Concentration Of Enteric Viruses Research Articles

DETERMINISTIC VERSUS STOCHASTIC QUANTITATIVE MICROBIAL RISK ASSESSMENT MODELS FOR WASTEWATER IRRIGATION OF FOOD CROPS

Reuse of wastewater to irrigate food crops is being practiced in many parts of the world and is becoming more commonplace as the competition for, and stresses on, freshwater resources intensify. But there are risks associated with wastewater irrigation, including the possibility of transmission of pathogens causing infectious disease, to both workers in the field and to consumers buying and eating produce irrigated with wastewater. To manage these risks appropriately we need objective and quantitative estimates of them. This is typically achieved through one of two modelling approaches: deterministic or stochastic. Each parameter in a deterministic model is represented by a single value, whereas in stochastic models probability functions are used. Stochastic models are theoretically superior because they account for variability and uncertainty, but they are computationally demanding and not readily accessible to water resource and public health managers. We constructed models to estimate risk of enteric virus infection arising from the consumption of wastewater-irrigated horticultural crops (broccoli, cucumber and lettuce), and compared the resultant levels of risk between the deterministic and stochastic approaches. Several scenarios were tested for each crop, accounting for different concentrations of enteric viruses and different lengths of environmental exposure (i.e. the time between the last irrigation event and harvest, when the viruses are liable to decay or inactivation). In most situations modelled the two approaches yielded similar estimates of risk (within 1 order-of-magnitude). The two methods diverged most markedly, up to around 2 orders-of-magnitude, when there was large uncertainty associated with the estimate of virus concentration and the exposure period was short (1 day). Therefore, in some circumstances deterministic modelling may offer water resource managers a pragmatic alternative to stochastic modelling, but its usefulness as a surrogate will depend upon the level of uncertainty in the model parameters.

Current Methods for Extraction and Concentration of Enteric Viruses from Fresh Fruit and Vegetables: Towards International Standards

Virus-contaminated soft fruits or vegetables are increasingly identified as causes of foodborne viral illness. Noroviruses and hepatitis A virus are the most common pathogens in viral infections transmitted by these kinds of foods. To improve microbiological detection and monitoring and to increase insights into the contribution of fruits and vegetables to foodborne viral transmission, sensitive, reliable, and standardized methods are needed. More studies on virus detection methods for foods are being published, but validated consensus protocols are not yet available. In this paper, different methodologies are reviewed critically. The use of process controls and internal amplification controls is discussed.

ウイルスに関する安全性確保のための下水再生処理法の選定方法

In this study, we attempted to quantify the risk of enteric virus infection due to wastewater reuse, and to clarify the necessary reclamation methods to ensure safety. To assess the potential risk associated with the use of reclaimed wastewater in various reuse applications, some exposure scenarios were assumed, and enteric virus concentrations in secondary effluents were monitored at ten wastewater treatment plants for 2 years. The virus concentrations in secondary effluents distributed according to a log normal curve. Annual infection risks (r) corresponding to the scenarios were calculated using the Monte Carlo method. Then, assuming a number of virus removal efficiencies (x), annual infection risks (r) were calculated and the relationship between virus removal efficiency and annual infection risk (r=f (x)) was obtained for each scenario. The necessary virus removal efficiency (x0), satisfying the assumed acceptable annual risk (r0) under the given scenario, was calculated using r=f (x). A virus removal method satisfying the virus removal efficiency (x0) was chosen using the existing data of enteric virus removal efficiencies of several reclamation methods.

A simple method for isolation of enteric viruses (noroviruses and enteroviruses) in water

A simple and improved protocol for the isolation and detection of noroviruses (‘Norwalk-like viruses’, NLVs) and enteroviruses in ground- and drinking water is described. An improved procedure was developed for concentration of enteric viruses from water, whereby viruses are directly lysed after filtration on a negatively charged membrane. As the method is free from possible recovery losses during usual rinsing, elution, flocculation or concentration steps prior to RNA extraction, a high sensitivity and reliability is achieved. Detection was carried out by using a modified reverse-transcription polymerase chain reaction described previously and agarose gel electrophoresis. The overall detection sensitivity of our method by spiking 1 l of water was 0.1 PFU for polioviruses (Sabine 1). For Norwalk-like viruses ggII (Lordsdale), the detection limit is clearly lower compared to older protocols with elution and concentration steps (concentration of viral particles in positive stool samples were not known). Another simple protocol was used to isolate NLVs from contaminated stool samples.

