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Bovine contaminants of vaccine cell cultures

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Appl Microbiol Biotechnol. 2005 Sep;68(4):456-66. Epub 2005 Oct 26. Related Articles, Links
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Microbiological control in stem cell banks: approaches to standardisation.

Cobo F, Stacey GN, Hunt C, Cabrera C, Nieto A, Montes R, Cortes JL, Catalina P, Barnie A, Concha A.

Stem Cell Bank of Andalucia (Spanish Central Node), Hospital Universitario Virgen de las Nieves, Avda Fuerzas Armadas, 2, 18014, Granada, Spain.

The transplant of cells of human origin is an increasingly complex sector of medicine which entails great opportunities for the treatment of a range of diseases. Stem cell banks should assure the quality, traceability and safety of cultures for transplantation and must implement an effective programme to prevent contamination of the final product. In donors, the presence of infectious micro-organisms, like human immunodeficiency virus, hepatitis B virus, hepatitis C virus and human T cell lymphotrophic virus, should be evaluated in addition to the possibility of other new infectious agents (e.g. transmissible spongiform encephalopathies and severe acute respiratory syndrome). The introduction of the nucleic acid amplification can avoid the window period of these viral infections. Contamination from the laboratory environment can be achieved by routine screening for bacteria, fungi, yeast and mycoplasma by European pharmacopoeia tests. Fastidious micro-organisms, and an adventitious or endogenous virus, is a well-known fact that will also have to be considered for processes involving in vitro culture of stem cells. It is also a standard part of current good practice in stem cell banks to carry out routine environmental microbiological monitoring of the cleanrooms where the cell cultures and their products are prepared. The risk of viral contamination from products of animal origin, like bovine serum and mouse fibroblasts as a "feeder layer" for the development of embryonic cell lines, should also be considered. Stem cell lines should be tested for prion particles and a virus of animal origin that assure an acceptable quality.

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PMID: 16012832 [PubMed - indexed for MEDLINE]

Vet Rec. 2004 Oct 30;155(18):563-4. Related Articles, Links
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Assessment of the risk of transmission of vaccine viruses by using insufficiently cleaned injection devices.

Makoschey B, Beer M.

Virological R & D Department, Intervet International, Wim de Korverstraat 35, NL-5831 AN Boxmeer, The Netherlands.

PMID: 15559423 [PubMed - indexed for MEDLINE]

Vet Microbiol. 2004 Sep 8;102(3-4):131-40. Related Articles, Links
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Comparison of the sensitivity of in vitro and in vivo tests for detection of the presence of a bovine viral diarrhoea virus type 1 strain.

Antonis AF, Bouma A, de Bree J, de Jong MC.

Division of Infectious Diseases and Food Chain Quality, Animal Sciences Group, Wageningen University and Research Centre (WUR), P.O. Box 65, Lelystad AB8200, The Netherlands.

Veterinary vaccines are usually tested for the absence of contaminants. However, the quality control does not always imply that vaccines are not contaminated as, for example, illustrated by the bovine herpes virus 1 (BHV1) vaccine used in The Netherlands in 1999 that contained a small amount of bovine viral diarrhoea virus (BVDV1). Thousands of cows were vaccinated with BHV1 vaccine batches, and the question arose as to whether these small amounts of BVDV1, most likely not detected with in vitro tests, could have infected cattle. More in general, the question was whether the outcome of the in vitro tests, i.e. the in vitro infectivity, was indicative for the infectivity for cattle, i.e. the in vivo infectivity. We therefore carried out in vitro experiments to determine the sensitivity of a BVDV1 isolation assay. In addition, we performed two animal experiments, in which we estimated the lowest dose needed to infect calves with BVDV1. We extrapolated the experimental in vitro and in vivo results from a tissue culture infectious dose (TCID50) to a cattle infectious dose (CID50). We observed a partial response in the calves inoculated with this dose: four out of six calves turned out to be infected. In the tissue culture test, all 20 samples tested negative. The response in vivo, however, was not significantly higher than the in vitro response, which implies that no difference in susceptibility was observed between the animal test and the tissue culture test. Based on the results in our experiments, some cattle may have been infected with BVDV1 after the application of the contaminated BHV1 vaccine during the vaccination campaign. The question remains that how many cattle received contaminated vaccine, and became infected with BVDV1.

