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Biomedical subjects

C P Gerba

Publications and source records attributed to C P Gerba.

At least 73 records · Page 4Linked to original sources

Detection of Cryptosporidium in water by using polypropylene cartridge filters.

Members of the genus Cryptosporidium are protozoan parasites that cause gastroenteritis in humans and animals and appear to be spread largely by the fecal-oral route. A method was developed for the concentration and detection of Cryptosporidium oocysts in water to assess their occurrence in the environment and potential for waterborne disease transmission. This method was developed by using spun polypropylene cartridge filters. Optimal conditions for concentration, filter elution, filter porosity, and detection were determined. Fluoresceinated monoclonal antibodies were used for oocyst detection. Experiments also were conducted to study the effect of flow rate, low oocyst numbers, and the addition of detergents on recovery and retention of oocysts. The method that was developed was sensitive enough to detect oocysts at levels of less than 1 per liter. Using this method, we isolated Cryptosporidium oocysts from secondarily treated sewage.

Coccidia↗

Occurrence of Cryptosporidium oocysts in sewage effluents and selected surface waters.

An existing method for the detection of Cryptosporidium oocysts in water was modified to investigate oocyst prevalence in large volumes of water. Surface waters and sewage effluents were filtered, eluted from the filter, and concentrated using centrifugation. The resultant pellet was then homogenized, sonicated, and placed on a sucrose gradient to separate oocysts from the sediment. The uppermost gradient layer was then examined by immunofluorescence using a labeled monoclonal antibody. Using this technique, average numbers of oocysts detected in raw and treated sewage were 5.18 X 10(3) and 1.30 X 10(3)/L, respectively. Filtered sewage effluents had significantly lower numbers of oocysts (10.0/L). These data show that sand filtration may reduce the concentrations of this parasite in waste waters. Highly variable oocyst numbers were encountered in surface waters. Since Cryptosporidium oocysts are frequently present in environmental waters, they could be responsible for waterborne outbreaks of disease.

Animals↗

Use of geostatistics to predict virus decay rates for determination of septic tank setback distances.

Water samples were collected from 71 public drinking-water supply wells in the Tucson, Ariz., basin. Virus decay rates in the water samples were determined with MS-2 coliphage as a model virus. The correlations between the virus decay rates and the sample locations were shown by fitting a spherical model to the experimental semivariogram. Kriging, a geostatistical technique, was used to calculate virus decay rates at unsampled locations by using the known values at nearby wells. Based on the regional characteristics of groundwater flow and the kriged estimates of virus decay rates, a contour map of the area was constructed. The map shows the variation in separation distances that would have to be maintained between wells and sources of contamination to afford similar degrees of protection from viral contamination of the drinking water in wells throughout the basin.

Coliphages↗

Adsorption of viruses to charge-modified silica.

The purpose of this study was to provide a clearer understanding of virus adsorption, focusing specifically on the role of electrostatic interactions between virus particles and adsorbent surfaces. The adsorption of poliovirus 1, reovirus types 1 and 3, and coliphages MS-2 and T2 to colloidal silica synthetically modified to carry either positive or negative surface charge was evaluated. Adsorption experiments were performed by combining virus and silica in 0.1-ionic-strength buffers of pH 4.0, 6.4, and 8.5. Samples agitated for specified adsorption periods were centrifuged to pellet adsorbent particles plus adsorbed virus, and the supernatants were assayed for unadsorbed virus. All viruses adsorbed exclusively to negatively charged silica at pH values below their isoelectric points, i.e., under conditions favoring a positive surface charge on the virions. Conversely, all viruses adsorbed exclusively to positively charged silica at pH values above their isoelectric points, i.e., where virus surface charge is negative. Viruses in near-isoelectric state adsorbed to all types of silica, albeit to a lesser degree.

Adsorption↗

Virus persistence in groundwater.

More than 50% of the outbreaks of waterborne disease in the United States are due to the consumption of contaminated groundwater. An estimated 65% of the cases in these outbreaks are caused by enteric viruses. Little, however, is known about the persistence of viruses in groundwater. The purpose of this study was to determine whether measurable chemical and physical factors correlate with virus survival in groundwater. Groundwater samples were obtained from 11 sites throughout the United States. Water temperature was measured at the time of collection. Several physical and chemical characteristics, including pH, nitrates, turbidity, and hardness, were determined for each sample. Separate water samples were inoculated with each of three viruses (poliovirus 1, echovirus 1, and MS-2 coliphage) and incubated at the in situ groundwater temperature; selected samples were also incubated at other temperatures. Assays were performed at predetermined intervals over a 30-day period to determine the number of infective viruses remaining. Multiple regression analysis revealed that temperature was the only variable significantly correlated with the decay rates of all three viruses. No significant differences were found among the decay rates of the three viruses, an indication that MS-2 coliphage might be used as a model of animal virus survival in groundwater.

