glass-wool-filters-for-concentrating-waterborne-viruses-agricultural

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 ...

Recent studies on the contamination of groundwater with human enteric viruses have focused on public water systems, whereas little is known about the occurrence of viruses in private household wells. The objective of the present study was to estimate the incidence of viruses in Wisconsin household wells located near septage land application sites or in rural subdivisions served by septic systems. Fifty wells in seven hydrogeologic districts were sampled four times over a year, once each season. Reverse transcriptase PCR (RT-PCR), followed by Southern hybridization, was used to detect enteroviruses, rotavirus, hepatitis A virus (HAV), and Norwalk-like viruses (NLVs). In addition, cell culture was used to detect culturable enteroviruses. Companion water samples were collected for total coliforms, Escherichia coli, fecal enterococci, F-specific RNA coliphages, nitrate, and chloride analyses. Among the 50 wells, four (8%) were positive for viruses by RT-PCR. Three wells were positive for HAV, and the fourth well was positive for both rotavirus and NLV in one sample and an enterovirus in another sample. Contamination was transient, since none of the wells was virus positive for two sequential samples. Culturable enteroviruses were not detected in any of the wells. Water quality indicators were not statistically associated with virus occurrence, although some concordance was noted for chloride. The present study is the first in the United States to systematically monitor private household wells for virus contamination and, combined with data for public wells, provides further insight on the extent of groundwater contamination with human enteric viruses.

Incidence of enteric viruses in groundwater from household wells in Wisconsin.

Die Inzidenz von enteralen Viren im Grundwasser aus Brunnen Haushalt in Wisconsin.

This study documents the first reported transmission of Salmonella enterica serotype Typhimurium definitive type 104 (DT104) to premature fraternal twins via their mother's breast milk. When premature twin neonates developed severe enteritis in the neonatal intensive care unit (NICU), stool samples and the mother's breast milk were cultured for the presence of Salmonella. Antibacterial susceptibility patterns were determined. Semiquantitative organism abundance data were retrospectively gathered on 54 stored breast milk samples collected on 34 different days using a rapid, real-time polymerase chain reaction (PCR) methodology (LightCycler PCR). Fecal samples from other infants in the NICU at that time were also tested. Pulsed-field gel electrophoresis (PFGE) was used to assess the genetic composition of the isolated organisms. The twins' neonatal stools and mother's breast milk cultures revealed a resistance pattern (R-type) to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline. LightCycler PCR analysis of sequential breast milk samples confirmed this to be the likely source of transmission. In the subsequent outbreak investigation, none of the NICU surveillance fecal samples proved positive for this organism. The genetic composition of organisms isolated from the maternal breast milk was indistinguishable from those isolated from neonatal specimens as determined by PFGE. Antibiotic susceptibility tests coupled with PFGE patterns suggested that these Salmonella isolates were DT104. Because the prevalence of DT104 infections is rising in the United States, neonatologists should be aware of breast milk as a potential mode of transmission.

Transmission of Salmonella enterica serotype typhimurium DT104 to infants through mother's breast milk.

Die &#xDC;bertragung von Salmonella enterica Serotyp Typhimurium DT104 bei S&#xE4;uglingen durch Muttermilch.

Virginiamycin use in poultry selects for Enterococcus faecium with cross-resistance to quinupristin-dalfopristin, a drug for vancomycin-resistant E. faecium in humans. We conducted an epidemiologic study of poultry exposures as risk factors for human carriage of quinupristin-dalfopristin-resistant E. faecium.

Use of streptogramin growth promoters in poultry and isolation of streptogramin-resistant Enterococcus faecium from humans.

Virginiamycin Einsatz bei Gefl&uuml;gel w&auml;hlt f&uuml;r Enterococcus Faecium mit Kreuzresistenz zu Quinupristin-Dalfopristin, ein Medikament f&uuml;r Vancomycin-resistente E. Faecium beim Menschen. Wir haben eine epidemiologische Studie von Gefl&uuml;gel Ausstellungen als Risikofaktoren f&uuml;r menschliche Bef&ouml;rderung von Quinupristin-Dalfopristin-resistente E. Faecium durchgef&uuml;hrt.

