Concentration of enteroviruses from estuarine water.

Pleated cartridge filters readily adsorb viruses in estuarine water at low pH containing aluminum chloride. Adsorbed viruses are efficiently recovered by treating filters with glycine buffer at high pH. By using these procedures, it was possible to recover approximately 70% of the poliovirus added to 400 liters of estuarine water in 3 liters of filter eluate. Reconcentration of virus in the filter eluate in small volumes that are convenient for viral assays was more difficult. Reconcentration methods described previously for eluates from filters that process tap water or treated wastewater were inadequate when applied to eluates from filters used to process estuarine water containing large amounts of organic compounds. Two methods were found to permit efficient concentration of virus in filter eluates in small volumes. In both methods, virus in 3 liters of filter eluate was adsorbed to aluminum hydroxide flocs and then recovered in approximately 150 ml of buffered fetal calf serum. Additional reductions in volume were achieved by ultrafiltration or hydroextraction. By using these procedures 60 to 80% of the virus in 3 liters of filter eluate could be recovered in a final volume of 10 to 40 ml.     

Concentration of enteroviruses from estuarine water.

Pleated cartridge filters readily adsorb viruses in estuarine water at low pH containing aluminum chloride. Adsorbed viruses are efficiently recovered by treating filters with glycine buffer at high pH. By using these procedures, it was possible to recover approximately 70% of the poliovirus added to 400 liters of estuarine water in 3 liters of filter eluate. Reconcentration of virus in the filter eluate in small volumes that are convenient for viral assays was more difficult. Reconcentration methods described previously for eluates from filters that process tap water or treated wastewater were inadequate when applied to eluates from filters used to process estuarine water containing large amounts of organic compounds. Two methods were found to permit efficient concentration of virus in filter eluates in small volumes. In both methods, virus in 3 liters of filter eluate was adsorbed to aluminum hydroxide flocs and then recovered in approximately 150 ml of buffered fetal calf serum. Additional reductions in volume were achieved by ultrafiltration or hydroextraction. By using these procedures 60 to 80% of the virus in 3 liters of filter eluate could be recovered in a final volume of 10 to 40 ml.     

Effects of larvae ontogeny, turbidity, and turbulence on prey attack rate and swimming activity of Atlantic herring larvae

The effect of turbulence, light level, and ontogeny on herring larva's attack rate and swimming activity was tested in a previous study. However, during larval seasons (spring and autumn), water clarity is frequently impaired by alga blooms, which also most probably will affect larva feeding rate. Therefore, this study was to investigate the effects of turbidity, turbulence, and ontogeny on the attack rate and swimming activity of herring larvae. By adding diatomaceous earth (DE) to the water, three turbidity levels were established: 0, 35, and 80 Jackson Turbidity Unit [JTU; which coincide with a beam attenuation (c) of 0.1, 2.5, and 4.8 m⁻¹, respectively]. An unfavourable (8×10⁻⁶ W/kg) and a favourable turbulence level (1×10⁻⁶ W/kg) were chosen based on results from the earlier study. The results show that intermediate turbidity (35 JTU) has a positive effect on the attack rate of smaller larvae (20 mm), while high turbidity (80 JTU) has a negative effect on attack rate of all tested larvae size groups. In general, attack rate was lower at the highest turbulence compared to the low level, independent of turbidity level. However, there was one exception, when turbidity was at the highest, the largest larvae (29 mm) seemed to gain from feeding in the highest turbulence level. The overall activity level was higher in the presented study than in the earlier study without turbidity. The favourable turbidity level (35 JTU) coincides with turbidity levels normally found at the equivalent depth during spring and autumn blooms in the area of where the experimental larvae originate. In addition, turbidity's effect on light absorbtion and how it influences the maximum feeding depth of the larva are discussed.     

