RESUMEN
To select a tentative standard method for detection of viruses in sludge the American Society for Testing and Materials D19:24:04:04 Subcommittee Task Group initiated round robin comparative testing of two procedures that, after initial screening of several methodologies, were found to meet the basic criteria considered essential by the task group. Eight task group member laboratories agreed to perform round robin testing of the two candidate methods, namely, The Environmental Protection Agency or low pH-AlCl3 method and the Glass or sonication-extraction method. Five different types of sludge were tested. For each particular type of sludge, a single laboratory was designated to collect the sludge in a single sampling, make samples, and ship it to the participating laboratories. In most cases, participating laboratories completed all the tests within 48 h of sample arrival. To establish the reproducibility of the methods, each laboratory tested each sludge sample in triplicate for the two candidate virus methods. Each processed sludge sample was quantitatively assayed for viruses by the procedures of each individual round robin laboratory. To attain a more uniform standard of comparison, a sample of each processed sample from all laboratories was reassayed with one cell line and passage number by a single laboratory (Environmental Protection Agency Environmental Monitoring and Support Laboratory, Cincinnati, Ohio). When the data were statistically analyzed, the Environmental Protection Agency method was found to yield slightly higher virus recoveries for all sludge types, except the dewatered sludge. The precisions of both methods were not significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Compuestos de Aluminio , Cloruros , Enterovirus/aislamiento & purificación , Aguas del Alcantarillado , Aluminio , Cloruro de Aluminio , Análisis de Varianza , Concentración de Iones de Hidrógeno , Técnicas Microbiológicas/normas , SonicaciónRESUMEN
Air drying of raw sludge caused inactivation of indigenous viruses. A gradual loss of infectivity occurred with the loss of water until the solids content reached about 80%. A more rapid decline of viral infectivity occurred with further dewatering.
Asunto(s)
Desecación , Poliovirus/crecimiento & desarrollo , Aguas del Alcantarillado , Virus/crecimiento & desarrollo , AireRESUMEN
Five general methods for recovering indigenous viruses from raw wastewater sludge were compared. Each method included elution, concentration, and disinfection steps. The elution method, found to consistently yield the greatest viral recovery, was a two-phase technique that involved blending sludge with Freon. Other methods, including two being tested as American Society for Testing Materials tentative standard methods, were less effective. Viral recoveries were generally greater (sometimes much greater) if samples were concentrated by high-speed centrifugation rather than by organic flocculation with 3% beef extract. Three cell lines were used to measure viral recoveries by the plaque assay. The efficiency of recovery was greatest on BGM cells, followed by RD and MA-104 cells.
Asunto(s)
Aguas del Alcantarillado , Virus/aislamiento & purificación , Línea Celular , Centrifugación , Floculación , Métodos , Ensayo de Placa Viral , Virus/crecimiento & desarrollo , Microbiología del AguaRESUMEN
A preliminary study was carried out on evaluating a flow-through gauze sampler for its efficiency in recovering virus from both fresh and seawater. An attenuated type 1 poliovirus was used as the working model. When tap water was sampled, the amounts of virus adsorbed by the gauze pads were very small, about 2% of the total number of virus particles flowing through the device. The virus adsorption and recovery increased to 15 to 19% when seawater was sampled. Addition of NaCl to tap water produced a much better effect on virus adsorption and recovery by this device, i.e., 47% of the total virus particles in each sample. The best viral elution from the pads was obtained by using buffer solution of pH 8.0 to 9.0 containing a small amount of animal serum. Repeated elutions from the pads were necessary to recover the most virus although the first eluate contained approximately 50% of the adsorbed virus. Further development of this device appears warranted, because of (i) the simplicity of the procedure, (ii) its capability of sampling large volume of water, (iii) the low cost of collecting samples, and (iv) the feasibility of obtaining a rough quantitative assessment of viral pollutants in water examined.
Asunto(s)
Poliovirus/aislamiento & purificación , Microbiología del Agua/instrumentación , Contaminación del Agua , Adsorción , Animales , Bovinos , Gossypium , Concentración de Iones de Hidrógeno , Sueros Inmunes , Métodos , Modelos Biológicos , Agua de Mar , Cloruro de SodioRESUMEN
A study was carried out to further evaluate the practicability of viral depuration by assaying individual shellfish. The Northern quahaug and a strain of the type 1 attenuated poliovirus were used as the working model. Two types of depuration systems were employed: the small experimental tanks and a pilot-size tank with a capacity of approximately 24 bushels (836 liters) of shellfish. Volumes of the individual shellfish samples were found uniform throughout the experiments when a prior selection for the weight of the shellfish was made. There was also no significant difference in volumes of the individual samples during the course of depuration (24 to 96 hr). Under controlled hydrographic conditions, however, the uptake of virus in individual shellfish varied considerably. In general, the individual variability reached 10- to 100-fold. This wide variation would explain the variability of viral contents obtained in pooled samples during depuration as reported previously. During a later phase of depuration, although a great majority of shellfish were free of the virus, a few still harbored minimal amounts of contaminants. The presence of virus in some of the shellfish after various periods of depuration would, theoretically, be obscured by the pooling of the sampled shellfish. Further examination of the negative samples by assaying larger quantities than those routinely used revealed that a few still contained virus. To simulate naturally polluted shellfish as closely as technically possible, shellfish were polluted with minimal amounts of virus. The shellfish were cleansed more rapidly by the depuration process than were those polluted with more virus. Since the naturally polluted shellfish were shown to contain less virus than those studied in the laboratory, it is anticipated that the former type of shellfish may be cleansed more readily by this process within a reasonable period of time. Justification for a field trial of depuration in this country is presented.