Verwendung von Streptogramine Wachstumsf&ouml;rderer in Gefl&uuml;gel und Isolierung von Streptogramine-resistente Enterococcus Faecium vom Menschen.

Microsporidia are obligate intracellular pathogens capable of infecting humans. There is credible evidence to suggest that microsporidial infections may be transmitted through consumption of spores in contaminated water; however, methods to detect this pathogen have not been standardized and microsporidia occurrence studies have not been conducted. Concentration of spores by continuous flow centrifugation (CFC), purification using immunomagnetic separation (IMS), and detection by either microscopy or real-time polymerase chain reaction (PCR) were evaluated for detection of Encephalitozoon intestinalis spores in seeded water samples. Recovery efficiency of CFC using microscopic detection ranged from 38.7-75.5% in filtered tap water. Using an indirect IMS method, 78.8-90.2% of seeded spores were recovered in ultrapure water (18 M Omega); however, the lack of a specific monoclonal antibody and the presence of other particulates interfered with the IMS assay in some turbid samples. Despite low recovery efficiencies and the detectable presence of PCR inhibitors in each of the samples, a combination of CFC concentration, indirect IMS, and real-time PCR produced a positive test result in six of ten natural water samples (turbidity 0.1-28.9 NTU) at a seeding level of 50 spores/L.

Development of a method for the detection of waterborne microsporidia.

Microsporidia sind intrazellul&auml;re Erreger infizieren Menschen f&auml;hig verpflichten. Es gibt glaubw&uuml;rdige Hinweise darauf, dass Microsporidial, dass Infektionen durch den Verzehr von Sporen in verunreinigtes Wasser &uuml;bertragen werden k&ouml;nnen; jedoch Methoden, diese Erreger zu erkennen sind nicht genormt und Microsporidia vorkommen Studien nicht durchgef&uuml;hrt worden. Konzentration der Sporen durch kontinuierliche Zentrifugieren (CFC), Reinigung verwenden Immunomagnetic Trennung (IMS) und der Erkennung durch Mikroskopie oder Real-Time Polymerase-Kettenreaktion (PCR) zum Nachweis von Encephalitozoon Intestinalis Sporen in ges&auml;ten Wasserproben bewertet wurden. Restitutionsg&uuml;te FCKW mit mikroskopischen Nachweis reichte von 38,7-75,5 % in gefilterte Leitungswasser. Einen indirekten IMS Methode, wurden 78,8-90,2 % der ges&auml;ten Sporen erholt in Reinstwasser (18 M Omega); mischte der Mangel an einem spezifischen monoklonalen Antik&ouml;rper und das Vorhandensein von anderen Partikeln mit der IMS-Assay in einigen tr&uuml;ben Proben. Trotz niedrigen Erholung Effizienz und der nachweisbaren Anwesenheit von PCR-Inhibitoren in jedem der Beispiele, eine Kombination aus CFC-Konzentration, indirekte IMS und Echtzeit-PCR produziert ein positives Testergebnis in sechs von zehn nat&uuml;rliche Wasserproben (Tr&uuml;bung 0.1-28,9 NTU) auf einer seeding 50 Sporen/L.

Entwicklung einer Methode zum Nachweis von Wasserwegen Microsporidia.

