303
can become exposed to infectious organisms or envi-
ronmentally hazardous agents. Regulations relating to
the transportation of biological agents within Europe
have been recently updated (e.g. CHIPS II 1994, UK;
EC Commission Directive 92/1-3/EEC) and for global
transportation by air IATA also have special regula-
tions (DSM). Whatever the country of origin the person
despatching cultures should remember that they have
a responsibility to ensure that the recipient is aware of
any hazards relating to each culture and that the recipi-
ent laboratory has appropriate facilities and staff to use
the culture safely.
Conclusions
The hazards posed to the environment by the acci-
dental release of baculovirus expression vectors can
be put into perspective by the results obtained from
experiments in which AcNPV was released deliber-
ately into the field (Bishop et al
.,
1992). Polyhedrin
positive viruses will persist in soil and on leaf surfaces
for periods comprising weeks and months. Howev-
er, polyhedrin negative viruses (similar to those used
as expression vectors) do not survive in similar sit-
uations. In consequence, accidental release of bac-
ulovirus expression vectors poses negligible hazard.
The risk of such a release will largely depend on the
skill of the operators. This does not take into account
the hazard posed by the recombinant product which is
being made by the virus-infected insect cell. Synthe-
sis of a mammalian-specific toxin, of course, would
require particularly careful manipulation of the virus-
infected cell culture.
The fact that insect cell lines represent an undefined
risk, in terms of carriage of adventitious agents means
that their containment should be maintained at a mini-
mum of the European containment level 2. Where the
tissue of origin has a high risk of infection with human
pathogens or where cells may have been used in a virus
culture laboratory then appropriate testing is advisable.
Careful risk assessment respecting the scale of work
and whole procedures (in addition to individual assess-
ment of agents and reagents) will ensure safe working
conditions for laboratory staff. If applied properly safe-
ty procedures will also succeed in encouraging clean,
efficient and well documented work procedures which
are synonymous with the economical use of time and
resources and good science.
References
ACDP (1990) Advisory committee on dangerous pathogens accord-
ing to hazard and categories of containment. 2nd Edition. HMSO
Books, London, UK.
Bishop DHL, Cory JS & Possee RD (1992) The use of genetically
engineered virus insecticides to control insect pests. In: Release
of genetically engineered and other organisms: 137–146. Day M
& Fry JC (eds) Cambridge University Press, Cambridge, UK.
BS5726 British Standards Parts l–4, HMSO Books, London, UK.
Chadee DD & Le Maitre (1990) Ants: Potential mechanical vectors
of hospital infections in Trinidad. Trans. Roy. Soc. Trop. Med.
Hyg.
84:
297.
CHIPS II. Chemicals (Hazards Information and Packaging for Sup-
ply) Regulations 1994. HSE books, Sudbury, UK.
DelGiudice RA & Gardella RS (1984) Mycoplasma infection of cell
cultures: Effects, incidence and detection. In: In vitro Monograph
5. Uses and standardisation of vertebrate cell culture: 104–115.
Tissue Culture Association, Gaithersburg, USA.
DSM Information on current international transportation and air-
freight regulations for infectious organisms is available from the
DSM, Mascheroder Weg 1b, D–38124 Braunschweig, Germany.
Erickson GA, Bolin SR & Landgraf JG (1991) Viral contamination
of fetal bovine serum used for tissue culture: Risks and concerns.
Dev. Biol. Stand. 75: 173–175.
Fotedar R, Shriniwas UB & Verma A (1991) Cockroaches
(
Blattella
germanica
)
as carriers of microorganisms of medical importance
in hospitals. Epidemiol. Infect. 107: 181–187.
Frommer W, Archer L & Boon B et al. (1993) Recommendations for
safe work with animal and human cell cultures concerning poten-
tial human pathogens: safe biotechnology (5)
.
Applied Microbi-
ology and Biotechnology 39: 141–147.
Hirumi H (1974) Mycoplasma and viral contamination of insect cell
cultures. Proceedings of the US. Japan cooperative conference on
invertebrate tissue culture: Applications in fundamental research,
Tokyo, Japan.
Hirumi H (1976) Viral, microbial and extrinsic cell contamination
of insect cell cultures. In: Invertebrate Tissue Culture Research
Applications: 233–268. K Maramorosch (ed.) Academic Press,
New York, USA.
Kuzio JA, Jaques R & Faulkner P (1989) Identification of p74 a gene
essential for virulence of baculovirus occlusion bodies. Virology
173:
759–763.
Ng ML & Westaway EG (1980) Establishment of persistent infec-
tions by flaviviruses in Aedes albopictus cells. In: Invertebrate
systems in vitro: 389–402. Kurstak E, Maramorosch K & Duben-
dorfer A (eds) Elsevier/North Holland, Amsterdam.
Plus N (1980) Further studies on the origin of the endogenous viruses
of Drosophila melanogaster cell lines. In: Invertebrate systems
in vitro: 435–439. Kurstak E, Maramorosch K & Dubendorfer A
(eds) Elsevier/North Holland, Amsterdam.
Rady MH, Abdel-Raouf N, Labib I & Merdan AI (1992) Bacterial
contamination of the housefly, Musca domestica
,
collected from
four Ciaro hospitals. J. Egypt. Soc. Parasitol. 22: 279–288.
Stacey GN & Sheeley HJ (1994) Have biosafety issues in cell culture
been overlooked. J. Chem. Tech. Biotechnol. 61: 95–96.
Steiner T & McGarrity G (1983) Mycoplasma Infection of Insect
cell cultures. In vitro 19: 672–682.
Vaughn JL (1991) Insect cells: Adventitious agents. Dev. Biol. Stand.
76: 319–324.