 23% (v/v), the deicer ethanol is 2% (v/v), and the solvent is water.
Example 33:
the disinfectant of the present embodiment is used for disinfecting bacillus cereus at a temperature of-40 ℃ to 0 ℃, and compared with the disinfectant of the embodiment 30, the deicer is glycol, and the content of the deicer is 10% (v/v).
Example 34:
the disinfectant of the present embodiment is used for disinfecting bacillus cereus at a temperature of-40 ℃ to 0 ℃, and compared with the disinfectant of the embodiment 30, the deicer is ethylene glycol and the content of the deicer is 32% (v/v).
Example 35:
the disinfectant of the present embodiment is used for disinfecting bacillus cereus at a temperature of-40 ℃ to 0 ℃, and compared with the disinfectant of the embodiment 30, the deicer is ethylene glycol and the content of the deicer is 55% (v/v).
Example 36:
the disinfectant of the present embodiment is used for disinfecting bacillus cereus at a temperature of-40 ℃ to 0 ℃, and compared with the disinfectant of the embodiment 30, the deicer is ethylene glycol and the content of the deicer is 5% (v/v).
Example 37:
the disinfectant formulations of examples 1-36 were used to perform disinfection experiments on vehicles on the bank of Manchurian ports, wherein the temperatures of the experiments of examples 1-36 are listed in Table 1, the disinfection duration for propagating bacteria and viruses was 15min, and the disinfection duration for spores was 60 min.
The breeding bacteria are selected from Escherichia coli and Staphylococcus aureus, the viruses are selected from Newcastle Disease Virus (NDV), and the spores are selected from Bacillus cereus.
The experimental method for disinfecting the breeding bacteria and the spores comprises the following steps:
will dyeBacterial sheet (10) with specific bacteria5～106CFU/piece) is placed in an autoclaved sterile filter paper bag, the mouth of the bag is tightened, and different parts of the vehicle to be sterilized are placed. And after the disinfection is finished, counting the viable bacteria and calculating the sterilization rate. According to the regulation of the veterinary drug evaluation center 992-No. 101-animal disinfectant identification technical specification (trial) of Ministry of agriculture, the sterilization rate reaches 99.9 percent, and the disinfectant can be evaluated to be effective.
The preparation method of the breeding bacterium and spore bacterium tablet comprises the following steps:
(1) filter paper of size 10mm by 10mm was used as support material. After the carrier is sterilized by high-pressure steam, the carrier is infected by a drop-dyeing method.
(2) Adjusting the bacterial content of the bacterial suspension to about 1 × 10 by using a turbidimeter8CFU/mL～5×108CFU/mL, spreading the carrier filter paper in a clean sterile plate, dropwise adding 20 mu L of bacterial liquid by a liquid transfer machine, and staining the bacteria by a drop staining method. Placing in a constant temperature incubator at 37 ℃ or drying at room temperature for later use.
(3) The number of recovered bacteria per bacterial piece (carrier) should be 5X 105CFU/sheet-5X 106The bacterial concentration of the CFU/disc, all discs in the same batch test must be the same.
The preparation method of the reproductive bacterium and spore bacterium suspension comprises the following steps:
(1) preparation of bacterial propagule suspension
a. Opening the strain tube in aseptic operation, adding appropriate amount of nutrient broth culture medium with capillary suction tube, and slowly blowing and sucking for several times to melt and disperse strain. Taking a test tube containing 5.0 mL-10.0 mL of nutrient broth culture medium, dripping a little strain suspension, and culturing at 37 ℃ for 18 h-24 h. Taking the bacterial suspension cultured in the 1 st generation by using an inoculating loop, streaking and inoculating the bacterial suspension on a nutrient agar culture medium plate, and culturing for 18-24 h at 37 ℃. Or taking one strain from the microbank, inoculating the strain on a plate, culturing for 18-24 h at 37 ℃, selecting a typical colony in the 2 nd generation culture, inoculating the typical colony on a nutrient agar slant, and culturing for 18-24 h at 37 ℃, thus obtaining the 3 rd generation culture.
b. Taking a 3 rd to 6 th generation nutrient agar culture medium to culture a fresh slant culture for 18h to 24h, sucking 3.0mL to 5.0mL of diluent (generally TPS (sodium hydrosulfite) and normal saline for acidified water) by using a 5.0mL suction pipe, adding the diluent into a slant test tube, repeatedly blowing and sucking, after bacterial lawn is washed, transferring a washing solution into another sterile test tube, and mixing for 20s by using an electric mixer or shaking for 80 times on a palm to uniformly mix the bacteria.
c. The bacteria concentration of the primarily prepared bacteria suspension can be primarily determined by using a bacteria turbidimeter, and then the bacteria suspension is diluted to the required concentration by using a diluent.
d. The bacterial propagule suspension should be stored in a refrigerator at 4 ℃ for later use without being used overnight.
