Presence Of Human Enteric Viruses Research Articles

Prevalence and quantitative assessment of foodborne viruses on the imported mussels in Polish market

The aim of this study was an assessment of the prevalence of human enteric viruses such as norovirus (NoV) and hepatitis A and E virus (HAV, HEV) in mussels sold on the Polish market. In addition, the presence of human (hAdV) and swine (pAdV) adenoviruses as indicators of faecal contamination of shellfish was also studied. In total, 246 shellfish samples representing 41 batches of blue and Mediterranean mussels were purchased from Polish seafood wholesalers. Shellfish were processed according to the method in ISO 15216-2:2019 followed by isolation of viral nucleic acids using NucliSens®Magnetic Extraction Reagents (bioMérieux). For virus detection several duplex real-time PCR protocols with incorporated internal amplification controls (IAC) were used. The presence of human enteric viruses was shown in 42 shellfish samples (17.1%). The most frequently detected were NoV (41 samples; 16.7%) followed by HEV (1 sample; 0.4%). None of the samples was positive for HAV, hAdV or pAdV. 15 shellfish batches (36.6%) were virus positive. By the use of Bayesian inference, a true seasonal and country related virus prevalence in shellfish was estimated. NoV GI was more frequently detected in Dutch mussels (18.9%; 95% CI: 6.7%–34.0%), while NoV GII was more frequent in Danish mussels (17.1%; 95% CI: 5.8%–32.5%). However significant differences in virus prevalence in individual and shellfish batches between producing countries were not shown. A seasonal virus distribution pattern in shellfish samples was observed. Lower level of NoV contamination was found in mussels harvested during spring-summer (0.0%–1.3%, 95% CI: 0.1%–4.8%) compared to the fall-winter season (13.0%–20.7%, 95% CI: 6.1%–31.2%). For shellfish batches, a similar seasonal correlation was only observed for NoV GII. The monitoring of marketed shellfish would be an essential tool allowing a reduction of the number of viral food-borne outbreaks.

Town scale response of water viral communities to town source surface water contamination with hydrochemical parameters

This study aims to explore the relationship between human enteric viruses found in town-scale surface water sources and certain chemical contaminants present in the water. From October 2010 to April 2012, water samples were collected and analyzed using a combination of biophysical and molecular techniques to detect the presence of human adenoviruses (HAdV) and human enteroviruses (HEV) as well as chemical parameters as predictors for virus survival. The concentrations of 12 chemical contaminants were found to be within WHO-recommended limits. The study found positive and negative associations between viral genome detection and four out of the 12 metal and nonmetal analytes. Specifically, there was a correlation between Cd and HAdV genome detection (rho = 0.146, p = 0.032) and between Pb and Fe with HEV (rho = 0.156, p = 0.022) and (rho = 0.148, p = 0.029), respectively. For nonmetals, phosphates were slightly negatively correlated to HEV (rho = 0.174, p = 0.010). The results of the study did not provide support for the hypothesis of an association between the presence of human enteric viruses and the levels of twelve chemical contaminants.

Wastewater-Based Epidemiology of Enteric Viruses and Surveillance of Acute Gastrointestinal Illness Outbreaks in a Resource-Limited Region.

Acute gastrointestinal illness (AGI) continues to be an important cause of morbidity and mortality among all ages. This study applied the principles of wastewater-based epidemiology for the preventive identification of potential outbreaks of acute viral gastroenteritis and hepatitis A by analyzing the presence of human enteric viruses in influents of small municipal wastewater treatment plants (WWTPs) handling domestic sewage, together with public health reports of acute diarrheal and hepatitis A disease in Costa Rica during 2013. Raw wastewater samples were collected during four seasonal periods with different rainfall levels. The presence of five human enteric viruses (rotavirus A, norovirus GI, norovirus GII, enterovirus, and hepatitis A virus) was studied by endpoint and real-time polymerase chain reaction in influents of five WWTPs. Cases of AGI were analyzed using historical public health reports of endemic levels and quartile ranges for each administrative and territorial area where the WWTPs are located and for its surrounding counties. A tendency for a higher rate of positive viral tests was present 1 week before an increase of AGI cases. Epidemiological weeks categorized as Outbreak (above the 75th percentile) and Success (below the 25th percentile) showed statistically significant differences in terms of positive viral test rates (Wilcoxon test, P = 0.05). Virological monitoring of wastewater in small WWTPs is an appropriate model for epidemiological surveillance of diarrheal and hepatitis A diseases in low- and middle-resource countries.