Bacteriophages↗

Endotoxin removal by charge-modified filters.

The effects of positively charged nylon and depth (cellulose-diatomaceous earth) filters on endotoxin removal from various solutions were evaluated. The charged filter media removed significant amounts of Escherichia coli and natural endotoxin from tap water, distilled water, sugars, and NaCl solutions; no significant removal of endotoxin was observed with negatively charged filter media. The extent of removal was influenced by pH, the presence of salts, and organic matter. Such media may be useful for the control of endotoxins in raw-product water or solutions used to prepare parenteral drug products or in other fluids where endotoxin control is desired.

Endotoxins↗

Agarose isoelectrofocusing of intact virions.

A convenient and accurate method for determining the isoelectric points of intact virions is described. Tritium-labeled poliovirus 1 (strains Brunhilde and LSc-2) and echovirus 1 (isolates V239, V248, V212, R115 and 4CH-1) were successfully focused into sharp bands at their respective isoelectric points using a thin-layer agarose isoelectric focusing system. In situ detection of labeled virus bands in the agarose was by fluorography. Freezing and thawing of virus samples prior to isoelectric focusing did not alter their respective isoelectric points.

Enterovirus B, Human↗

Virus movement in soil during saturated and unsaturated flow.

Virus movement in soil during saturated and unsaturated flow was compared by adding poliovirus to sewage water and applying the water at different rates to a 250-cm-long soil column equipped with ceramic samplers at different depths. Movement of viruses during unsaturated flow of sewage through soil columns was much less than during saturated flow. Viruses did not move below the 40-cm level when sewage water was applied at less than the maximum infiltration rate; virus penetration in columns flooded with sewage was at least 160 cm. Therefore, virus movement in soils irrigated with sewage should be less than in flooded groundwater recharge basins or in saturated soil columns. Management of land treatment systems to provide unsaturated flow through the soil should minimize the depth of virus penetration. Differences in virus movement during saturated and unsaturated flow must be considered in the development of any model used to simulate virus movement in soils.

Poliovirus↗

Effect of ionic composition of suspending solution on virus adsorption by a soil column.

The effect of various electrolytes on the adsorption of poliovirus was measured in 250-cm-long soil columns with ceramic samplers at different depths. Viruses suspended in deionized water moved much farther through the soil than those suspended in tap water, whereas movement in sewage water was intermediate. The salt content of the tap water and sewage water promoted virus adsorption, but evidently the organic compounds in sewage retarded adsorption. When viruses were suspended in chloride solutions of K, Na, Ca, and Mg, virus adsorption increased as the cation concentration and valence increased. The depth of virus penetration was related to the ionic strength of the solutions. Virus penetration data for NO(3), SO(4), and H(2)PO(4) salts of K, Na, and Ca indicated that other anions were more effective than Cl in promoting virus adsorption. Also, NH(4) was more effective than other cations in limiting the penetration depth of viruses. It seems that ions composed of radicals are more effective than ions composed of single atoms in promoting virus adsorption. Al was the most effective ion in limiting virus penetration, probably owing to flocculation of the viruses. Adding AlCl(3) concentrations to secondary sewage effluent to provide an Al concentration of 0.1 mM reduced the virus penetration depth to 40 cm. These studies show that the ionic composition of the suspending solutions must be considered in predicting virus penetration depths, and it may be practical to add low concentrations of a flocculating agent such as AlCl(3) to sewage water to limit virus movement through very porous soils.

Journal Article↗

Seasonal occurrence of rotavirus in sewage.

A seasonal distribution was observed for rotaviruses in sewage by using indirect immunofluorescence. Levels were low from May through September and generally higher during winter and spring. In contrast, no seasonal pattern was observed for total enteroviruses. Limitations of the indirect immunofluorescence assay and enzyme immunoassay for environmental samples are discussed.

Enterovirus↗

Comparison of microporous filters for concentration of viruses from wastewater.

The 1-MDS Virosorb filter and the 50S and 30S Zeta-plus filters, all with a net positive charge, were compared with the negatively charged Filterite filter for concentration of naturally occurring coliphages and animal viruses from sewage effluent. When Filterite filters were used, the effluent was adjusted to pH 3.5 and AlCl3 was added before filtration to facilitate virus adsorption. No adjustment was required with the positively charged filters. Sets of each filter type were eluted with 3% beef extract (pH 9.5) or eluted with 0.05 M glycine (pH 11.5). A maximum volume of 19 liters could be passed through 142-mm diameter Filterite filters before clogging, whereas only 11, 11, and 15 liters could be passed through the 1-MDS, 50S, and 30S filters, respectively. For equal volumes passed through the filters, coliphage recoveries were 14, 15, 18, and 37% in primary effluent and 40, 97, 50, and 46% in secondary effluent for the Filterite , 1-MDS, 50S, and 30S filters, respectively. No statistically significant difference was observed in the recovery of animal viruses among the filters from secondary effluent, whereas in the Filterite and 50S filters, higher numbers of viruses from primary effluent were recovered than in the 1-MDS and 30S filters in two of three collections. Glycine was found to be a less-efficient eluent than beef extract in the recovery of naturally occurring viruses.