PMID: 15327789 [PubMed - indexed for MEDLINE]


Sci Cult (Lond). 2004 Mar;13(1):75-88. Related Articles, Links

Intersecting discourses: MMR vaccine and BSE.

Wilson C.

School of Humanities, Faculty of Arts, Education, and Social Sciences, University of Western Sydney, Locked Bag 1797, South Penrith Distribution Centre, NSW 1797, Australia.

PMID: 15971371 [PubMed - indexed for MEDLINE]


Biologicals. 2003 Sep;31(3):203-8. Related Articles, Links
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Bovine viral diarrhoea virus antigen in foetal calf serum batches and consequences of such contamination for vaccine production.

Makoschey B, van Gelder PT, Keijsers V, Goovaerts D.

Virological R&D Department, Intervet International b.v., Wim de Korverstraat 35, NL-5831 AN, Boxmeer, The Netherlands.

A protocol to test foetal calf serum (FCS) for contamination with bovine viral diarrhoea virus (BVDV) is described. Following this protocol, which combines cell culture methods and detection of pestivirus RNA, seven batches of FCS were tested. Infectious BVDV was detected in four of those batches. One of the remaining batches contained a relatively high number of non-infectious BVDV particles. A sample of this batch was formulated with aluminium hydroxide and aluminium phosphate as adjuvant into an experimental vaccine preparation. This product was injected twice into BVDV seronegative cattle with a 4 week interval. Blood samples taken 4 weeks after the second application were negative for BVDV specific antibodies. Our data stress that detection of BVDV RNA is not sufficient for a complete risk assessment on FCS. Discrimination between infectious and non-infectious BVDV is essential. This can only be achieved by cell culture methods.

PMID: 12935809 [PubMed - indexed for MEDLINE]


Vet Res Commun. 2003 Oct;27(7):577-89. Related Articles, Links
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Experimental infection of calves with bovine viral diarrhoea virus type-2 (BVDV-2) isolated from a contaminated vaccine.

Falcone E, Cordioli P, Tarantino M, Muscillo M, Sala G, La Rosa G, Archetti IL, Marianelli C, Lombardi G, Tollis M.

Istituto Superiore di Sanita, Laboratorio di Medicina Veterinaria, Viale Regina Elena 299, 00161 Rome, Italy.

A non-cytopathic strain of BVDV-2 was isolated from a batch of live infectious bovine rhinotracheitis (IBR) vaccine, and inoculated intranasally into four 3-month-old calves. Severe signs of disease developed by days 4 and 6 in three of the calves, free of BVDV and antibodies to BVDV, that had been exposed to the virus. These calves survived the acute phase of the infection and progressively recovered. BVDV was consistently isolated, or the respective viral RNA was detected, in the buffy coats from blood samples collected starting from days 2 or 4 up to days 11 or 14 after the experimental infection. Viral RNA was also detected in sera from these infected calves until the presence in the serum of virus neutralizing antibodies was demonstrated. By contrast, the only calf having pre-existing neutralizing antibodies to BVDV at the start of the study was protected from the disease. No virus was detected at any time after experimental inoculation of this calf. Genomic characterization of the BVDV-2 isolated in cell cultures, or detected in sera from the experimentally infected animals, revealed 100%, homology in the nucleotide sequence with the BVDV-2 detected as a contaminant of the live IBR virus vaccine. These findings provided evidence of the infective nature of the contaminant BVDV-2 and of its potential to generate disease outbreaks when inoculated into susceptible animals.

PMID: 14609269 [PubMed - indexed for MEDLINE]
Br J Haematol. 2003 Jul;122(1):3-9. Related Articles, Links

The pathophysiology of variant Creutzfeldt-Jacob disease: the hypotheses behind concerns for blood components and products.

