A Field Technique Measuring Virus Decay and Potential Aerosol Hazard from Wastewater Sprinkler Irrigation

The increased use of domestic wastewater for irrigation purposes has stimulated a growing practice of sprinkler irrigating from oxidation ponds and other domestic wastewaters. Aerosols generated from these sprinkler irrigation systems may contain potentially hazardous pathogens. Subsequently, the aerosols can contain infective viruses which can be carried through the air to surrounding populations. Thus, a public health hazard can be created by sprinkler irrigating domestic wastewater. This study is an investigation of a means by which the virus decay rate of viruses in aerosols and the potentials hazard of sprinkler irrigation aerosol clouds may be examined. A means of injection of a human and animal virus stimulant, MS-2 bacteriophage, is described. In addition, the factors which are known to effect the survival of viruses in aerosols are discussed. The ambient air factors that are known to effect virus survival and which are discussed include relative humidity, air temperature, solar radiation, and aerosol age. The suspending fluid factors that are known to effect virus survival and which are discussed include dissolved inorganic salt contact, dissolved organic content, filterable solids and pH. The decay rate of the aerosolized MS-2 was measured with an all-glass impinger (AGI-30) when the wind velocity and distance of the sampler from the sprinkler system, and the initial and final concentrations of aerosolized MS-2 virus were known. The aerosol hazard of a domestic wastewater sprinkler irrigation system is defined in terms of the likelihood of infective aerosol particles to be inhaled and penetrate the human lung. Thus, the aerosol hazard is a function of aerosol particle size. The aerosol particle size distribution of the infective aerosol cloud was measured wit the Andersen sampler. A high and homogeneous concentration of the virus in the wastewater was insured by using pressure differentials in the sprinkler irrigation delivery line. By injecting the MS-2 virus into the line at a constant rate along with a tracer, Bacillus subtilus var niger (Bacillus globigii) spores, the decay rate of the virus during airborne exposure to environmental factors could be determined. The decay rate was determined assuming the environmental factors had no affect on the concentration of the tracer. It was proposed that the environmental engineer, after knowing the virus decay rates under varying environmental conditions, can define buffer zones which would be required around sprinkler irrigation sites. The buffer zone would reduce the possibility of contaminating humans by prohibiting access. The usefulness of the filed technique was demonstrated and the MS-2 was found to undergo a 33.3 percent decay per minute in the dark (no solar radiation) at 33 percent relative humidity, 70.6 percent dissolved organic material, 29.4 percent dissolved inorganic salts, 30.1 mg/1 filterable solids, and 17 degree C air temperature. The aerosol cloud resulting from the spray irrigation process appeared potentially hazardous because the Andersen sampler collected 84.2 percent of the virus infective droplets in the size range that could be inhaled

Studies on Viruses in Water

A new procedure for the detection of viral antigens in fecal material was developed. The test is performed by first diluting a fecal sample with phosphate buffered saline to give a liquid consistency. The pH is then adjusted to 8.5-9.0 and the solids are allowed to settle for five minutes. Supernatant fluid from above the fecal sediment is placed on the upper surface of a well of an inverted Immulon microtiter plate and incubated for one hour at 37 degrees C to allow virus to adsorb to the plastic. The Immulon plate is then washed three times with a Tween 20 solution and dried. Adsorbed virus is stained with fluorescein labled antiviral antibody containing Evan\u27s Blue dye. The stained preparations are examined by epi-fluorescence microsopy for the presence of viral aggregates and virus-containing cellular membranes. The test is applied in a continuous water monitoring procedure that can be used to upplement methods in which infectious viruses are isolated from water. In another study a protamine sulfate procedure for concentrating and an immunofluorescent cell procedure for assaying infectious virus (IV, reovirus that is infectious without proteolytic enzyme treatment), and potentially infectious virus (PIV, enzyme enhanceable reovirus) from polluted waters have been developed. The presence of PIV inthe environment had not previously been investigated. In following these procedures, protamine sulfate concentratiosn of 0.005 percent for the first precipitation of the sample, and 0.0025 percent for the second were used. With these protamine concentrations and 0.25 percent fetal bovine serum, IV and PIV are concentrations over 500-fold from river water inoculated with virus. Virus recoveries are between 80 and 100 percent. The IV and PIV fractions are assayed respectively before and after treatment with 200 ug fo chymotrypsin per millileter. When PIV is precipitated from river water, and the precipitate is dissolved and stored at 20 degrees C as a protamine-virus concentrate, only 5 percent of the viral infectivity is lost after 14 days. Therefore, reovirus can be precipitated from water at the sampling site, and only the protamine concentrate needs to be taken to the laboratory to be examined for virus content. When reoviruses are treated with chlorine, PIV is more resistant to inactivation thatn IV, and PIV appears to be at least as resistant to chlorination as poliovirus and coxsackievirus A-2. Granular media filtration systems (i.e., sand, anthracite coal and sand; anthracite coal; sand and garnet) are ineffectual in the removal of the acteriophage MS 2 from water when used as in-line direct filters. Batch assays have indicated a 93 percent reduction of MS 2 can occur when polyelectrolytes are added to the water. In addition, alum concentrations of 20, 30, 40, and 50 mg/1 remove 80 to 98 percent of the virus by precipitation. No reduction of MS 2 was observed at alum concentrations from 1 to 10 mg/1