Available filtration methods to concentrate waterborne viruses are either too costly for studies requiring large numbers of samples, limited to small sample volumes, or not very portable for routine field applications. Sodocalcic glass wool filtration is a cost-effective and easy-to-use method to retain viruses, but its efficiency and reliability are not adequately understood. This study evaluated glass wool filter performance to concentrate the four viruses on the U.S. Environmental Protection Agency contaminant candidate list, i.e., coxsackievirus, echovirus, norovirus, and adenovirus, as well as poliovirus. Total virus numbers recovered were measured by quantitative reverse transcription-PCR (qRT-PCR); infectious polioviruses were quantified by integrated cell culture (ICC)-qRT-PCR. Recovery efficiencies averaged 70% for poliovirus, 14% for coxsackievirus B5, 19% for echovirus 18, 21% for adenovirus 41, and 29% for norovirus. Virus strain and water matrix affected recovery, with significant interaction between the two variables. Optimal recovery was obtained at pH 6.5. No evidence was found that water volume, filtration rate, and number of viruses seeded influenced recovery. The method was successful in detecting indigenous viruses in municipal wells in Wisconsin. Long-term continuous filtration retained viruses sufficiently for their detection for up to 16 days after seeding for qRT-PCR and up to 30 days for ICC-qRT-PCR. Glass wool filtration is suitable for large-volume samples (1,000 liters) collected at high filtration rates (4 liters min(-1)), and its low cost makes it advantageous for studies requiring large numbers of samples.

Concentration of enteroviruses, adenoviruses, and noroviruses from drinking water by use of glass wool filters.

Verf&uuml;gbare Filtrierung Methoden Wasserwege Viren konzentrieren sind f&uuml;r Studien, die eine gro&szlig;e Anzahl von Proben, beschr&auml;nkt auf kleinen Probenvolumina zu kostspielig oder nicht sehr portabel f&uuml;r Routine-Feld-Anwendungen. Sodocalcic Glaswolle Filtration ist eine kosteng&uuml;nstige und einfach zu bedienende Methode, Viren zu behalten, aber seine Effizienz und Zuverl&auml;ssigkeit sind nicht ausreichend verstanden. Diese Studie ausgewertet Glaswolle Filterleistung der US Environmental Protection Agency-Kontaminanten-Kandidatenliste, d.h. die vier Viren konzentrieren, Coxsackievirus, schmerzhafte, Norovirus, Adenoviren, sowie und Polioviren. Total Virus Zahlen zur&uuml;ckgewonnen wurden gemessen durch quantitative reverse Transkription-PCR (qRT-PCR); infekti&ouml;se Labornetz wurden mit integrierten Zellkultur (ICC) - qRT - PCR quantifiziert. Erholung Effizienz durchschnittlich 70 % f&uuml;r Polioviren und 14 % f&uuml;r Coxsackievirus B5, 19 % f&uuml;r schmerzhafte 18, 21 % f&uuml;r Adenoviren 41 29 % f&uuml;r Norovirus. Virus-Stamm und Wasser-Matrix betroffen Recovery, mit bedeutenden Interaktion zwischen den beiden Variablen. Optimalen Regeneration wurde bei pH 6.5 erhalten. Keine Beweise ergab, dass die Wassermenge, Filtrationsrate und Anzahl der Viren beeinflusst Wiederherstellung ausges&auml;t. Die Methode war erfolgreich bei der Aufdeckung von indigener Viren in st&auml;dtischen Brunnen in Wisconsin. Langfristige kontinuierliche Filtration beibehalten Viren ausreichend f&uuml;r ihre Erkennung f&uuml;r bis zu 16 Tage nach der Ansaat qRT-PCR und bis zu 30 Tagen f&uuml;r ICC-qRT-PCR. Glaswolle Filtration ist geeignet f&uuml;r gro&szlig;volumige Proben (1.000 Liter) zu hohe Preise (4 Liter min(-1)), und seine niedrigen Kosten macht es vorteilhaft f&uuml;r Studien, die gro&szlig;e Anzahl von Proben erfordern.

Konzentration von Enteroviren, Adenoviren und Noroviren aus dem Trinkwasser von Glaswolle-Filter.