Animals↗

Detection of enteric viruses in treated drinking water.

The occurrence of viruses in conventionally treated drinking water derived from a heavily polluted source was evaluated by collecting and analyzing 38 large-volume (65- to 756-liter) samples of water from a 9 m3/s (205 X 10(6) gallons [776 X 10(6) liters] per day) water treatment plant. Samples of raw, clarified, filtered, and chlorinated finished water were concentrated by using the filter adsorption-elution technique. Of 23 samples of finished water, 19 (83%) contained viruses. None of the nine finished water samples collected during the dry season contained detectable total coliform bacteria. Seven of nine finished water samples collected during the dry season met turbidity, total coliform bacteria, and total residual chlorine standards. Of these, four contained virus. During the dry season the percent removals were 25 to 93% for enteric viruses, 89 to 100% for bacteria, and 81% for turbidity. During the rainy season the percent removals were 0 to 43% for enteric viruses, 80 to 96% for bacteria, and 63% for turbidity. None of the 14 finished water samples collected during the rainy season met turbidity standards, and all contained rotaviruses or enteroviruses.

Coliphages↗

Simple field method for concentration of viruses from large volumes of water.

A pressure spray tank was adapted to supply positive pressure for processing water samples for concentrating viruses with microporous filters under field conditions. This low-cost system allows water to be processed in locations where electric current is not readily available or where light-weight portable equipment is required.

Fresh Water↗

Concentration of viruses from tap water and sewage with a charge-modified filter aid.

Positively charged microporous filters have been shown to have definite advantage over negatively charged filters for concentrating viruses from water as they eliminate the need of water conditioning (acidification and multivalent cation addition) prior to filtration. The present study was designed to evaluate the use of a charge-modified (Zeta plus) filter aid (AMF CUNO, Meriden, CT) for the concentration of viruses (Poliovirus-1, Coxsackievirus B-3 and ECHO virus-7) from large volumes of tap water. Charge-modified filter aid could efficiently adsorb polio, coxsackie and ECHO viruses from water. The adsorbed viruses from both tap water and secondary sewage were most efficiently eluted with 4% beef extract - 0.5 M NaC1, pH 9.5. The efficiency of recovery of poliovirus, coxsackievirus and ECHO virus from 10- to 20-1 vols. of tap water ranged between 32 and 98%, 67 and 100% and 69 and 100%, respectively. This technique appears to have distinct advantages over previous methods as it requires low cost equipment (47 mm polypropylene filter housing) and materials, is simple and easy to handle, can filter large volumes of water (greater than or equal to 20 1) with good recoveries at ambient pH and can be efficiently used to monitor viral water quality.

Adsorption↗

Viradel method for detection of rotavirus from seawater.

This report describes the use of a virus adsorption-elution (Viradel) technique for the concentration and detection of rotavirus from seawater. Simian rotavirus SA-11 was used as a model virus because it can be readily propagated and plaqued in vitro. The virus adsorbed optimally to fiberglass epoxy filters when seawater was adjusted to pH 3.5 and aluminum chloride added to a concentration of 1.0 mM. Adsorbed virus was eluted with 6% beef extract at pH 10.5 and was further concentrated to a smaller volume by a modification of the organic flocculation method. Using this method, in conjunction with an indirect immunofluorescence test, rotavirus was detected in sewage-contaminated seawater.

Adsorption↗

Bacteriophage concentration from water by filter chromatography.

The efficiency of an electropositive filter for membrane chromatography of viruses was examined using coliform phages T1, T4, lambda and Salmonella phage P22. Phages diluted in dechlorinated tap water were adsorbed to filters at neutral pH and eluted by 3% beef extract in 0.05 M glycine buffer at selected alkaline pH values. With exception of lambda phage, which displayed erratic adsorption behavior at any pH, all bacteriophages studied, adsorbed to filters with an efficiency of 97-100% at pH values ranging between 6.0 and 8.0. Each phage was readily eluted at alkaline pH levels. Maximal elution (86.2%) of T1 phage and lambda phage (79%) occurred at pH 10, while T4 and Salmonella phages were eluted most efficiently at pH 11 at values of 91.7 and 81.9%, respectively. The resolving power of the filter was such that individual phages within the same virus group (T1 and T4 phage) could be eluted at pHs differing by only one unit.

Adsorption↗