Monitoring of human enteric virus and coliphages throughout water reuse system of wastewater treatment plants to irrigation endpoint of leafy greens

One solution to current water scarcity is the reuse of treated wastewater. Water reuse systems have to be examined as a whole, including the efficacy of water-reclamation treatments and the operation steps from the wastewater inlet into the WWTP to the irrigation endpoint, including the irrigated crop. In this study, the monitoring of human enteric viruses and coliphages were assessed in two water reused systems. The presence of hepatitis A virus (HAV) and human noroviruses genogroups I and II (GI and GII) were analyzed by real-time RT-PCR (RT-qPCR) in water (n = 475) and leafy green samples (n = 95). Total coliphages were analyzed by the double-layer agar plaque technique. The prevalence of HAV in water samples was very low (c.a. 2%), mostly linked to raw sewage, while for leafy green samples, none was positive for HAV. In leafy greens, prevalence of norovirus was low (less than 5–6%). The highest reductions for norovirus were observed in samples taken from the water reservoirs used by the growers near the growing field. The virus die-off during water storage due to solar radiation could be considered as an additional improvement. Reclamation treatments significantly reduced the prevalence and the counts of noroviruses GI and GII and coliphages in reclaimed water. However, the coliphage reductions (c.a. 5 log) do not comply with the specifications included in the new European regulation on reclaimed water (≥6.0 log). Correlations between noroviruses GI and GII and coliphages were found only in positive samples with high concentrations (>4.5 log PFU/100 mL). A high percentage of samples (20–25%) negative for total coliphages showed moderate norovirus counts (1–3 logs), indicating that coliphages are not the most suitable indicator for the possible presence of human enteric viruses.

Effects of Contamination of Freshwater Habitat With Common Heavy Metals and Anions on the Prevalence of Human Adenoviruses and Enteroviruses

The occurrence and survival of enteric viruses in open surface waters can be impacted by a host of factors including fecal emission levels, seasonal variations, virus stability and the physicochemical parameters. In this research, we aimed to document the association between contaminations of water samples with human enteric viruses (adenoviruses and enteroviruses) from a freshwater lake with variations in chemical contaminants. We collected 216 water samples from October 2010 to April 2012, from a 4 km stretch along Lake Victoria (LV) basin in Homa Bay town located in the western region of Kenya. The samples were analyzed for the existence of human adenoviruses (HAdV) and human enteroviruses (HEV), using the nested PCR (nPCR). We also assessed in the water samples the levels of twelve chemical contaminants consisting of six heavy metal elements and six anions. About 8.3 % of the samples were found to be contaminated with the enteric viruses. The concentrations of the 12 chemical contaminants were found to be largely within the WHO suggested limits. Most of the chemical contaminants were not related to the detection rates of the viruses from the statistical analysis. However, some positive and negative associations between the viral genome's detection and the chemical concentrations were established for only three metals (Fe, Pb, Cd) and the PO43− Radical. Cd had a weak positive significant relationship with HAdV (rho = 0.146, p = 0.032) while Pb and Fe had a weak positive significant relationship with HEV genome detection (rho = 0.156, p = 0.022) and (rho = 0.148 and p = 0.029) respectively. There was a modest negative relationship between phosphate ions and HEV (rho = −0.174, p = 0.010). The results of our study do not provide support for the hypothesis of an association between the presence of human enteric viruses and the levels of twelve chemical contaminants.

Occurrence and reduction of F-specific RNA bacteriophage genotypes as indicators of human norovirus at a wastewater treatment plant.