Septic systems that are built in compliance with regulations are generally not expected to be the cause of groundwater borne disease outbreaks, especially in areas with thick vadose zones. However, this case study demonstrates that a disease outbreak can occur in such a setting and outlines the combination of epidemiological, microbiological, and hydrogeological methods used to confirm the source of the outbreak. In early June 2007, 229 patrons and employees of a new restaurant in northeastern Wisconsin were affected by acute gastroenteritis; 6 people were hospitalized. Epidemiological case-control analysis indicated that drinking the restaurant's well water was associated with illness (odds ratio = 3.2, 95% confidence interval = 0.9 to 11.4, P = 0.06). Microbiological analysis (quantitative reverse transcription-polymerase chain reaction) measured 50 genomic copies per liter of norovirus genogroup I in the well water. Nucleotide sequencing determined the genotype as GI.2 and further showed the identical virus was present in patrons' stool specimens and in the septic tank. Tracer tests using dyes injected at two points in the septic system showed that effluent was traveling from the tanks (through a leaking fitting) and infiltration field to the well in 6 and 15 d, respectively. The restaurant septic system and well (85-m deep, in a fractured dolomite aquifer) both conformed to state building codes. The early arrival of dye in the well, which was 188 m from the septic field and located beneath a 35-m thick vadose zone, demonstrates that in highly vulnerable hydrogeological settings, compliance with regulations may not provide adequate protection from fecal pathogens.

Norovirus outbreak caused by a new septic system in a dolomite aquifer.

Septische Systeme, die in &Uuml;bereinstimmung mit Bestimmungen basieren sollen im Allgemeinen nicht die Ursache des Grundwassers Seuchenausbr&uuml;che, vor allem in Gebieten mit dicken phreatische Zonen getragen werden. Diese Fallstudie zeigt jedoch, dass ein Krankheitsausbruch, in einer solchen Umgebung auftreten kann und die Kombination aus epidemiologischen, mikrobiologische und hydrogeologische Methoden verwendet umrei&szlig;t, um die Quelle des Ausbruchs zu best&auml;tigen. Anfang Juni 2007 waren 229 G&ouml;nner und Mitarbeiter ein neues Restaurant im nord&ouml;stlichen Wisconsin durch akute Magen-Darm betroffen; 6 Personen wurden ins Krankenhaus eingeliefert. Epidemiologische Fall-Kontroll-Analyse zeigte, dass das Restaurant gut Trinkwasser Krankheit zugeordnet (Odds Ratio = 3.2, 95 % Konfidenzintervall = 0,9 bis 11,4, P = 0,06). Mikrobiologische Analyse (quantitative reverse Transkription-Polymerase-Kettenreaktion) gemessen 50 genomische Kopien pro Liter Norovirus Genogroup ich in das Brunnenwasser. Nukleotid-Sequenzierung bestimmt der Genotyp als GI.2 und weitere zeigten, dass das identische Virus in Schutzherren Hocker Exemplare und in die Kl&auml;rgrube vorhanden war. Tracer-Tests, die Verwendung von Farbstoffen, die an zwei Stellen in der septischen System injiziert ergaben, dass Abwasser aus den Tanks (durch eine undichte Armatur) und Unterwanderung Feld zum Brunnen in 6 bis 15 d, bzw. unterwegs war. Das Restaurant septischen System und gut (85 m Tiefe, in einem gebrochenen Dolomit-Aquifer) entsprach sowohl staatliche Bauvorschriften. Die fr&uuml;he Ankunft der Farbstoff im Brunnen, der unterhalb einer 35-m dicken phreatische Zone, und 188 m vom septischen Bereich wurde, zeigt, dass in besonders gef&auml;hrdeten Hydrogeologische Einstellungen, Einhaltung von Vorschriften nicht ausreichenden Schutz vor f&auml;kale Erreger bieten kann.

Norovirus Ausbruch verursacht durch ein neues septischen System in einem Dolomit-Aquifer.