Burthem J, Roberts DJ.

Department of Biomedical Sciences, University of Manchester Institute of Science and Technology, Manchester, UK.

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PMID: 12823340 [PubMed - indexed for MEDLINE]

Biologicals. 2002 Dec;30(4):289-96. Related Articles, Links
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Detection and characterization of pestivirus contaminations in human live viral vaccines.

Studer E, Bertoni G, Candrian U.

Official Medicines Control Laboratory Biologika and R&D Unit, Division of Biologicals, Swiss Federal Office of Public Health, P.O. Box 3003, Bern, Switzerland.

In view of the use of potentially contaminated foetal calf serum (FCS) in cell cultures pestiviruses may be present in live viral vaccines. Thirty-six lots of human live viral vaccines produced by three manufacturers were tested for the presence of pestiviruses. Bovine viral diarrhoea virus (BVDV) RNA was detected in 33% of the vaccine lots. All positive results were caused by the mumps component of a single manufacturer. Partial sequences of the 5' untranslated region of BVD viral RNA were determined. The sequences were closely related to that of the NADL strain of BVDV. The amount of BVDV RNA in the vaccines was determined by real-time RT-PCR using the LightCycler. Between 3.3*10(2) and 6.2*10(5) RNA copies per dose were found to be present in the vaccine samples.Additionally, culture tests were done with FCS and human diploid cells used in the vaccine production of the manufacturer whose vaccines were positive by PCR. All attempts to detect virus antigen in MRC-5 human diploid cells or to infect these cells with BVDV failed. This suggests that BVDV RNA detected in human live viral vaccines represents passive carry over of BVDV from contaminated FCS rather than active virus replication in human diploid cells. Our results indicate that contamination with BVDV of FCS used in vaccine production does not appear to be of immediate concern to human health. Furthermore, our results indicate that gamma-irradiation of FCS destroys BVDV particles and is also effective in preventing the presence of BVDV RNA in the vaccines. Copyright 2002 The International Association for Biologicals. Published by Elsevier Science Ltd. All rights reserved.

PMID: 12421586 [PubMed - indexed for MEDLINE]
Am J Health Syst Pharm. 2002 Feb 1;59(3):254-60; quiz 261-3. Related Articles, Links
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Implications of prion-induced diseases for animal-derived pharmaceutical products.

Erstad BL.

Department of Pharmacy Practice and Science, College of Pharmacy, University of Arizona, 1703 E. Mabel Street, Tucson, AZ 85721-0207, USA.

The implications of prion-induced diseases for the use of medications that theoretically could harbor the infectious pathogens are discussed. Prions have been identified as protein particles that lack nucleic acids. There is evidence that prions cause the transmissible neurodegenerative diseases known as transmissible spongiform encephalopathies. Of these diseases, bovine spongiform encephalopathy (BSE) and the human spongiform encephalopathy to which it has been linked, new variant Creutzfeldt-Jakob disease (CJD), have generated the most attention. The first cases of new variant CJD appeared in Britain in the mid-1990s. Ingestion of prion-infected beef remains the only known cause of new variant CJD. No cases of BSE or new variant CJD have been documented in the United States. The time from exposure to the development of clinical sequelae appears to be about 10 years. The median duration of illness is 14 months, and the outcome is invariably death. There is no treatment; currently the only available approach is prevention. There is no reliable method of predicting the number of new cases that might occur because of lack of definitive information on the efficiency of transmission from animals to humans and the number of people currently infected and at risk for infection. The infectivity of medications and plasma fractionation products containing material from cattle with BSE is unknown, but the risk is believed to be very low. No cases of such transmission have been identified. Guidelines to keep the risk of transmission via medications low have been promulgated by FDA, and further research is warranted. There have been no reports of medications or plasma fractionation products being contaminated with the prions that cause new variant CJD. Ongoing vigilance and research are appropriate, however.

