Biological Consulting Services of North Central Florida, Inc.

Our 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 or Filterite filters) samples are processed according to EPA  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 ASTM method D4994-89 or Standard Methods 9510B (Standard Methods for the Examination of Water and Waste Water, 20th Edition, APHA).

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

Cell Culture

Currently, we use any of following cell strains for the detection of viable enteric viruses:

Buffallo Green Monkey (BGM) and RD (ATCC# CCL-136) - for standard pan enteric virus analysis for monitoring.

MA104 (ATCC# CRL-2378.1) – enteroviruses, rotaviruses, reoviruses,

BS-C-1 (ATCC# CCL-26) – enteroviruses, Hepatitis A,

FRhK-4 (ATCC# CRL-1688) – Hepatitis A,

MRC-5 (ATCC# CCL-171) – coronaviruses, enteroviruses,

Additionally Caco-2 and PLC/PRF/5 Cell lines are also used in conjunction to the BGM cell lines in monitoring for enteric viruses in all ASTM and EPA/ICR protocols

Background Enteric Virus Information

Human enteric viruses represent a diverse group.  Most of them are non-enveloped RNA viruses.  Viral families represented include picornaviruses, rotavirus, Norwalk and Norwalk-like viruses.  Adenovirus is the sole DNA virus.  Coronaviruses and reoviruses have also been linked to enteric disease.

Symptoms range from gastroenteritis to more life threatening diseases such as myocarditis and aseptic meningitis.  Rotavirus and Norwalk virus are two of the common agents that cause viral gastroenteritis.  Mortality from this condition can be very significant in developing countries.  The mechanisms for their pathogenesis are quite varied but diarrhea is largely due to the loss of cell lining in the intestinal tract.  High concentrations of virus in excess of 10⁸ per gram of diarrheal stool is often observed.

Transmission is generally considered to occur through the oral/fecal route.  However, other routes such as food, contaminated water, or surface contact can occur.  A potentially important mode of transmission, particularly in developing countries, is by waterborne transmission.  Contaminated vegetables and fruits are also an increasing concern.

Relatively little is known about many of the viruses in this group because many enteric viruses are difficult to culture or are not cultureable and produce diseases that are not readily identifiable (i.e. Have symptoms that are common with other pathogens).  Molecular methods to detect enteric viruses are becoming more common.

The impact of diarrheal disease on a worldwide scale is very significant.  The burden of disease and death falls disproportionately on preschool-age children.  Deaths are attributed to severe dehydration secondary to loss of fluids from diarrhea and vomiting.  Failure to thrive has been reported in children in developing countries where infections may be exacerbated by malnutrition or be synergistic with other enteropathogens.

List of common viruses that we have worked with:

Adenovirus - Non-enveloped, double-stranded DNA virus, 60-90 nm. May cause gastroenteritis.

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

Coronavirus - Enveloped, plus-sense RNA virus, 80-200 nm. Causes primarily a upper respiratory disease. However, the virus has been found in the stool of individuals with diarrhea.

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 (type A and B), Echovirus, Enterovirus 68-71. 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.

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.