RU2534356C2 - Method of quality control of disinfection of poultry premises - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to the field of quality control of disinfection. The method comprises disinfection of premises, selection of test objects which are used as a suspension Bacillus subtilis of the strain ATCC PTA-6737 (PB6) sorbed on the carrier - the filter paper with an adhesive surface part. The test objects are placed in the premises of disinfection and added to the nutrient medium. The tubes are heated to 60-80°C and cultured at 37°C for 12-24 hours followed recording the result by recording the growth of bacteria after 12 hours. Clouding of the nutrient medium to form a mucous film is indicative of inadequate quality of disinfection, absence of clouding and formation of a film after 24 hours is indicative of the satisfactory disinfection.

EFFECT: invention enables to simplify the quality control of disinfection of premises.

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Description

The invention relates to veterinary sanitation, and in particular to methods for controlling the quality of aerosol and gas disinfection of poultry rooms.

A known method of monitoring the quality of disinfection, based on microbiological methods, proposed by A.A. Polyakov in 1957 (Polyakov A.A. Once again about the theory and practice of veterinary disinfection / A.A. Polyakov, A.V. Kulikovsky // - Veterinary . - 1989. - No. 2. - S.19-23). The criterion of this method is the presence or absence of Escherichia coli group bacteria in the studied objects.

Also, the quality of disinfection can be determined by the absence of staphylococci (Rules for the disinfection and disinfestation of objects of state veterinary supervision "(approved by the Ministry of Agriculture of the Russian Federation on July 15, 2002 N 13-5-2 / 0525)).

These methods do not allow to reliably confirm the absence of more stable microflora at the facility, because These test microorganisms, in terms of resistance to disinfectants, belong to the second group, and most pathogenic microorganisms that are important for poultry farming, in terms of resistance, belong to the second and third groups: pathogens of salmonellosis, influenza, bursal disease, reovirus tenosynovitis, smallpox of birds.

The prototype of the proposed method is a biotest based on the destruction of spores of the strain Bacillus stearothermophilus VKM B-718 applied to the test object (Sanitary rules SP 1.2.011-94. Goskomsanepidnadzor Russia. M., 1994, p.138). However, the cultivation of this microorganism requires expensive nutrient media and a temperature of 55 ° C.

The objective of the proposed solution is to expand the arsenal of ways to control the quality of disinfection of premises for poultry.

The problem is solved in that in a method for controlling the quality of disinfection of poultry facilities, including placing test objects on a carrier before disinfection, with the test microorganism spores Bacillus subtilis applied according to the invention, the Bacillus subtilis ATCC- strain is used as a test microorganism PTA 6737 (PB6).

The invention also consists in the fact that filter paper with an adhesive surface is used as a carrier.

The invention also consists in the fact that the cultivation is carried out at 37 ° C for 1 day on meat and peptone broth. The method is as follows:

Materials and equipment: thermostat 37 ° С, water bath 70-80 ° С, drying cabinet 70 ° С, 10 G centrifuge, shaker, shredder, bacteriological tubes with sterile MPB, filter paper, double-sided adhesive tape, physiological saline.

Protocol for obtaining a test object:

1. Build tank. masses: cultivate Bacillus subtilis PB6 on BCH using a shaker for aeration and increasing the rate of accumulation of the tank. mass at 37 ° C - 2 days.

2. Translation of the vegetative form into a spore: anaerobic conditions - 1 day.

3. Washing and concentration of spores: centrifuge a suspension of spores in a nutrient medium at 10 G for 20 minutes. Remove the supernatant and resuspend the natant in distilled water in a ratio of 1: 100.

4. Sorption of spores on a carrier - immerse the “blue ribbon” filter paper in a spore suspension, dry in an oven at 60-70 ° С.

5. Production of test strips: apply strips of double-sided adhesive tape 1 cm wide with an interval of 5 cm onto the carrier sheet and pass through the shredder in the transverse direction.

6. Storage: at 2-8 ° C and relative humidity not more than 20% - 2 years.

Samples for the study: the test objects subjected to disinfection should be taken after removal or inactivation of the disinfectant for a period of 0-6 hours.

