Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (13): 2906-2915.doi: 10.3864/j.issn.0578-1752.2021.13.018

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

The Ability of Acidic Calcium Sulfate to Kill Common Clinical Pathogenic Microorganisms

FU XiaLi(),ZHENG ZiFang,MA ZhiQian,XU LeLe,LI ZhiWei,LI Yang,XIAO ShuQi,LI Shuang()   

  1. College of Veterinary Medicine, Northwest Agricultural and Forestry University, Yangling 712100, Shaanxi
  • Received:2020-06-01 Revised:2020-12-30 Online:2021-07-01 Published:2021-07-12
  • Contact: Shuang LI E-mail:1579459296@qq.com;lishuang2006001@126.com

Abstract:

【Objective】The purpose was to study the effect of disinfectant acid calcium sulfate (ACS) on the killing of microorganisms, and to provide basic data and theoretical basis for disinfectant prevention and control of human and animal diseases.【Method】After incubating ACS with 3% lecithin and 5% Tween-80 as phosphate buffer neutralizer for a period of time, Escherichia coli, Staphylococcus aureus, Salmonellaand porcine reproductive and respiratory syndrome virus (PRRSV) were added respectively for a period of time, We spread the culture mixture with bacteria on nutrient agar plates and cultured at 37℃ biochemical incubator for 18-24 h, the mixture with virus was inoculated on the cell plates and incubate in 37℃ cell incubator for a certain period of time to evaluate the neutralizing effect of the neutralizer. The comparison settings and evaluation criteria were as follows: disinfectant mixed with bacterial suspension or virus suspension (group 1), the mixture of disinfectant and bacterial suspension or virus suspension was mixed with neutralizer (group 2), the mixture of neutralizer and disinfectant was mixed with bacterial suspension or virus suspension (group 3), the mixture of sterile hard water and bacterial suspension or virus suspension was mixed with neutralizer (group 4), the mixture of sterile hard water and bacterial suspension or virus suspension was mixed with PBS (group 5), sterile hard water mixed with PBS (group 6). The evaluation criteria of the neutralizer effect in the bacteria killing test were as follows: group 1 had a very small amount of bacterial growth or aseptic growth; group 2 had bacterial growth, which was significantly less than that of groups 3, 4, and 5, but more than the group 1; the number of bacteria in group 3, 4,5 was close to that of the positive control group, no bacterial growth in group 6. All three repeated tests met the above conditions, and the results were consistent, it was determined that the selected neutralizer and concentration were appropriate. The evaluation criteria of the neutralizer effect in the virus inactivation test were as follows: group 1 had very little virus growth or no virus growth; group 2 had virus growth and was significantly less than that of groups 3, 4, and 5, but more than group 1. The growth of viruses in groups 3, 4, and 5 was similar to the original inoculation; the cells in group 6 grew normally. The results of the three repeated tests were consistent, and it was determined that the selected neutralizer and concentration were appropriate. We used the suspension quantitative sterilization method and the method of determining the virus titer to evaluate the elimination effect of ACS on the above-mentioned bacteria and viruses. We diluted the ACS 200, 300, 400, 500, 600, and 700 times to interact with the above-mentioned bacteria or virus for different time and added neutralizer for neutralization, then spread the bacterial mixture on nutrient agar plates to evaluate the killing effect of ACS by calculating the number of colonies, and determined the virus titer in the virus mixture to evaluate the killing effect of ACS on the virus.【Result】 The selected neutralizer can effectively neutralize the residual effects of ACS 200-fold dilution on bacteria and viruses, and the neutralizer was non-toxic to bacteria, viruses and cells. When ACS interacted with the bacteria for 0 h, it was neutralized immediately. The sterilization rate ofEscherichia coli at the maximum dilution of 300 times was 100%, the sterilization rate of Staphylococcus aureusat the maximum dilution of 600 times was 100%, and when the maximum dilution was 700 times, the sterilization rate of Salmonella was 100%. When ACS was diluted to 700 times, it was neutralized one day or six days after treatment with the above bacteria, and the sterilization rate was 100%. In addition, when ACS was diluted to 200 times, it was neutralized 60 minutes after treatment with PRRSV, and no virus titer was detected.【Conclusion】 When ACS diluted 700 times withEscherichia coli,Staphylococcus aureus and Salmonella for one day or more, it can produce better killing effect; when ACS diluted 200 times with PRRSV for 60 minutes, it can completely kill the virus, which will provide strong data support for the selection of disinfectants in the farms and provide reference for the prevention and control of epidemic diseases.

