Viruses are obligate intracellular parasites that can only replicate inside living host cells. There are several methods for cultivating or growing viruses, including inoculation into animals, embryonated eggs, and cell/tissue culture. Inoculation into animals allows study of viral replication and disease but is expensive and difficult. Embryonated eggs are widely used as they are inexpensive and support growth of many viruses. Tissue/cell culture has replaced eggs for many viruses as it is more convenient and sensitive. Various cell types, including primary cultures, continuous cell lines, and explant cultures can support viral growth. Detection of viral growth involves monitoring for cytopathic effects, hemadsorption, interference, transformation, or metabolic inhibition.
This document discusses various methods for cultivating and detecting viruses. It begins by describing three primary purposes of virus cultivation: to isolate viruses from clinical samples, conduct research on viral structure and effects on host cells, and produce viruses for vaccines. It then explains three main cultivation methods: animal inoculation using mice or monkeys, embryonated egg inoculation, and tissue culture using primary cells or continuous cell lines. Detection methods for viruses in cell culture include observing cytopathic effects, hemadsorption, metabolic inhibition, interference, and immunofluorescence.
Chickenpox is caused by the varicella-zirus, an enveloped virus 200nm in diameter and 300nm in length with linear, double-stranded segmented DNA. It spreads through coughing, sneezing or sharing food/drink and causes skin lesions and fever. Treatment can be done at home with calamine lotion but serious cases require seeing a doctor. The chickenpox vaccine can prevent it, especially for young children, and having it once makes one immune from getting it again.
The document discusses host and virus interaction, including the stages of viral infection and types of virus-cell interaction. It provides details on:
1. The stages of viral infection include primary infection of cells at the site of entry, viremia where the virus enters the bloodstream, and secondary infection of other organs.
2. Virus-cell interaction can be cytocidal, causing cell death, or non-cytocidal where cells survive infection. Persistent and latent infections allow long-term virus maintenance in cells.
3. Common sites of viral entry into the host include respiratory, urogenital, ocular, and skin routes, which viruses access by exploiting breaches in barriers.
Viruses require living host cells to replicate and are cultivated using various methods. They can be grown in laboratory animals, embryonated eggs, or tissue/cell cultures. Animal cultivation allows studying pathogenesis but is expensive and difficult. Eggs are inexpensive and facilitate growth of viruses like influenza in specific areas. Tissue cultures, especially primary cultures and diploid cell lines, support growth of many viruses and are widely used. Isolated viruses are identified using techniques observing cytopathic effects, hemagglutination, interference, or immunofluorescence. Cell cultures are now the primary method for isolating and identifying viruses.
The document discusses methods for cultivating viruses. It states that viruses require living host cells to support their replication, as they are obligate intracellular parasites. The primary purposes of cultivating viruses are to isolate and identify viruses, demonstrate viral presence for diagnosis, and allow research on viral structure and effects. Common cultivation methods discussed include cell lines, embryonated eggs, and experimental animals. Specific tissues or sites within eggs are used depending on the virus. Cytopathic effects and other indicators help identify viruses grown through cultivation.
General virology - Introduction, by Dr. Himanshu KhatriDrHimanshuKhatri
This document provides an overview of general virology. It discusses the properties of viruses including their inability to replicate without host cell machinery and resistance to antibiotics. Viruses have a protein coat called a capsid that protects their nucleic acids. They can be enveloped or non-enveloped. The viral life cycle involves attachment, penetration, uncoating, biosynthesis of proteins and nucleic acids, assembly, and release. Viruses can mutate or undergo recombination. They are classified based on having DNA or RNA and examples of virus families in each category are provided along with their key properties.
RhabdoVirus is a single stranded, linear, negative sense, non-segmented RNA virus that is enveloped and bullet shaped. It multiplies in the cytoplasm. Rabies virus is an example. Rabies virus enters through bites from rabid animals and is carried in the saliva, depositing at the wound site. If untreated, 50% of those exposed will develop rabies as the virus multiplies in muscles, connective tissue and nerves before infecting the central nervous system. Symptoms include headache, fever, sore throat, nervousness, confusion, pain or tingling at the bite site, hallucinations, hydrophobia, paralysis, and eventually coma and death. Diagnosis involves antigen detection
This document discusses various methods for cultivating and detecting viruses. It begins by describing three primary purposes of virus cultivation: to isolate viruses from clinical samples, conduct research on viral structure and effects on host cells, and produce viruses for vaccines. It then explains three main cultivation methods: animal inoculation using mice or monkeys, embryonated egg inoculation, and tissue culture using primary cells or continuous cell lines. Detection methods for viruses in cell culture include observing cytopathic effects, hemadsorption, metabolic inhibition, interference, and immunofluorescence.