F-specific RNA bacteriophages (FRNAPHs) have been suggested as good indicators of the presence of human enteric viruses in water treatment facilities. The occurrence and reduction of norovirus (NoV) and FRNAPH genotypes in wastewater treatment plants (WWTPs) have been well studied; however, the relationship between these genotypes in WWTPs has not been fully elucidated. Thus, we aimed to investigate the occurrence and reduction of FRNAPH genotypes in an attempt to identify NoV indicators in a WWTP via a 1-year survey. All FRNAPH and NoV genotypes were detected in WWTP influents at high rates (71-100%), including the infectious FRNAPH genotype IV (GIV), which has been rarely detected in previous studies. The reductions of FRNAPH GII and NoV GII during wastewater treatment indicated a relationship between the two (r = 0.69, P < 0.01), and the mean values were not significantly different. These results suggested that FRNAPH GII could be used as an appropriate indicator of NoV GII during wastewater treatment. FRNAPH GI was also found to be an appropriate indicator of viral reduction because of its high resistance to wastewater treatment compared with the other FRNAPH and NoV genotypes; therefore, it can be considered as a worst-case scenario organism.

Total coliforms as an indicator of human enterovirus presence in surface water across Tianjin city, China

BackgroundEnteric viruses in surface water pose considerable risk to morbidity in populations living around water catchments and promote outbreaks of waterborne diseases. However, due to poor understanding of the correlation between water quality and the presence of human enteric viruses, the failure to assess viral contamination through alternative viral indicators makes it difficult to control disease transmission.MethodsWe investigated the occurrence of Enteroviruses (EnVs), Rotaviruses (HRVs), Astroviruses (AstVs), Noroviruses GII (HuNoVs GII) and Adenoviruses (HAdVs) from Jinhe River over 4 years and analyzed their correlation with physicochemical and bacterial parameters in water samples.ResultsThe findings showed that all target viruses were detected in water at frequencies of 91.7% for HAdVs, 81.3% for HuNoVs GII, 79.2% for EnVs and AstVs, and 70.8% for HRVs. These viruses had a seasonal pattern, which showed that EnVs were abundant in summer but rare in winter, while HAdVs, HRVs, AstVs, and HuNoVs GII exhibited opposite seasonal trends. Pearson correlation analysis showed that total coliforms (TC) was significantly positively correlated with EnVs concentrations while no consistent significant correlations were observed between bacterial indices and viruses that precipitate acute gastroenteritis.ConclusionsTaken together, the findings provide insights into alternative viral indicators, suggesting that TC is a potentially promising candidate for assessment of EnVs contamination. However, it failed to predict the presence of HAdVs, HRVs, AstVs, and HuNoVs GΙΙ in surface water across the city of Tianjin.

Presence of Human Enteric Viruses, Protozoa, and Indicators of Pathogens in the Bagmati River, Nepal.

Quantification of waterborne pathogens in water sources is essential for alerting the community about health hazards. This study determined the presence of human enteric viruses and protozoa in the Bagmati River, Nepal, and detected fecal indicator bacteria (total coliforms, Escherichia coli, and Enterococcus spp.), human-fecal markers (human Bacteroidales and JC and BK polyomaviruses), and index viruses (tobacco mosaic virus and pepper mild mottle virus). During a one-year period between October 2015 and September 2016, a total of 18 surface water samples were collected periodically from three sites along the river. Using quantitative polymerase chain reaction, all eight types of human enteric viruses tested—including adenoviruses, noroviruses, and enteroviruses, were detected frequently at the midstream and downstream sites, with concentrations of 4.4–8.3 log copies/L. Enteroviruses and saliviruses were the most frequently detected enteric viruses, which were present in 72% (13/18) of the tested samples. Giardia spp. were detected by fluorescence microscopy in 78% (14/18) of the samples, with a lower detection ratio at the upstream site. Cryptosporidium spp. were detected only at the midstream and downstream sites, with a positive ratio of 39% (7/18). The high concentrations of enteric viruses suggest that the midstream and downstream regions are heavily contaminated with human feces and that there are alarming possibilities of waterborne diseases. The concentrations of enteric viruses were significantly higher in the dry season than the wet season (p < 0.05). There was a significant positive correlation between the concentrations of human enteric viruses and the tested indicators for the presence of pathogens (IPP) (p < 0.05), suggesting that these IPP can be used to estimate the presence of enteric viruses in the Bagmati River water.

