Re: Nonobacteria (contaminant of fetal bovine serum)

[Guest guest]

Sheri, I'm going to pass this around to some! When I heard Dr.

Wakefield speak at the conference. He said that fetal bovine serum

was found in the urine of the autisitc children that were in his

study.

> This information was posted on another list I'm on (nothing to do

with

> vaccines)

>

> Has anyone heard of this? Anyone have time to research more??????

> Magda - with Informed Parent just met with a group in France, one

of who

> has tons of information on animal contamination of vaccines -

Magda - can

> you get anything in writing in English or ask about this? Jim

West -

> what's your take on this? Anyone else?

> Sheri

>

>

> Fetal Bovine Serum - Varicella Vax, some oral polio vax, Merck

reported it

> was in MMR to Ray Gallup , and in the former Rotavirus

vaccine.......

>

> At first this seems like an ad........but........read all the way

thru and

> go to links

>

> " As reported May 23rd, 2001</b> at the 101st General Meeting of the

> American Society for Microbiology, Nanobacteria has been found to

be a

> contaminant in previously assumed-to-be-sterile medical products,

> specifically IPV Polio Vaccine. Most human biologicals and

vaccines are

> made in fetal bovine serum, a medium that is known to be

contaminated

> with nanobacteria. In order to prevent this problem in the

future, human

> biological products must be made in Nano-Free Culture medium

(filtered

> first through 20 nanometer filters,Gamma-Irradiated with 150

megarads and

> then heated to 90 degrees Centigrade for at least an hour to kill

any

> nanobacteria present) "

>

> http://www.nanobaclabs.com/PageDisplay.asp?p1=6578

>

> excerpts......

> " The term " Nanobacteria " is short for it's scientific genus &

> species name " Nanobacterium sanguineum " , a Latin scientific term

for blood

> nanobacteria. Nanobacteria are " nano " -sized in that they are

from

> 20-200 nanometers in size (a nanometer is 1 billionth of a

meter. A

> nanometer is the width of ten hydrogen atoms side-to-side!) and are

the

> smallest known self-replicating bacteria. They are from the Archaea

Family

> of bacteria, known for their primitive pleomorphic lifestyles "

>

> " Nanobacteria infection by Nanobacterium sanguineum is an " emerging

> infectious disease " meaning that it is newly discovered and

that the

> diseases it cause are being researched and further described. Its

DNA, RNA

> and Lipopolysaccaride profiles have been accurately mapped by

multiple

> scientific researchers at many universities worldwide.

Nanobacteria are

> not nice bugs and have absolutely no known positive benefits to

humans. The

> discoverers of nanobacteria, Drs. Ciftcioglu & Kajander developed

antigen &

> antibody diagnostic blood testing for nanobacterial infections that

we

> offer as the " NanobacTEST " . NanobacLabs has developed safe and

effective

> nanobiotic prescription treatments "

>

> " Nanobacteria are extremely small, slowly growing bacteria that

can be

> cultured from the blood of humans and mammals.</b> Their size

is

> 20-200 nanometers....when compared to " regular " bacteria,

Nanobacteria are

> 1/100 to 1/1,000 the size, allowing them to easily move

around into

> other cells and invade them. Nanobacteria cause apoptosis

(cell

> death) when exposed to human cells or other bacteria. They can cause

> alteration of RNA and DNA gene-expression patterns of cells

they

> infect.....this can lead to genetic alteration, abnormal cell

growth and

> proliferation. When compared to other bacteria, Nanobacteria

grow

> very, very slowly, only reproducing every 3 days…..where " regular "

> bacteria reproduce in minutes or hours. Nanobacteria cannot be

grown in

> standard culture media and can only be grown in mammalian blood

or serum.

> Nanobacteria were discovered in 1988 by Nobel Prize Nominees

Dr. Neva

> Ciftcioglu, PhD and Olavi Kajander, MD, PhD as

a " contaminant " in

> mammalian cell cultures that kept killing the mammalian cells

> (apoptosis) in their mammalian cell culture research. They

have been

> researching nanobacterial pathophysiology for nearly 14 years now

and are

> the worldwide experts on nanobacterial basic science. They are

currently

> guiding and teaching researchers all over the world. NanobacLabs is

the

> world leader in the research and development of prescription

NANOBIOTICS

> that eradicate pathological calcification and nanobacterial

infections "

>

>

> " As reported May 23rd, 2001</b> at the 101st General Meeting of the

> American Society for Microbiology, Nanobacteria has been found to

be a

> contaminant in previously assumed-to-be-sterile medical products,

> specifically IPV Polio Vaccine. Most human biologicals and

vaccines are

> made in fetal bovine serum, a medium that is known to be

contaminated

> with nanobacteria. In order to prevent this problem in the

future, human

> biological products must be made in Nano-Free Culture medium

(filtered

> first through 20 nanometer filters,Gamma-Irradiated with 150

megarads and

> then heated to 90 degrees Centigrade for at least an hour to kill

any

> nanobacteria present) "

>

> ********

> http://www.uku.fi/~kajander/

>

> http://www.uku.fi/~kajander/fatal.html

> Mol. Biol. Cell, Suppl., Vol. 7, (1996): 517a

> Fatal (fetal) bovine serum: discovery of Nanobacteria

> E. Olavi Kajander, Ilpo Kuronen and Neva Ciftcioglu

> Department of Biochemistry and Biotechnology, University of Kuopio,

> FIN-70211 Kuopio, Finland

>

> A new potential threat for blood and blood prouducts, cell culture,

cell

> and tissue banking and biotechnology has been discovered:

Nanobacterium

> sanguineum gen. et sp. nov.. These self-replicating ultrafilterable

> bacteria were isolated from over 80% of commercial ÔsterileÕ fetal

and

> newborn bovine sera and are thus the most common contaminant

present in

> cell cultures. Growth occured in vitro under cell culture

conditions (with

> or without mammalian cells) but not under anaerobic conditions.

