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Size Range
Measure in nm = nanometers (1 nm = 1 thousandth of a micrometer or 1 millionth of a mm). Smallest have the size of ribosome (25-30 nm). More than 2,000 bacterial viruses could fit into an average bacterial cell. Bacteriophages average 65 x 200 nm. More than 50 million polioviruses could be accommodated by an average human cell. Animal viruses range in size from the small parvoviruses (20 nm = 0.02 m in diameter). Poxviruses can be as large as bacteria (up to 450 nm in length). Vaccinia (210 nm). Some cylindrical viruses (TMV) are relatively long (800 nm = 0.8 m) but narrow in diameter (15 nm = 0.015 m).
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Host cells are often 1,000 times the volume of a virus infecting it. 1/10 to 1/3 of bacterial size.
Cultivation of Viruses
Initially it was believed that viruses, as obligatory intracellular parasites, could not replicate in any cell-free medium, no matter how complex. This dictum was nullified in 1991 by the demonstration that infectious poliovirus could be synthesized in a cell-free extract of human cells incubated with viral RNA. Despite this finding, most work on viruses is done in: Cultured cells Embryonated eggs Laboratory animals
Poliovirus multiplied in cultured cells not of neuronal origin (1949). This Nobel Prize discovery led the way to:
The propagation of many other viruses in cultured cells. The discovery of new viruses. The development of viral vaccines (poliomyelitis, measles, and rubella). Studies on biochemistry and molecular biology. Resolution of 3D structural composition of viral particles.
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Consist of several cell types and have a limited cell span, no more than 5 or 10 cell divisions. Mostly derived from monkey kidney, human embryonic amnion, kidney, or foreskin; and chicken or mouse embryos. Used in vaccine production (e.g. attenuated poliovirus vaccine strains are propagated in primary monkey kidney cells.
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Often are abnormal in chromosome morphology and number (aneuploid), and can be tumorigenic (e.g. in mice). Can be propagated indefinitely and, like diploid cell strains, they retain viability after being frozen at -70 to -196C. Can also be maintained in suspension cultures, in which cells are continuously stirred by a spinning magnet.
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Formation of viral crystals. Duplication of membranes. Clumps of ribosomes in virions. Enteroviruses and herpes viruses can cause cytopathic effects in 1 to 2 days and destroy the cell monolayer in 3 days. Cytomegalovirus, rubella and some adenoviruses may not produce cytopathic effects for several weeks. Many viruses multiply in cells without causing an obvious cytopathic effect (members of Arenaviridae, Paramyxoviridae and Reoviridae).
Embryonated Eggs
Before the advent of cell culture, many viruses were propagated in embryonated chicken eggs. At 5 to 14 days after fertilization, a hole is drilled in the shell and virus is injected into the site appropriate for its replication. This method is used for the influenza virus, mumps virus, Newcastle virus and avian adenovirus. Robust yields have led to their widespread use in research labs and for vaccine production.
Laboratory Animals
In the early 1900s it was necessary to maintain virus stocks by continuous passage of virus from animal to animal. It led to the selection of viral mutants. Cell culture has large supplanted the use of animals for propagating viruses, but some
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viruses cannot be grown in cell culture. Hepatitis C virus replicates only in chimpanzees. Many viruses that cause human gastroenteritis, such as Norwalk virus, can not be grown in cell culture. Understanding how the immune system or any complex organ system reacts to a virus can not be achieved without research on living animals. The development of viral vaccines, antiviral drugs, and diagnostic tests has also benefitted from research on viral diseases in experimental animals. Examples Polio virus in monkeys led to an understanding of the basis of the disease and the development of the polio vaccine. The development of the hepatitis B vaccine was due to experimental studies with chimpanzees. Animals are increasingly being discarded for the following reasons: 1. Breeding and maintenance of animals infected with pathogenic viruses are expensive. 2. Whole animals are complex systems, in which it is sometimes difficult to discern events. 3. Results obtained are not always reproducible, due to host vaccination. 4. They are rapidly being overtaken by modern science cell culture and molecular biology.
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