A molecular survey of farmed and edible snails for the presence of human enteric viruses: Tracking of the possible environmental sources of microbial mollusc contamination

Snails have been grown or harvested for human consumption for centuries. Similarly to tissues or meat of other food animals, snail meat should be free from microbiological hazards including viruses. The aim of the study was to investigate the presence of human enteric viruses (hepatitis A and E virus, norovirus genogroup I, II and human adenovirus) in farmed and free-living snails intended for human consumption. In order to find the possible sources of snail contamination with viruses, the quality management system operating in the most widely-adopted snail farming system in Poland was reviewed. Additionally, the process hygiene criteria were checked through testing of snail meat and soil samples for bacterial contamination. Snails of genus Cornu and Helix were collected from two heliciculture farms and a purchase point. They were subjected to bacteriological (n = 180) and viral examination (n = 60). In the case of soil samples (n = 8) they were taken from different areas along the breeding cycle of the snails. For virus detection several real-time PCR protocols with an incorporated target-specific internal amplification control (IAC) were used. Determinations of the total aerobic bacteria count (TBC), number of Enterobacteriaceae (ENC) and enterococci (ECC) were performed using the international reference methods. Human enteric viruses were not detected in any of the tested snail samples. TBC for raw snail meat ranged from 5.1 up to 6.6 log cfu/g and ENC values did not exceed 4.8 log cfu/g. The ECC counts were below 2.0 log cfu/g. The overall microbiological status of snail meat indicated good hygiene and appropriate environmental conditions being maintained during snail breeding. TBC for soil samples was in the range of 6.0–7.7 log cfu/g indicating a low or moderate level of contamination. Only the soil samples from the plots used at the fattening stage were characterised by high ECC content increasing the risk of virus occurrence. In contrary to the sporadic finding of ECC, the ubiquitous presence of bacteria from the Enterobacteriaceae family in all types of soil samples was shown. A good microbial quality of snail meat resulted from the implemented GMP/GHP programmes and food safety management practices. This strategy appeared sufficient in reduction of microbiological risk and in prevention of virus contamination in snails. The study showed that the risk of foodborne virus infection due to consumption of snails and related products is considered negligible.

First detection of papillomaviruses and polyomaviruses in swimming pool waters: unrecognized recreational water-related pathogens?

Viral outbreaks associated with swimming pools have been described worldwide. The objective of this study was to examine the extent of viral contamination in indoor and outdoor swimming pools. Pools were examined for the presence of human enteric viruses (adenovirus, norovirus and enterovirus) and nonenteric viruses (papillomavirus and polyomavirus-BK, JC, KI, WU and Merkel cell). Bacteriological parameters were also evaluated. The analysed pool waters met microbiological quality standards. Enteric viruses were not detected. On the other hand, papillomaviruses (HPV8, 12, 23, 25, 120 and unclassified HPVs) and polyomaviruses (JC and Merkel cell polyomaviruses) were detected in 9/14 samples (64%). The number of HPV DNA copies in pool waters, measured by quantitative Real-time PCR, ranged from 1.27E+04 to 1.13E+05/10L. Results show that a variety of nonenteric viruses may be discharged in pool waters by various secretions and excretions from infected individuals or asymptomatic carriers. To the best of our knowledge, this is the first report on human papillomaviruses and polyomaviruses in swimming pools. The likelihood that these viruses can be transmitted by recreational activities deserves to be explored in future studies.

Preliminary Study on Norovirus, Hepatitis A Virus, Escherichia coli and their Potential Seasonality in Shellfish from Different Growing and Harvesting Areas in Sardinia Region.

