Adams, V. Dean

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Utah State University DigitalCommons@USU Reports Utah Water Research Laboratory January 1981 Sulfur Dioxide Treatment of Secondary sewage: Effect on Viruses V. Dean Adams Follow this and additional works at: https://digitalcommons.usu.edu/water_rep  Part of the Civil and Environmental Engineering Commons, and the Water Resource Management Commons Recommended Citation Adams, V. Dean, "Sulfur Dioxide Treatment of Secondary sewage: Effect on Viruses" (1981). Reports. Paper 659. https://digitalcommons.usu.edu/water_rep/659 This Report is brought to you for free and open access by the Utah Water Research Laboratory at DigitalCommons@USU. It has been accepted for inclusion in Reports by an authorized administrator of DigitalCommons@USU. For more information, please contact digitalcommons@usu.edu. SULFUR DIOXIDE TREATMENT OF SECONDARY SEWAGE: EFFECT ON VIRUSES by V. Dean Adams Report to International Environmental, Inc, Salt Lake City, Utah Submitted by Utah Water Research Laboratory Utah State University Logan, Utah 84322 August 1981 VIRAL INACTIVATION • OBJECTIVE • METHODS AND PROCEDURES RESULTS AND DISCUSSION CONCLUSIONS FUTURE STUDIES REQUIRED REFERENCES TABLE OF CONTENTS Pag~ 1 4 4 6 9 9 10 SULFUR DIOXIDE TREATMENT OF SECONDARY SEWAGE: EFFECT ON VIRUSES VIRAL INACTIVATION Recently there has been considerable concern regarding virus inactivation Ln wastewater using techniques such as chlorination, ozonation, ultraviolet light, etc. Some of the parameters required for inactivation of viral type microorganisms are shown in Table 1. Engineering aspects of chlorine contactor design have been reported by Longley (1978) and he concluded that conventional wastewater chlorination practice produced negligible viral inactivation at doses up to 17 mg/l. Use of turbulent mixing and plug flow contact chambers seem to improve treatment efficiency. It has been observed that the residual-contact time combinations for viral inactivation are greater in lake water than in distilled water and that water hardness or an associated water quality parameter, influenced sensitivity (Wedemeyer et al. 1978). Improved efficiency by using ultrasonic treatment during chlorination of f2 bacteriophage has been shown by Zotova et al. (1977) who attributed it to the destruction of viral aggregates. Repeated cycles of chlorination, dechlorination, and exposure of surviving progency (Poliovirus Type I (LSc» at pH 7 resulted in apparently more resistant viral cultures (Bates et al. 1977). Farooq et al. (1978) reviewed the literature on engineering design of ozone disinfection reactors and proposed contact times and residuals for appropriate operation. Small bubble size improved microorganism-bubble contact and increased gas-liquid interfacial area thus improving disinfection. Low pH enhanced ozone stability Table 1. Reported parameters for inactivation of viral type microorganisms. Organism Echovirus Type II Infectious Pancreatic Necrosis Virus (IPNV) IPNV IPNV Medium Disinfectant 10% fetal calf Chlorine serum ? Chlorine Distilled water Chlorine Distilled water Ozone Infectious Hematopoietic Distilled water Chlorine Necrosis Virus (IHNV) IHNV Poliovirus I and II Coxsackievirus A9 & B5 Echovirus 1 and 5 SV40 and KilhamRat Virus 6 Enteroviruses Poliovirus I Coxsackievirus Type B5 Distilled water ? Wastewater Sewage Buffer plus fetal calf serum I Ozone Chlorine Ozone Methylene blue Chlorine Contact Time (min) 0.5 5.0 1.0 1.0 0.5 0.5 0.3-3.4 1.5-96 10 5 0.5 Dose (D) or Residual (R) mgtl 1200 (D) 4 (D) 0.1 (R) 0.01 (D) 0.1 (R) 0.01 (D) 0.5 (R) pH 6 0.5 (R) pH 10 3.0 (D) 670 nm light 20 w/m2 pH 10 600-800 mg/l Criterion 3.5 logs Inactivation 4.5 logs Inactivation ? ? ? ? 2 logs Inactivation ? Reference Drulak et al. (1978b) Elliott and Amend (1978) Wedemeyer et al. (1978) " II " Engelbrecht et al. (1978) Evison (1978) Gerba et al. (1977) 99.999% Drulak et 'ala Inactivation (1978a) N and thus improved inactivation efficiency, but inactivation by ultra-violet radiation catalysis was not significant. 