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PubMed2001

Adsorption and survival of faecal coliforms, somatic coliphages and F-specific RNA phages in soil irrigated with wastewater.

This study was carried out to compare the adsorption and survival of faecal coliforms, somatic coliphages and F-specific RNA phages in soil irrigated with wastewater. Adsorption isotherms showed that 3-10x more faecal coliforms than somatic coliphages were adsorbed from wastewater onto soil. The adsorption behavior of F-specific RNA phages was intermediate between those of these two microorganisms. In wastewater, the inactivation factor of somatic coliphages at 8-22 degrees C was 5-7 lower than those of faecal coliforms. F-specific RNA phages have a decrease close to faecal coliforms. In soil, at temperatures of 8-22 degrees C and at moistures of 15-35%, somatic coliphages survived longer than the two other microorganisms. These results seemed to be confirmed by the soil column experiments. The rate of inactivation of all microorganisms was lower in soil than in wastewater and depended extensively on soil temperature and moisture content. Survival was optimal at low temperature (8 degrees C) and low moisture content (15%). Thus, somatic coliphages seemed to be a better indicator of faecal contamination than faecal coliforms under our experimental conditions and based only on the two criteria tested (survival and adsorption). Somatic coliphages were able to contaminate the soil over greater distances and survive better in both wastewater and soil than faecal coliforms. These results need to be confirmed by studies on several soil columns using different kinds of soil and different kinds of wastewater.

Adsorption↗

PubMed2002

Removal of indigenous coliphages and fecal coliforms by a novel sewage treatment system consisting of UASB and DHS units.

A novel sewage treatment system, which consists of an upflow anaerobic sludge blanket (UASB) pre-treatment unit and the following downflow hanging sponge (DHS) unit for polishing up the UASB effluent, was developed as a cost-effective and easy-maintenance sewage treatment system for developing countries. A long-term experiment with actual sewage was conducted in order to evaluate its treatment efficiency of organic substances, nutrients, and pathogen indicator microorganisms such as total coliphages, F+-specific RNA coliphages (RNA coliphages), and fecal coliforms. The main objective of this paper is to investigate the removal efficiency of those indicator microorganisms by the UASB-DHS combined system. The results obtained from the continuous flow experiment indicated a fairly promising removal of the indicator microorganisms, i.e., the log10 reductions of total coliphages, RNA coliphages, and fecal coliforms (based on sewage and DHS effluent) achieved were 2.01 log, 2.02 log, and 2.57 log, respectively. The UASB-DHS combined system was superior to the conventional activated sludge process in the reduction of fecal coliforms, but in the reductions of total and RNA coliphages, the system showed somewhat less removal efficiency. The vertical reducing patterns of the indicator microorganisms along the DHS reactor were also discussed.

Bacteria, Anaerobic↗

PubMed2005

Occurrence and correlations between coliphages and anthropogenic viruses in the Massachusetts Bay using enrichment and ICC-nPCR.

We evaluated a two-step enrichment procedure to detect coliphages and an integrated cell culture-nested polymerase chain reaction (ICC-nPCR) to detect human astrovirus, enteroviruses, rotavirus and adenovirus type 40 and 41 in marine water samples collected by the Massachusetts Water Resource Authority (MWRA). MWRA has been monitoring its receiving waters for coliphages, anthropogenic viruses and indicator bacteria in order to evaluate the impact of Boston's Deer Island Sewage Treatment Plant discharge. Coliphages and enteric viruses were originally assayed using single agar overlay and most probable number cell culture (MPN) methods, respectively. Reanalysis of these samples for enteric viruses by ICC-nPCR demonstrated that 46% were positive for at least one virus compared with 23% with the MPN method. Use of the enrichment method showed a 47% increase in the detection of male specific and somatic coliphages compared with the single agar overlay method. Correlations between the presence of coliphages, enteric viruses and indicator bacteria were based on proximity to the treatment plant discharge, seasonal variations and site levels. The presence of enteric viruses was significantly correlated to coliphages but not to indicator bacteria. Preliminary comparative results demonstrate that effective and efficient monitoring of anthropogenic contamination can be achieved using these more sensitive and specific techniques.