Edible lamellibranch molluscs can be involved in foodborne disease and infections of varying severity. They are filter feeding animals able to retain and concentrate in their organism bacteria, parasites, viruses and biotoxins marine algae present in their external environment. Major shellfish harvesting and relaying areas from different areas in Sardinia region were defined and studied by analysing different physicochemical parameters in the water and the levels of Escherichia coli (E. coli), Norovirus (NoVs) genogroup I (NoVGI), NoVs genogroup II (NoVGII) and hepatitis A virus (HAV) in the shellfish harvested and farmed from 2009 to 2011. During that period the identification of the viral agents was carried out by one step real-time reverse transcriptase-polymerase chain reaction and Escherichia coli according to ISO TS 16649-3:2005 standard method. A total of 1266 shellfish samples were tested for NoVGI, NoVGII, HAV and faecal indicators. Norovirus contamination was found in 337 samples (26.6%); only one sample of mussels was positive for HAV (0.08%); while E. coli prevalence was 3.8% in shellfish. The probability of observing shellfish samples positive for NoVs, HAV and E. coli presence was associated with harvesting, growing and relaying areas, period of sampling, environmental parameters, animal species (P<0.05). Although the higher prevalence rate of human enteropathogenic viruses was found in the winter period, we did not observe a significant relationship between the effect of seawater temperature (seasonality) and NoVs presence all over the study period; in fact, according to statistical analysis, the presence of human enteric viruses does not appear to be related to water temperature.

Presence of Human Enteric Viruses in the Stools of Healthy Malawian 6‐Month‐Old Infants

The acquisition of intestinal microbiota is essential for infants who are also in close contact with intestinal viruses. We assayed the presence of human enteric viruses in the faeces of 44 healthy breast-fed 6-month-old infants from rural Malawi. Half of the infants tested harboured enteroviruses, although the infants had no gastric symptoms, suggesting a viral community mainly composed of human asymptomatic enteroviruses.

Sewage pollution in urban stormwater runoff as evident from the widespread presence of multiple microbial and chemical source tracking markers

The concurrence of human sewage contamination in urban stormwater runoff (n=23) from six urban catchments across Australia was assessed by using both microbial source tracking (MST) and chemical source tracking (CST) markers. Out of 23 stormwater samples human adenovirus (HAv), human polyomavirus (HPv) and the sewage-associated markers; Methanobrevibacter smithii nifH and Bacteroides HF183 were detected in 91%, 56%, 43% and 96% of samples, respectively. Similarly, CST markers paracetamol (87%), salicylic acid (78%) acesulfame (96%) and caffeine (91%) were frequently detected. Twenty one samples (91%) were positive for six to eight sewage related MST and CST markers and remaining two samples were positive for five and four markers, respectively. A very good consensus (>91%) observed between the concurrence of the HF183, HAv, acesulfame and caffeine suggests good predictability of the presence of HAv in samples positive for one of the three markers. High prevalence of HAv (91%) also suggests that other enteric viruses may also be present in the stormwater samples which may pose significant health risks. This study underscores the benefits of employing a set of MST and CST markers which could include monitoring for HF183, adenovirus, caffeine and paracetamol to accurately detect human sewage contamination along with credible information on the presence of human enteric viruses, which could be used for more reliable public health risk assessments. Based on the results obtained in this study, it is recommended that some degree of treatment of captured stormwater would be required if it were to be used for non-potable purposes.

Norovirus and Other Human Enteric Viruses in Moroccan Shellfish

The aim of this study was to evaluate the presence of human enteric viruses in shellfish collected along the Mediterranean Sea and Atlantic Coast of Morocco. A total of 77 samples were collected from areas potentially contaminated by human sewage. Noroviruses were detected in 30 % of samples, with an equal representation of GI and GII strains, but were much more frequently found in cockles or clams than in oysters. The method used, including extraction efficiency controls, allowed the quantification of virus concentration. As in previous reports, results showed levels of contamination between 100 and 1,000 copies/g of digestive tissues. Sapoviruses were detected in 13 % of samples mainly in oyster and clam samples. Hepatitis A virus was detected in two samples, with concentrations around 100 RNA copies/g of digestive tissues. Only two samples were contaminated with enterovirus and none with norovirus GIV or Aichi virus. This study highlights the interest of studying shellfish samples from different countries and different production areas. A better knowledge of shellfish contamination helps us to understand virus levels in shellfish and to improve shellfish safety, thus protecting consumers.

