Date: 21 October 2003
by William A. Croft, DVM, Ph.D.
Dr. Croft works for Environmental Diagnostic Group Inc. of Madison, Wisc.,
investigating human diseases expressed within the environment. He received his
B.S. in physiology from the University of Wisconsin in Madison and a Doctor of
Veterinary Medicine from the University of Minnesota. Dr. Croft then received a
Ph.D. in Medical Pathology from the University of Wisconsin, Madison, Wisc.
The National Institutes of Health (NIH), the National Cancer Institute and the
U.S. Food and Drug Administration (FDA) have accepted me as a medical
pathologist. Medical pathologists are the most qualified to study human diseases,
establish methods to diagnose them, and determine safety, as well as the most
qualified to assist the Health and Human Secretary in protecting the public from
disease.
I have been involved in the study of human disease and poisoning as medical
pathologist and toxicologist for over 33 years, and also have been accepted as a
medical pathologist in United States and Canadian courts as an expert witness. I
have provided medical pathology reviews to the U.S. government, FDA, U.S.
Environmental Protection Agency (EPA) and NIH. I have been awarded
$900,000 in highly competitive research grants from NIH to use animals for the
study of human disease, and have used over 10,000 rats, guinea pigs, hamsters,
fish, and pigs, as well as bovine and equine animals, to study human disease. I
published the first peer-reviewed paper concerning trichothecene mycotoxicosis
from mold-contaminated buildings in the North American continent (Croft et al.,
1986). The most referenced article on mold to date, it has been referenced 130
times in peer-reviewed articles.
Dr. Gots is a medical doctor and has a Ph.D. degree in toxicology, a research
degree. However, Dr. Gots cannot do research on humans directly and he does
not have a veterinary medicine degree. He is not qualified to use animals to
study human diseases nor is he a pathologist. When scientific work is published
or presented and there is a need for explanation about the observations reported,
scientists attempt to reproduce the work. Dr. Gots is not qualified to reproduce
my work. Therefore, Dr. Gots is not qualified to review my paper, “Clinical
confirmation of trichothecene mycotoxicosis in patient urine.”
In his critique, Dr. Gots listed what he considered to be some key errors in my
research, followed by other purported errors. I will address those points here.
“No controls.”
The study was designed simply to determine whether trichothecene mycotoxins
were being excreted in the urine of humans exposed to different levels of
mycotoxins. The weanling Sprague-Dawley rats used in my study were chosen
because of their sensitivity to trichothecene mycotoxins. Rats are good models
for pathological tissue examination using light microscopy, the basic means to
study human disease. Pathology is the basis of all human disease and each
disease-causing agent imprints a “fingerprint” within the tissue of humans or
animals (Cheville, 1976). Weanling rats are about 1,000 times more sensitive in
detecting trichothecene mycotoxins than a Gas Chromatograph (GC), Mass
Spectrometer, which needs 1 ppm (parts per million) to determine identification
of a mycotoxin, where as weanling rats can detect mycotoxins at ppb (parts per
billion) levels.
In the study, two rats were used as negative controls, receiving the control
solution 50 percent ethyl alcohol intraperitoneally, to compare to the
mycotoxin-exposed animals. Two negative control rats were all that were needed
because of the severe pathology changes versus the control animals.
“Croft asserts in his paper that certain urine extraction methods precipitate
trichothecenes.”
The urine test is an extraction method and does not involve precipitation. This is
part of research methodology, for I have accomplished many extractions from
moldy media and used other solvents. Data was reported in detail so that it could
be repeated by another scientist. The choice for this extraction was ethyl acetate
based on solubility. Since this mycotoxin was observed in animals’ urine, it was
a logical step to check for its presence in human urine. Ethyl acetate is
commonly used as a mycotoxin solvent in extractions on mold-contaminated
media.
The fingerprint of trichothecene mycotoxicosis was identified in humans with the
consumption of toxic grain and was described by Forgacs and Carll, 1962. The
test method for the study was to expose weanling rats to the extracted mycotoxin
material and observe the generated pathology to determine if the same causative
agent or fingerprint was present. The study rats displayed the same pathology
fingerprint, which clearly establishes the extraction of trichothecene mycotoxins
from patients exposed to toxic mold conditions and satisfies Koch’s Postulants.