(2) Preparation of bacterial spore suspension
a. Opening the strain tube in aseptic operation mode, adding appropriate amount of nutrient broth culture medium with capillary suction tube, and blowing and sucking gently for several times to melt and disperse strain. Taking a test tube containing 5.0 mL-10.0 mL of nutrient broth culture medium, dripping a little strain suspension, and culturing at 37 ℃ for 18 h-24 h. Inoculating the bacterial suspension cultured in the 1 st generation with an inoculating loop, inoculating on a nutrient agar slant, and culturing at 37 ℃ for 18-24 h. And (3) selecting typical colonies in the culture of the 2 nd generation, inoculating the typical colonies on a nutrient agar slant, and culturing at 37 ℃ for 18-24 h to obtain the culture of the 3 rd generation.
b. Taking the nutrient agar culture medium of the 3 rd to 6 th generation to culture a fresh slant culture for 18h to 24h, taking a colony by using an inoculating loop to inoculate the colony on the nutrient agar slant culture, and placing a slant test tube in a constant temperature incubator at 37 ℃ to culture for 5d to 7 d.
c. And (3) taking a little of lawn by using an inoculating loop, coating the lawn on a glass slide, fixing, staining by a gram staining method, and performing microscopic examination under a microscope (oil microscope). When the spore formation rate is 95% or more, the following treatment is performed. Otherwise, the mixture is kept standing at room temperature for a certain time until the spore formation rate is reached, and then the following treatment is carried out.
d. Sucking 3.0-5.0 mL of physiological saline by using a 5.0mL suction tube, adding into the inclined test tube, repeatedly blowing and sucking, and washing off the lawn. The aspirated bacterial suspension was concentrated in a sterile Erlenmeyer flask containing glass beads and shaken for 5 min.
e. The Erlenmeyer flask is placed in a water bath at 45 ℃ for 24 hours to ensure that the bacteria are subjected to autolysis and are dispersed into single spores.
f. The spore suspension is filtered through sterile cotton or gauze to remove the agar clot.
g. The spore suspension is placed in a sterile centrifuge tube and centrifuged for 30min at the speed of 3000 r/min. Discarding the supernatant, adding distilled water to blow and suck the spores again, and repeating the step for 3 times.
h. The cleaned spore suspension is put into a triangular flask containing a proper amount of small glass beads, and water bath is carried out at 80 ℃ for 10min to kill residual bacterial propagules. After cooling to room temperature, shaking, storing in a refrigerator at 4 ℃ for later use. The effective service life is half a year.
i. When the spore suspension is used, viable bacteria should be cultured and counted firstly.
The method for counting the live bacteria of the sterilized bacterial tablets comprises the following steps:
c. and (3) putting the bacterial sheet into a sterile PE tube containing 1.0mL of neutralizing agent after the test bacteria and the disinfectant interact for each set time, and mixing the bacterial sheet in an electric mixer for 20s or shaking and knocking the bacterial sheet on a palm for 80 times to elute the bacteria on the bacterial sheet into the neutralizing solution.
d. The number of viable bacteria was measured by the viable bacteria culture counting method by sucking 100. mu.L of the above-mentioned washing solution.
e. Before the sprayer is replaced with the disinfectant with new concentration, the original residual disinfectant is cleaned and replaced with the disinfectant with new concentration.
f. And (4) setting a positive control for directly using the infectious microbe carrier as viable count.
g. All test specimens were incubated at 37 ℃ in an incubator and the final count was observed after 24h incubation.
h. The test was repeated 3 times, and the number of viable bacteria (CFU/disc) of each group was calculated and converted into a logarithmic value to calculate the sterilization rate.
The sterilization rate calculation method comprises the following steps:
the sterilization rate or the kill index is used as an evaluation criterion. According to the standard sterilization rate (Pt) of Ministry of agriculture (sterilization technical specification), the sterilization rate reaches 99.9 percent, and the calculation formula is as follows:
sterilization rate at time of sterilization t (Pt) [ (n)0-nt/n0)]×100％
In the formula n0The number of live bacteria is before disinfection or in a control group; n istThe number of live bacteria after disinfection or experimental groups.
The results of the sterilization experiments are shown in Table 1.
Example 38:
the corrosiveness of metals was determined using the disinfectant of example 31.
Method step
(1) Soaking in surface active detergent for 10min, removing oil, cleaning, grinding with 120 # abrasive paper to remove oxide layer on the surface of the metal sheet, and washing with tap water. The diameter, thickness, and pore diameter (to the nearest 0.1mm) of the metal sheet were measured, respectively. Degreasing again by using anhydrous acetone or anhydrous ethanol. Drying in a constant temperature incubator at 50 deg.C for 1h, weighing after the temperature is reduced to room temperature, weighing each metal sheet for 3 times to 0.1mg, and taking the average value as the weight before test. When weighing, the metal sample wafer can not be directly touched by hands when wearing clean gloves.