Disinfection Quality Control Protocol:

1. Place the test objects in the disinfection object in advance or immediately before it begins.

2. Disinfect the object with an aerosol or gas.

3. After removal or inactivation of the disinfectant, the test objects are taken aseptically as a whole, separating them with sterile forceps, or partially, cutting off the end of the strip with sterile scissors and placed in bacteriological tubes with BCH.

4. Warm up test tubes with objects in the BCH in a water bath at 80 ° C for 20 minutes to reduce the risk of secondary contamination.

5. Cultivate the selected material at 37 ° C for 12-24 hours. After 12 hours, we can ascertain the growth, and after 24 - its absence.

6. Interpret the result of the study depending on the absence or presence of growth of Bacillus subtilis (clouding of the MPB with the formation of a mucous membrane on the surface of the nutrient medium) as negative or positive.

Tests of the disinfection quality control method in a biotest with B.subtilis spores of strain ATCC-РТА 6737 (РВ6) in comparison with reference methods were carried out with volumetric disinfection of poultry houses for floor and cell methods for keeping broiler chickens using the apparatus for generating aerosol AG-UD-2 and the drug "Virocid" 2% at the rate of 1 l / 40 m (Table 1) and the apparatus "AIST-2M" with formalin at the rate of 20 ml / m (Table 2) with an exposure of 12 hours.

Bacterial contamination of the premises was monitored after unloading from the premises of the bird before the stage of mechanical cleaning (washing) of the premises and after washing using reference methods: detection of bacteria of the Escherichia coli group and the genus Staphylococcus, and after disinfection using the test method. 20 samples of material were taken from the surfaces of the room (swabs from the 10 * 10 cm section or test objects) from different levels vertically and horizontally for bacteriological examination.

The results of the study were taken into account by reference methods 2 days after sowing, and the test subject 1 day later.

Table 1 Disinfection with the device AG-UD-2 with the drug "Virocid" 2% at the rate of 1 l / 40 m with an exposure of 12 hours Bacteria of the Escherichia coli group, positive samples,% Staphylococcus spp., Positive samples,% Biotest with spores of B. subtilis, positive samples,% Broiler Floor Housing, 20 samples before washing and disinfection 85 one hundred Did not After washing 35 80 Did not After disinfection 0 5 10 Broiler cage housing, 20 samples before washing and disinfection 90 one hundred Did not After washing 40 80 Did not After
disinfection 0 10 fifteen

The data presented indicate that the proposed method for controlling the quality of disinfection can be used to control volumetric aerosol disinfection using the apparatus AG-UD-2 and disinfectant Virocid at a concentration of 2% at a saturation of 1 l / 40 m with an exposure of 12 hours.

table 2 Disinfection with AIST-2M using formalin at a rate of 20 ml / m ³ with an exposure of 12 hours and aerosol neutralization of 5% aqueous ammonia at a rate of 15 ml / m Bacteria of the Escherichia coli group, positive samples,% Staphylococcus spp., Positive samples,% Biotest with spores of B.subtilis, positive samples,% Broiler cage housing, 20 samples before washing and disinfection 75 one hundred Did not After washing thirty 85 Did not After disinfection 0 0 0 Broiler cage housing, 20 samples before washing and disinfection 95 one hundred Did not After washing 45 75 Did not After disinfection 0 0 0

The data presented also indicate that the proposed method for controlling the quality of disinfection can be used to control volumetric aerosol disinfection using the AIST-2M apparatus with formalin at a rate of 20 ml / m ³ with an exposure of 12 hours and neutralization with an aerosol of 5% aqueous ammonia at a rate of 15 ml / m ³ .

The data presented in the tables indicate a greater, in comparison with the reference methods, sensitivity and reliability of the method, and the use of the cultivation mode at 37 ° C for 1 day on BCH provides a research speed of 1 day.

Claims (1)

A method for controlling the quality of disinfection of poultry facilities, which includes disinfection of the premises, selection of test objects, which are a suspension of Bacillus subtillis strain ATCC-PTA 6737 (PB6) on a carrier - filter paper with an adhesive surface, placing test objects in a nutrient medium, heating test tubes with the test objects in a water bath to 60-80 ^° C, cultivation at 37 ^° C for 12-24 hours followed by considering the result by recording the growth of bacteria through 12 hours - the nutrient medium to form a blurred slizis nd indicates poor film as disinfection, no cloud point and film formation at 24 hours- of satisfactory disinfection.