We thank William T. Eckert for narrating the video. Development of the glass wool protocol was part of the Wisconsin Water And Health Trial for Enteric Risks (WAHTER Study), funded by US EPA STAR Grant R831630. Alaska samples were collected by A. Reeves, A. Ramey, and B. Meixell with financial support from USGS. Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the U.S. Government.

1. Preparing the Glass Wool
<ol>
 <li> Before and after making each batch of filters, sterilize the work area with 10% bleach solution. </li>
 <li> Put on gloves and gown. Sterilize a bucket by autoclaving at 121&deg;C and 15 psi for at least 20 minutes. Place the glass wool in the sterile bucket. </li>
 <li> Saturate the glass wool with reverse osmosis water and let soak for 15 minutes. </li>
 <li> Drain the reverse osmosis water from the bucket.</li>
 <li> Saturate the glass wool with 1 M HCl and let soak fo...

<ol>
	<li>US Environmental Protection Agency. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+1623:+Cryptosporidium+and+Giardia+in+Water+by+Filtration/IMS/FA.">Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA.</a> EPA 815-R-05-002. , (2012).</li><li>Cashdollar, J. L., Dahling, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+method+to+re-use+electropositive+cartridge+filters+for+concentrating+viruses+from+tap+and+river+water.">Evaluation of a method to re-use electropositive cartridge filters for concentrating viruses from tap and river water.</a> J. Virol. Methods. 132, 13-17 (2006).</li><li>Lambertini, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concentration+of+enteroviruses,+adenoviruses,+and+noroviruses+from+drinking+water+by+use+of+glass+wool+filters.">Concentration of enteroviruses, adenoviruses, and noroviruses from drinking water by use of glass wool filters.</a> Appl. Environ. Microbiol. 74, 2990-2996 (2008).</li><li>Spackman, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+real-time+reverse+transcription+PCR+assay+for+Type+A+influenza+virus+and+the+avian+H5+and+H7+hemagglutinin+subtypes.">Development of a real-time reverse transcription PCR assay for Type A influenza virus and the avian H5 and H7 hemagglutinin subtypes.</a> J. Clin. Microbiol. 40, 3256-3260 (2002).</li><li>Environment Agency. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimisation+of+a+new+method+for+detection+of+viruses+in+groundwater.">Optimisation of a new method for detection of viruses in groundwater.</a> Report No. NC/99/40. , (2000).</li><li>Vilagin&#233;s, P., Sarrette, B., Husson, G., Vilaginés, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glass+wool+for+virus+concentration+at+ambient+water+pH+level.">Glass wool for virus concentration at ambient water pH level.</a> Water Sci. Technol. 27, 299-306 (1993).</li><li>Vivier, J. C., Ehlers, M. M., Grabow, W. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+enteroviruses+in+treated+drinking+water.">Detection of enteroviruses in treated drinking water.</a> Water Res. 38, 2699-2705 (2004).</li><li>Powell, K. L., Sililo, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Enteric virus detection in groundwater using a glass wool trap. In: Groundwater: Past Achievements and Future Challenges. , 813-816 (2000).</li><li>Hunt, R. J., Borchardt, M. A., Richards, K. D., Spencer, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+sewer+source+contamination+of+drinking+water+wells+using+tracers+and+human+enteric+viruses.">Assessment of sewer source contamination of drinking water wells using tracers and human enteric viruses.</a> Environ. Sci. Technol. 44, 7956-7963 (2010).</li><li>van Heerden, J., Ehlers, M. M., Heim, A., Grabow, W. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence,+quantification+and+typing+of+adenoviruses+detected+in+river+and+treated+drinking+water+in+South+Africa.">Prevalence, quantification and typing of adenoviruses detected in river and treated drinking water in South Africa.</a> J. Appl. Microbiol. 99, 234-242 (2005).</li><li>Vilagin&#233;s, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Round+robin+investigation+of+glass+wool+method+for+poliovirus+recovery+from+drinking+water+and+sea+water.">Round robin investigation of glass wool method for poliovirus recovery from drinking water and sea water.</a> Water Sci. Technol. 35, 445-449 (1997).</li><li>Gantzer, C., Senouci, S., Maul, A., Levi, Y., Schwartzbrod, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enterovirus+genomes+in+wastewater:+concentration+on+glass+wool+and+glass+powder+and+detection+by+RT-PCR.">Enterovirus genomes in wastewater: concentration on glass wool and glass powder and detection by RT-PCR.</a> J. Virol. Methods. 65, 265-271 (1997).</li><li>Borchardt, M. A., Jokela, W. E., Spencer, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen+losses+in+surface+water+runoff+from+dairy+manure+applied+to+corn+fields.">Pathogen losses in surface water runoff from dairy manure applied to corn fields.</a> , (2011).</li><li>Deboosere, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+validation+of+a+concentration+method+for+the+detection+of+influenza+A+viruses+from+large+volumes+of+surface+water.">Development and validation of a concentration method for the detection of influenza A viruses from large volumes of surface water.</a> Appl. Environ. Microbiol. 77, 3802-3808 (2011).</li><li>Lambertini, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virus+contamination+from+operation+and+maintenance+practices+in+small+drinking+water+distribution+systems.">Virus contamination from operation and maintenance practices in small drinking water distribution systems.</a> J. Water Health. 9, 799-812 (2011).</li></ol>

Glass wool filters have been used by several research teams3,5,6 to concentrate human enteric viruses from a variety of water sources such as finished drinking water7, groundwater8,9, surface water10, sea water11, wastewater12, and agricultural runoff13. Here we report the filters are also effective in concentrating avian influenza virus as well as the bacterial and protozoan pathogens Salmonella enterica (serovar Typhimurium) and C...