Microbial quality assessment of household greywater

A monitoring program was undertaken to assess the microbial quality of greywater collected from 93 typical households in Melbourne, Australia. A total of 185 samples, comprising 75 washing machine wash, 74 washing machine rinse and 36 bathroom samples were analysed for the faecal indicator Escherichia coli. Of these, 104 were also analysed for genetic markers of pathogenic E coli and 111 for norovirus (genogroups GI and GII), enterovirus and rotavirus using RT-PCR. Enteric viruses were detected in 20 out of the 111 (18%) samples comprising 16 washing machine wash water and 4 bathroom samples. Eight (7%) samples were positive for enterovirus, twelve (11%) for norovirus genogroup GI, one (1%) for norovirus genogroup GII and another (1%) for rotavirus. Two washing machine samples contained more than one virus. Typical pathogenic E. coli were detected in 3 out of 104 (3%) samples and atypical enteropathogenic E. coli in 11 (11%) of samples. Levels of indicator E. coli were highly variable and the presence of E. coli was not associated with the presence of human enteric viruses in greywater. There was also little correlation between reported gastrointestinal illness in households and detection of pathogens in greywater.

Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

An estimated 884 million people worldwide do not have access to an improved drinking water source, and the microbial quality of these sources is often unknown. In this study, a combined tangential flow, hollow fiber ultrafiltration (UF), and real-time PCR method was applied to large volume (100 L) groundwater (N = 4), surface water (N = 9), and finished (i.e., receiving treatment) drinking water (N = 6) samples for the evaluation of human enteric viruses and bacterial indicators. Human enteric viruses including norovirus GI and GII, adenovirus, and polyomavirus were detected in five different samples including one groundwater, three surface water, and one drinking water sample. Total coliforms and Escherichia coli assessed for each sample before and after UF revealed a lack of correlation between bacterial indicators and the presence of human enteric viruses.

Virus occurrence in municipal groundwater sources in Quebec, Canada

A 1 year study was undertaken on groundwater that was a source of drinking water in the province of Quebec, Canada. Twelve municipal wells (raw water) were sampled monthly during a 1 year period, for a total of 160 samples. Using historic data, the 12 sites were categorized into 3 groups: group A (no known contamination), group B (sporadically contaminated by total coliforms), and group C (historic and continuous contamination by total coliforms and (or) fecal coliforms). Bacterial indicators (total coliform, Escherichia coli, enteroccoci), viral indicators (somatic and male-specific coliphages), total culturable human enteric viruses, and noroviruses were analyzed at every sampling site. Total coliforms were the best indicator of microbial degradation, and coliform bacteria were always present at the same time as human enteric viruses. Two samples contained human enteric viruses but no fecal pollution indicators (E. coli, enterococci, or coliphages), suggesting the limited value of these microorganisms in predicting the presence of human enteric viruses in groundwater. Our results underline the value of historic data in assessing the vulnerability of a well on the basis of raw water quality and in detecting degradation of the source. This project allowed us to characterize the microbiologic and virologic quality of groundwater used as municipal drinking water sources in Quebec.