IMPROVED METHOD FOR CONCENTRATION OF GIARDIA, CRYPTOSPORIDIUM, AND POLIOVIRUS FROM WATER

ABSTRACT Methods for the concentration of enteric viruses and the protozoan parasites, Giardia and Cryptosporidium, from drinking water currently require the use of two different types of filters. Electropositive or electronegative microporous filters (0.2–0.45 µm nominal porosity) are used for the collection of enteroviruses, while polypropylene spun-fiber filters (1 µm porosity) and small pleated cartridge filters are used for the collection of protozoan parasites from water. Since the filter mechanically traps the protozoa by size exclusion, a microporous filter with an appropriately small nominal porosity could possibly be used for co-collection of both protozoa and enteroviruses. This study compared the concentration efficiencies of a polypropylene fiber cartridge (DPPPY) filter and two different microporous filters (Filterite and 1MDS) with poliovirus (type 1), with respect to their ability to concentrate Giardia and Cryptosporidium from water. Giardia cysts and Cryptosporidium oocysts were added to 400 l of either tap water or tertiary treated wastewater and passed through the test filter. The protozoa were eluted from the polypropylene filter by hand-washing in a detergent solution. Viruses and protozoa were eluted from the microporous filter by two consecutive back-washes with a 1.5% beef extract, 0.1% Tween 80 solution. The eluent was then centrifuged to remove the parasites and the supernatant assayed for viruses. The overall efficiency was greater for the Filterite filter (40.4% for Giardia; 36.6% for Cryptosporidium) when compared to the spun fiber filter (10.1% for Giardia; 16.0% for Cryptosporidium). The Filterite filters were easier and faster to process than the polypropylene spun fiber filters. There was no significant difference in the recovery of protozoa from 1MDS and DPPPY filters. Co-collection of viruses and protozoan parasites from water onto the same filter is possible and can reduce the time and cost of routine water monitoring.

A new and simple method for concentration of enteric viruses from water

A new type of electropositive filter media particle was tested to adsorb bacteriophage f 2 and enteric viruses from tap water. 3×nutrient broth (pH 7.2) was used to elute the adsorbed viruses, and the eluate was reconcentrated by polyethylene glycol ( M W 6000) precipitation with a final concentration of 10% (wt./vol.). The adsorption of bacteriophage was reliable and efficient, and not affected by the pH value, temperature, turbidity and organic materials in water. This method gave a recovery of Polio 1 virus 96.0% for small-volume tap water; 88.7% for large-volume water; and gave a comparable recovery of HAV, Coxsackie B 3 and Echo 7 from tap water. The concentration method need not acidify virus-containing water, add exogenous multivalent cation salts, or require expensive equipment.

Estimating the safety of wastewater reclamation and reuse using enteric virus monitoring data

The reliability of wastewater reclamation and reuse to meet a given annual risk of infection, considering the variability of enteric virus concentrations, has been investigated. Two concepts related to the reliability and safety of wastewater reclamation and reuse are presented. The first is reliability, defined as the probability that the risk of infection from enteric viruses in reclaimed wastewater does not exceed an acceptable risk. The second is based on the expectation of the acceptable annual risk in which the exposure to enteric viruses may be estimated stochastically by numerical simulation. To assess the potential risks associated with the use of reclaimed wastewater in various reuse applications, four exposure scenarios were tested: golf course irrigation, food crop irrigation, recreational impoundments, and groundwater recharge. Past monitoring data on enteric virus concentrations in unchlorinated secondary effluents in California were used. Because enteric virus concentrations in unchlorinated secondary effluents were found to vary over a wide range, characterizing their variability was found to be extremely important. The reliability criterion of meeting the less than 10−4 annual risk of infection (less than or equal to one infection per 10 000 population per year) at least 95% of the time was used to assess the safety of using reclaimed wastewater in the four different exposure scenarios. The methodologies used in this study should be refined, based on a larger enteric virus database developed using standardized field and laboratory protocols.