Their

> doubling time was 1-5 days. They were culturable also in protein and

> lipid-free medium beyond three years with monthly passages. Colony

> formation on solid media was poor. The agent multiplied in culture

with

> serum in a logarithmic mode that could be prevented with

aminoglycoside

> antibiotics, EDTA, cytosine arabinoside and gamma irradiation. They

showed

> procaryotic structure with specific antigens detectable by

monoclonal

> antibodies, were generally mobile, coccoid with a diameter of 200

to 300 nm

> in serum, stained poorly with bacteriological stains, were very

resistant

> to antibiotics and passed through 100 but not 50 nm filters. Their

> aminoterminal protein sequences were novel and reproducible.

Considerable

> evidence suggested presence of nontraditional DNA. They incorporated

> radiolabelled uridine into DNA. 16S rRNA gene sequence results

place them

> in alpha-2 subgroup of Proteobacteria which includes Brucella, also

> pathogens of mammalians with preference to the fetus. This new

agent causes

> cytotoxicity and senescence in many cultured cell lines by

apoptotic cell

> death and growth arrest.

>

> *******

> http://www.uku.fi/~kajander/threat.html

> (pictures and graphs at the site)

>

> Vaccines 97, Brown & al ed.,Cold Spring Harbor Laboratory Press,

New York,

> 1997

> A New Potential Threat in Antigen and Antibody Products:

Nanobacteria

> Neva Ciftcioglu, Ilpo Kuronen, Kari Akerman, Erkki Hiltunen, Jukka

> Laukkanen and E. Olavi Kajander

> Department of Biochemistry and Biotechnology, University of Kuopio,

> FIN-70211 Kuopio, Finland.

>

>

>

>

> Several vaccines are currently being produced by using cultured

mammalian

> cells. Microbiological sterility of such vaccines is of great

importance

> since several examples indicate potential safety hazards in vaccines

> contaminated with unknown organisms. Fetal bovine serum (FBS) used

as a

> supplement in cell culture is a known safety risk (Hodgson, 1995).

> Obviously, not all of the risk factors of FBS are yet known and

thus cannot

> be controlled. It is commonly known that only about 10% of FBS

batches

> support cell cloning well (Liddel and Cryer, 1991) but the reasons

for this

> have remained unclear. As with many other cell culturers, we faced a

> problem about 10 years ago of poorly thrivingo cells not

attributable to

> any known contaminant. In this report, we describe the discovery of

a new

> bacterium from mammalian blood and blood products, tentatively

named as

> Nanobacterium sanguineum gen. et sp. nov., and show that this agent

is

> common and harmful.

>

>

>

>

>

> DISCUSSION

> Culture and Diagnosis of Nanobacteria

> The discovery of Nanobacteria came about because we had a problem

with cell

> cultures namely vacuolized cells (Fig. 1A) and poorly thriving

cultures

> without any contaminant detectable by standard methods. Transmission

> electron microscopy (TEM) made from these poorly thriving cell

cultures

> indicated the presence of internalized procaryotic organisms (Fig.

1B).

> That their source was the commercial " sterile " FBS was proven by

> gamma-irradiating all the culture components (Table 1). This

experiment

> also indicated that sterile culture media for detection of new

organisms

> can be made by using gamma-irradiated serum as a supplement. The new

> organisms passed through 100 nm (but not 50 nm) filters and were

called

> nanobacteria, since no other bacteria are known that can pass

through

> filters with such small pores. This ability to pass through such

small-pore

> filters was most remarkable since they were shown to have a cell

wall and

> yet were able to surpass the filterablity of cell-wall-less

bacteria. They

> were unculturable in microbiological media but could be cultured

under cell

> culture conditions (with or without mammalian cells, CO2 5-10%).

These

> minute generally coccoid organisms had a diameter of 200 to 300 nm

in

> serum, and their size increased during the culture due to the

production of

> a very thick cell envelope (Fig. 1C, D). The thick and calcified

envelope

> made them visible even by light microscopy. The doubling time of

> nanobacteria was 1-5 days (Fig. 2). Their multiplication could be

detected

> by specific ELISA, optical density, microscopic counting, SDS-PAGE

or

> methionine and uridine incorporation, and the multiplication could

be

> prevented with high doses of aminoglycoside antibiotics, EDTA,

cytosine

> arabinoside and gamma-irradiation. Considerable evidence suggested

the

> presence of nontraditional DNA. 16S rRNA gene sequence results

(data will

> be published elsewhere) placed them into the alpha-2 subgroup of

> Proteobacteria which includes Brucella(which are also pathogens of

> mammalians with preference to the fetus) and Bartonella.