3 Parrella et al. (1977) reported that chlorine dioxide was more effective than chlorine in wastewater disinfection containing Poliovirus Type I (Sabin) and bacteriophage T4. Dryden et al. (1979) studied the removal of viruses in model systems for tertiary wastewater treatment (coagulation, sand filtration, direct filtration, and carbon adsorption followed by chemical disinfection with either chlorine or ozone). All systems produced virus reductions of at least five orders of magnitude. Berg et al. (1979) compared chlorine dioxide with chlorine as a dis-infectant for wastewater effluents and found that both were effective in reducing VLrus levels. The efficiency of chlorine as a raw wastewater disinfectant was found to be dependent on pH, temperature, time, demand, and mixing; however, the quantitative prediction of performance was stated to be difficult (Irving 1980). Hajenian and Butler (1980) have found that bacteriophage f2 was more sensitive at low pH than at high pH, whereas no consistent effect of pH on the sensitivity of poliovirus to chlorine could be noted. Ozone inactivation of six viruses in a continuous flow reactor was studied with a 40 fold difference existing between the most resistant and the most sensitive virus (Roy 1979). Keswick (1979) has found that BreI was two to three times more effective at inactivating poliovirus than chlorine in pH 6 buffer. In three studies, chlorine dioxide has been found to be a more effective disinfectant than chlorine against bacteria and viruses in wastewater (Longley et al. 1980, Berg et al. 1980, Aieta et al. 1980). 4 Many resources and much effort has been devoted to viral inactiva-tion studies due to potential serious health implications associated with viral contamination. As can be seen from the brief literature survey~ many different types of virus microorganisms under a variety of conditions have been treated with the most commonly used disinfectants. Because of the complex nature of wastewater and the "stamina" of virus microorganisms there is no clear or decisive inactivation pathway or an ultimate disinfectant which can classically destroy all pathogens. Thus there is still a tremendously active research effort taking place to solve many of the unanswered questions regarding viral inactivation. OBJECTIVE Determine the effect of sulfur dioxide treatment of secondary sewage containing 1) reovirus and 2) poliovirus. METHODS AND PROCEDURES The first set of experiments was conducted using reovirus. Nine beakers each containing 1 liter of secondary sewage were divided into three groups with each set of conditions being monitored Ln triplicate. The secondary sewage was collected (prior to chlorination) from the Hyrum wastewater treatment plant, Hyrum~ Utah. Three sets of conditions were established. One set of three samples termed "control" received no further treatment. Another set of three samples labeled "pH control" were brought to pH 2.5 by the addition of HCl. The "S02" group of three samples received 802 to the extent of 500 mg/l which resulted in pH of 2.5. 5 One ml of reovirus type II stock (previously assayed at 2 x 109 virus per ml) was added to each sample. The addition of virus to the sample marked time zero. The samples were mixed at 20 rpm throughout the experiment using a standard jar test apparatus. One ml samples were taken from each of the nine flasks at various time intervals. Each 1.0 ml sample was added to 1.0 ml of 0.15 M phosphate buffer pH 7.2 in a wet ice bath and maintained at approximately 4°C until assayed later that day. The assay for reovirus was the immunofluorescent cell count procedure utilizing Madin Darby Bovine Kidney cells (Spendlove 1967). Preliminary experiments were run to determine satisfactory buffers and dilutions that would allow assay of the treated sewage on mammalian cell cultures. The reproducibility of this assay is ~ 20 percent (Barnett 1975) when sufficient numbers of infected cells are counted. A second set of experiments was also run. The study was designed to evaluate concentrations of 500 mg/l S02 and 1000 mg/l S02 in secondary wastewater spiked with reovirus. The response of both infectious reovirus (IV) and potentially infectious reovirus (PIV) were monitored. The