F-specific RNA bacteriophages are adequate model organisms for enteric viruses in fresh water

Culturable enteroviruses were detected by applying concentration techniques and by inoculating the concentrates on the BGM cell line. Samples were obtained from a wide variety of environments, including raw sewage, secondary effluent, coagulated effluent, chlorinated and UV-irradiated effluents, river water, coagulated river water, and lake water. The virus concentrations varied widely between 0.001 and 570/liter. The same cell line also supported growth of reoviruses, which were abundant in winter (up to 95% of the viruses detected) and scarce in summer (less than 15%). The concentrations of three groups of model organisms in relation to virus concentrations were also studied. The concentrations of bacteria (thermotolerant coliforms and fecal streptococci) were significantly correlated with virus concentrations in river water and coagulated secondary effluent, but were relatively low in disinfected effluents and relatively high in surface water open to nonhuman fecal pollution. The concentrations of F-specific RNA bacteriophages (FRNA phages) were highly correlated with virus concentrations in all environments studied except raw and biologically treated sewage. Numerical relationships were consistent over the whole range of environments; the regression equations for FRNA phages on viruses in river water and lake water were statistically equivalent. These relationships support the possibility that enteric virus concentrations can be predicted from FRNA phage data.

Pig erythrocyte ghost cells used for concentration of enteric viruses from experimentally contaminated clinical specimens

A simple, rapid and efficient procedure of virus concentration from urine, cerebrospinal fluid (CSF), and feces was developed. Pig erythrocyte ghost cells were used to adsorb and elute such viruses as poliovirus LSC-1, echovirus 6, and human rotavirus (clinical isolate). In urine and CSF, the adsorption efficiency range was 80-100% and elution was from 85% to greater than 100%. In addition, poliovirus LSC-1 was used as an experimental model to examine this procedure under various clinical conditions, such as calcium, glucose, amino acids, and urea at high concentrations. These were added to normal urine specimens to simulate pathological conditions. The results suggest that pig erythrocyte ghost cells are an efficient matrix for adsorption and elution of enteric viruses found in clinical specimens of urine, CSF, and feces. This method might be useful in virus concentration from clinical specimens and for preparative microscopy and other clinical laboratory methods that require subsequent virus concentration.

Wound fiberglass depth filters as a less expensive approach for the concentration of viruses from water.

Wound fiberglass depth cartridge filters (25.4 cm) with a nominal porosity of 1 micron were used to concentrate viruses from large volumes of surface water. They were found to be an excellent, less expensive alternative to the 0.2-micron pleated cartridge filters normally used for the concentration of enteric viruses from water. More than 99% of experimentally seeded poliovirus was adsorbed to these filters when the pH of the water was adjusted to pH 3.5 and aluminium chloride was added to a final concentration of 0.001 M, as recommended for electronegative filters. In comparative recovery of indigenous viruses from river water, similar results were obtained with two 1-micron or a 3-microns + 0.2-micron filter combination. The cost of the two 1-micron filters is about Can. $26, while it is about Can. $58 for the other combination.

Coliphages as indicators of enteric viruses in activated sludge

The use of indigenous coliphage as indicators of enteric viruses during activated sludge treatment of domestic wastewater was examined. Enteric viruses, coliphage attacking three strains of Escherichia coli and a number of wastewater parameters were monitored in the primary effluent, secondary effluent and secondary sludge of a wastewater treatment plant. Removal of viruses during secondary treatment appeared to be the result of rapid adsorption of influent virions to mixed liquor suspended solids. Adsorption was followed by inactivation of the viruses during aeration of the mixed liquor. A larger proportion of influent enteric viruses than coliphage was recovered from the secondary sludge. This suggests that activated sludge treatment was less antagonistic towards enteric viruses than towards the coliphage. Coliphage plaques of less than or equal to 1 mm in diameter were composed of large viruses (approx. 100 nm in diameter). Plaques greater than 3 mm in diameter appeared to be the result of host infection by a much smaller virus (approx. 45 nm in diameter). Plaques between 1 and 3 mm in diameter were composed of both small and large coliphages. Coliphage concentrations could not be correlated with enteric virus concentrations in either primary or secondary effluent. However, it was found that coliphage giving rise to plaques greater than 3 mm in diameter were positively related with enteric viruses in the secondary effluent. This result suggests that this group of coliphage may serve as an indicator of the efficacy of activated sludge treatment of enteric viruses.