> Nanobacteria were isolated from more than 80% of commercial FBS and

newborn

> bovine sera and are the most common contaminant present in cell

cultures.

> In addition, we isolated nanobacteria from the blood of 4% of

medical

> students at our university. Positive identification of nanobacteria

> involved growth in cell culture medium with typical growth rate and

optical

> properties, specific stainability with Hoechst 33258 using the high

dye

> concentration and positive immunoassay results with

immunofluorescence

> and/or ELISA using monoclonal anti-nanobacteria antibodies.

>

>

>

>

> Cytotoxicity of Nanobacteria

> Nanobacteria are cytopathic in cell cultures and invade mammalian

cells in

> a distinctive manner: They trigger cells that are not normally

phagocytic

> to engulf them. These novel organisms are one of the causes for cell

> vacuolization, poor thriving and unexpected cell lysis, problems

often

> encountered in mammalian cell culture. Several mammalian fibroblast

lines

> were cultured in MEM medium as described previously (Kajander et

al.,

> 1990), and were infected with nanobacteria. Electron microscopy and

FITC

> staining with specific monoclonal antibodies indicated that

nanobacteria

> were bound on the surface of the fibroblasts (Fig. 1E-G). We

concluded that

> they were internalized either by receptor-mediated endocytosis or

by a

> closely related pathway. After the internalization, fibroblasts

showed

> apoptotic abnormalities and died if subjected to a high dose (>100

> nanobacteria/cell).

>

>

>

>

> Different Growth Phases of Nanobacteria

> Washed nanobacteria added to serum-free medium grew very slowly as

> evidenced by increase in their numbers and protein level and were

firmly

> attached to the culture plates. These cultures progressed to large

> multicellular formations covered by layers of a firm protective

material

> several micrometers thick (Fig. 1H). After addition of sterile

serum, the

> layer disappeared, with typical small coccoid nanobacteria later

appearing

> in the same cultures with the mobile, larger D-shaped ones (Fig.

1I).

> Specific monoclonal antibodies indicated the presence of the same

antigenic

> sites in both D-shaped and coccoid nanobacteria, and their 16S rRNA

gene

> sequences were 98% identical.

>

>

>

>

> How can Cell Culture be Possible with Nanobacteria-contaminated

Fetal

> Bovine Serum?

> Although more than 80% of cell culture serum batches are

contaminated with

> nanobacteria, many cell culturers have not faced this problem with

their

> cell cultures. We have experienced a major problem with

nanobacteria in

> cell culture only when they are present at high concentrations

relative to

> cells. This can occure typically in cell cloning and in long-term

> experiments where mammalian cells do not multiply. Internalization

of

> numerous nanobacteria by a cell results in cytotoxicity.

Importantly, most

> cell lines multiply faster than nanobacteria. Thus, cytotoxic

> concentrations may be avoided.

>

>

>

>

> Why is Nanobacteria a Potential Threat?

> Nanobacteria can cause a chronic infection in laboratory animals

and in

> humans. The agent could be isolated from blood of one peron for 5

years

> despite the presence of antibody. When nanobacteria were injected

into

> rabbits, the agent was initially isolated from urine and then from

> cerebrospinal fluid after one year. Nanobacteria multiply very

slowly and,

> if pathogenic in humans, might cause slow chronic autoimmune-like

disorders

> (compare with leprosy or brucellosis). Sofar, there are no chronic

> bacteraemia known that would not be harmful. Thus, the posibility

that

> nanobacteria may be present in vaccines made with cell culture, or

in

> gammaglobulin or other antibody preparations, must be controlled.

>

>

>

>

> SUMMARY AND CONCLUSIONS

> Nanobacteria are novel microorganisms that are not detectable with

present

> sterility testing methods, but they are detectable with new culture

and

> immunomethods. They are commonly present in bovine and blood

products and

> thus in cell cultures and antigens, including vaccines derived

therefrom,

> and may be present in antibody and gammaglobulin products.

Nanobacteria are

> a potential risk because of their cytotoxic properties and ability

to

> infect fetuses, and thus their pathogenicity should be scrutinized.

>

>

>

>

> ACKNOWLEDGMENTS

> We thank E. Tahvanainen, H. Martikainen, A. Pelttari and T. Ojanen

for

> their valuable help in microbiological, microscopic, and chemical

> techniques. P. Mäenpää, T. Eloranta, J. Kärjä and O. Lindqvist

contributed

> valuable ideas in discussions. The work was supported by the

Academy of

> Finland, University of Kuopio, Finland Technology Center, Juho

Vainio

> Foundation and Savo High Technology Foundation.

>

>

>

>

> REFERENCES

> Hodgson, J. 1995. To treat or not to treat: That is the question

for serum.

> BioTechnology 13: 333.

> Kajander, E. O., R. J. Harvima, L. Kauppinen, K.K. Akerman, H.

Martikainen,

> R. L. Pajula, and S. O. Kärenlampi. 1990. Effects of

selenomethionine on

> cell growth and on S-adenosylmethionine metabolism in cultured

malignant

> cells. Biochem. J. 267: 767.

> Liddel, J. E., and A. Cryer. 1991. in A practical guide to

monoclonal

> antibodies, p. 25. Wiley, New York.

> Figure 1. Ultrastructure of nanobacteria and their interaction with

> fibroblasts.