Comparative reduction of Norwalk virus, poliovirus type 1, F+ RNA coliphage MS2 and Escherichia coli in miniature soil columns

Norwalk-like viruses (NLVs) are important agents of waterborne illness and have been linked to several groundwater-related outbreaks. The presence of human enteric viruses, in particular the presence of NLVs, is difficult to detect in the environment. Consequently, surrogate organisms are typically used as indicators of viruses from faecal contamination. Whether traditional bacterial indicators are reliable indicators for viral pathogens remains uncertain. Few studies have directly compared mobility and reduction of bacterial indicators (e.g. coliforms, Escherichia coli) and other surrogate indicators (coliphages) with pathogenic human viruses in soil systems. In this study the mobility and comparative reduction of the prototype NLV, Norwalk Virus (NV), was compared to poliovirus 1 (PV1), a bacterial indicator (E coli, EC) and a viral indicator (coliphage MS2) through miniature soil columns. Replicate, 10 cm deep, miniature columns were prepared using three soils representing a range of soil textures (sand, organic muck, and clay). Columns were initially conditioned, then incubated at 10-14 degrees C, dosed twice weekly for 8 weeks with one column pore volume of virus-seeded groundwater per dose, followed by 8 weeks of dosing with one column pore volume per dose of unseeded, simulated rainwater. Columns were allowed to drain after each dosing until an effluent volume equivalent to an applied dose was collected. Column effluents and doses were assayed for all viruses and EC. Rapid mobility with minimal reduction was observed for all organisms in the sand. Similar reductions were observed in organic muck for most organisms but NV showed a greater reduction. No organisms were shown to pass through the clay columns. Elution of viruses, in particular PV1, from the columns was gradual. After cessation of microbe dosing, E. coli was less detectable than viruses in column effluents and, therefore, unreliable as a virus indicator.

Sunlight inactivation of human enteric viruses and fecal bacteria

Three human enteric viruses (poliovirus, echovirus, coxsackievirus) suspended in seawater or buffer were stable for 6 hr in the absence of sunlight but were inactivated at the same rate in the presence of sunlight. Under summer sunlight conditions, at least 3 logs of these viruses were inactivated by one-hit kinetics while under winter sunlight conditions only 1 log of these viruses was inactivated by two-hit kinetics. Under these same conditions, 6 logs of E. coli were inactivated within 1 hr by one-hit kinetics under summer and winter conditions. In comparison, E. faecalis was inactivated by two-hit kinetics and only 2.5 logs of inactivation were observed after 4 hr of exposure to winter sunlight. Since human enteric viruses are considerably more resistant to sunlight inactivation than E. coli and moderately more resistant than E. faecalis, marine recreational water quality standards should be based on concentrations of enterococci and not on coliform bacteria. Since the mechanism and rate of inactivation of coliphage and human enteric viruses are similar, coliphages appear to be the best indicator for the presence of human enteric viruses in recreational waters, especially coastal waters where abundant sunshine is available.

Incidence of enteroviruses in Mamala Bay, Hawaii using cell culture and direct polymerase chain reaction methodologies

The consequence of point and nonpoint pollution sources, discharged into marine waters, on public recreational beaches in Mamala Bay, Hawaii was evaluated using virus cell culture and direct reverse transcriptase-polymerase chain reaction (RT-PCR). Twelve sites, nine marine, two freshwater (one stream and one canal), and one sewage, were assessed either quarterly or monthly for 1 year to detect the presence of human enteric viruses. Water samples were concentrated from initial volumes of 400 L to final volumes of 30 mL using Filterite electronegative cartridge filters and a modified beef extract elution procedure. Cell culture was applied using the Buffalo Green Monkey kidney cell line to analyze samples for enteroviruses. Positive samples were also evaluated by RT-PCR, using enterovirus-specific primers. Levels of RT-PCR inhibition varied with each concentrated sample. Resin column purification increased PCR detection sensitivity by at least one order of magnitude in a variety of sewage outfall and recreational marine water samples but not in the freshwater canal samples. Using cell culture, viable enteroviruses were found in 50 and 17% of all outfall and canal samples, respectively. Samples were positive at beaches 8% of the time. These data illustrate the potential public health hazard associated with recreational waters. Using direct PCR, viruses were detected at the outfall but were not found in any beach or canal samples, in part, owing to substances that inhibit PCR. Therefore, conventional cell culture is the most effective means of detecting low levels of infectious enteroviruses in environmental waters, whereas direct RT-PCR is rendered less effective by inhibitory compounds and low equivalent reaction volumes.