Enteric Bacterial and Viral Pathogens and Indicator Bacteria in Hard Shell Clams

Enteric bacteria and virus levels were determined in hard shell clams, Mercenaria mercenaria, harvested from areas open or closed for commercial shellfishing on the basis of total coliform levels in water. Four pairs of open and closed stations were sampled seasonally over a 1-year period. Enteric viruses were isolated from 3 of 13 100-g clam samples from open beds and 6 of 15 samples from closed beds. Salmonella was found in 1 of 15 samples from closed areas, but not in any samples from open areas. No Shigella or Yersinia were isolated from clams taken from either open or closed beds. Levels of Vibrio parahaemolyticus, an indigenous estuarine microorganism, were similar in clams from open and closed areas. No statistically significant difference was found in the occurrence of enteric viruses in clams from open and closed areas. Product-moment correlations between concentrations of enteric viruses and bacteria in clams or water demonstrated no statistically significant correlations between virus concentrations in clams and total coliforms or fecal coliforms in water or total coliforms, fecal coliforms, fecal streptococci or aerobic plate counts in clams.

Evaluating adsorbent filter performance for enteric virus concentrations in tap water

Under varying operating conditions, the performance limits of an electropositive adsorbent filter were equivalent to those of a conventional electronegative adsorbent filter. Performance limits were determined with respect to adsorption flow rate (hydraulic flux), tap water process volume (hydraulic load), stability of adsorbed virus during storage, and ability to concentrate several different enteric viruses.

Relationships between environmental factors, bacterial indicators, and the occurrence of enteric viruses in estuarine sediments.

Current standards for evaluation of the public health safety of recreational and shellfish-harvesting waters are based upon bacteriological analysis, but do not include an evaluation of the number of viruses. The objective of this study was to determine the occurrence of enteric viruses in estuarine sediments and to find a relationship, if any, between the presence of viruses in seawater or sediment or both and various biological and physicochemical characteristics of the environment. Viruses were found in greater numbers in sediment than in overlying seawater on a volume basis. Several types of enteroviruses were isolated: coxsackievirus types A16, B1, and B5, echovirus type 1, and poliovirus type 2. On several occasions, viruses were isolated from sediments when overlying seawaters met bacteriological water quality standards for recreational use. Statistical analysis of the relationship between viruses in seawater or in sediment and other variables measured yielded only one significant association: the number of viruses in sediment was found to be positively correlated with the number of fecal coliforms in sediment. No other physical, chemical, or biological characteristic of seawater or sediment that was measured showed statistically significant association with viral numbers. No correlation was found between bacterial indicators and virus in the overlying waters. The data indicated that evaluation of the presence of bacteria and viruses in sediment may provide additional insight into long-term water quality conditions and that indicator bacteria in water are not reflective of the concentration of enteric viruses in marine waters.

Concentration of enteric viruses in wastewater and effluent: a two year survey

A 2 year survey has been carried out on the wastewater and effluent of the Haifa treatment plant in order to gather information on the concentration and composition of the enteric viral population. The results obtained showed (a) the enteric viruses are present in wastewater and effluent all year round. (b) Their numbers fluctuate and are usually lower during the winter. (c) The highest numbers have been recorded between April and September with peaks occurring in wastewater during the month of July (106,000 PFU 100 ml −1) and in effluent in August (94,000 PFU 100 ml −1). During the months January through March the most prevalent viruses isolated were Polio I and ECHO 4.

Concentration of enteric viruses from water with lettuce extract.

A method for recovering enteroviruses, adenovirus, and reovirus from water with lettuce extract is described. Lettuce extract at pH 8.5 was added to the sample and the pH was reduced stepwise with hydrochloric acid to 4.0 to 4.5. The flocculent lettuce-extract particles, and adsorbed virus, were readily removed from solution by low-speed centrifugation. Electron microscopy suggests that, under conditions suitable for adsorption, virus particles are coated with the lettuce-extract colloid.

Concentration of Enteric Viruses from Water with Lettuce Extract

A method for recovering enteroviruses, adenovirus, and reovirus from water with lettuce extract is described. Lettuce extract at pH 8.5 was added to the sample and the pH was reduced stepwise with hydrochloric acid to 4.0 to 4.5. The flocculent lettuce-extract particles, and adsorbed virus, were readily removed from solution by low-speed centrifugation. Electron microscopy suggests that, under conditions suitable for adsorption, virus particles are coated with the lettuce-extract colloid.