> (A) Perinuclear vacuolization of an infected 3T6 cell under phase-

contrast

> microscope;

> ( TEM image of a nanobacterium engulfed by a BHK cell;

> © cultured coccoid nanobacteria (bars 200 nm).

> (D) SEM image of nanobacteria attached to culture vessel;

> (E) nanobacteria attached to a fibroblast surface (arrow shows the

surface

> of the fibroblast; bars 1 µm).

> (F) Indirect immunofluorescence staining of cultured healthy 3T6

cells with

> a monoclonal antibody (8/0) against nanobacteria;

> (G) 3T6 cells inoculated with nanobacteria;

> (H) TEM of a nanobacterial population in a serum-free culture

(arrow shows

> a D-shaped nanobacterium in this population);

> (I) D-shaped nanobacteria after culture in serum-containing medium

(bars 1

> µm).

> Figure 2. Growth-curve of nanobacteria. As a control, gamma-

irradiated FBS

> was used. At each time point, samples from triplicate incubations

were

> taken, frozen and analyzed by turbidometer at the end of the

experiment.

> Turbidometer units are means of three measurements from 1/6

dilutions of

> cultures.

>

>

> Table 1. The Effect of 60Co Gamma-Irradiation of Culture Components

on

> Multiplication of Nanobacteria

>

>

> Culture Multiplication

> FBS

> RPMI +

> FBS

> *RPMI +

> *FBS

> RPMI -

> *FBS

> RPMI

> nanobacteria or FBS +

> *FBS

> RPMI

> *nanobacteria or * FBS -

>

>

> The material marked with asterisk (*) received a sterilization dose

of 3

> megarads during 16 h at room temperature. Cultures were established

using

> 10 % serum and nanobacterial counts were followed for 4 weeks.

>

> ********

> http://www.idreview.co.uk/pdf/pdf-1/11E1.PDF

>

> " Another intriguing subject is that of the putative nanobacteria

studied by

> a Finnish group. Present inhuman and bovine sera, they might have

> contaminated many biological preparations and have spread in human

> populations. As they induce deposition of calcium salts,they may be

> involved in diseases involving such depositions, such as renal

lithiasis

> and atherosclerotic plaques, or even neuro-degenerative diseases.

Their

> minimal size (200 nM) precludes conventional packaging of a length

of DNA

> sufficient to code for an autonomous microorganism. But it is

possible that

> their genetic information is encoded in a modified, more compact

DNA.In

> conclusion, our fight against emerging diseases has just begun. We

should

> always be vigilant against the resurgence of known infectious germs

and the

> emergence of new agents. More than ever, a worldwidenetwork of

sentinel

> laboratories and a coordinated multidisciplinary effort in

biomedical

> research are required for our survival. "

> --------------------------------------------------------

> Sheri Nakken, R.N., MA

> Vaccination Information & Choice Network, Nevada City CA & Wales UK

> $$ Donations to help in the work - accepted by Paypal account

> vaccineinfo@b...

> (go to http://www.paypal.com) or by mail

> PO Box 1563 Nevada City CA 95959 530-740-0561 Voicemail in US

> http://www.nccn.net/~wwithin/vaccine.htm

> ANY INFO OBTAINED HERE NOT TO BE CONSTRUED AS MEDICAL OR LEGAL

ADVICE. THE

> DECISION TO VACCINATE IS YOURS AND YOURS ALONE.

> Well Within's Earth Mysteries & Sacred Site Tours

> http://www.nccn.net/~wwithin

> International Tours, Homestudy Courses, ANTHRAX & OTHER Vaccine

Dangers

> Education, Homeopathic Education

> CEU's for nurses, Books & Multi-Pure Water Filters

[Guest guest]

> " As reported May 23rd, 2001</b> at the 101st General Meeting of the

> American Society for Microbiology, Nanobacteria has been found to be a

> contaminant in previously assumed-to-be-sterile medical products,

> specifically IPV Polio Vaccine.

Press release from the meeting.

http://www.asmusa.org/pcsrc/gm2001/33749.htm

101st General Meeting of the American Society for Microbiology

May 20-24, 2001, Orlando, Florida

For more information on any presentation at the 101st General Meeting conta=

ct Jim Sliwa, ASM Communications at jsliwa@....

Another debate in viral vaccines: nanobacterial contamination

(Session 078/Y , Paper Y-3)

Neva Ciftcioglu

University of Kuopio

Kuopio,

Finland

35817163054

neva.ciftcioglu@...

Vaccines are an indispensable weapon in the battle to prevent disease, both=

in humans and animals. Provision and protection of the vaccine supply is so=

essential that no possible pathogen is beyond suspicion. So it is with nano=

bacteria (NB), a potential infectious agent so new that science is still deb=

ating its existence as a " living organism " . We analysed 6 veterinary vaccine=

s and 3 inactivated human polio vaccines produced in cell culture for NB. We=

report that 3 of 6 European veterinary vaccines contained NB. Of the 3 dist=

inct lots of polio vaccine from European manufacturers, 2 lots from manufact=

urer-1 were NB-positive and 1 lot from manufacturer-2 was NB-negative. These=

results suggest that not all lots of vaccine contain detectable NB. The pub=

lic health risk, if any, from nanobacteria is yet to be defined, but nanobac=

teria have been found in kidney stones, liver and kidney cyst fluids, and im=

plicated in kidney stone formation.

