#136 - Risk Assessment Impossible
#138 - Sewage sludge (Biosolids) is a potential killer

D. Strauch in his 1991 paper, "Survial of pathogenic micro-organisms and parasite in extreta,
manure and sewage sludge" reported that two groups of researchers had found that
pathogenic disease organisms will be taken up inside the food crops." "Salmonella has
survived in forest stands between 424 and 820 days" " most pathogenic agents can survive
the treatment process" and the sewage treatment process causes some of the pathogenic
disease organisms to be absorbed or enclosed in faecal particles during the treatment
process." (1991)

Lewis found,  "The water repellent lubricants such as silicon and petroleum products cover
the pathogens and prevent them from being found by standard test methods. It was only
when he dissolved the lubricants with acetone or other solvents, that the pathogens showed
up in tests." "The problem of pathogen detection in sludge, according to Lewis, "is that the
sewage treatment process changes the outside crust of the aggregates in sludge and only
the pathogens on the outside of the aggregates are measured by standard tests." He says
that most of the microbes are trapped inside the aggregates

According to microbiologists, some bacteria form an endospore, a cell type developed from
the vegetative bacterial cell through a sequence of morphological changes. Although the
vegetative cell of bacteria is usually killed by heat and disinfectant, the endospore is resistant
to agents that kill the vegetative cell (heating, drying, freezing, chemicals, and radiation).  
Nester, Roberts, Pearsall and McCarthy (1978)

Both treated and control animals exhibited signs of discomfort immediately after dosing.
These signs were manifest as frequent examinations of the injection site and increased
activity. However, fifteen to Jhirty minutes after dosing, the Boston sludge-treated animals
began to show signs of effects on the central nervous system. Orientation was affected to
some degree in all groups as animals rotated repeatedly ii) both clockwise or counter-
clockwise directions; one animal (dosed at 32.9 mg/kg) turned over continuously for a period
of four to six minutes. Additionally,  motor coordination  and  gait were  adversely affected by
the Boston sludge extract in nearly  all the animals. Animals dosed with Dallas sludge extract
showed few of the signs of toxicity of the Boston sludge extract groups and were more nearly
like control animals

No gross lesions were observed in any animals which could be associated with the test
materials. Additionally, no treatment effects were seen in gross or relative organ weights.

Interactions of pathogens and irritant chemicals in land-applied sewage sludges (biosolids). A
prevalence of Staphylococcus aureus infections of the skin and respiratory tract was found.
Approximately 1 in 4 of 54 individuals were infected, including 2 mortalities[deaths]
(septicaemia, pneumonia).

J Water Health 03 (2005) 249-257

Induction of Escherichia coli and Salmonella typhimurium into the viable but nonculturable
state following chlorination of wastewater
James D. Oliver, Maya Dagher and Karl Linden
Department of Biology, University of North Carolina at Charlotte, Charlotte, NC, 28223, USA,
Tel: 704-687-8516, Fax: 267-327-2083, E-mail:

Department of Biology, University of North Carolina at Charlotte, Charlotte, NC, 28223, USA,
Tel: 704-687-8516, Fax: 267-327-2083, E-mail:

Department of Civil and Environmental Engineering, Duke University, Durham, NC, 27708-
0287, USA


We examined the effects of chlorine disinfection on Escherichia coli and Salmonella
typhimurium in secondary-treated wastewater to determine whether such treatment might
induce these bacteria into the viable but nonculturable (VBNC) state. In this state, cells lose
culturability but retain viability and the potential to revert to the metabolically active and
infectious state. To examine the effects of chlorination on cells in different physiological
states, cells from the logarithmic and stationary phases, or nutrient starved, or grown in
natural wastewater, were studied. Isogenic cells with and without plasmids were also
examined. Whereas a mixture of free and combined chlorine, as occurs under typical
wastewater disinfection, was found to be rapidly lethal to most cells, regardless of their
physiological state or plasmid content, c. 104 of the original 106 cells ml-1 did survive in the
VBNC state. While we were not successful in resuscitating these cells to the culturable state,
the presence of such nonculturable cells in treated wastewater offers a potential public health

McGowan's Note---assuming the following---28406 mL in a cubic foot and 27 cubic feet per
ton---on the average, then a ton of sludge contains, using the above author's findings---
7,669,680,000 viable but non culturable E. coli. But they did not look at persisters. Also
assuming that many discharges from hospitals have enough antibiotics in the effluent to
cause resistance, the level of persisters may go up a 1000 fold. These combined results may
see a very serious, but previously undiscussed issue on the risks of land applied sewer

In a recent meeting of our task-group, one of the members, a wastewater engineer, raised
the question relating to land application of composted biosolids. Composting raises the level
of manipulation of biosolids to that of a manufactured product, often incorporating green-
waste, i.e., trimmings from various forms of vegetation. The essence of the question related
to the survival of pathogens, hence the underlying issue of surviving MDRB. The question
went something like this---"If Staphyloccus aureus were found dead, did that mean that the
problem was solved?" The corollary--- was it dead or merely in the viable but non-culturable
(VNC) state? Was it a classic persister? (see below). Further, this says nothing for uptake of
released naked DNA.
Additionally, during the above noted meeting, I had mentioned that there is now strong
medical evidence that about ½ of the non-hospital but community acquired skin infections in
the Greater Los Angeles area are now MRSA. MRSA stands for methicillin resistant
Staphylococcus aureus.