Virus in Water. II. Evaluation of Membrane Cartridge Filters for Recovering Low Multiplicities of Poliovirus from Water

The efficiency of a Millitube MF cartridge filter, a membrane filter, for recovery of poliovirus from 100-gal volumes of both fresh (tap) and estuarine water was determined. In the high multiplicity of virus input-output experiments, recovery of 97% or greater of input virus was achieved in both types of water when the final concentration of divalent cation as Mg(2+) was 1,200 mug/ml and the pH was 4.5. Virus was effectively eluted from the membrane cartridge with 5x nutrient broth in 0.05 M carbonate-bicarbonate buffer at pH 9.0. Four elutions of 250 ml each were used. In the low multiplicity of virus input-output experiments under the same cationic and pH conditions, up to 67% of the input virus was recovered when the virus was further concentrated from the eluates by the aqueous polymer two-phase separation technique. The volume reduction was 126,000-190,000 to 1. The use of the combined techniques, i.e., membrane adsorption followed by aqueous polymer two-phase separation, provided a highly sensitive, simple, and remarkably reliable sequential methodology for the quantitative recovery of poliovirus occurring at multiplicities as low as 1 to 2 plaque-forming units per 5 gal of water.     

Influence of visual conditions on foraging and growth of juvenile fishes with dissimilar sensory physiology

The influence of turbidity on foraging and growth of young-of-the-year (YOY) of two percid species with relatively dissimilar sensory physiology, perch Perca fluviatilis and pikeperch Sander lucioperca was tested. A littoral mysid, Neomysis integer , was used as prey. Functional response was investigated indoors at two levels of turbidity, 3 NTU (clear) and 25 NTU (turbid). Growth and consumption rate were studied both during day and night in outdoor experiments at similar turbidity levels. Pikeperch were not affected by turbid conditions in either of the experiments. Foraging of perch, on the other hand, was significantly negatively affected by higher turbidity in the functional response experiments and during the night-time in the outdoor experiments. As opposed to pikeperch, consumption rates of perch decreased markedly during nights. Perch also grew more slowly in the treatments with turbid water. The dissimilar reactions of the two species indicate that sensory physiological adaptations and foraging behaviour are important factors that partly can explain disparate reactions of YOY fishes to the level of turbidity.     

Cryptosporidium and giardia recoveries in natural waters by using environmental protection agency method 1623

Relatively few studies have examined recoveries from source waters by using Environmental Protection Agency method 1623 with organism spike doses that are environmentally realistic and at turbidity levels commonly found in surface waters. In this study, we evaluated the filtration capacities and recovery efficiencies of the Gelman Envirochek (standard filter) and the Gelman Envirochek high-volume (HV) sampling capsules under environmental conditions. We also examined the performance of method 1623 under ambient conditions with matrix spike experiments using 10 organisms/liter. Under turbid conditions, the HV capsule filtered approximately twice the volume filtered by the standard filter, but neither could filter 10 liters without clogging. In low-turbidity waters, oocyst, but not cyst, recoveries were significantly higher when the HV capsule was used. In turbid waters, organism recoveries were lower than those in nonturbid waters and were not significantly different for the different filters. When the HV capsule was used, Cryptosporidium recoveries ranged from 36 to 75%, and Giardia recoveries ranged from 0.5 to 53%. For both organisms, recoveries varied significantly by site. Turbidity could explain variation in Giardia recoveries (r(2) = 0.80) but not variation in Cryptosporidium recoveries (r(2) = 0.16). The inconsistent recoveries across sites suggested that the background matrix of the ambient water affected recovery by method 1623. A control sample collected at the height of the winter rainy season detected one organism, highlighting the difficulty of using this method to accurately measure pathogen abundance under natural conditions. Our findings support the use of the HV filter under field conditions but suggest that designing a cost-effective and statistically valid monitoring program to evaluate sources and loads of protozoan pathogens may be difficult.     

Concentration of enteroviruses from large volumes of turbid estuary water.