Key words: acid calcium sulfate, microbial, neutralizing agent, killing effect

Table 1

The colony number of each test bacteria after neutralization by quantitative germicidal test neutralizer(cfu/mL)"

分组
Grouping
大肠杆菌
Escherichia coli
金黄色葡萄球菌
Staphylococcus aureus
沙门氏菌
Salmonella
1 1120 2500 1900
2 24300 36500 13800
3 35900000 27400000 20200000
4 32600000 23500000 20200000
5 37800000 25900000 23800000
6 0 0 0

Fig. 1

Effects of neutralizer, disinfectant and neutralization products on the growth of mack-145 cells A: the growth state of MARC-145 cells with normal medium only; B: growth state of MARC-145 cells with neutralizing agent; C: the growth state of MARC-145 cells with disinfectant added; D: MARC-145 cell growth state with neutralization product added"

Fig. 2

Growth of Escherichia coli in plates A: positive control, and the growth of Escherichia coli immediately neutralized with ACS of different dilution ratios; B: positive control, and the growth of Escherichia coli neutralized by neutralizing agent after 1 day of action with ACS with different dilution ratios; C: positive control, and the growth of Escherichia coli neutralized with neutralizer after 6 days of action with ACS with different dilution ratios "

Table 2

Killing rate of ACS with different dilution ratio to Escherichia coli "

试验菌
Test bacteria
稀释倍数
Dilution multiple
杀灭率 Killing rate (%)
0(h) 1(d) 6(d)
大肠杆菌
Escherichia coli
200 100 100 100
300 100 100 100
400 99.65 100 100
500 99.30 100 100
600 97.25 100 100
700 91.54 100 100

Fig. 3

Growth of Staphylococcus aureus in the plates A: positive control, as well as the growth of Staphylococcus aureus which was immediately neutralized with ACS with different dilution ratios; B: positive control, and the growth of Staphylococcus aureusneutralized with neutralizing agent after 1 day of treatment with ACS with different dilution ratios; C: positive control, and the growth ofStaphylococcus aureusneutralized with neutralizer after 6 days of action with ACS with different dilution ratios "

Table 3

Killing rate of Staphylococcus aureus by ACS with different dilution ratio "

试验菌
Test bacteria
稀释倍数
Dilution multiple
杀灭率 Killing rate (%)
0(h) 1(d) 6(d)
金黄色葡萄球菌
Staphylococcus aureus
200 100 100 100
300 100 100 100
400 100 100 100
500 100 100 100
600 100 100 100
700 98.32 100 100

Table 4

The killing rate of different dilution ACS to Salmonella "

试验菌
Test bacteria
稀释倍数
Dilution multiple
杀灭率 Killing rate (%)
0(h) 1(d) 6(d)
沙门氏菌
Salmonella
200 100 100 100
300 100 100 100
400 100 100 100
500 100 100 100
600 100 100 100
700 100 100 100

Fig. 4

Growth of Salmonella in plates A: positive control, and the growth of Salmonella immediately neutralized with ACS of different dilution ratios; B: positive control, and the growth of Salmonella which was neutralized with a neutralizer after 1 day of action with ACS with different dilution ratios; C: positive control, and the growth of Salmonella neutralized with a neutralizer after 6 days of action with ACS with different dilution ratios "

Table 5

The value of virus titer pair after the interaction with ACS with different dilutions"

稀释倍数 Dilution multiple 200 300 400 500 600 700
病毒滴度对数值 Virus titer value 0.00 2.94 4.56 4.63 4.69 4.75

Table 6

Mean inactivation value of ACS to PRRSV with different dilution ratio"

稀释倍数 Dilution multiple 200 300 400 500 600 700
平均灭活对数值 Average inactivation value 7.13 4.19 2.57 2.50 2.44 2.38
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