In the early 1990's, Drs. Olavi Kajander and Neva Ciftcioglu, University of=

Kuopio, Finland discovered a minute self replicating agent, which was named=

" nanobacteria " , in fetal bovine serum (FBS) that mediates mineral (calcium =

phosphate) formation under conditions found in blood and urine. NB was subse=

quently found to contaminate cell cultures widely used in research and to pa=

ss through filters commonly used to sterilize vaccines. Several vaccines are=

produced in mammalian cell cultures that were initially or are currently cu=

ltured in media supplemented with FBS. Thus, the question arose, " Could the =

NB found in some batches of FBS contaminate some vaccines " ?

To detect nanobacteria in these 9 vaccines, the Kuopio University team cult=

ured the vaccines in routine tissue culture medium known by prior testing to=

be free of NB and examined the original vaccines and cultured vaccines by e=

lectron microscopy, and immunologic tests for NB antigens (immunofluorescent=

staining, dot blot assay and ELISA). NB were found 3 of 6 vet vaccine produ=

cts and 2 of 3 human vaccine products prepared using FBS. A second question =

arose, " Even if NB are present in the culture broth, why were they not remov=

ed during the purification and sterilizing steps of vaccine production " ?

Cell culture mediated viral vaccine production requires the use of cells to=

propagate large amounts of virus, which is then filtered to remove contamin=

ates and cells present in the propagation step. The viruses passing through =

the filters are then further processed and inactivated prior to use. NB are =

in the same size range as certain large viruses, and may adhere to viruses t=

hus escaping purification steps using filtration. NB are also resistant to m=

any disinfecting chemicals and antibiotics. NB can exist intracellularly and=

grow in the absence of FBS. At this time it is not known if FBS added to th=

e production cell lines or NB in the cells lines themselves are the source o=

f NB contamination in these cases.

The Center for Biologics Evaluation and Research (CBER) and the U.S. Food a=

nd Drug Administration (FDA) learned earlier this year that some vaccines we=

re manufactured with bovine–derived materials obtained from countries where =

bovine spongiform encephalopathy (BSE; mad cow disease) is prevalent or wher=

e a substantial risk for BSE exists. Creutzfeldt-Jakob disease (vCJD), the h=

uman equivalent of BSE, has been attributed to, among other possibilities, e=

ating beef products from cattle infected with the prion agent of BSE. Althou=

gh there is no evidence to date that cases of vCJD are related to the use of=

vaccines, interestingly a vaccine lot was withdrawn because of being expose=

d to FBS from the country where high risk for this prion disease exists.

The study summarised in the first paragraph will be presented in the 101st =

ASM Meeting in Orlando in May 21, 2001 (1.00-2.30pm). The Kuopio team's mess=

age to the scientific society in this presentation will be: More research is=

needed to understand nanobacteria and their role in human and animal diseas=

es. To err on the side of safety is no vice; just as prions and viruses were=

viewed with scepticism before they were completely characterised, so concep=

ts of nanobacteria (and the scientists who study them) will be knocked aroun=

d until more is known about NB. Vaccines should be regarded as safe and esse=

ntial to human health in the modern age until they are proven otherwise.

Related Literature:

NB have been linked to kidney stones (Ciftcioglu et al., Kidney Internation=

al 56:1893-8,1999), as a kidney stone causative agent ( Cuerpo et al.,=

Arch Esp Urol 53:291-303, 2000), and in polycystic kidney disease (Hjelle e=

t al, Kidney International 57: 2360-2374, 2000).

For a recent review of nanobacteria, (Kajander, Ciftcioglu, -Hjelle, =

Hjelle. Current Opinions in Nephrology and Hypertension 2001 in press).

[Guest guest]

This may have been posted before. After reading it again I noticed a

mention of the parvo virus. This is the virus they reported in the

rash that was happening in some of the 27 states that I posted the

other day. Wonder if these children were at the age of catching up on

vaccines, or the older children receiving the required Hep B.?

I only posted part of the PDF. For the full version use the URL

posted.

http://www.jeffsutherland.org/complementary/vaccine_contamination_mcre

arden.pdf

There is now heightened concern that this virus and others can cross

species lines, creating new strains as they adapt to their new hosts,

and this would include passage of the virus to and from humans.

Whether the human strain of BVDV causes overt illness is uncertain,

because physicians may be uninformed and not even be looking for this

virus. It may be useful however, to compare the infection patterns in

cattle. They can be persistently infected at a low level for their

entire life with a non-pathogenic strain of the virus. Under these

conditions, they consistently create and shed virus into the

surrounding environment, which then infects other animals. The virus

can nonetheless become lethal to the animal if it mutates, with the

new form also causing " visible cell damage and death " in cultured

conditions (10). The animal succumbs to gradual or acute

deterioration of the gastrointestinal mucous lining, which produces

diarrhea and its eventual demise. However, mutated virus is not

always necessary to provoke debilitating illness and death, and

ordinary virus can be isolated from the cow's pancreas, adrenal

glands, and pituitary glands (11); the virus has also been documented

as causing serious pulmonary illness (12). A study describes an

outbreak of disease

among goats due to a vaccine contaminated with a bovine pestivirus;

oddly, these animals experienced reproductive failure and lesions to

the central nervous system (13). So, can these disease symptoms in

varied organs and tissues also occur in humans when they carry this

virus short or longterm?