A method is described for the efficient concentration of viruses from large volumes of highly turbid estuary water. Virus in acidified seawater in the presence of aluminum chloride is adsorbed to a 10-in. (about 25.4 cm) fibreglass depth cartridge and 2- and 0.65-micron epoxy-fibreglass filters in series. This filter series is capable of efficiently adsorbing enteroviruses from 50 U.S. gallons (about 190) of estuary water of varying salinity and turbidity. Adsorbed viruses were eluted from the filters with glycine buffer (pH 11.5) and the eluate reconcentrated by using a precipitate formed by the addition of ferric chloride. Viruses were eluted from this precipitate with fetal calf serum. Using this procedure, four different enteroviruses in 50 gallons (about 190) of estuary water were concentrated 9 000- to 12 000-fold with an overall efficiency of 41%.     

Glass wool filters for concentrating waterborne viruses and agricultural zoonotic pathogens

The key first step in evaluating pathogen levels in suspected contaminated water is concentration. Concentration methods tend to be specific for a particular pathogen group, for example US Environmental Protection Agency Method 1623 for Giardia and Cryptosporidium, which means multiple methods are required if the sampling program is targeting more than one pathogen group. Another drawback of current methods is the equipment can be complicated and expensive, for example the VIRADEL method with the 1MDS cartridge filter for concentrating viruses. In this article we describe how to construct glass wool filters for concentrating waterborne pathogens. After filter elution, the concentrate is amenable to a second concentration step, such as centrifugation, followed by pathogen detection and enumeration by cultural or molecular methods. The filters have several advantages. Construction is easy and the filters can be built to any size for meeting specific sampling requirements. The filter parts are inexpensive, making it possible to collect a large number of samples without severely impacting a project budget. Large sample volumes (100s to 1,000s L) can be concentrated depending on the rate of clogging from sample turbidity. The filters are highly portable and with minimal equipment, such as a pump and flow meter, they can be implemented in the field for sampling finished drinking water, surface water, groundwater, and agricultural runoff. Lastly, glass wool filtration is effective for concentrating a variety of pathogen types so only one method is necessary. Here we report on filter effectiveness in concentrating waterborne human enterovirus, Salmonella enterica, Cryptosporidium parvum, and avian influenza virus.     

Survival of Poliovirus in Flowing Turbid Seawater Treated with Ultraviolet Light

The effectiveness of a model ultraviolet (UV) radiation unit for treating flowing turbid seawater contaminated with poliovirus was determined. At a turbidity of 70 ppm, the observed survival ratios ranged from 1.9 x 10(-3) (99.81% reduction) to 1.5 x 10(-4) (99.98% reduction) at flow rates ranging from 25 to 15 liters/min; no virus was recovered at flow rates of 10 and 5 liters/min. At a turbidity of 240 ppm, the observed survival ratios ranged from 3.2 x 10(-2) (96.80% reduction) to 2.1 x 10(-4) (99.98% reduction) at flow rates ranging from 25 to 5 liters/min. As expected, turbidity had an adverse influence on the effectiveness of UV radiation; however, by adjusting the flow rate of the seawater through the treatment unit, adequate disinfection was shown to be predictable.     

Effect of turbidity on chlorination efficiency and bacterial persistence in drinking water.

To define interrelationships between elevated turbidities and the efficiency of chlorination in drinking water, experiments were performed to measure bacterial survival, chlorine demand, and interference with microbiological determinations. Experiments were conducted on the surface water supplies for communities which practice chlorination as the only treatment. Therefore, the conclusions of this study apply only to such systems. Results indicated that disinfection efficiency (log10 of the decrease in coliform numbers) was negatively correlated with turbidity and was influenced by season, chlorine demand of the samples, and the initial coliform level. Total organic carbon was found to be associated with turbidity and was shown to interfere with maintenance of a free chlorine residual by creating a chlorine demand. Interference with coliform detection in turbid waters could be demonstrated by the recovery of typical coliforms from apparently negative filters. The incidence of coliform masking in the membrane filter technique was found to increase as the turbidity of the chlorinated samples increased. the magnitude of coliform masking in the membrane filter technique increased from less than 1 coliform per 100 ml in water samples of less than 5 nephelometric turbidity units to greater than 1 coliform per 100 ml in water samples of greater than 5 nephelometric turbidity units. Statistical models were developed to predict the impact of turbidity on drinking water quality. The results justify maximum contaminant levels for turbidity in water entering a distribution system as stated in the National Primary Drinking Water Regulations of the Safe Drinking Water Act.     