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PMID: 11862637 [PubMed - indexed for MEDLINE]

Curr Opin Neurol. 2002 Jun;15(3):333-8. Related Articles, Links
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Neurological adverse events associated with vaccination.

Piyasirisilp S, Hemachudha T.

Division of Neurology, Department of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.

Public tolerance to adverse reactions is minimal. Several reporting systems have been established to monitor adverse events following immunization. The present review summarizes data on neurologic complications followingcination, and provides evidence that indicates whether they were directly associated with the vaccines. These complications include autism (measles vaccine), multiple sclerosis (hepatitis B vaccine), meningoencephalitis (Japanese encephalitis vaccine), Guillain-Barre syndrome and giant cell arteritis (influenza vaccine), and reactions aftermaterials)] are discussed.

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PMID: 12045734 [PubMed - indexed for MEDLINE]

Tijdschr Diergeneeskd. 2001 Mar 15;126(6):189-90. Related Articles, Links

[Detection of bovine virus diarrhea virus in a live bovine herpes virus 1 marker vaccine.]

[Article in Dutch]

Bruschke CJ, Paal HA, Weerdmeester K.

Instituut voor Dierhouderij en Diergezondheid, ID Lelystad, Postbus 65, 8200 AB Lelystad.

In February 1999, 12 Dutch herds were vaccinated with a live bovine herpesvirus 1 vaccine from which bovine virus diarrhea virus (BVDV) could be isolated. All vaccine batches that were on the Dutch market and that had not yet reached the expiry date were tested for BVDV. In total, seven of 82 batches tested were found positive. Batch numbers TX3607, VB3914, VB3915, VB4046, TW3391, and TV3294 were positive for BVDV type 1, and batch number WG4622 was positive for BVDV type 2. This latter batch induced clinical signs of BVDV in an animal experiment with susceptible animals.

PMID: 11285638 [PubMed - indexed for MEDLINE]


Anasthesiol Intensivmed Notfallmed Schmerzther. 2001 Feb;36(2):79-89. Related Articles, Links

[Transmissible spongiform encephalopathies--anesthesiologic and intensive care management]

[Article in German]

Menges T, Langefeld TW, Krumholz W, Hempelmann G.

Abteilung Anaesthesiologie und Operative Intensivmedizin Justus-Liebig-Universitat, Giessen.

The transmissible spongiform encephalopathies (TSE) are known to affect humans and various animals. The bovine spongiform encephalopathy (BSE) and the human Creutzfeldt-Jacob disease (CJD) are among the most notable degenerative disorders caused by prions. Considering the BSE epidemic and the description of a new variant of Creutzfeldt-Jacob disease (nvCJD), which is probably related to bovine spongiform encephalopathy, TSE have recently gained a lot of public attention. Although the causative factors (prions, viruses) are still under discussion, none of the present concepts are explanatory for all aspects of the human CJD. CJD may present as a sporadic, genetic, or infectious illness and there is now considerable concern that bovine prions may have been passed to humans. To exclude transmission of CJD via medical products and instruments, the effectiveness of cleaning, disinfection and sterilization procedures must be firmly established. This manuscript presents an overview to anaesthesiology and intensive care medicine of recommended inactivation procedures and assessed these procedures in the light of the inactivation of prions.

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PMID: 11269018 [PubMed - indexed for MEDLINE]

JAMA. 2001 Feb 7;285(5):532. Related Articles, Links
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From the Centers for Disease Control and Prevention. Public Health Service recommendations for the use of vaccines manufactured with bovine-derived materials.

[No authors listed]

PMID: 11236765 [PubMed - indexed for MEDLINE]
Biologicals. 2000 Mar;28(1):41-6. Related Articles, Links
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Evaluation of vaccines, interferons and cell substrates for pestivirus contamination.

Audet SA, Crim RL, Beeler J.

Laboratory of Pediatrics and Respiratory Viral Diseases, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Bethesda, Maryland 20892, USA.