<table border="1">
<tbody>
 <tr>
 <td>Name of reagent or item</td>
 <td> Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>Hydrochloric acid</td>
 <td><a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>A144-500</td>
 </tr>
 <tr>
 <td height="24">Sodium hydroxide</td>
 <td><a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>BP359-212</td>
 </tr>
 <tr>
 <td>Phosphate Buffered Saline 
 Sodium chloride
 
 Potassium phosphate-dibasic
 
 Potassium phosphate-monobasic</td>
 <td>
 
 <a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a> 
 <a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a> 
 <a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>
 
 BP358-212
 
 BP363-500
 
 BP362-500</td>
 </tr>
 <tr>
 <td>Sodium hypochlorite
 i.e., household bleach</td>
 <td><a href="http://www.clorox.com" target="_blank">The Clorox Co.</a></td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Sodium thiosulfate, anhydrous</td>
 <td><a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>S 475-212</td>
 </tr>
 <tr>
 <td>Beef extract, desiccated</td>
 <td> <a href="http://www.bd.com" target="_blank">Becton, Dickinson and Company</a></td>
 <td>211520</td>
 </tr>
 <tr>
 <td>Glycine</td>
 <td><a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>G46-500</td>
 </tr>
 <tr>
 <td>Oiled sodocalcic glass wool
 
 Or
 
 R-11 unfaced fiberglass insulation </td>
 <td> <a href="http://www.isover.com" target="_blank">Isover</a>
 
 <a href="http://www.jm.com" target="_blank">Johns Manville</a></td>
 <td>Bourre 725 QN </td>
 </tr>
 <tr>
 <td>Polypropylene mesh</td>
 <td> <a href="http://www.industrialnetting.com" target="_blank">Industrial Netting</a></td>
 <td>xN4510</td>
 </tr>
 <tr>
 <td>2"x4" Sch 80 PVC threaded pipe nipple</td>
 <td> <a href="http://www.grainger.com" target="_blank">Grainger</a></td>
 <td>6MW35</td>
 </tr>
 <tr>
 <td>2" Sch 40 PVC cap</td>
 <td><a href="http://www.grainger.com" target="_blank">Grainger</a></td>
 <td>5WDW3</td>
 </tr>
 <tr>
 <td>Male adapter nylon fitting (1/2"x1/2")</td>
 <td><a href="http://www.usplastic.com" target="_blank"> US Plastic Corp.</a></td>
 <td>62178</td>
 </tr>
 <tr>
 <td>Sample bottles for eluate- 1 liter</td>
 <td><a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>03-313-4F</td>
 </tr>
 <tr>
 <td>60 mL syringe</td>
 <td><a href="http://www.fishersci.com" target="_blank">Fisher Scientific</a></td>
 <td>NC9661991</td>
 </tr>
 <tr>
 <td>pH strips</td>
 <td><a href="http://www.whatman.com" target="_blank">Whatman</a></td>
 <td>2614 991</td>
 </tr>
 <tr>
 <td>Prefilter, Polypropylene, 10 inch cartridge, 10 &mu;m</td>
 <td>McMaster-Carr</td>
 <td>4411K75</td>
 </tr>
 <tr>
 <td>Prefilter housing</td>
 <td>Cole-Parmer</td>
 <td>S-29820-10</td>
 </tr>
</tbody>
</table>

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 Cryptosporidium1, 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 viruses2. 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.

Watch this Scientific Journal Video about Glass Wool Filters for Concentrating Waterborne Viruses and Agricultural Zoonotic Pathogens at JoVE.com

Glass Wool Filters for Concentrating Waterborne Viruses and Agricultural Zoonotic Pathogens

Glass wool filters have been used to concentrate waterborne viruses by a number of research groups around the world. Here we show a simple approach for constructing glass wool filters and demonstrate the filters are also effective in concentrating waterborne viral, bacterial and protozoan pathogens.

The key first step in evaluating pathogen levels in suspected contaminated water is concentration.  Concentration methods tend to be specific for a ...

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

Research

JoVE Journal

Immunology and Infection

17.9K visualizações.  United States Geological Survey. Os filtros de lã de vidro têm sido utilizados para concentrar os vírus pela água por um número de grupos de pesquisa em todo o mundo. Aqui, mostramos uma abordagem simples para a construção de filtros de lã de vidro e demonstrar os filtros são também eficazes na concentração waterborne virais, agentes patogénicos bacterianos e protozoários.