treatment and assay procedures were essentially the same as above. For this experiment the infectious reovirus was prepared by treating potentially infectious reovirus Type II with trypsin (Sigma type IX) at 100 mg/ml for 1 hour at 37°C. A third set of experiments was run using poliovirus. Similar conditions and experimental set up as for the pre-vious experiments were used. The secondary sewage was spiked with poliovirus to yield a final concentration of about 107 poliovirus per liter, which is 100 fold higher than what is usually found in raw sewage from large metropolitan 6 areas. The pH of the sewage was 7.3, the pH after addition of sulfurous acid was 2.6. RESULTS AND DISCUSSION The data from the first experimental run are given in Table 2. The data indicate a 90 percent reduction in reovirus by S02 treatment for 40 minutes. This is somewhat less inactivation than was observed following pH adjustment to 2.5 with HCl. A non-spiked sewage sample was assayed in parallel with these reovirus spiked samples. The non-spiked sample did not contain reovirus at levels sufficient to be detected by our assay procedure. The data for the second experimental run are presented in Table 3. These data are in approximate agreement with the data of previous experiments and they do not indicate any significant difference between the response of the two forms of reovirus to S02 treatment. After 40 minutes of treatment with S02 at 500 mg/l, 13 percent of the PIV remained and 27 percent of the IV remained. Using 1,000 mg S02 per liter left 10 percent of the PIV and 17 percent of the IV. The results of the S02 treatment of poliovirus in secondary wastewater are shown in Table 4. The S02 treatment clearly inactivated the poliovirus beyond the degree observed following simple pH adjustment. The inactivation was rapid, having plateaued at the first time point (Le. 5 min.) and resulted in 97 percent of the infectious virus being removed. In summary, the third set of experiments using the poliovirus was the most encouraging. Disinfection conditions could possibly be optimized to achieve even a better inactivation of the poliovirus. Table 2. Effect of S02 treatment on reovirus viability Ln secondary sewage effluent. Sample Treatment Viable Reovirus Present in Sample at Selected Times After Addition of Reovirus. (Time in Minutes) 1 5 15 30 40 Control (no 2.5xl06 2.1xl06 adjustments) pH Control (pH to 4xl04 7xl04 2.5 with HC!) S02 (500 mg/l) 7xl05 4xl05 4xl05 2xl05 Table 3. Effect of S02 treatment on reOVLrus viability in secondary sewage ef fl uent • Sample Treatment Control Cont rol pH 2.4 Control pH 2.4 Control S02 500 mg/l; pH 2.4 S02 500 mg/l; pH 2.4 pH 2.1 Control pH 2.1 Control Viable Reovirus Present in Sample After Addition of S02 (Time in minutes). 1 5 15 30 40 PIVa 7. 7xl07 8.6xl07 IVb 4.0xl07 4.0xl07 PIV 7.7xl07 2.2xl07 IV 3.9xl07 2.2xl07 PIV 4.1xl07 3.3xl07 2.0xl07 1.Oxl07 IV 2.5xl07 1.4xl07 1. 5xl07 1.1xl07 PIV 9.8xl07 4.1x107 IV 3.0xl07 1. Oxl07 7 S02 1,000 mg/l pH 2.4 PIV 3.4xl07 2.6xl07 1.4xl07 a.8xl07' S02 1,000 mg/l pH 2.4 IV 1.6xl07 1. 2xl07 1.lxl07 0.7xl07 apIV--potentially infectious virus, double shelled form which must be enzyme activated just before assay. bIV--infectious virus, single shelled form. Table 4. Effect of S02 treatment on poliovirus viability ~n secondary sewage. Sample Treatment 1 min Control (no adjust- 102+12 ments pH 7.3) pH Control 48+20 (pH 2.6 with HCl) S02 at 500 mg/l (pH 2.6) Viable Poliovirusa,b,c/ml at Selected Treatment Times 5 min 15 m~n 30 min 1.8+0.6 3.2+1.4 2.6+1.6 aplaque forming units xlO-2 • 40 min 114+22 38+14 2.6+1.2 bThe assay system for poliovirus was a plaque assay using a methyl cellulose overlay on monkey kidney cells. cThe standard deviation on the triplicate runs ~s indicated as plus/minus. 8 CONCLUSIONS Conclusions reached from this study are: 1) Reovirus (PIV or IV) was inactivated with 802 treatment by up to 90 percent under the conditions used. 2) Poliovirus was inactivated by 97 percent with 500 mg/l S02 treatment of wastewater spiked with poliovirus. 