Pestiviruses are potential contaminants of biological products produced in bovine or porcine cells or manufactured via processes using animal-derived raw materials such as bovine serum. In order to investigate possible contamination of products including those manufactured and/or licensed in the US, 38 lots of viral vaccines and five lots of interferon alpha (IFNalpha) were tested by reverse transcriptase polymerase chain reaction (RT-PCR) for the presence of bovine viral diarrhoea virus (BVDV). All vaccines and interferons were negative for contaminating BVDV RNA when tested by RT-PCR, with the exception of an experimental live viral vaccine that had been produced in BVDV contaminated rabbit kidney cells. Cell lines commonly used to produce biological products and vaccines were experimentally infected with the NADL strain of BVDV to determine if they were permissive for virus replication. MRC-5 and WI-38 cells were not infected. In contrast, Vero, CHO and CEF cells showed evidence of pestivirus infection. Taken together these data suggested that currently licensed viral vaccines were unlikely to be contaminated with pestiviruses. However, cell banks derived from non-human primate, hamster or rabbit kidney cell lines, or cultures of primary chick embryo fibroblasts, may be infected with BVDV if exposed to pestivirus contaminated raw materials during manufacture.

PMID: 10799055 [PubMed - indexed for MEDLINE]
Vaccine. 1999 Oct 14;18(5-6):387-8. Related Articles, Links

Bovine viral diarrhea disease associated with a contaminated vaccine.

Falcone E, Tollis M, Conti G.

Publication Types:

PMID: 10636817 [PubMed - indexed for MEDLINE]

Dev Biol Stand. 1999;99:3-8. Related Articles, Links

Benefits and risks due to animal serum used in cell culture production.

Wessman SJ, Levings RL.

USDA, APHIS, VS, Center for Veterinary Biologics-Laboratory, Ames, Iowa, USA.

Infection with bovine viral diarrhoea virus (BVDV) and other viruses is frequent in the bovine population. In utero infection leads to virus and antibody contamination of foetal and other serum used in cell culture production. The use of contaminated cells for vaccine production may result in contaminated vaccines, which may lead to seroconversion or disease in the vaccinated animal. Contaminated serum or cell cultures may also interfere with the diagnosis of viral infections. Methods for the detection of BVDV and other viruses in serum, cell cultures, seed viruses and vaccines at the CVB-L, and the frequency of detection are described. Reasons for continued use of serum in cell culture production, and the risks of using serum, are discussed.

PMID: 10404869 [PubMed - indexed for MEDLINE]
Dev Biol Stand. 1999;99:41-4. Related Articles, Links

Bovine sera used in the manufacture of biologicals: current concerns and policies of the U.S. Food and Drug Administration regarding the transmissible spongiform encephalopathies.

Asher DM.

Laboratory of Method Development, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, MD 20852-1448, USA.

Since 1993, consistent with its statutory responsibility to ensure that regulated products are safe, pure, and free of << extraneous organisms, >> the United States Food and Drug Administration (FDA) has requested that, with certain exceptions, bovine-derived materials from animals born in or residing in countries where bovine spongiform encephalopathy has occurred, should not be used to manufacture products intended for humans. FDA's Center for Biologics Evaluation and Research (CBER) has specifically recommended that serum used to produce biologicals be obtained from sources << certified to be free from contaminants and adventitious agents, such as the agent responsible for the production of Bovine Spongiform Encephalopathy. >> The United States Department of Agriculture (USDA) has prohibited importation of such serum for use in products. FDA staff are aware that bovine blood, including foetal blood, and placental tissues and fluids that might contaminate foetal serum have not been found to contain the infectious agent of BSE, and that those tissues are considered by most authorities to have little risk for transmitting disease to humans or animals. However, studies of BSE have been limited in size and sensitivity, and several experimental studies of scrapie and CJD in rodents found their blood to be infectious. In addition, a recent unpublished study of BSE (requiring confirmation) reported finding infectivity in the bone marrow of cattle. Possible transmission of BSE from cows to calves, although unlikely to constitute a major mode for maintaining the BSE outbreak, has also not been rigorously ruled out. Considering the special nature of biological products, especially of vaccines intended for widespread use in children, it seems prudent for U.S. regulatory authorities to continue current conservative policies that discourage or prohibit the use of bovine serum from countries with BSE.