A cursory examination of the literature indicates this may be

occurring. One revealing study tells us " faeces from children under 2

years old who had gastroenteritis that could not be attributed to

recognised enteric pathogens were examined…for Pestivirus antigens.

Such antigens were detected in 30 of 128 episodes of gastroenteritis…

The diarrhoeal disease in children excreting Pestivirus antigens

resembled that in other children except that it was more commonly

associated with signs and symptoms of respiratory inflammation. " (14)

There are also concerns regarding a pattern of pestivirus infection

in infacts born with microcephaly, a condition wherein the head or

cranial capacity is unusually small (15, 16).

Scientists from the USDA National Veterinary Services Laboratory

describe the situation quite clearly, and give an indication of the

seriousness of the problem: " The high frequency of virus and antibody

detection in individual animal or small pool samples suggests that

any large pool of unscreened sera will be contaminated. Infection of

cell cultures with BVDV can lead to interference with the growth of

other viruses. Vaccine produced on contaminated cells may in turn be

contaminated, leading to seroconversion or disease in the vaccine.

The safety, purity, and efficacy of viral vaccines require BVDV

testing of ingredients, cell substrates and final product. " (17)

And here is a similar statement from a New York Blood Center: " Bovine

viral diarrhea virus, whose small virion size does not allow 100%

assurance of its removal by filtration, may potentially contaminate

every lot of commercially produced fetal bovine serum. " (18) In

reality though, how much of this particular viral contaminant has

trickled into humans? Well, in spite of manufacturers and regulatory

agencies claiming efficacy of their testing procedures, one 2001

study found 13% of human MMR, polio, or Streptococcus pneumoniae

vaccines tested

positive for pestivirus RNA (19). And another researcher

observes, " serum antibodies against BVDV have been detected in

approximately 30% of human population who had no contact with

potentially infected animals. " (16) Also, " pestiviruses adapted to

human cell cultures may be harmful because serious BVDV infections in

humans have been frequently suggested…The BVDV persistently infected

in cell cultures used for vaccine productions have been shown to be a

source of contamination in live virus vaccines. It is, therefore,

prerequisite to examine pestivirus contamination in cell cultures to

avoid secondary infectio ns in humans as well as in animals. " (20)

Continuous immortal cell lines

This same scientist brings up another important issue. Because many

medical-use biological products (including vaccines) are now being

cultured or produced on what is called " continuous " cell lines (i.e.,

these are cell cultures consisting of " immortal " or cancerous types

of cells because they have no limits on how many times they can

divide), there is concern that viral contamination of these cell

lines with a pathogen like bovine viral diarrhea virus, could spread

cancer-promoting material into the human recipient. How could this

happen? Briefly, it works like this. The virus (which in this case

has a single strand of RNA for its genome) is capable of

incorporating RNA from the cells in which it has been cultured, into

its own genome. If any contaminant RNA virus is present in a culture

that contains immortal cancerous cells, this virus can easily mutate

to include unwanted oncogenic material, which can then get passed

into the biological product intended for human medical use (16).

Were you aware that biological products, including some common

vaccines (for instance, polio and rabies), are being produced

on " continuous " immortal cell lines? Manufacturers, scientists, and

agencies will often assure us that these cells themselves are

not " tumorigenic " , i.e., they do not cause tumors per se. A closer

look however, shows this is not always the case. While lab culturing

may indicate that these types of cells are not immediately changing

to overt tumor cells, it is now wellknown in the scientific community

that after these cells have been repeatedly cultured a certain number

of times, something causes them to convert to a cancerous state (21).

This journal article summary addresses the issue in regards to Vero

cells, which is a continuous cell line coming from the African green

monkey, and is commonly used in vaccine production. It states, " One

of the current criteria for evaluating the acceptability of cell

lines for use in vaccine

production is lack of tumorigenicity. Vero cells represent an example

of a class of cells known as continuous cell lines. They were derived

from African green monkey kidney, and their growth properties and

culture characteristics have many advantages over other cell

substrates for use in vaccine production. We have tested Vero cells

for tumorigenicity in nude mice and in a human muscle organ culture

system, and found a significant increase in their tumorigenic

potential with

increasing passage numbers. Cells at passage 232 and higher produced

nodules in all nude mice inoculated. " (22) [The term " passage " in this

context means the number of times a cell line has been cultured].

There is another very important issue reported in studies that is

evidently being largely ignored as regards long-term vaccine effects

and safety. There is obvious evidence that in the lab, continuous

immortal cell lines react differently between one type of animal

species and another (21, 23). As an example, tissue from one species

will allow the immortal cell to induce a cancerous change more

quickly, in comparison to tissue from a different species. These

results then beg the following

questions. How extensively have these continuous cell lines been

tested on human tissues, and would the results vary from one type of

tissue to another? And what happens over the long term…if an immortal

cell from a vaccine culture makes its way into the final vaccine

product, does it keep dividing in the human body? Another scenario

might suggest the tumor-promoting portion of its DNA inserting into a

viral genome, which then gets injected into the body…what happens at

that

point?

Furthermore, given the evidence that closely-related animal species

(as an example, various species of monkeys) react differently to

immortal cells, do we also need to consider that any one vaccine

intended for all humans might ultimately react differently among the

various races, ethnic groups, and sexes? And what are the effects of

the vaccine contaminants on persons with immune

depression, on the elderly, or on infants?