Base Sequence↗

PubMedDate not supplied

[Coliphages, viral indicators of contamination].

This paper illustrates the importance of coliphages, which are specific bacteriophages for Escherichia coli strains, as viral indicators, and suggests the usefulness of a routine coliphage search in the detection of viral contamination. The samples studied consisted of 57 surface water, (46 of sea water and 11 of river water), 2 of mud taken from biological treatment plants and 3 samples of sludge. The results revealed that coliphages can be considered valid indicators of viral contamination as they show a good response to the general standards established for indicators. In particular: they are found in sludge (both untreated and treated) and in surface water (both river and sea) in higher concentrations than those of enterovirus; they show greater resistance than bacterial indicators to disinfection as, in different experimental conditions, the drop in the level does not exceed orders of magnitude they are relatively simple to identify, and are easily quantifiable by means of the MPN method, with the result that they can be used for the management of water resources. As regards the possible use of coliphages for the control of the necessary qualitative requisites of sea water for bathing, the relation with fecal coli enables us to make an assessment of coliphages corresponding to 100 fecal coli/100 ml. This levels is equivalent to 0.95 so that a suitable guide line for water for bathing purposes could for example be established at 10 coliphages/1. Although coliphages may be considered good indicators of viral contamination this clearly does not mean that they can replace classic bacterial indicators but rather, that they can provide useful additional information, particularly in those cases where a rapid drop in bacterial content may conceal high risk health and hygiene conditions.

Bacteria↗

PubMed1993

[Inactivation of bacteria and coliphages in surface water highly polluted by secondary effluent and purified by flocculation and filtration by means of UV irradiation at a pilot plant scale].

The water of a channel in Berlin which is highly polluted by municipal sewage effluent is treated at the phosphate elimination plant (PEP) Tegel by flocculation and filtration in order to reduce eutrophication in the following Lake Tegel. The elimination of bacteria and coliphages in the effluent of the PEP was investigated in a scale pilot UV irradiation reactor installed at the outlet of the PEP Tegel. The influence of technical parameters such as flow rate and the arrangement of 23 UV lamps in the reactor on the inactivation was tested. The UV irradiation dose was calculated 119 mJ/cm2 and 49 mJ/cm2 at a flow rate of 50 m3/h and 120 m3/h, respectively and for an irradiation zone of 97.5 cm. The colony count of bacteria and concentrations of coliform organisms, E. coli, and feacal streptococci as well as the plaque forming units of coliphages in the influent of the UV reactor were reduced 2-3 lg units by an irradiation dose of 119 mJ/cm2. These elimination was found being only one lg unit at a UV irradiation dose of 49 mJ/cm2. The concentration of E. faecalis and Coliphages f2 seeded into the influent of the UV reactor decreased after UV irradiation by 119 mJ/cm2 by 2-4 lg units and 1-2 power of magnitude, respectively. A UV dose of 49 mJ/cm2 caused only a 90% elimination of E. faecalis and a 75% inactivation of Coliphages f2. Due to heterogenous distribution and the different retention period of the inflowing water in the irradiation zone, the inactivation of E. faecalis and Coliphages f2 was unequal. Both test organisms decreased in the middle of the reactor up to 2 lg units more than at the sides of the reactor. The hygienic-microbiological quality of a secondary effluent from sewage treatment plants can be improved by a combination of flocculation-filtration and UV irradiation due to their additive elimination effect. However, this UV reactor, which was tested under field conditions can only ensure the inactivation of bacteria and coliphages in the pretreated effluent, if more homogenous distribution of the inflowing water can be achieved. Further, the water must be irradiated by a higher UV dose.

Coliphages↗

PubMed1980

Identification of the major adherence ligand of Klebsiella pneumoniae in the receptor for coliphage T7 and alteration of Klebsiella adherence properties by lysogenic conversion.