Publication Types:

PMID: 10404874 [PubMed - indexed for MEDLINE]

Dev Biol Stand. 1991;75:177-81. Related Articles, Links

Bovine viral diarrhea virus contamination of nutrient serum, cell cultures and viral vaccines.

Levings RL, Wessman SJ.

National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, USDA, Ames, IA 50010.

Bovine viral diarrhea virus (BVDV) infection is common in the bovine population. Infection in utero leads to virus and antibody contamination of the fetal bovine serum used in cell cultures. These contaminants can interfere with diagnosis of viral infection. 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. Methods for detection of BVDV in nutrient serum, cell cultures, seed viruses, and viral vaccines, and the frequency of their detection at the National Veterinary Services Laboratories are discussed.

PMID: 1665461 [PubMed - indexed for MEDLINE]
Dtsch Tierarztl Wochenschr. 1990 Feb;97(2):63-5. Related Articles, Links

Attempts to characterize bovine viral diarrhea virus isolated from cattle after immunization with a contaminated vaccine.

Kreeft HA, Greiser-Wilke I, Moennig V, Horzinek MC.

Regional Institute for Veterinary Health, Gouda, The Netherlands.

Bovine viral diarrhea virus (BVDV) was isolated from 28 animals with a history of immunization against respiratory disease with a vaccine contaminated with BVDV. The vaccine-derived parental virus strain and the 28 isolates were analyzed using 10 monoclonal antibodies (MAbs) directed against different epitopes and antigenic domains on the major envelope glycoprotein of BVDV. None of the isolates displayed a reaction pattern identical with the parental virus. Instead, seven different reaction patterns (#A-G) emerged. Circumstantial evidence indicated that six of these were vaccine related whereas in one case (pattern #F) the origin of the isolate was unclear. The results indicated that BVDV rapidly changed during animal passages and that the tracing of the vaccine contaminant using Mabs was impossible.

PMID: 2155768 [PubMed - indexed for MEDLINE]
Dev Biol Stand. 1989;70:59-66. Related Articles, Links

Detection and elimination of adventitious agents in continuous cell lines.

Erickson GA, Landgraf JG, Wessman SJ, Koski TA, Moss LM.

National Veterinary Services Laboratories, Ames, IA 50010.

The National Veterinary Services Laboratories (NVSL) routinely monitors continuous cell lines (CCL's) used for veterinary biologicals and diagnostic virology. All veterinary biologicals produced in CCL's must follow the master seed concept which limits the use of the master seed CCL to up to 20 passages beyond the passage level characterized and deposited at NVSL. All CLL's are evaluated for the presence of adventitious agents such as mycoplasma, bovine viral diarrhea virus, and other bacteria and viruses. Previously, CCLs were evaluated for tumorigenicity by the Syrian hamster cheek pouch method; however, this procedure has now been eliminated. The adventitious agents most frequently detected in CCL's have been bovine viral diarrhea virus and mycoplasma. Our laboratory has consistently found that the source of bovine viral diarrhea contamination of CCLs has been the use of contaminated fetal bovine cell culture enrichment serum. Gamma irradiation at 2.5-3.5 megarads at -40 degrees C of carefully screened fetal bovine serum has been used in the Diagnostic Virology Laboratory for over 10 years. If the irradiated serum is used at a final concentration of 10 percent, there is no untoward effect on cell susceptibility for virus propagation or cell culture growth. Gamma irradiation has also been demonstrated to be a very efficient inactivator of mycoplasma. Specific conditions utilized by our laboratory to preserve fetal bovine serum cell culture growth factors while eliminating adventitious bovine viral diarrhea virus will be presented.

PMID: 2759356 [PubMed - indexed for MEDLINE]

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