A letter from the FDA to vaccine manufacturers dated as recently as

March 2001 shows that this issue regarding immortal cell lines is

still of concern. It states, " In general, CBER [Center for Biologics

Evaluation and Research] currently views Vero cells as an acceptable

substrate for viral vaccines, but has residual concerns…CBER

recommends that all products derived from Vero cells be

free of residual intact Vero cells. If your manufacturing process

does not include a validated filtration step or other validated

procedure to clear residual intact Vero cells from the product,

please incorporate such a procedure into your manufacturing

process. " (24) It is now 16 years after the WHO gave a go-ahead (in

1986) to use continuous cell lines for vaccine production (25), and

yet there are very basic safety questions not resolved by the

manufacturers, agencies, and scientific community, much less the

finer details (26, 27). One 1991 study reports: " Cell substrate DNA

was shown to be an abundant contaminant in the clarified preparations

of the Sabin type 1, 2 and 3 poliovaccines produced on a continuous

cell line " (28). Another indicates that immortal cell lines showed 100-

times greater number of DNA recombination events compared to normal

cells (29). As one researcher states, " Using neoplastic cell lines as

substrates for vaccine development could inadvertently result in

viral-viral or viral-cellula r interactions whose biological

consequences are unclear…viral-viral and viral-cellular interactions

can result in the generation of new retroviruses with pathological

consequences. " (30). We note the term " neoplastic " means the quality

of having an abnormal growth characteristic. There is an even

stronger statement dating back to 1990. A scientist in the field

writes, " The present concern is for safety of vaccines made using

transformed or neoplastic mammalian cells that may contain endogenous

contaminating viruses or integrated gene sequences from oncogenic

viruses.

There is also concern for use of plasmid vectors employing promoter

elements from oncogenic viruses. The principal concern for safety

lies with retention of residual DNA in the vaccine, especially since

induction of cancer is a single -cell phenomenon, and a single

functional unit of foreign DNA integrated into the host cell genome

might serve to induce cell transformation as a single event or part

of a series of multifactorial events. Current proposed standards for

vaccines would permit contamination with up to 100 pg [picograms] of

heterologous DNA per dose. This is equivalent to about 10

(8) `functional lengths' of DNA. Total safety would seem to require

complete absence of DNA from the product. " (31)

Please note that 10(8) means 10 to the power of 8, or

100,000,000 " functional lengths " of DNA are allowed per dose of

vaccine . Is there something wrong with this picture? How long will

the general public be subjected to these vaccine products that

according to this information, are nowhere near safe? It has taken,

for instance, approximately forty years for the scientific community

to finally

acknowledge that we have a serious problem as a result of the

contaminatio n of polio vaccines with simian virus 40 (SV40) in the

late 1950s-early 1960s. There has been previous evidence of some

human brain and other tumors containing this virus (32, 33), but the

medical community has been slow to acknowledge a definitive link

between SV40 and cancer in humans. However, two independent research

teams have recently found this virus present in 43% of cases of non-

Hodgkins lymphoma (34, 35). Another study found it present in 36% of

brain tumors, 16% of healthy blood cell samples, and 22% of healthy

semen samples (36). And strangely, SV40 has now been found to infect

children (37). Considering that children of this era, are not

supposed to be receiving the virus

via the vaccine contamination route, this would therefore imply that

SV40 is being transmitted from one human to another, in ways not

previously known.

Other simian viruses may also be contaminating the (Vero) monkey cell

lines used for vaccine production. One example from the literature

cites the contamination presence of SV20, which is a oncogenic simian

adenovirus (38). Simply put, are we in a state of denial that

vaccines are ultimately transmitting viruses, DNA,

and proteins into humans from foreign animal sources (and possibly

unhealthy human sources), and that this may be strongly contributing

to the incredible upsurge in cancers and serious chronic diseases?

Are these foreign animal genes altering your DNA? Furthermore, given

that viral presence can sometimes take years to manifest actual

disease symptoms, and then considering the tendencies of

health-related agencies and corporations towards short-term solutions

and profits, will we ever truly know the long-term consequences until

it is too late?

Other bovine viruses Another contaminating virus found in the calf

serum used for vaccine production is bovine polyomavirus

(polyomaviruses are strongly ssociated with cancer); one pertinent

article is titled " Bovine polyomavirus, a frequent contaminant of

calf serum " (39). Other contaminants include a virus from the

parvovirus family (40); another study cites " virus-like particles "

and " mycoplasma-like agents " in 68% and 20% of the samples,

respectively (41); and yet another mentions the presence of

infectious bovine rhinotracheitis virus (aka bovine herpesvirus 1),

and parainfluenza-3 virus in addition to the common BVDV (42). An

interesting report from 1975 not only affirms the presence

of these viruses in calf serum, and mentions the additional presence

of bovine enterovirus-4, but also tells us that 25% of serum lots

that were pre-tested by the suppliers and " considered to be free of

known viral contaminants " were actually contaminated with bovine

viruses (43). It should be obvious that any bovine blood-borne virus

(including serious retroviruses such as bovine leukemia virus, bovine

visna virus, and bovine immunodeficiency virus) could ultimately end

up in human or animal vaccines via the use of calf serum in the

manufacturing process.