We have studied the adherence of both laboratory and wild-type Klebsiella pneumoniae strains, isolated from sputum, urine, and stool samples, to human buccal and intestinal and urinary tract epithelial cells. Of 32 unencapsulated strains, 30 adhered to all epithelial cells tested. Four K. pneumoniae strains lysogenic for AP3, a phage which causes conversion to resistance of coliphages T3, T7, and phi I, were all unable to adhere to epithelial cells. One of these strains was cured from phage infection and became capable of adhering, Spontaneous mutants resistant to coliphage T7, as well as K. pneumoniae K59-sensitive cells preadsorbed with inactivated T7 particles, did not adhere to epithelial cells. All strains capable of adhering were able to adsorb coliphage T7 and T3, whereas all nonadhesive strains were not. AP3-like prophages were induced from 7 of 12 nonadhesive Klebsiella strains. A laboratory strain which was able to adhere was lysogenized with 2 of these phages. In both cases, the strain lost its ability to adsorb coliphages T3, T7, and phi I and to adhere to human epithelial cells. All K. pneumoniae adhesive strains agglutinated yeast cells, whereas the nonadhesive strains did not. Competition studies have shown that D-mannose and concanavalin A prevented adherence to human epithelial cells, yeast agglutination, and adsorption of coliphage T7 to K. pneumoniae cells. It is concluded that in K. pneumoniae adherence to epithelial cells is mediated by the receptor for coliphages T7 (and T3), which in turn recognizes D-mannose in the receptors it binds.

Cell Adhesion↗

PubMed1964

EFFECTS OF ENVIRONMENTAL FACTORS ON THE SURVIVAL OF AIRBORNE T-3 COLIPHAGE.

Experiments were conducted to determine the effects of storage temperatures, relative humidity, and additives on the survival of aerosolized Escherichia coli phage T-3. The aerosol stability of the coliphage, calculated as per cent recovery, was not affected by storage at 10 or -70 C for up to 4 months. However, an increase in aerosol decay rate of coliphage stored at 10 C was observed. The effect of humidities ranging from 20 to 90% relative humidity was studied, and it was observed that humidities lower than 70% relative humidity significantly reduce the survival of airborne coliphage. The effect of various compounds on the aerosol decay rate of T-3 coliphage was studied at 50 and 85% relative humidity. Addition of dextrose in 0.1 M concentrations to the disseminating fluid significantly reduced aerosol decay rate at 50% relative humidity without affecting the decay at 85%. Addition of spermine, spermidine-phosphate, thiourea, galacturonic acid, and glucosaminic acid, individually or in combination, had no effect on aerosol decay rates. The use of deuterium oxide as the suspending fluid for dissemination had no effect on aerosol stability of the coliphage.

Aerosols↗

PubMed2006

Multiplex quantitative real-time reverse transcriptase PCR for F+-specific RNA coliphages: a method for use in microbial source tracking.

It is well documented that microbial contamination of coastal waters poses a significant risk to human health through recreational exposure and consumption of shellfish. Identifying the source of microbial contamination (microbial source tracking) plays a dominant role in enabling effective management and remediation strategies. One method used to determine the source of the contamination is quantification of the ratio of the four subgroups of F+-specific RNA coliphages (family Leviviridae) in impacted water samples. Because of typically low concentrations in the environment, enrichment assays are performed prior to detection, even though differential replication rates have been reported. These assays are also compromised by differential loss of phage infectivity among subgroups after release into the environment, thus obscuring the initial ratio. Here, a culture-independent multiplex real-time reverse transcriptase-PCR (RT-PCR) protocol for the simultaneous quantification of all four subgroups of F+-specific RNA coliphages using novel primer sets and molecular beacons is presented. This assay is extremely sensitive, achieving detection with as few as 10 copies of isolated coliphage RNA, and is linear for a minimum of six orders of magnitude. During survival experiments, the real-time RT-PCR technique was able to quantify coliphages in seawater when culture-based double agar layer assay failed. While infectivity was lost at different rates at the subgroup level, decay constants in seawater, calculated using the real-time RT-PCR estimates, did not vary among subgroups. The accurate determination of the in situ concentration of F+-specific RNA coliphages using this method will facilitate more effective remediation strategies for impacted environments.