This study investigated the source, transport, and occurrence of human enteric viruses in municipal well water, focusing on sanitary sewer sources. A total of 33 wells from 14 communities were sampled once for wastewater tracers and viruses. Wastewater tracers were detected in four of these wells, and five wells were virus- positive by qRT-PCR. These results, along with exclusion of wells with surface water sources, were used to select three wells for additional investigation. Viruses and wastewater tracers were found in the groundwater at all sites. Some wastewater tracers, such as ionic detergents, flame retardants, and cholesterol, were considered unambiguous evidence of wastewater. Sampling at any given time may not show concurrent virus and tracer presence; however, given sufficient sampling over time, a relation between wastewater tracers and virus occurrence was identified. Presence of infectious viruses at the wellhead demonstrates that high-capacity pumping induced sufficiently short travel times for the transport of infectious viruses. Therefore, drinking-water wells are vulnerable to contaminants that travel along fast groundwater flowpaths even if they contribute a small amount of virus-laden water to the well. These results suggest that vulnerability assessments require characterization of "low yield-fast transport" in addition to traditional "high yield-slow transport", pathways.

Assessment of sewer source contamination of drinking water wells using tracers and human enteric viruses.

Diese Studie untersuchte die Quelle, Transport und Vorkommen des menschlichen enterischen Viren in st&auml;dtische Brunnenwasser, Konzentration auf Kanal Quellen. Insgesamt 33 Brunnen aus 14 Gemeinden wurden f&uuml;r Abwasser-Tracer und Viren einmal abgetastet. Abwasser-Tracer in vier von diesen Brunnen erkannt, und f&uuml;nf Brunnen wurden Virus-positiven durch qRT-PCR. Diese Ergebnisse zusammen mit Ausschluss der Wells mit Oberfl&auml;chenwasser Quellen dienten drei Brunnen f&uuml;r weitere Untersuchungen ausw&auml;hlen. Viren und Abwasser Taster wurden in das Grundwasser an allen Standorten gefunden. Einige Abwasser-Tracer, wie Ionischen Detergentien, Flammschutzmittel und Cholesterin, galten eindeutige Beweise f&uuml;r Abwasser. Probenahme zu einem bestimmten Zeitpunkt kann nicht gleichzeitige Virus und Tracer Pr&auml;senz zeigen; Da ausreichend Probenahme im Laufe der Zeit, wurde jedoch eine Beziehung zwischen Abwasser-Tracer und Virus-auftreten identifiziert werden. Infekti&ouml;se Viren untersucht bei der Bohrlochkopf zeigt, dass die Pumpen von hoher Kapazit&auml;t ausreichend kurze Fahrzeiten f&uuml;r den Transport von infekti&ouml;sen Viren induzierte. Trinkwasser-Brunnen sind daher anf&auml;llig f&uuml;r Verunreinigungen, die schnelle Grundwasser Flowpaths durchfahren, auch wenn sie eine kleine Menge von Virus-beladene Wasser zum Brunnen beitragen. Diese Ergebnisse legen nahe, dass Schwachstellen Charakterisierung von &quot;geringe Ausbeute-schnell-Transport&quot; neben der traditionellen &quot;hohe Rendite-langsam Transport&quot;, Wege erfordern.

Bewertung der Kanalisation Quelle Kontamination von Trinkwasser-Brunnen mit Tracer und menschlichen enterischen Viren.

We tested the association of common events in drinking water distribution systems with contamination of household tap water with human enteric viruses. Viruses were enumerated by qPCR in the tap water of 14 municipal systems that use non-disinfected groundwater. Ultraviolet disinfection was installed at all active wellheads to reduce virus contributions from groundwater to the distribution systems. As no residual disinfectant was added to the water, any increase in virus levels measured downstream at household taps would be indicative of distribution system intrusions. Utility operators reported events through written questionnaires. Virus outcome measures were related to distribution system events using binomial and gamma regression. Virus concentrations were elevated in the wells, reduced or eliminated by ultraviolet disinfection, and elevated again in distribution systems, showing that viruses were, indeed, directly entering the systems. Pipe installation was significantly associated with higher virus levels, whereas hydrant flushing was significantly associated with lower virus levels. Weak positive associations were observed for water tower maintenance, valve exercising, and cutting open a water main. Coliform bacteria detections from routine monitoring were not associated with viruses. Understanding when distribution systems are most vulnerable to virus contamination, and taking precautionary measures, will ensure delivery of safe drinking water.

