Biological Consulting Services of North Central Florida, Inc.

Your source for accurate biological testing.

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Environmental Virology ICR / EPA1615

 

Human enteric viruses are often present in domestic sewage, treated wastewater, recreational surface waters and biosolids intended for land application. The detection of viable viruses in a sample is indicative of the definite presence of fecal pollution and of human pathogens.

List of common viruses that we have worked with:

Calicivirus: Non-enveloped, plus-sense RNA virus, 26-32 nm. Major cause of viral gastroenteritis.

Enteroviruses: All in the picornavirus family. Non-enveloped, plus-sense RNA viruses, 22-30 nm. Viruses of human origin within this group include, Poliovirus, Coxsackievirus (types A and B), Echovirus, Enterovirus 68-71.

Hepatitis A virus: Also in the picornavirus family. Non-enveloped, plus-sense RNA virus. Major cause of viral hepatitis.

Reovirus: Non-enveloped, segmented double-stranded RNA virus, 65-75 nm. Can be found in water or sewage. Possible enteric pathogen.

Rotavirus: Non-enveloped, segmented double-stranded RNA virus, 65-75 nm.  Major cause of viral gastroenteritis.

These viruses can be transmitted through the oral-fecal route and many have the ability to target the central nervous system.  Enteroviruses can produce a large number of clinical symptoms depending on the type of virus. These include, myalgia, pancreatitis, diabetes, myocarditis, hand-foot-and-mouth disease, conjunctivitis, and gastroenteritis among others.

 

BCS Laboratory is completely equipped for environmental virology. We utilize innovative methodologies for the detection, enumeration, and identification of enteric viruses. We also provide high titer viral stocks for decontamination/inactivation testing. Our virologists are fully trained in the latest cell culture and molecular techniques to ensure the most reliable and informative results.  We provide routine analysis to numerous private, state and government agencies, and universities.

Water, biosolids, and filter (1MDS, NanoCeram, or Filterite filters) samples are processed according to EPA ICR methodology (US EPA ICR microbial laboratory manual; EPA/600/R-95/178 and the US EPA MANUAL OF METHODS FOR VIROLOGY EPA/600/4-84/013 ), or EPA 1615, or ASTM method D4994-89, or Standard Methods 9510B (Standard Methods for the Examination of Water and Waste Water, 20th Edition, APHA).  BCS holds accreditation for analysis of matrices for the above methods.

Additionally, we provide bacteriophage or Coliphage testing as a surrogate to enterovirus testing.  This analysis provides a low cost prediction of the efficacy of the process of enterovirus transport and/or inactivation.

Method for the Recovery and Assay of Total Culturable Viruses from Sludge EPA/625/R-92/013 appendix H and Standard Practice for the Recovery of Viruses from Wastewater Sludges; ASTM D4994-89. Elution, concentration and viable enteroviruses enumeration of sample (biosolids) by cell culture analysis on Buffalo Green Monkey cells, Rhabdosarcoma cells, and MA 104 cells.

Detection of Enteric Viruses in Water and Wastewater (Standard Methods 9510A-G).

Virus recovery, concentration, and enumeration from small volumes of water as per Standard Methods for the Examination of Water and Wastewater, 21th edition, APHA, AWWA, WEF, 2012.

US EPA ICR EPA/600/R-95/178 and EPA/600/4-84/013

Cultivable enterovirus detection: Recovery of enteric viruses from 1MDS or Filtrite filters and enumeration by cell culture on Buffalo Green Monkey cells, Rhabdosarcoma cells, and MA 104 cells as per ICR-EPA rule.

EPA/600/R-10/181 (2012) Method 1615 Measurement of Enterovirus and  Norovirus Occurrence in Water by Culture and RT-qPCR

EPA Method 1615 provides culture and molecular procedures for detecting human enteroviruses, human noroviruses and mammalian orthoreoviruses (culture procedure only) in water. The cell culture procedure detects enterovirus and orthoreovirus species that are capable of infecting and producing cytopathic effects (CPE) in the Buffalo Green Monkey kidney (BGM) cell line.  Although this cell line is considered a "gold standard" for detection of infectious waterborne viruses, noroviruses and a number of enteroviruses do not replicate in BGM cells and thus cannot be detected by cell culture. There is no established cell line for detection of infectious human noroviruses. The molecular procedure incorporated into EPA Method 1615 detects the noroviruses and enteroviruses, including those enteroviruses that do not replicate on BGM cells.  However, as a molecular procedure, it does not distiguish infectious from noninfectious viruses and therefore does not provide any information on viability and thus infectivity of the species detected.

Enteric virus sample collection kit: Sample collection kits can be requested and shipped to customers for sampling. The sampling kit includes a cooler and icepacks with container(s) for sludge samples. For water samples, the sampling kit includes a cooler and icepacks, filter(s), housing, tubing, and a flow meter.

 

 

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