Chickenpox is caused by the varicella-zirus, an enveloped virus 200nm in diameter and 300nm in length with linear, double-stranded segmented DNA. It spreads through coughing, sneezing or sharing food/drink and causes skin lesions and fever. Treatment can be done at home with calamine lotion but serious cases require seeing a doctor. The chickenpox vaccine can prevent it, especially for young children, and having it once makes one immune from getting it again.
The document discusses host and virus interaction, including the stages of viral infection and types of virus-cell interaction. It provides details on:
1. The stages of viral infection include primary infection of cells at the site of entry, viremia where the virus enters the bloodstream, and secondary infection of other organs.
2. Virus-cell interaction can be cytocidal, causing cell death, or non-cytocidal where cells survive infection. Persistent and latent infections allow long-term virus maintenance in cells.
3. Common sites of viral entry into the host include respiratory, urogenital, ocular, and skin routes, which viruses access by exploiting breaches in barriers.
Viruses require living host cells to replicate and are cultivated using various methods. They can be grown in laboratory animals, embryonated eggs, or tissue/cell cultures. Animal cultivation allows studying pathogenesis but is expensive and difficult. Eggs are inexpensive and facilitate growth of viruses like influenza in specific areas. Tissue cultures, especially primary cultures and diploid cell lines, support growth of many viruses and are widely used. Isolated viruses are identified using techniques observing cytopathic effects, hemagglutination, interference, or immunofluorescence. Cell cultures are now the primary method for isolating and identifying viruses.
The document discusses methods for cultivating viruses. It states that viruses require living host cells to support their replication, as they are obligate intracellular parasites. The primary purposes of cultivating viruses are to isolate and identify viruses, demonstrate viral presence for diagnosis, and allow research on viral structure and effects. Common cultivation methods discussed include cell lines, embryonated eggs, and experimental animals. Specific tissues or sites within eggs are used depending on the virus. Cytopathic effects and other indicators help identify viruses grown through cultivation.
General virology - Introduction, by Dr. Himanshu KhatriDrHimanshuKhatri
This document provides an overview of general virology. It discusses the properties of viruses including their inability to replicate without host cell machinery and resistance to antibiotics. Viruses have a protein coat called a capsid that protects their nucleic acids. They can be enveloped or non-enveloped. The viral life cycle involves attachment, penetration, uncoating, biosynthesis of proteins and nucleic acids, assembly, and release. Viruses can mutate or undergo recombination. They are classified based on having DNA or RNA and examples of virus families in each category are provided along with their key properties.
RhabdoVirus is a single stranded, linear, negative sense, non-segmented RNA virus that is enveloped and bullet shaped. It multiplies in the cytoplasm. Rabies virus is an example. Rabies virus enters through bites from rabid animals and is carried in the saliva, depositing at the wound site. If untreated, 50% of those exposed will develop rabies as the virus multiplies in muscles, connective tissue and nerves before infecting the central nervous system. Symptoms include headache, fever, sore throat, nervousness, confusion, pain or tingling at the bite site, hallucinations, hydrophobia, paralysis, and eventually coma and death. Diagnosis involves antigen detection
Virology is the study of viruses and their relationship with hosts. Viruses are acellular organisms that can only replicate inside host cells. They have nucleic acid genomes and use host cell machinery to assemble new viral particles. Viruses come in a variety of shapes and sizes, and some have envelopes derived from host cell membranes. They enter host cells, express their genes, replicate their genomes, assemble new viral particles, and exit host cells to infect new targets. Viruses are cultivated using various methods including cell cultures, embryonated eggs, and animal models to study viral replication and pathogenesis.