The influence of turbidity on juvenile marine fishes in estuaries. part 1. field studies at Lake St. Lucia on the southeastern coast of Africa

Lake St. Lucia, the largest estuarine system in Africa (325 km²), was chosen as the field study area for a 3.5-yr ((1980)–83) investigation into relationships between water turbidity and estuarine fish distribution. The variety of habitats, from clear water, open sandy shores to shallow muddy substrata and turbid waters, together with high species diversity (108 species) rendered the area suitable for this study. The relationships between fish distribution and environmental factors were monitored by monthly seine netting of fishes at seven sites representative of the range of conditions in St. Lucia. Simultaneously, water turbidity, salinity, and temperature were recorded.The possible influences of substratum type and food availability were also investigated by using recently published data on invertebrate benthos and Zooplankton distributions. Published data were also used to determine the diet of the common fish species. The results showed that the distribution of juveniles of the 20 commonest fish species were statistically correlated only with water turbidity, water temperature, and food availability. The correlation with temperature was related to seasonal not spatial temperature patterns.Turbidity and food type influences were difficult to separate but exceptions were the anchovyThryssa vitrirostris (Gilchrist and Thompson) and the soleSolea bleekeri Boulenger which occurred only in turbid water despite the widespread occurrence of their prey, andGenes acinaces Bleeker,G. rappi (Barnard), andG. fllamentosus Cuvier, all of which occurred only in clear water although the greatest densities of their bivalve prey were in turbid waters. Similarly, the sparidsRhabdosargus holubi (Steindachner) andR. sarba (Forsskal) were distributed according to turbidity and not their preferred foods.Principal component analysis with a minimum spanning tree plot and a canonical correlation test showed that the fish fauna could be divided into five groups according to their occurrence in various turbidities. These were: clear water species (e.g. Gerreidae) in < 10 NTU, clear to partially turbid species (e.g.Liza dumerilii (Steindachner) andL. macrolepis (Smith)) in < 50 NTU, intermediate turbidity species (e.g.Valamugil cunnesius (Valenciennes) andLeiognathus equula (Forsskal)) in 10–80 NTU, turbid-water species (e.g.Elops machnata (Forsskal) andThryssa vitrirostris) in > 50 NTU, and species indifferent to turbidity (e.g.Acanthopagrus berda (Forsskal) andTeraponjarbua (Forsskal). It is, therefore, suggested that turbidity plays a significant roˆle, either singly, or in combination with other variables in determining the distribution of juvenile marine fishes in estuaries.     

Comparative Study of Four Microporous Filters for Concentrating Viruses from Drinking Water

Four microporous virus-absorbent filter media for recovering low levels of virus from 380 liters of drinking water were compared. In addition two of the filter media were compared with 1,900 liters of drinking water. The filter media evaluated were MF nitrocellulose membranes (293 mm), AA Cox M-780 epoxy-fiberglass-asbestos disks (267 mm), K-27 yarn-wound fiberglass cartridges + AA Cox M-780 disks (127 mm), and Balston epoxy-fiberglass tubes (24.5 by 63.5 mm). The filters were used to concentrate seeded poliovirus from 380 liters of finished drinking water. Sodium thiosulfate was added to the drinking water to neutralize chlorine, and hydrochloric acid was added to adjust the pH to 3.5. Virus was eluted from the filters with glycine-NaOH buffer at pH 11.5. In terms of virus recovery efficiency, the filter media ranked Balston greater than Cox 267-mm greater than MF 293-mm congruent to K-27 + Cox 127-mm, but differences were slight. The Balston filters and holders were also superior to the other systems in terms of size, weight, cost, and handling factors. Experiments with 2- and 8-mum porosity Balston filters showed no statistically significant difference in virus recovery. Virus was readily detected by the Balston and the MF 293-mm systems at input levels of 12 to 22 PFU/1,900 liters. Preliminary experiments indicated that an elution pH lower than 11.5 may be satisfactory.     