3) There is a definite inactivation effect of the 802 on poliovirus when complared to the pH control. FUTURE STUDIES REQUIRED It is important for future studies to: 1) Optimize S02 inactivation conditions of viral pathogens. 9 2) Design and carry out more in-depth study which will give a statis-tical significance to viral inactivation by S02 treatment of wastewater. 3) Investigate the effects of S02 on other selected health hazard related pathogens. 4) Determine "the inactivation mechanism of viruses by 8°2 _ 10 REFERENCES Aieta, E. M., et al. 1980. Dose-response comparisons for chlorine and chlorine dioxide in disinfecting wastewater. Water Chlorination: Environmental Impact and Health Effects, 3, 697. Barnett, B. B., et al. 1975. Can. J. Compo Med. 39:462-465. Bates, R. C., et al. 1977. Development of poliovirus having increased resistance to chlorine inactivation. Appl. Environ. Microbiol. 34, 849. Berg, J. D., et al. 1979. Effectiveness of chlorine dioxide as a wastewater disinfectant. pp. 61-70, In: Progress in Wastewater Disinfection Technology. A. D. Venosa (Ed.) Berg, J. D., et al. 1980. Comparison of viral and bacterial indicators of disinfection in wastewater with chlorine dioxide and chlorine. Water Chlorination: Environmental Impact and Health Effects, 3, 711. Drulak, M., et al. 1978a. The relative effectiveness of commonly used disinfectants in inactivation of Coxsackievirus B5. Jour. Hyg. 81, 389. Drulak, M., et al. 1978b. The relative effectiveness of commonly used disinfectants 1n inactivation of Echovirus II. Jour. Hyg. (Camb.), 81, 77. Dryden, F. D., et al. treatment systems. 1979. Virus removal in advanced wastewater Jour. Water Poll. Control Fed., 51, 2098. Elliott, D. G., and F. D. Amend. 1978. Efficacy of certain dis-infectants against infectious pancreatic nercrosis virus. Jour. Fish Biol. 12, 4, 277. Englebrecht, R. S., et al. 1978. Virus sens1t1v1ty to chlorine disinfec-tion of water supplies. U.S. EPA EPA-600/2-78-123. Evison, L. M. 1978. Inactivation of enteroviruses and couphages with ozone in waters and wastewaters. Progr. Water Technol. (G. B • ), 10, 365. Farooq, S., et al. 1978. Process considerations in design of ozone contact or for disinfection. Jour. Environ. Eng. Div. Proc. Amer. Soc. civil Eng., 104, 835. Gerba, C. P., et al. 1977. Application of photodynamic oxidation to the disinfection of tapwater, seawater, and sewage and sewage contaminated with poliovirus. Photochem. Photobiol., 26, 499. Hajenian, H. G., and M. Butler. 1980. municipal effluent by chlorine. Inactivation of viruses in Jour. Hyg. 84, 63. Irving, T. E. 1980. Sewage chlorination and bacterial regrowth. Technical Report TR-132, Water Research Centre, Medmenham, Engl. Keswick, B. H. 1979. Bromine chloride as an alternative disinfectant to chlorine of human enteric viruses and other pathogens 1n water and wastewater. PhD Thesis, University of Hawaii. Longley, K. E. 1978. Turbulence factors in chlorine disinfection of wastewater. Water Res. (G.B.) 12, 813. Longley, K. E., et al. 1980. dioxide as disinfectants. 2098. Comparison of chlorine and chlorine Jour. Water Poll. Control Fed. 52, Parrella, A., et al. 1977. Antiviral act1v1ty of and sodium hypochlorite: Comparative study. Inquinamento Deputazion (It.), 6, 412. chlorine dioxide Ing. Ambientale: Roy, D. 1979. Inactivation of enteroviruses by ozone. PhD Thesis, University of Illinois at Urbana-Champaign. Spendlove, R. S. 1967. Methods of virology Vol. 3. pp. 475-520. K. Maramorosch and H. Koprowski, Eds. Academic Press, New York. U.S. Environmental Protection Agency. Development Report No. 600/9-79-9. 1979. Office of Research and 018, Cincinnati, Ohio. 11 Wedemeyer, G. A., et al. 1978. Survival of the salmonid viruses infectious hematopoietic necrosis (IHNV) and infectious pancreatic necrosis (IPNV) in ozonated, chlorinated and untreated waters. Jour. Fish Res. Bd. Can., 35, 875. Zotova, V. I., et al. 1977. Effects of ultrasonic treatment on chlorination of tap water with respect to viruses. Gig i Sanit. (USSR). 9. 99. 

Sulfur Dioxide Treatment of Secondary sewage: Effect on Viruses

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Sulfur Dioxide Treatment of Secondary sewage: Effect on Viruses

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