(2) The disinfectant used in the test is prepared according to the highest concentration of the disinfectant used for soaking the test sample. During soaking, each metal sheet needs to be soaked in 200mL of disinfectant.
(3) The metal sample is suspended in the disinfectant by plastic wires and marked with a number and date by a label. Soaking for 72h at one time. If the disinfectant contains volatile or unstable effective components, the disinfectant is replaced according to the situation until the disinfectant is soaked for 72 hours.
(4) Each metal was tested in 3 coupons per test. When soaking, if each sample of the same metal is separated by more than 1cm, the soaking can be carried out in the same container (containing 600mL of disinfectant).
(5) After soaking for a prescribed time, the metal sheet is removed, rinsed with tap water and then removed with a brush or other flexible implement. If corrosion products remain unremoved, the following removal method can be used as described in GB 10124:
copper sheet: soaking in hydrochloric acid solution (500mL, 36-38% hydrochloric acid and distilled water to 1000mL, the specific gravity of hydrochloric acid is 1.19) at room temperature for 1-3 min.
Carbon steel sheet: boiling in sodium hydroxide solution containing zinc powder 200g/L for 5-30 min.
Aluminum sheet: soaking in chromic anhydride phosphoric acid solution (20 g of chromic anhydride, 500mL of phosphoric acid, and distilled water to 1000 mL. the specific gravity of phosphoric acid is 1.69), heating to 80 deg.C, and maintaining for 5-10 min. If not cleaned, the glass can be immersed in a nitric acid (specific gravity of 1.42) solution for 1min at room temperature.
Stainless steel: soaking in 60 ℃ nitric acid solution (66-68% nitric acid 100mL and distilled water to 1000mL) for 20 min. Or soaking in 70 deg.C ammonium citrate solution (150 g ammonium citrate with distilled water to 1000m L) for 10-60 min.
(6) Removing corrosion products from the metal sample, cleaning, sucking surface water by using filter paper, placing the metal sample in a plate filled with the filter paper, placing the plate in a constant-temperature incubator at 50 ℃, drying for 1h, clamping by using tweezers, and weighing on a balance respectively after the temperature of the metal sample is reduced to room temperature. The average value of 3 weighed times was used as the weight after the test.
When weighing, the same as before the test, clean gloves should be worn without touching the sample directly with the hand (the same below).
(7) When the sample is used to remove the corrosive substance by chemical method, a corresponding blank control is needed to be set up so as to correct the error. The blank control sample was surface treated, cleaned and weighed as the test group sample, but without disinfectant soak. The samples of the test group were then subjected to chemical treatment, water rinsing, drying, weighing, and the average loss of weight was calculated.
(8) A contrast that the stainless steel sheet is soaked in distilled water is required in the whole test process, and the weight difference before and after soaking is less than 0.3 mg. Otherwise, after finding the cause, all tests are redone.
(9) Expressed as the average of the metal corrosion rate (R), the loss of weight value of the blank control coupon should be subtracted from the calculation. The calculation formula is as follows:
wherein R is corrosion rate, mm/a (mm/year); m is the weight of the metal sheet before the test, g; m istWeight of the sheet metal after the test, g; m iskFor chemical treatment to remove the weight loss value g of the corrosion product sample wafer, and for those without chemical removal treatment in the test, the formula is calculated by deleting mkA value; s is the total surface area of the metal sheet, cm2(ii) a t is test time, h; d is the density of the metal material in kg/m3。
Grading standard of corrosivity
TABLE 1 Sterilization test results Table
Of examples 1-28, the formulations of examples 1-3, examples 5-7, examples 9-11, examples 13-15, examples 17-19, examples 21-23, examples 25-27 were formulated according to the claimed formulations, the formulations of examples 4, 8, 12, 16, 20, 24, 28 were outside the claimed scope of the invention, and the amount of deicing agent was below the claimed level; as is clear from the above table, the disinfectant effects of the disinfectants of examples 1 to 3, examples 5 to 7, examples 9 to 11, examples 13 to 15, examples 17 to 19, examples 21 to 23 and examples 25 to 27 can meet the requirement of the veterinary drug evaluation center 1992 No. 101 of the agricultural (herding) letters of the Ministry of agriculture (Processary of agriculture) on the identification technical Specification of disinfectant for animals (trial), and have excellent killing effects on reproductive bacteria and viruses; the disinfectant effects of the examples 4, 8, 12, 16, 20, 24 and 28 do not meet the regulation of No. 101 & lt & gt veterinary disinfectant identification technical Specification (trial) of the agricultural ministry of veterinary drug review center 1992, and the disinfectant has poor killing effect on reproductive bacteria and viruses;
of examples 29-36, the formulations of examples 29-31, examples 33-35 were formulated according to the claimed formulations, the formulations of examples 32, 36 were outside the claimed scope of the invention, and the amount of deicing agent was below the claimed range; as is clear from the above table, the disinfecting effects of the disinfectants of examples 29-31 and examples 33-35 can meet the requirements of the veterinary drug evaluation center 1992 No. 101 of the agricultural (herdsmedicine) of the technical Specification for identifying the disinfectant for animals (trial implementation), and have excellent killing effect on spores; the disinfectant effects of the examples 32 and 36 are not in accordance with the regulation of No. 101 of the letter "veterinary disinfectant identification technical Specification" (trial) of agricultural (herding medicine) of the animal remedy center 1992 of Ministry of agriculture, and the disinfectant effect on spores is poor.