Viruses cannot be grown on culture media like bacteria, but require living host cells. They can be cultivated in vivo in whole organisms or in vitro in cell cultures. For bacteriophages, a bacterial lawn is used where plaques indicate viral growth. Common methods of culturing animal viruses involve embryonated eggs, live animals, and cell cultures. Cell cultures are now most widely used and include primary cells, diploid cells, and continuous cell lines that can be maintained indefinitely. Organ cultures also allow culturing while preserving cell morphology but are less commonly used now. Virus cultivation is important for diagnosing diseases, producing vaccines and research.
Viruses can only be grown within living host cells. The document discusses three main methods for cultivating viruses: inoculation into animals, embryonated eggs, and tissue culture. It provides details on each method, including commonly used animal and egg types, inoculation sites, advantages and limitations. Tissue culture involves growing viruses in cultured cells, including primary cultures that can only grow briefly, diploid cell strains for limited passages, and continuous cell lines that can divide indefinitely.
This document discusses Leptospira, a pathogenic spirochete bacteria that causes leptospirosis. It is a zoonotic disease transmitted through contact with infected animal urine. Leptospira has a thin, coiled morphology and grows slowly in culture. It can penetrate skin or mucous membranes, causing a mild flu-like illness or the potentially fatal Weil's disease characterized by jaundice and kidney damage. Diagnosis involves culture, serology, or ELISA testing of blood, urine or CSF. Treatment is with penicillin or doxycycline. Prevention involves controlling rodent populations, vaccinating livestock, and taking protective measures during high risk activities like farming.
Staphylococci are Gram positive cocci that commonly cause localized suppurative lesions. Staphylococcus aureus is an important pathogenic species that can cause a variety of infections like food poisoning, toxic shock syndrome, and nosocomial infections. S. aureus has developed resistance to many antibiotics like penicillin through production of beta-lactamases. Methicillin resistant S. aureus strains are a major concern as they are resistant even to methicillin and related antibiotics.
Obligate intracellular, unable to self-replicate.
Once inside living cells, viruses induce the host cell to synthesize virus particles.
The genome is either DNA or RNA (single or double stranded).
Viruses do not have a system to produce ATP.
Viruses range in size from 25 to 270 nm.
Viral tropism!!
The classification of viruses is based on nucleic acid type, size and shape of virion, and presence or absence of an envelope.
Viral Structure
I . Virion is the entire viral particle.
2. Capsid is the protein coat that encloses the genetic material.
3. Capsomer is the protein subunit that makes up the capsid.
4. Nucleocapsid is composed of the capsid and genetic material.
5. The envelope is the outer coating composed of a phospholipid bilayer, which is composed of viral-encoded glycoproteins and sometimes viral encoded matrix proteins. The envelope is derived from a host cell's membrane.
Some viruses use the plasma membrane, whereas others use endoplasmic reticulum, Golgi, or nuclear membranes. Naked nucleocapsids are viruses with no envelopes.
1. The document discusses the structure and classification of several important animal viruses: rhabdovirus, influenza virus, paramyxovirus, hepatitis B virus, and retroviruses.
2. It provides information on their genetic material, morphology from electron micrographs, and proteins. Rhabdovirus is bullet-shaped with a helical nucleocapsid. Influenza virus is spherical with segmented negative-sense RNA.
3. Paramyxovirus is spherical and polymorphic with a helical nucleocapsid. Hepatitis B virus is spherical and enveloped with circular DNA. Retroviruses have spherical cores with two copies of positive-sense RNA and the enzyme reverse transcript
This document discusses the Reoviridae family of viruses. It provides information on Rotavirus, including that it is the most common cause of severe diarrhea in infants and children worldwide, with about 500,000 child deaths annually. Rotavirus has a double-stranded RNA genome and infects the intestinal tract. It spreads easily between young children and causes symptoms like vomiting, diarrhea, and dehydration. While there is no specific treatment, prevention through vaccination is available.
This document discusses virus isolation and cultivation. It explains that viruses require living cells to replicate and the primary purposes of cultivation are to isolate viruses from clinical samples, conduct research, and produce vaccines. Viruses can be cultivated in experimental animals, embryonated eggs, or tissue culture. Tissue culture is now most commonly used and involves growing viruses in primary cells, diploid cell strains, or continuous cell lines. The document describes different tissue culture methods and how viral growth can be detected using cytopathic effects, hemadsorption, interference, transformation, and microscopy.