Animals↗

PubMed1980

Concentration of coliphages from large volumes of water and wastewater.

Membrane filter adsorption-elution technology has been extensively used for the concentration and detection of animal viruses from large volumes of water. This study describes the development of positively charged microporous filters (Zeta Plus) for the concentration of coliphages from large volumes of water and wastewater. Four different coliphages were studied: MS-2, phi X174, T2, and T4. Positively charged microporous filters were found to efficiently adsorb these coliphages from tap water, sewage, and lake water at neutral pH. Adsorbed viruses were eluted with a 1:1 mixture of 8% beef extract and 1 M sodium chloride at pH 9. Using this method, coliphages could be concentrated from 17-liter volumes of tap water with recoveries ranging from 34 to 100%. Coliphages occurring naturally in raw and secondarily treated sewage were recovered with average efficiencies of 56.5 and 55.0%, respectively. This method should be useful in isolation of rare phages, the ecology of phages in natural waters, and the evaluation of water quality.

Coliphages↗

PubMed1988

Incidence of coliphage in potable water supplies.

Samples of drinking water from different sources in greater Cairo, Egypt, and bottled drinking water were tested for total coliform, fecal coliform, and coliphage populations. Of the 147 samples tested, 4 samples were positive for both total coliforms and coliphage, 65 samples were negative for total coliforms, fecal coliforms, and coliphage, and 78 samples were positive for coliphage and negative for total coliforms and fecal coliforms. The incidence of coliphage in these potable water supplies reflects the probability of human pathogenic virus survival in these waters also.

Coliphages↗

PubMed1992

Rates of inactivation of waterborne coliphages by monochloramine.

A sophisticated water quality monitoring program was established to evaluate virus removal through Denver's 1-million-gal (ca. 4-million-liter)/day Direct Potable Reuse Demonstration Plant. As a comparison point for the reuse demonstration plant, Denver's main water treatment facility was also monitored for coliphage organisms. Through the routine monitoring of the main plant, it was discovered that coliphage organisms were escaping the water treatment processes. Monochloramine residuals and contact times (CT values) required to achieve 99% inactivation were determined for coliphage organisms entering and leaving this conventional water treatment plant. The coliphage tested in the effluent waters had higher CT values on the average than those of the influent waters. CT values established for some of these coliphages suggest that monochloramine alone is not capable of removing 2 orders of magnitude of these specific organisms in a typical water treatment facility. Electron micrographs revealed one distinct type of phage capable of escaping the water treatment processes and three distinct types of phages in all.

Chloramines↗

PubMed2004

Molecular detection and genotyping of male-specific coliphages by reverse transcription-PCR and reverse line blot hybridization.

In recent years, there has been increased interest in the use of male-specific or F+ coliphages as indicators of microbial inputs to source waters. Sero- or genotyping of these coliphages can also be used for microbial source tracking (MST). Among the male-specific coliphages, the F+ RNA (FRNA) viruses are well studied, while little is known about the F+ DNA (FDNA) viruses. We have developed a reverse line blot hybridization (RLB) assay which allows for the simultaneous detection and genotyping of both FRNA as well as FDNA coliphages. These assays included a novel generic duplex reverse transcription-PCR (RT-PCR) assay for FRNA viruses as well as a generic PCR for FDNA viruses. The RT-PCR assays were validated by using 190 field and prototype strains. Subsequent DNA sequencing and phylogenetic analyses of RT-PCR products revealed the classification of six different FRNA clusters, including the well-established subgroups I through IV, and three different FDNA clusters, including one (CH) not previously described. Within the leviviruses, a potentially new subgroup (called JS) including strains having more than 40% nucleotide sequence diversity with the known levivirus subgroups (MS2 and GA) was identified. We designed subgroup-specific oligonucleotides that were able to genotype all nine (six FRNA, three FDNA) different clusters. Application of the method to a panel of 351 enriched phage samples from animal feces and wastewater, including known prototype strains (MS2, GA, Q beta, M11, FI, and SP for FRNA and M13, f1, and fd for FDNA), resulted in successful genotyping of 348 (99%) of the samples. In summary, we developed a novel method for standardized genotyping of F+ coliphages as a useful tool for large-scale MST studies.