The disinfectant of example 31 was used to soak copper, aluminum, carbon steel and stainless steel sheets for 72h, and the metal sheets were weighed before and after soaking, respectively, to calculate the corrosion rate of the methomyl disinfectant on the metal sheets (fig. 1). Comparing the corrosion rates of 4 different metal sheets, the methomyl disinfectant has no corrosion to the copper sheets and the stainless steel sheets; the corrosion rate of the aluminum sheet reaches 0.029mm/a, while the corrosion rate of the carbon steel sheet is the fastest of the 4 metal sheets, and the corrosion rate reaches 0.107 mm/a. Therefore, according to the corrosion rate, the sterilization combination of the methomyl forest has no corrosion to the copper sheets and the stainless steel sheets basically (R is less than 0.01), has slight corrosion to the aluminum sheets (R is more than 0.0100 and less than 0.100), and has moderate corrosion to the carbon steel sheets (R is more than 0.100 and less than 1.00). It can be seen that the disinfectant formulation of example 31 is less corrosive to metals, whereas the concentration of methomyl in example 31 is higher, and the remaining disinfectant formulations containing a lower concentration of methomyl have lower metal corrosivity, so that the claimed disinfectant can be used as a low temperature disinfectant formulation for the disinfection of port sites in northern regions.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (11)
1. The disinfectant for the methomyl forest suitable for low-temperature use is characterized in that the use temperature is-40-0 ℃, and the components of the disinfectant are the methomyl forest, a deicing agent and a solvent which are mixed in proportion.
2. A methomyl disinfectant suitable for low temperature use as claimed in claim 1, wherein the concentration of methomyl is 0.1-0.5% (v/v) and the solvent is water.
3. The disinfectant for methomyl suitable for low temperature use according to claim 2, wherein the use temperature is-20 to 0 ℃, the deicer is ethanol, and the content of the deicer is 5 to 41% (v/v).
4. The disinfectant for methomyl suitable for low temperature use according to claim 2, wherein the use temperature is-20 to 0 ℃, the deicer is glycol, and the content of the deicer is 10 to 50% (v/v).
5. The disinfectant for methomyl suitable for low temperature use according to claim 2, wherein the use temperature is-20 to 0 ℃, the deicer is propylene glycol, and the content of the deicer is 5 to 50% (v/v).
6. A methomyl disinfectant suitable for low temperature use as claimed in claim 2, wherein the use temperature is-20 to 0 ℃, and the deicer is sodium chloride in an amount of 0.3 to 23 wt%.
7. A methomyl disinfectant suitable for low temperature use according to claim 2, wherein said deicer is ethanol at a temperature of-40 to-20 ℃ and the content of said deicer is 35 to 64% (v/v).
8. A methomyl disinfectant suitable for low temperature use according to claim 2, wherein said deicer is ethylene glycol or propylene glycol at a temperature of-40 ℃ to-20 ℃ and the content thereof is 35-55% (v/v).
9. A methomyl disinfectant suitable for low temperature use as claimed in claim 1, wherein the methomyl concentration is 15-30% (v/v) and the solvent is water.
10. A methomyl disinfectant suitable for low temperature use according to claim 9, wherein said deicer is ethanol in an amount of 5-64% (v/v).
11. A methomyl disinfectant suitable for low temperature use according to claim 9, wherein said deicer is ethylene glycol in an amount of 10-55% (v/v).
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| CN112753699A (en) * | 2020-12-30 | 2021-05-07 | 中国疾病预防控制中心环境与健康相关产品安全所 | Low-temperature chlorine-containing disinfectant and preparation and application thereof |
| CN114794143A (en) * | 2021-01-21 | 2022-07-29 | 苏州倍爱尼生物技术有限公司 | Hydrogen peroxide disinfectant suitable for use in low temperature environment above-24 DEG C |
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