Contamination of calf serum with certain bovine herpesviruses, and

the possible implication for human health, deserves a bit of

scrutiny. It is known that bovine herpesvirus-1 replicates easily in

a human embryo cell line called WI-38 (44). It is also known that

bovine herpesvirus-4 is quite " persistent " in calf serum, and has a

wide host range, including human cells (45). In fact, this

particular virus strongly replicates in two human embryonic cell

lines, WI-38 and MRC-5, enough so to prompt one author to give these

details and a warning: " PCR [polymerase chain reaction] detected a

10,000-times-higher level of BHV-4 [bovine herpesvirus-4] DNA… the

supernatant indicated a 100-fold increase of infectious particles.

Since this is the first bovine (human herpesvirus 8 and

Epstein-Barr virus rela ted) herpesvirus which replicates on human

cells in vitro, the danger of possible human BHV-4 infection should

not be ignored. " (46)

The clincher to this possible contamination, is that these same human

cell lines WI-38 and MRC-5 are two of the most common human cell

lines used to manufacture viral vaccines, (for example - rubella,

chickenpox, smallpox) and these cell lines are of course, commonly

nurtured with calf serum.

Contaminants from chicken sources Some viral vaccines are produced by

growing the virus in chicken eggs. Common human vaccines manufactured

by this method include influenza, mumps, measles, yellow fever, and

others. Like the vaccines that include bovine-source materials, those

derived from chicken embryo culture are plagued with some very

serious viral contamination problems.

Avian leukosis virus (aka avian leukemia virus or ALV) is a

retroviral pathogen that infects large segments of the modern poultry

industry, is present in commercial chickens and eggs, and thus

exposes humans on a consistent basis (47). An interesting virus in

the sense that it can be considered a " parent " , it easily transforms

into a dizzying array of related viruses by hijacking one of numerous

cancer-related gene segments from its host, and inserting it into its

own genome.

Furthermore, it has the additional capability of inserting itself

into the host (including human) genome, hiding out so to speak, and

causing cancerous cell transformation from that location. There is

now much scientific literature available that describes the various

active mechanisms of this and other cancer-associated

viruses (48). Viruses that originate from the " parent " avian leukosis

virus, include the potent Rous sarcoma virus, Rous-associated

viruses, avian myeloblastosis virus, avian myelocytoma virus, avian

erythroblastosis virus, Fujinami sarcoma virus, etc. One group of

researchers studying the mechanism of ALV writes, " Serial passaging

of a retrovirus that does not carry an oncogene on such

cultures leads with a high frequency to the emergence of new viruses

that have transduced oncogenes… " (49). In other words, given the right

growth conditions, ALV can easily transform into other closely

related viruses that are known to be cancer-related. Just how common

is this avian leukosis virus in viral vaccines? The first evidence of

contamination came to light in the 1960s when yellow fever vaccine

was found to contain it (50). Since that time, it is common knowledge

in the industry that this virus (or components thereof) still linger

in human and animal vaccines (51). Indeed, the respected Fields

Virology text (year 2001 edition) states, " At the present time,

vaccines produced by some of the world's 12 manufacturing institutes

are contaminated with avian leukosis virus " (52). One point that

researchers in this field do agree upon, are the presence of ALV,

avian endogenous virus, avian reticuloendotheliosis virus (another

poultry retrovirus), and also an enzyme called reverse transcriptase

(a component of retroviruses) in final vaccine products intended for

human use, especially the mumps, measles, yellow fever, and influenza

vaccines (53, 54, 55). What they do not agree upon are the effects on

humans in terms of transmission, infection, and possible subsequent

disease. A recent study coming out of the U.S. CDC (Centers for

Disease Control), which analyzed frozen blood serum samples from

children

that had received MMR vaccinations, reports no avian viral presence

in these samples (56).

And yet, we see reports from other researchers that make us question

the results of that study. As is often the case with viruses, some

strains will show particular affinities for certain types of tissues

or growth conditions, and ALV is no exception (57). One researcher

makes the effort to explain, " Because of the difficulty in infecting

mammalian cells in vitro with these viruses, it is generally held

that they do not infect humans…Our results show that exposed poultry

workers and subjects with no occupational exposure to these viruses

have antibodies in their sera specifically directed against ALSV

[Avian leucosis/sarcoma viruses]… Further investigation into whether

these findings mean that virus has been integrated into the human

genome is needed, to assess the public health implications of these

results. " (58). He also explains in another article, that given the

known behavior of these viruses in mammalian cellular culture, a

blood serum test will not always provide the correct evidence of

viral presence in the human body (47). In other words, does the virus

(or viral antibodies) need to be actively present in the blood stream

at the time of the blood draw? What if the viral particles have

retreated into other tissues? Thus the CDC study mentioned above may

not have presented an accurate assessment of viral presence, or long-

term effects from the numerous ALVassociated " offspring " viruses.

> This information was posted on another list I'm on (nothing to do

with

> vaccines)

>

> Has anyone heard of this? Anyone have time to research more??????

> Magda - with Informed Parent just met with a group in France, one

of who

> has tons of information on animal contamination of vaccines -

Magda - can

> you get anything in writing in English or ask about this? Jim

West -

> what's your take on this? Anyone else?

> Sheri