Dr. Edo McGowan
11/30/2006
Subject: [California] Title-22 and its defects, the spinach issue and naked DNA
When we wanted to look at "treated" wastewater from the reclamation plant in Salinas, we were generally thwarted.
When we ask this of others investigating the spinach outbreak, there has been little apparent interest in things other
than animal sources. Some of us have speculated on the reasons for this. Here, then is an addition to that speculation.
There is a lot of money and political horsepower in agriculture’s use of genetically modified crops and animals. If EPA
did any kind of work on antibiotic resistance they would run immediately into the following issue. Most crops and other
systems that use genetically engineered material contain constructs that are selected because they contain spliced
antibiotic resistant information. The spliced in information is not necessarily removed during the production process,
thus it accompanies the product.
Most of the engineered naked nucleic acids and constructs have either never existed in nature, or if they have, not in
such large amounts. They are, by definition, xenobiotics -- substances foreign to nature -- with the potential to cause
harm.
At least in Europe, there is no regulation governing the release of naked nucleic acids into the environment. Many novel
constructs are incorporated into transgenic micro-organisms and animal cell cultures for commercial drug production,
and into crops, livestock, fish and other aquatic organisms for food, animal feed, and other purposes. These constructs
are therefore greatly amplified, and at the same time introduced into foreign genomes where recombination with host
genes and the genes of the host’s viral pathogens may readily occur. Transgenic wastes containing large amounts of
free or potentially free transgenic DNA are being released unregulated into the environment via microorganisms in liquid
waste, and all killed microorganisms and cells containing transgenic DNA as solid waste.
The lack of regulation of naked/free nucleic acids is based largely on the assumption, now proven to be erroneous, that
naked/free nucleic acids would be rapidly broken down in the environment and in the digestive system of animals.
Actually Fred Griffith, in 1928, demonstrated this but no one at the time would recognize his finding.
The standard lab tests used in ascertaining the health implications for reclaimed water have recently been questioned
(actually there have been questions for a very long time). We know that Title-22 lab tests do not demonstrate the VBNC
microbes and coliforms are but one of innumerable bacteria that can enter this state. This test for indicator bacteria
says absolutely nothing of viruses and these can be a worrisome and persistent source of health risk. Further as noted
herein, there are issues of naked DNA/RNA and its uptake and transfer between pathogens and then its reassortment in
these pathogens and within the host, thus leading to novel combinations of risk. None of this is well discussed in these
investigations, if even discussed at all.
For those of you interested in this area, the following are worth a read.
… , persistence, transfer: an update on current knowledge on GMs and the fate of their recombinant DNA - group of 2 »
B Tappeser, M Jäger, C Eckelkamp - TWN Biotechnology & Biosafety Series, 1999 - biotech-info.net
... As efficiency of inactivation of naked DNA depends on different factors like
temperature, it varies with the season and type of treatment plant under study ...
Cited by 10 - Related Articles - View as HTML - Web Search
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ORIGINAL ARTICLE Survival of Listeria monocytogenes and Enterococcus faecium in sludge evaluated by real-time
PCR and culture methods N. Wery1, A.-M. Pourcher2, V. Stan3, J.-P. Delgenes1, F. Picard-Bonnaud2 and J.-J. Godon1
Abstract
Aims: This study evaluates the behaviour in spiked sludge of a pathogenic bacteria, Listeria monocytogenes, by cultural
and molecular techniques, and compares its survival with the one of a faecal indicator, Enterococcus faecium.
Methods and Results: Listeria monocytogenes strain Scott A and E. faeciumT were followed for 17 days after inoculation
in sludge. Kinetics of survival depended on the bacteria and on the technique used [most probable number method,
direct plate count or real-time quantitative PCR (qPCR)]. The concentration of L. monocytogenes decreased rapidly
regardless of the technique, but the decrease was much more dramatic with culture techniques than with qPCR. On the
contrary, the concentrations of culturable E. faeciumT were stable.
Conclusions: The results suggest that the cells of L. monocytogenes strain Scott A might have entered a viable, but
nonculturable (VBNC) status, whereas cells of the indicator bacteria, E. faeciumT, maintained themselves better and
stayed culturable.
Significance and Impact of the Study: The difference of survival kinetics in the sludge of a faecal indicator (E. faecium)
and a pathogenic bacterium (L. monocytogenes) may be linked to the fact that they either enter or do not enter into a
VBNC status.
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Prevalence, quantification and typing of adenoviruses detected in river and treated drinking water in South Africa , J.
van Heerden1,3, M.M. Ehlers1,2, A. Heim3 and W.O.K. Grabow1
Abstract
j. van heerden, m.m. ehlers, a. heim and w.o.k. grabow. 2005.
Aims: Human adenoviruses (HAds), of which there are 51 serotypes, are associated with gastrointestinal, respiratory,
urinary tract and eye infections. The importance of water in the transmission of HAds and the potential health risks
constituted by HAds in these environments are widely recognized. Adenoviruses have not previously been quantified in
river and treated drinking water samples. In this study, HAds in river water and treated drinking water sources in South
Africa were detected, quantified and typed.
Methods and Results: Adenoviruses were recovered from the water samples using a glass wool adsorption-elution
method followed by polyethylene glycol/NaCl precipitation for secondary concentration. The sensitivity and specificity of
two nested PCR methods were compared for detection of HAds in the water samples. Over a 1-year period (June 2002
to July 2003), HAds were detected in 5·32% (10/188) of the treated drinking water and 22·22% (10/45) of river water
samples using the conventional nested PCR method. The HAds detected in the water samples were quantified using a
real-time PCR method. The original treated drinking water and river water samples had an estimate of less than one
copy per litre of HAd DNA present. The hexon-PCR products used for typing HAds were directly sequenced or cloned
into plasmids before sequencing. In treated drinking water samples, species D HAds predominated. In addition,
adenovirus serotypes 2, 40 and 41 were each detected in three different treated drinking water samples. Most (70%) of
the HAds detected in river water samples analysed were enteric HAds (serotypes 40 and 41). One HAd serotype 2 and
two species D HAds were detected in the river water.