Animals↗

PubMed2004

Comparison of coliforms and coliphages as tools for assessment of viral contamination in river water.

The aim of the study was to evaluate the presence of pathogenic viruses in the Moselle River and to compare the usefulness of thermotolerant coliforms and somatic coliphages as tools for river water quality assessment in terms of viral contamination. Thermotolerant coliforms and somatic coliphages were enumerated by standardized methods in 170 samples of river water drawn from five sampling sites along the Moselle River (eastern France). BGM cell culture and integrated cell culture-reverse transcription-PCR DNA enzyme immunoassay were used to determine the presence of pathogenic viral genome (Enterovirus and Norovirus genogroup II [GGII]) and infectious Enterovirus spp. in 90 1-liter samples. No infectious Enterovirus spp. were isolated, but Enterovirus and Norovirus GGII genomes were detected in 38% of the samples. Norovirus GGII genome was mostly detected in winter, whereas Enterovirus genome was mostly detected in summer and fall. Somatic coliphages appeared to be less sensitive to higher river water temperature than thermotolerant coliforms. Furthermore, the number of river water samples positive for pathogenic viral genome increased with increasing concentration of somatic coliphages, whereas coliform concentration was unrelated to viral genome contamination. Consequently somatic coliphages, which are less sensitive to environmental factors than thermotolerant coliforms in river water, would provide a promising tool for assessment of river water quality in terms of fecal and viral pollution.

Coliphages↗

PubMed2004

Usefulness of monitoring tropical streams for male-specific RNA coliphages.

The objective of this study was to evaluate the usefulness of monitoring streams in Hawaii for FRNA coliphages as a reliable indicator of sewage contamination. This study was undertaken as a result of our previous findings that monitoring streams in Hawaii for traditional faecal indicator bacteria (faecal coliform, Escherichia coli, enterococci) was not useful in determining when streams are contaminated with sewage, because environmental (soil) sources rather than sewage accounted for the high concentrations of faecal bacteria in streams. Two perennial streams, sewage and soil samples were monitored for traditional faecal indicator bacteria (faecal coliform, E. coli, enterococci) and FRNA coliphages. The results showed that sewage treatment processes and disinfection drastically reduced the concentrations of traditional faecal indicator bacteria but FRNA coliphages were still present in significant concentrations in the treated sewage effluents. These results indicate that monitoring sewage effluents and environmental waters for only traditional faecal indicator bacteria may not be adequately protective of human health effects. Ambient concentrations of traditional faecal indicator bacteria in soil and streams of Hawaii were consistently high but consistently low for FRNA coliphages, indicating that monitoring streams of Hawaii for FRNA coliphages can be used to determine when streams are contaminated with sewage.

Adolescent↗

PubMed1981

Coliphages as ecological indicators of enteroviruses in various water systems.

The occurrence of coliphages and enteroviruses in a variety of water systems in Czechoslovakia was monitored for two years. Two host strains of Escherichia coli bacteria were used to test 1161 water samples for the presence of bacteriophages. These strains were polyvalent hosts for a broad spectrum of morphologically distinct coliphages, and their use thus gave quantitative data on the degree of viral pollution in any given water sample. Ninety-two water samples were tested in parallel for the presence of enteroviruses, by using a flocculation method to concentrate the viruses followed by isolation in cultures of a buffalo green monkey (BGM) kidney continuous cell line. The enterovirus and coliphage recovery rates showed similar differences when waters with different levels of pollution were compared. Seasonal fluctuations of both the coliphage and enterovirus (mostly poliovirus) levels in river water were demonstrated by statistical analysis of the data collected. The levels increased in the winter and sharply declined in the summer months as the river water temperature increased. Chemical pollution did not seem to influence the survival of either the coliphages or the enteroviruses in the observed rivers.