Antipredator behaviour of 0+ year Perca fluviatilis: effect of vegetation density and turbidity

In this study, the combined influence of vegetation density and water turbidity on habitat utilization of a prey fish, 0+ year perch Perca fluviatilis , under predation risk (pike, Esox lucius ) was investigated. The vegetated habitat was overall preferred over the open habitat in the presence of a predator. The level of turbidity, and to a lesser extent vegetation density, however, influenced the response of 0+ year perch. The use of the vegetated habitat was lower in very turbid than in clear and turbid conditions, suggesting reduced antipredator behaviour in very turbid water. The effect of vegetation density on antipredator behaviour was only present in clear water, where the use of a structural refuge decreased with increasing vegetation density. No such effect was observed in turbid and very turbid water. The results showed that the structuring role of vegetation or habitat complexity may diminish with increased turbidity. The observed masking effect of turbidity suggests that predator‐prey interactions in vegetated habitats are more complex than what has generally been thought.     

Growth of pike larvae: effects of prey, turbidity and food quality

We studied experimentally the effects of turbidity and prey composition on pike larval growth and hypothesized that pike larval growth varies with turbidity and food quality. We reared the first-feeding pike larvae (Esox lucius) in laboratory tanks with (1) clear or (2) turbid water provided with zooplankton rations from (3) an inner and (4) an outer archipelago site. The sites differ in physical features, salinity, eutrophication status, zooplankton community structure and density. Pike larvae showed the highest weight increase in clear water with zooplankton from the outer site and the poorest weight increase in turbid water with zooplankton as prey from the inner site. Our fatty acid analysis revealed that unsaturated fatty acid levels were highest in the outer site. The relative percentage of copepods was also higher in the outer site. This study supports the hypothesis that turbidity weakens the ability of pike larvae to capture certain prey. Further, zooplankton community composition matters in turbid water, but is not a primary factor in clear water.     

Effects of bentonite clay solids on poliovirus concentration from water by microporous filter methods

To determine whether suspended solids interfere with enteric virus recovery from water by microporous filter methods, the effects of bentonite clay solids at a concentration of 10 nephelometric turbidity units on the recovery of poliovirus type 1 from seeded, activated carbon-treated, filtered tap water were studied. Volumes (500 ml) of virus-laden water at pH 5.5 or 7.5, with and without 50 mM MgCl2, were filtered through 47-mm-diameter, electropositive (Virosorb 1MDS) and electronegative (Filterite) filters that had been pretreated with Tween 80 to minimize direct virus adsorption to filter surfaces. Bentonite solids enhanced virus retention on both types of filters, even under conditions in which viruses were not solids associated. However, bentonite solids also interfered with elution of retained viruses when eluting with 0.3% beef extract-50 mM glycine (pH 9.5). Under some conditions, overall virus recoveries were lower from water with bentonite solids than from solids-free control water. The results of this study indicate that clay turbidity can interfere somewhat with virus recovery by current microporous filter methods.     

Effects of bentonite clay solids on poliovirus concentration from water by microporous filter methods.

To determine whether suspended solids interfere with enteric virus recovery from water by microporous filter methods, the effects of bentonite clay solids at a concentration of 10 nephelometric turbidity units on the recovery of poliovirus type 1 from seeded, activated carbon-treated, filtered tap water were studied. Volumes (500 ml) of virus-laden water at pH 5.5 or 7.5, with and without 50 mM MgCl2, were filtered through 47-mm-diameter, electropositive (Virosorb 1MDS) and electronegative (Filterite) filters that had been pretreated with Tween 80 to minimize direct virus adsorption to filter surfaces. Bentonite solids enhanced virus retention on both types of filters, even under conditions in which viruses were not solids associated. However, bentonite solids also interfered with elution of retained viruses when eluting with 0.3% beef extract-50 mM glycine (pH 9.5). Under some conditions, overall virus recoveries were lower from water with bentonite solids than from solids-free control water. The results of this study indicate that clay turbidity can interfere somewhat with virus recovery by current microporous filter methods.     