Conclusions: Adenoviruses detected in river and treated drinking water samples were successfully quantified and typed.
The detection of HAds in drinking water supplies treated and disinfected by internationally recommended methods, and
which conform to quality limits for indicator bacteria, warrants an investigation of the risk of infection constituted by these
viruses. The risk of infection may have implications for the management of drinking water quality.
Significance and Impact of the Study: This study is unique as it is the first report on the quantification and typing of HAds
in treated drinking water and river water. This baseline data is necessary for the meaningful assessment of the potential
risk of infection constituted by these viruses.
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Prevalence, quantification and typing of adenoviruses detected in river and treated drinking water in South Africa , J.
van Heerden1,3, M.M. Ehlers1,2, A. Heim3 and W.O.K. Grabow1
Abstract
j. van heerden, m.m. ehlers, a. heim and w.o.k. grabow. 2005.
Aims: Human adenoviruses (HAds), of which there are 51 serotypes, are associated with gastrointestinal, respiratory,
urinary tract and eye infections. The importance of water in the transmission of HAds and the potential health risks
constituted by HAds in these environments are widely recognized. Adenoviruses have not previously been quantified in
river and treated drinking water samples. In this study, HAds in river water and treated drinking water sources in South
Africa were detected, quantified and typed.
Methods and Results: Adenoviruses were recovered from the water samples using a glass wool adsorption-elution
method followed by polyethylene glycol/NaCl precipitation for secondary concentration. The sensitivity and specificity of
two nested PCR methods were compared for detection of HAds in the water samples. Over a 1-year period (June 2002
to July 2003), HAds were detected in 5·32% (10/188) of the treated drinking water and 22·22% (10/45) of river water
samples using the conventional nested PCR method. The HAds detected in the water samples were quantified using a
real-time PCR method. The original treated drinking water and river water samples had an estimate of less than one
copy per litre of HAd DNA present. The hexon-PCR products used for typing HAds were directly sequenced or cloned
into plasmids before sequencing. In treated drinking water samples, species D HAds predominated. In addition,
adenovirus serotypes 2, 40 and 41 were each detected in three different treated drinking water samples. Most (70%) of
the HAds detected in river water samples analysed were enteric HAds (serotypes 40 and 41). One HAd serotype 2 and
two species D HAds were detected in the river water.
Conclusions: Adenoviruses detected in river and treated drinking water samples were successfully quantified and typed.
The detection of HAds in drinking water supplies treated and disinfected by internationally recommended methods, and
which conform to quality limits for indicator bacteria, warrants an investigation of the risk of infection constituted by these
viruses. The risk of infection may have implications for the management of drinking water quality.
Significance and Impact of the Study: This study is unique as it is the first report on the quantification and typing of HAds
in treated drinking water and river water. This baseline data is necessary for the meaningful assessment of the potential
risk of infection constituted by these viruses.
Gene Technology and Gene Ecology of Infectious Diseases
Mae-Wan Ho, Terje Traavik, Orjan Olsvik, Beatrix Tappeser, C. Vyvyan Howard, Christine von Weizsacker, George C.
Mcgavin
Abstract:
According to the 1996 WHO Report, the world is heading for a major crisis in public health as outbreaks of new and re-
emerging infectious diseases are striking at increasing frequencies within the past 10 to 15 years. The current strains of
pathogens are moreover, resistant to known treatments; some strains being resistant to all or nearly all drugs and
antibiotics. Horizontal gene transfer is now generally recognized to be responsible for the evolution of virulence and the
spread of drug and antibiotic resistances. Many pathogens have crossed species barriers, having acquired genes from
phylogenetically distant species that are involved in their ability to cause diseases. Recent findings document the
extremely wide scope of horizontal gene transfer and the extensive recombination between genetic material from
unrelated species that have contributed to the emergence of virulence and antibiotic resistances. The past 15 years
coincide with the development of genetic engineering biotechnology on a commercial scale. Genetic engineering
depends on designing vectors for cloning and transferring genes and involves artificially recombining and manipulating
genes from unrelated species and their viral pathogens, thereby enhancing the probability for horizontal gene transfer
and recombination. The urgent question which needs to be addressed is the extent to which genetic engineering
biotechnology, by facilitating horizontal gene transfer and recombination, is contributing to the resurgence of infectious,
drug-resistant diseases, and will continue to do so if allowed to proceed unchecked. An enquiry into the possible
contribution of genetic engineering biotechnology to the etiology of infectious diseases is all the more pressing in the
light of other relevant recent findings indicating that microorganisms genetically engineered for 'contained use' may not
be effectively contained. Thus, biologically 'crippled' strains of bacteria can survive in the environment to exchange
genes with other species; DNA released from cells is not readily broken down in the environment, thereby retaining the
ability to transform organisms; some viral DNA can be more infectious than the virus itself; and routine chemical
treatments for inactivating pathogenic microorganisms and viruses, before they are discharged into the environment,
may be ineffective, leaving a substantial percentage of pathogens in an active infectious state. The combination of the
different kinds of evidence is sufficiently compelling, especially in view of the precautionary principle, to warrant, at the
very least, an independent public enquiry into genetic engineering biotechnology and the etiology of infectious
diseases. Taylor & Francis. Volume 10, Number 1 / May 26, 1998 Pages: 33 - 59