Virus contamination from operation and maintenance events in small drinking water distribution systems.

Wir testeten die Vereinigung der gemeinsamen Veranstaltungen im Trinkwasser-Verteilersysteme mit Verunreinigung Haushalt Leitungswasser mit menschlichen enterischen Viren. Viren wurden von qPCR in das Leitungswasser der 14 kommunalen Systeme aufgelistet, die nicht desinfiziert Grundwasser verwenden. UV Desinfektion wurde bei allen aktiven Bohrlochk&ouml;pfe Virus Beitr&auml;ge von Grundwasser zu den Vertriebssystemen reduzieren installiert. Keine verbleibende Desinfektionsmittel zum Wasser hinzugef&uuml;gt wurde, w&auml;re die Erh&ouml;hungen Virus gemessenen flussabw&auml;rts am Haushalt H&auml;hne bezeichnend f&uuml;r Verteilung System eindringen. Dienstprogramm Operatoren berichtete Ereignisse durch schriftliche Frageb&ouml;gen. Virus ergebnisorientierten Ma&szlig;nahmen bezogen sich auf Verteilung System-Ereignisse mit einem Binom und Gamma Regression. Virus-Konzentrationen wurden erh&ouml;hte in den Brunnen, reduziert oder durch UV-Desinfektion beseitigt und wieder in Vertriebssystemen, zeigen, dass Viren die Systeme tats&auml;chlich direkt eingegeben wurden, erh&ouml;ht. Rohr-Installation war deutlich in Zusammenhang mit einem h&ouml;heren Virus, w&auml;hrend Hydrant Sp&uuml;len mit niedrigeren Ebenen der Virus deutlich verbunden war. Schwache positive Assoziationen wurden f&uuml;r Wasserturm Wartung, Ventil auszu&uuml;ben und eine Wasserleitung aufschneidet beobachtet. Coliforme Bakterien Erkennungen von routinem&auml;&szlig;igen &Uuml;berwachung wurden nicht mit Viren. Verst&auml;ndnis wenn Vertriebssysteme besonders anf&auml;llig f&uuml;r Virus-Kontamination und vorbeugende Ma&szlig;nahmen zu ergreifen sind sorgen f&uuml;r Lieferung von sauberem Trinkwasser.

Article	Year
Incidence of enteric viruses in groundwater from household wells in Wisconsin. Applied and environmental microbiology\| PubMed ID: 12571044	2003
Transmission of Salmonella enterica serotype typhimurium DT104 to infants through mother's breast milk. Pediatrics\| PubMed ID: 12777569	2003
Use of streptogramin growth promoters in poultry and isolation of streptogramin-resistant Enterococcus faecium from humans. The Journal of infectious diseases\| PubMed ID: 17041845	2006
Development of a method for the detection of waterborne microsporidia. Journal of microbiological methods\| PubMed ID: 17582531	2007
Concentration of enteroviruses, adenoviruses, and noroviruses from drinking water by use of glass wool filters. Applied and environmental microbiology\| PubMed ID: 18359827	2008
Norovirus outbreak caused by a new septic system in a dolomite aquifer. Ground water\| PubMed ID: 20199588	2011
Assessment of sewer source contamination of drinking water wells using tracers and human enteric viruses. Environmental science & technology\| PubMed ID: 20822128	2010
Virus contamination from operation and maintenance events in small drinking water distribution systems. Journal of water and health\| PubMed ID: 22048438	2011
Environmental health perspectives\| PubMed ID: 22659405	2012
Water research\| PubMed ID: 22682268	2012

Susan K. Spencer

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