Classification and cultivation of animal viruses

Animal viruses are infectious agents that replicate only inside animal cells, including those of humans. They contain either DNA or RNA as their genetic material and depend entirely on the host cell’s metabolic machinery for replication. For understanding their diversity, behavior, and medical significance, viruses are classified systematically and cultivated using living systems.

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2. Classification of Animal Viruses

Because viruses differ widely in structure and genome type, classification is based on several criteria rather than traditional biological taxonomy.

A. Basis of Classification

The main criteria used for classification of animal viruses include:

CriterionExample/Description
Type of nucleic acidDNA or RNA
Nature of nucleic acidSingle-stranded (ss) or double-stranded (ds)
Sense (polarity)+ sense or − sense (for ssRNA viruses)
Number of strandsSegmented or non-segmented genome
Capsid symmetryIcosahedral, Helical, or Complex
Presence/absence of envelopeEnveloped or Naked virus
Host rangeMammalian, avian, reptilian, etc.
Site of replication in host cellNucleus or cytoplasm
Mode of transmissionAirborne, vector-borne, sexual, etc.

3. Hierarchical Classification System

The International Committee on Taxonomy of Viruses (ICTV) provides the official framework for viral classification.
It recognizes the following taxonomic levels:

Realm → Kingdom → Phylum → Class → Order → Family → Genus → Species

Example:
Order: Mononegavirales
Family: Rhabdoviridae
Genus: Lyssavirus
Species: Rabies virus

4. Classification Based on Type of Genome

Viruses are broadly divided into DNA and RNA viruses, and further into enveloped and non-enveloped types.

A. DNA Animal Viruses

TypeFamilyExampleDisease Caused
Double-stranded DNA (dsDNA)AdenoviridaeAdenovirusRespiratory infections
HerpesviridaeHerpes simplex virus, Varicella-zoster virusCold sores, Chickenpox
PoxviridaeVariola virus, Vaccinia virusSmallpox, Cowpox
PapillomaviridaeHuman papillomavirus (HPV)Warts, Cervical cancer
HepadnaviridaeHepatitis B virusHepatitis B
Single-stranded DNA (ssDNA)ParvoviridaeParvovirus B19Erythema infectiosum (fifth disease)

B. RNA Animal Viruses

TypeFamilyExampleDisease Caused
Double-stranded RNA (dsRNA)ReoviridaeRotavirusInfantile diarrhea
Single-stranded RNA (+ sense)PicornaviridaePoliovirus, RhinovirusPolio, Common cold
TogaviridaeRubella virusGerman measles
FlaviviridaeDengue virus, Zika virus, Yellow fever virusDengue, Zika, Yellow fever
CoronaviridaeSARS-CoV, MERS-CoV, SARS-CoV-2COVID-19
Single-stranded RNA (− sense)OrthomyxoviridaeInfluenza virusInfluenza (Flu)
ParamyxoviridaeMeasles, Mumps virusMeasles, Mumps
RhabdoviridaeRabies virusRabies
FiloviridaeEbola virusHemorrhagic fever
Single-stranded RNA (diploid)RetroviridaeHIV (Human Immunodeficiency Virus)AIDS

C. Other Special Groups

TypeExampleFeature
ArbovirusesDengue, Zika, ChikungunyaArthropod-borne
Oncogenic virusesEpstein–Barr virus, HPVCause cancer
Enteric virusesPoliovirus, RotavirusTransmitted via fecal–oral route

5. The Baltimore Classification System

Proposed by David Baltimore (1971), this system classifies viruses based on the relationship between their genome type and mRNA production.

GroupGenome TypeExample
IdsDNAAdenovirus, Herpesvirus
IIssDNAParvovirus
IIIdsRNAReovirus
IV(+) ssRNAPoliovirus, Dengue virus
V(−) ssRNAInfluenza, Rabies virus
VIssRNA (RT; retrovirus)HIV
VIIdsDNA (RT)Hepatitis B virus

6. Cultivation of Animal Viruses

Since viruses require living host cells for replication, they cannot be grown on artificial media (like bacteria).
They are cultivated using living systems that provide host cells and metabolic support.

A. Purpose of Cultivation

  1. Isolation and identification of viruses from clinical samples
  2. Study of viral replication, morphology, and pathogenesis
  3. Preparation of vaccines
  4. Research on antiviral agents
  5. Serological and diagnostic testing

B. Methods of Cultivation

1. Animal Inoculation

  • One of the earliest and classical methods.
  • Involves inoculating virus samples into susceptible live animals (e.g., mice, rabbits, guinea pigs, monkeys).
  • Observations include symptoms, lesions, or death of the animal.

Advantages:

  • Demonstrates infectivity and pathogenicity.
  • Used for rabies virus and encephalitis viruses.

Disadvantages:

  • Expensive, ethical concerns, and variable results.

2. Embryonated (Fertilized) Egg Inoculation

  • Introduced by Goodpasture (1931).
  • Fertilized chicken eggs (7–12 days old) are used.
  • The virus is injected into specific parts of the egg.
Site of InoculationExample of Virus
Chorioallantoic membrane (CAM)Poxviruses, Herpesvirus
Allantoic cavityInfluenza, Newcastle disease virus
Amniotic cavityInfluenza virus
Yolk sacRickettsiae, some viruses (e.g., Herpesvirus)

Advantages:

  • Economical, sterile environment, easy to maintain.
  • Used extensively for vaccine production (e.g., Influenza vaccine).

3. Cell Culture Technique

The most widely used and reliable method for virus cultivation today.

It involves growing animal or human cells in controlled laboratory conditions.

Types of Cell Cultures:

TypeSourceLifespanExample/Use
Primary cell cultureNormal tissues (kidney, fibroblast)Short (few divisions)Used for rabies virus
Diploid cell strainHuman embryonic tissuesModerate (≈50 generations)Used for measles, polio
Continuous cell lineCancer cells (HeLa, Vero)ImmortalUsed for large-scale viral production, diagnostics

Detection of Virus Growth in Cell Culture

Viral infection produces characteristic Cytopathic Effects (CPEs):

Type of CPEDescriptionExample
Cell rounding/detachmentCells shrink, lose adherencePoliovirus
Syncytium formationFusion of cells → multinucleated giant cellsMeasles, RSV
Inclusion bodiesAccumulation of viral material in cytoplasm/nucleusNegri bodies (Rabies), CMV inclusions
HemadsorptionRBCs adhere to infected cellsInfluenza virus
Cell lysisDestruction of host cellsEnteroviruses

C. Other Advanced Methods

  1. Organ Culture: Small pieces of animal tissue (e.g., tracheal ring) maintained in vitro.
    Used for respiratory viruses.
  2. Molecular Cloning Systems: Recombinant DNA and plasmid-based replication of viral genes.
  3. Transgenic Animal Models: Mice genetically engineered to express viral receptors (for HIV, COVID-19 studies).

7. Preservation of Animal Viruses

For research and vaccine production, viruses are preserved by:

  • Lyophilization (freeze-drying)
  • Deep freezing (−70°C to −196°C)
  • Cryopreservation in liquid nitrogen

Table

FeatureDNA VirusesRNA Viruses
GenomeDNA (ss or ds)RNA (ss or ds)
Site of replicationNucleus (usually)Cytoplasm (usually)
ExamplesHerpesvirus, PoxvirusInfluenza, Rabies, HIV
CultivationEmbryonated eggs, cell cultureSame methods
Diagnostic markerIntranuclear inclusion bodiesCytoplasmic inclusion bodies

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