Human pathology presents the question, “are mycotoxins or another causative
agent responsible for the necrosis of neurons discovered in the autopsy on the
spinal cord?” An extraction of mycotoxins from the human urine sample was
performed, followed by dissection of the rat spinal cord tissues. The pathology
of the spinal cord dorsal and ventral motor neurons from the rat supported the
hypothesis that the poisoning effects of trichothecene mycotoxins were
responsible for the necrosis of neurons within the spinal cord of the human
subject. This means that these mycotoxins can cause Amyotrophic Lateral
Sclerosis within exposed humans. Each organ’s tissue within the exposed rats
and the human subjects matched the exact cellular fingerprint. It is not often that
the patient’s pathology can be clearly checked in animals as in this case,
therefore, making a stronger case for the extraction of trichothecene mycotoxins
within the urine and the associated pathology.
“Croft erroneously cited a test and misused a skin patch test (Higuchi et al.)
upon which [he] relied heavily … there is no established basis for the
assumption that mycotoxins affect alcohol dehydrogenase.”
It is well documented that trichothecene mycotoxins inhibit alcohol
dehydrogenase (Ueno, 1977). Alcohol dehydrogenase is a very common enzyme
(Casarett et al., 1995) as with other sulphydryl (SH) residues of SH enzymes, and
can be used to detect level of exposure in most cases as reported in the paper. I
did not rely on this test alone, but used it as another piece of the puzzle to help
establish exposure to trichothecene mycotoxins, especially in children who do
not drink alcohol. The test offers an option for the physician who suspects mold
exposure in his/her patients.
Higuchi et al., 1987, incorrectly used the term “aldehyde” dehydrogenase when
they should have used the term “alcohol” dehydrogenase. I am very familiar with
the naming of enzymes and have completed several research projects concerning
ethyl alcohol effects in humans. When the test is applied to patients, several
questions are asked to determine if there is a genetic deficiency of the enzyme or
medications or exposure to other toxic chemicals that can affect results of the
test.
“Croft went far beyond the test protocol of [Higuchi et al., and] … used a
different method and a different scoring system, protocols which have not been
validated.”a
Numbers are commonly assigned to signs/symptoms as a tool to quantify the
effects on a patient, enabling statistical analysis to be performed. In my study,
the numeric system allowed the clear comparison of numerical totals to levels of
exposure, indicating that one is working with the causative agents. Scores based
on signs/symptoms did correlate to level of exposure based on the amount of
mycotoxin extracted from the urine samples. Several doctors are now using this
numeric system for determining the disability of a whole person.
“[Croft et al.] give no references for their method [of extracting trichothecenes
from urine] and there is little reason to believe that trichothecenes were
extracted. Their conclusory statement that this was tested by thin layer
chromatography (TLC) is not properly discussed. They state that their TLC
results were consistent with those reported for trichothecenes, but which of the
trichothecenes?”
I have already discussed the extraction of the mycotoxins from urine. The TLC
was used to characterize that the extracted material was correctly identified as
trichothecenes and correlated to reports in the literature. The purpose of the
study was to see if trichothecene mycotoxins could be extracted. The test system
was the exposure to the rats, resulting in development of the fingerprint of the
causative agent for the disease. The question “which trichothecene” is academic
because all are very poisonous, and represents the next possible study
experiment.
SPECT scans can be a useful tool to help understand brain changes within
patients exposed to toxic mold. As a medical pathologist, I disagree with the
Society of Nuclear Medicine Brain Imaging Council and declare that SPECT
scans should be allowed because this procedure can be useful in medicine and
patient workups. Defense lawyers do not want such photographs presented to
juries because lay people can understand a photograph more easily than many
experts’ testimonies.
“The authors … state that urine samples left at room temperature for five
months showed no “putrefaction” and excessive protein at the bottom of the
container. How and why these constitute measures of the presence of
mycotoxins is merely stated with no support and no logical basis.”