Coliphages↗

PubMed2003

Effects of pH and temperature on the survival of coliphages MS2 and Qbeta.

The RNA F-specific coliphages, MS2 and Qbeta, have been used as virus indicators in water and wastewater studies. It is therefore useful to have a good understanding concerning the effects of environmental factors on their survival in order to choose an appropriate candidate for assessing microbial safety in relation to water quality management. The effects of pH and temperature on the survival of these two coliphages were investigated. MS2 survived better in acidic conditions than in an alkaline environment. In contrast, Qbeta had a better survival rate in alkaline conditions than in an acidic environment. The inactivation rates of both coliphages were lowest within the pH range 6-8 and the temperature range 5-35 degrees C. The inactivation rates of both coliphages increased when the pH was decreased to below 6 or increased to above 8. The inactivation rates of both coliphages increased with increasing temperature. Qbeta behaved peculiarly in extreme pH buffers, i.e. it was inactivated very rapidly initially when subjected to an extreme pH environment, although the inactivation rate subsequently decreased. In general, MS2 was a better indicator than Qbeta. However, within the pH range 6-9 and at temperatures not above 25 degrees C, either MS2 or Qbeta could be used as a viral indicator.

Environment↗

PubMed2002

Nucleotide sequence of a ssRNA phage from Acinetobacter: kinship to coliphages.

The complete nucleotide sequence of ssRNA phage AP205 propagating in Acinetobacter species is reported. The RNA has three large ORFs, which code for the following homologues of the RNA coliphage proteins: the maturation, coat and replicase proteins. Their gene order is the same as that in coliphages. RNA coliphages or Leviviridae fall into two genera: the alloleviviruses, like Q(beta), which have a coat read-through protein, and the leviviruses, like MS2, which do not have this coat protein extension. AP205 has no read-through protein and may therefore be classified as a levivirus. A major digression from the known leviviruses is the apparent absence of a lysis gene in AP205 at the usual position, overlapping the coat and replicase proteins. Instead, two small ORFs are present at the 5' terminus, preceding the maturation gene. One of these might encode a lysis protein. The other is of unknown function. Other new features concern the 3'-terminal sequence. In all ssRNA coliphages, there are always three cytosine residues at the 3' end, but in AP205, there is only a single terminal cytosine. Distantly related viruses, like AP205 and the coliphages, do not have significant sequence identity; yet, important secondary structural features of the RNA are conserved. This is shown here for the 3' UTR and the replicase-operator hairpin. Interestingly, although AP205 has the genetic map of a levivirus, its 3' UTR has the length and RNA secondary structure of an allolevivirus. Sharing features with both MS2 and Q(beta) suggests that, in an evolutionary sense, AP205 should be placed between Q(beta) and MS2. A phylogenetic tree for the ssRNA phages is presented.

3' Untranslated Regions↗

PubMed1994

Field evaluation of two colorimetric coliphage detection methods.

Two new methods for coliphage detection, a colorimetric agar-based (CAB) method and a liquid colorimetric presence-absence (LCPA) method, were compared to the coliphage method proposed by the American Public Health Association (APHA; Standard Methods for the Examination of Water and Wastewater, 18th ed., American Public Health Association, Washington, D.C., 1992). Both new methods are based on the induction of beta-galactosidase in Escherichia coli and the release of the enzyme through a lytic cell infection. The released enzyme then cleaves a chromogenic substrate which produces a colored reaction product. Ninety split water samples from four different sources were tested. A total of 52 samples were positive by the CAB method, 52 were positive by the LCPA method, and 53 were positive by the APHA method. Results indicated that (i) the CAB and LCPA methods were as sensitive in coliphage detection as the APHA method, (ii) both the CAB and LCPA methods were easier to read and interpret than the APHA method, and (iii) the CAB method detected more coliphages in a positive sample than the APHA method in two of the four types of water sources. Importantly, the rapid and simple LCPA method was as reliable and sensitive as either of the two agar-based methods in coliphage detection.

Journal Article↗