Multi antibiotic-resistance happens because small lengths of DNA, called transposons can
carry a variety of antibiotic resistance or virulence genes, are able to move from the bacterial
chromosome to plasmid or to a virus DNA (think phage) which can then be freely exchanged
between bacteria.

Harmless gut and soil bacteria have become reservoirs for multi resistance plasmids which
may be gained from pathogens or where there are other commensals that contained the
shared genetic information.

For example, Levy found that the resistance in gut bacteria of cattle moved to gut bacteria of
mice having access to the same area, then from the mice to pigs, chickens, and flies. He
notes a Dutch study that followed bacteria from animals to the human food chain and entered
the consumer’s kitchen. In other cited examples, he noted the distinct relationship between
MDRB in animals and thence to humans attending them, even though the humans used no
antibiotics or ate the animals. Levy’s work is not new. (Levy SB, MD. The Antibiotic Paradox.
New York, Plenum Press 1997). Rusin and Gerba have written on the transfer of pathogens
from common household surfaces via finger to mouth. Others have discussed dust as a
carrier of viable pathogens. Gerba has written extensively on the movement of pathogens in
sediment, their protection for long periods within sediments and the re-transport of viable
pathogens. The NRC in its 2002 report admonished EPA to look at off-site movement and
resistance. There is no evidence that this re-analysis has taken place, yet the World Health
Organization has raised the subject of resistance to a Global crisis.

Thus the current U.S. EPA Class B biosolids with its allowed fecal coliform counts of 2 X10/6
per gram may actually constitute a large aliquot when containing MDRB and applied to areas
with soil and water movement and animal or vector access. These bacteria are thus able to
colonize animals, including humans, through ingestion. There are indications within the
literature of E. coli O157:H7 being to travel up the vascular system in lettuce thus obviating
attempts to wash surfaces. Since lettuce is eaten raw, the risk should be clear to most
readers. Once ingested, the shiga containing plasmids may be transferable to normal flora,
thence later to pathogenic bacteria found in humans or animals, making later treatment with
particular antibiotics ineffective. Additionally, one finds that there is a remultiplication of
bacterial numbers within standing sludge, biosolids or compost (see Hassen below). Thus,
the current Part 503 limits on biosolid marker organisms may have little bearing on the
ultimate numbers.

During composting, the mesophiles (these function at normal body temperatures) can
transfer genetic information to thermophiles (these operate above the lethal fever
temperatures). The archaea, which are extreme thermophiles (these can take temperatures
above the boiling point of water), are recognized as a separate third domain of life together
with the bacteria and eucarya. Transfer of plasmids to bacteria from archaea, has been
demonstrated. Thus, in theory, it may be possible to develop a MDRB that can survive
temperatures found within composting.

Furthermore, there is experimental evidence that even when disrupted by radiation, these
ancient organisms can reassemble. This, from a theoretical perspective, then raises
questions of the eventual failure of pasteurization.Hassen, et al found that , gram-positive
bacteria, especially micrococcus, spores of bacilli, and fungal propagules survived, and
reached high concentrations in compost. Not only that, "the appearance of gram-negative
rods (opportunistic pathogens) during the cooling phase may represent a serious risk for the
sanitary quality of the finished product intended for agronomic reuse." (Bioresour Technol
2001 Dec;80(3):217-25)


J Water Health 03 (2005) 313-324

Quantitative detection of enteroviruses in activated sludge by cell culture and real-time RT-
PCR using paramagnetic capturing
D. Pusch, St. Ihle, I. Graeber, J. M. López-Pila and M. Lebuhn
Federal Environmental Agency (Umweltbundesamt), Department II 2.4. Microbiology and
Parasitology, Corrensplatz 1, Berlin,D-14195, Germany, Phone: +49-30-89031394, Fax: +49-
30-89031830, E-mail:

Federal Environmental Agency (Umweltbundesamt), Department II 2.4. Microbiology and
Parasitology, Corrensplatz 1, Berlin,D-14195, Germany, Phone: +49-30-89031394, Fax: +49-
30-89031830, E-mail:

Federal Environmental Agency (Umweltbundesamt), Department II 2.4. Microbiology and
Parasitology, Corrensplatz 1, Berlin,D-14195, Germany, Phone: +49-30-89031394, Fax: +49-
30-89031830, E-mail:

Federal Environmental Agency (Umweltbundesamt), Department II 2.4. Microbiology and
Parasitology, Corrensplatz 1, Berlin,D-14195, Germany, Phone: +49-30-89031394, Fax: +49-
30-89031830, E-mail:

Technical University of Munich, Institute of Water Quality Control and Waste Management,
Am Coulombwall, Garching,D-85748, Germany, E-mail:


We have compared in extracts of activated sludge the number of enteroviruses detectable
with buffalo green monkey (BGM) cell-cultures versus the number of enteroviral genomes
determined by reverse-transcription quantitative real-time PCR (RT-qPCR). In order to find
conditions adequate for quantifying enteroviral RNA isolated from (waste)water we have
investigated affinity capture of RNA with polystyrene beads (Dynabeads). The capture
efficiency strongly depended on the genomic region chosen for the affinity binding. Capture
of the RNA by its 3'-tail was most efficient (almost 100%); other regions within the genome
yielded variable but lower results. Indirect capture (first hybridization of the RNA to the
oligonucleotides, then attachment of the duplex molecules to the beads) was much more
efficient than direct capture (attachment of the oligonucleotides to the beads first, then
binding of the RNA), and resulted in RNA capture of maximally 60-80%. At least partly, this
was due to incomplete hybridization of the RNA to the complementary oligonucleotides. No
correlation was found between the number of cytopathic effects (CPE) determined by cell
culture and the number of genomes quantified by RT-qPCR; RT-qPCR values were
consistently much higher than the number of CPE. This points to overestimation of infectious
enteroviruses by RT-qPCR and/or underestimation by the cell culture approach.
New England Journal of Medicine
Volume 353:1245-1251  September 22, 2005  Number 12

Staphylococcus aureus Sepsis and the Waterhouse–Friderichsen Syndrome in Children

Patricia V. Adem, M.D., Christopher P. Montgomery, M.D., Aliya N. Husain, M.D., Tracy K.
Koogler, M.D., Valerie Arangelovich, M.D., Michel Humilier, M.D., Susan Boyle-Vavra, Ph.D.,
and Robert S. Daum, M.D.
Staphylococcus aureus has increasingly been recognized as a cause of severe invasive
illness. We describe three children who died at our institution after rapidly progressive clinical
deterioration from this infection, with necrotizing pneumonia and multiple-organ-system
involvement. The identification of bilateral adrenal hemorrhage at autopsy was characteristic
of the Waterhouse–Friderichsen syndrome, a constellation of findings usually associated with
fulminant meningococcemia. The close genetic relationship among the three responsible
isolates of S. aureus, one susceptible to methicillin and two resistant to methicillin,
underscores the close relationship between virulent methicillin-susceptible S. aureus and
methicillin-resistant S. aureus isolates now circulating in the community.

Sometimes it seems like the National Resource Defense Counsel (NRDC) is actually working
for the EPA. NRDC agreed not to sue EPA if ocean dumping was stopped and sludge was put
on land as a fertilizer. It parrots the EPA line that livestock waste is more dangerous than
sludge and it blames the states.

How States Fail to Prevent Pollution from Livestock Waste         
23] In 1993, cryptosporidium, a pathogen found in Milwaukee's drinking water, made 400,000
people sick and led to the deaths of more than 100 people. A suspected cause was dairy

.Health departments do investigate when large numbers of people are effected as was the
case in 1993, when Milwaukee suffered through a Cryptosporidium plague. Over 400,000
people were infected, 4000 were hospitalized and over 100 individuals died. Initially, the
outbreak was blamed on cattle manure
contaminating the water supply system. However, it would take over three years before the
health department identified the source of the infection. It was a human strain of

"the studies done on the Milwaukee outbreak of Cryptosporidium showed that it was due to
the human type coming from human fecal material and human-associated wastewater (Rose
et al., 2002). "

Cost of illness in the 1993 waterborne Cryptosporidium outbreak, Milwaukee,

To assess the total medical costs and productivity losses associated with the 1993
waterborne outbreak of cryptosporidiosis in Milwaukee, Wisconsin, including the average cost
per person with mild, moderate, and severe illness, we conducted a retrospective cost-of-
illness analysis using data from 11 hospitals in the greater Milwaukee area and epidemiologic
data collected during the outbreak. The total cost of outbreak-associated illness was 96.2
million US dollars: 31.7 million US dollars in medical costs and 64.6 million US dollars in
productivity losses. The average total costs for persons with mild, moderate, and severe
illness were 116 US dollars, 47 US dollars, and 7,808 US dollars, respectively. The potentially
high cost of waterborne disease outbreaks should be considered in economic decisions
regarding the safety of public drinking water supplies.        Emerg Infect Dis. 2003 Apr;9(4):

It is strange, but there is no cost included for the 400+ dead people.