Reporting what happens to the urine is part of researching the disease and is
important. The logical basis for no putrefaction is because trichothecene
mycotoxins are very anti-life, or antibiotic, in effects on biological life. The
mycotoxin kills the putrefaction bacteria, so the absence of the putrefaction
bacteria indicates the presence of the mycotoxin. I have observed this effect in
urine samples collected from patients over the last 20 years. The protein or
cellular debris on the bottom of the container is an indication of the amount of
poisoning taking place within the patient due to dead cells within the body from
the antibiotic effects of the mycotoxin.
Normal urine does not contain trichothecene mycotoxins
(http://www.mdsmetro.com/ patients/conditions/urinalysis.htm). My own urine
was used to confirm that, but also to determine the extraction method. The study
was not designed to determine the incidence or the frequency or background data
concerning trichothecene mycotoxins in urine. It has nothing to do with the
assay. We now know that trichothecene mycotoxins are indeed released in the
urine of toxic mold patients.
In this study, patients were studied and other interesting facts about the
mycotoxins’ effects were revealed. I recently discovered that the mycotoxin is
excreted from the body within 40 hours when in a totally clean environment (“a
walk away”), and the immune system will start to function in 14 days expressed
with the appearance of allergies and even anaphylaxis. Clothing is a carrier of
trichothecene mycotoxins and they cannot be washed from the contaminated
items. The use of bleach does not inactivate, but actually stimulates, mycotoxin
production because you are threatening the mold and the mold respond. Bleach
is used by the Defense Department to increase mycotoxin production.
Conclusion
I would like to thank the editor for giving me a chance to discuss Dr. Gots’
remarks. Medical doctors are not trained to do research and have problems
understanding pathology research. In seeking answers to help their patients,
medical doctors try to gather more information from a study than it was
designed for. The study has been peer-reviewed by several scientists for the Journal of En
Journal of Environmental Biology, and by numerous plaintiff and defense
lawyers attempting to find some problem. None was found.
The data also has been accepted in New York Court by Justice Louise Gans,
who accepted me as a pathologist and as an expert witness on the subject in a
Frye hearing in Marie Alston v. Henry Phipps Plaza South, et al., No.
116958/99-2001 (N.Y. Sup. Ct., N.Y. Cty.).
Finally, I have received positive feedback from prominent scientists who have
reviewed this study. The urine test is based on pathology and clearly establishes
that trichothecene mycotoxins are detected in human urine based on the
corresponding exposure. It is a useful tool for diagnosis, especially when
trichothecene vapors are released but spores cannot be detected. It can be used to
study the disease process in humans in the spinal cord, heart, lung, and organs of
the body and related pathology. Doctors can use the test to determine when
patients are free from mycotoxin exposure. The urine test helps doctors and
patients pinpoint the source of the contamination so that they can avoid the
contaminated environment.
PubMed Abstract
PubMed Abstract
Environmental Diagnostic Group Inc., Department of
Environmental Pathology and Toxicology, 521 Hilltop Drive,
Madison, Wisconsin 53711, USA. doccroft@hotmail.com
The investigations of four Cases involving mold-contaminated
buildings and human reaction to exposure, documents tests of
extracted urine containing trichothecene mycotoxins confirming
exposure and the diagnosis of mycotoxicosis in humans. In each of
four Cases, the urine demonstrated antibiotic activity, sulfuric acid
charring, and protein release. Urine was extracted using ethyl
acetate 40V/60V[EA]. Extracted mycotoxin spotted on (TLC)
displayed color and a range of (rf) between 0.2-0.6 using various
solvents. Extract was re-suspended using 50% ethanol V/V to inject
mycotoxins into weanling female Sprague-Dawley rats.
Degeneration and necrosis of the rat’s tissue followed. Koch’s
Postulates conditions were fulfilled by isolation of the causative
agent, the trichothecene mycotoxins and the reproduction of
disease. Examination of human tissue within the urine extraction
group confirms Koch’s Postulates and comparative pathology
confirms inhalation Mycotoxicosis, with severe necrosis of the
central nervous system and severe scarring within the lungs.
Extraction of mycotoxins from human patient urine is a very useful
confirmatory test to demonstrate exposure and identify
mycotoxicosis. Low concentrations (6%) of sodium hypochlorite
were ineffective against the activity of trichothecene mycotoxin.
The severity or stages of disease directly correlates the level of
exposure or poisoning (Patent Pending).
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