A simple and novel method for recovering adenovirus 41 in small volumes of source water

Aim: A new procedure was developed to recover adenovirus 41 in small volumes (1 l) of water samples based on adsorption, elution and evaporation. Methods and Results: One litre of source water seeded with adenovirus 41 was adjusted to pH 3·5 and filtered using a large pore size (8·0 μm) negatively charged membrane filter (SCWP, 47 mm diameter, made of mixed‐cellulose esters). Then, the filter was eluted using 4 ml of 1·5% beef extract plus 0·75% glycerol (pH 9·0). The eluate was reconcentrated to 0·1 ml or less volumes through evaporation assisted with air flow and heating at 55°C. Recovery of adenovirus 41 reached 55% under tested conditions and reduced filtration time by 85% in contrast to the widely used small pore size filter (0·45 μm pore size, 47 mm diameter). Reconcentration by evaporation achieved approx. 86·8% recovery from source water in approx. 1 h at no cost. Conclusion: The virus concentration method developed in this study is simple and cost‐effective and can be used to efficiently recover adenovirus 41 from turbid water samples. Significance and Impact of the Study: The procedure developed can be applied to detect adenovirus 41 in source water within hours of sampling. In addition, this is the first application of evaporation to concentrate viruses in water samples.     

Detection of virus in water: sensitivity of the tentative standard method for drinking water.

The sensitivity of several microporous virus-adsorbent media for reliably detecting low levels of poliovirus from 380 and 1,900 liters of drinking water by use of the tentative standard method was investigated. The virus-adsorbent media tested were (i) nitrocellulose membrane filters, (ii) epoxy-fiber glass-asbestos filters, (iii) yarn-wound fiber glass depth filters, and (iv) epoxy-fiber glass filter tubes. Virus was adsorbed to the filter media at pH 3.5 and eluted with glycine buffer, pH 11.5. The results from 44 samples demonstrated that poliovirus was detected with a 95% reliability at mean virus input levels of 3 to 7 plaque-forming units/380 liters when 1,900 liters of water was sampled. At mean virus input levels of less than 1 to 2 plaque-forming units/380 liters, the detection reliability was 66% in 76 samples when 1,900 liters of water was sampled. No significant difference in virus detection sensitivity was observed among the various virus adsorbent media tested. Overall virus recovery efficiency ranged from 28 to 42%, with a grand average of 35%. Members of the coxsackievirus groups A and B, echovirus, and adenovirus were also detected when 380 and 1,900 liters of water were sampled. These experimental observations attest to the sensitivity of the tentative standard method for detecting low levels of virus in large volumes of drinking water.     

Detection of virus in water: sensitivity of the tentative standard method for drinking water.

The sensitivity of several microporous virus-adsorbent media for reliably detecting low levels of poliovirus from 380 and 1,900 liters of drinking water by use of the tentative standard method was investigated. The virus-adsorbent media tested were (i) nitrocellulose membrane filters, (ii) epoxy-fiber glass-asbestos filters, (iii) yarn-wound fiber glass depth filters, and (iv) epoxy-fiber glass filter tubes. Virus was adsorbed to the filter media at pH 3.5 and eluted with glycine buffer, pH 11.5. The results from 44 samples demonstrated that poliovirus was detected with a 95% reliability at mean virus input levels of 3 to 7 plaque-forming units/380 liters when 1,900 liters of water was sampled. At mean virus input levels of less than 1 to 2 plaque-forming units/380 liters, the detection reliability was 66% in 76 samples when 1,900 liters of water was sampled. No significant difference in virus detection sensitivity was observed among the various virus adsorbent media tested. Overall virus recovery efficiency ranged from 28 to 42%, with a grand average of 35%. Members of the coxsackievirus groups A and B, echovirus, and adenovirus were also detected when 380 and 1,900 liters of water were sampled. These experimental observations attest to the sensitivity of the tentative standard method for detecting low levels of virus in large volumes of drinking water.