Largest viruses that infect vertebrates
Can be seen under light microscope
Poxvirus diseases are characterized by skin lesions – localized or generalized
Important diseases caused by poxviruses are-
Smallpox
Monkeypox
Cowpox
Tanapox
Molluscum contagiosum
The document defines various terminologies related to microbial pathogenicity and infection. It discusses terms like saprophytes, parasites, commensals, pathogens, opportunistic pathogens, infection, colonization, infestation, and more. It also classifies infections based on factors like source, clinical manifestation, epidemiological patterns, and more. Finally, it covers various mechanisms of microbial pathogenicity like adhesion, invasiveness, toxins, inhibition of phagocytosis, and more.
The document discusses the history and development of medical microbiology from its earliest concepts to modern times. It describes key contributions from Antonie van Leeuwenhoek, who first observed microorganisms under a microscope; Louis Pasteur, considered the father of microbiology; Joseph Lister, the father of modern surgery; and Robert Koch, the father of bacteriology. It was during the 1800s that microbiology emerged as a scientific discipline, aided by advances like staining techniques, pure culture isolation, and Koch's postulates for identifying microbes that cause disease. The early 1900s marked the discovery of viruses and antibiotics like penicillin. Overall, the document provides a comprehensive overview of the scientific milestones that established microbiology
Viruses can be cultivated within suitable host cells like bacteria to allow for viral replication. Bacteriophages are commonly grown by adding them to a culture of bacteria in a growth medium. The bacteriophages will multiply within the bacteria, destroying cells and releasing new viral particles. Viruses are also cultivated in tissue culture systems using cell lines grown in suspension. The virus infects the host cells, forcing them to produce new virus copies until the cells are lysed. Viral titers can be determined using plaque assays, where viral plaques are counted, or animal inoculation studies.
This presentation contains 45 slides on general virology comprises of topics on viral classification, transmission, pathogenesis, viral cytopathic effect, stages of viral infections, antiviral drugs and viral vaccines. It also have a slide noting an outline of laboratory diagnosis of viral infection. This power point presentation was designed for medical students, nurses and academicians teaching virology and microbiology in medical universities, schools or colleges.
Fungi are a separate kingdom from plants and animals that play an important role in decomposition. Dermatomycoses are fungal infections of the skin, hair, or nails caused by dermatophytes from three genera: Trichophyton, Microsporum, and Epidermophyton. Dermatophytes are transmitted through direct or indirect contact with infected skin or hair. Diagnosis involves microscopic examination of skin or hair samples in potassium hydroxide or culturing samples on growth media. Treatment depends on the severity but may include topical imidazole creams for mild cases or oral antifungals like griseofulvin for more severe or persistent infections.
Louis Pasteur and Robert Koch were two of the founders of bacteriology. Pasteur developed the process of pasteurization to prevent contamination and disproved spontaneous generation. He also discovered vaccines for anthrax, cholera, and rabies. Koch isolated pure bacterial cultures and invented techniques like the hanging drop method. He discovered the specific bacteria that cause anthrax, tuberculosis, and cholera and proposed Koch's postulates for identifying the microorganisms that cause diseases. Both scientists greatly advanced the germ theory of disease.
Sporulation is the process of spores formation by bacteria during unfavorable condition
Germination is the process of spores return to give vegetative cells under favorable condition.
Bacteriophage typing involves identifying viruses that infect bacteria up to the strain level. It is used to control infections and for epidemiological purposes. Strains are differentiated based on phenotypic and genotypic differences, known as 'typing'. Various typing methods exist, including phenotypic methods like biotyping, phage typing, serotyping, and genotypic methods like plasmid profiling, ribotyping, and pulse field gel electrophoresis. Each method has advantages and limitations in terms of typeability, reproducibility, discriminatory power, and ease of use. Molecular typing techniques are increasingly used for outbreak detection and epidemiological investigations.
Cultivation of viruses uhf copy - copyheena thakur
This document discusses viruses and methods for cultivating viruses. It describes viruses as obligate intracellular parasites that can only multiply inside living host cells. The three main methods for cultivating viruses discussed are inoculation of viruses into animals, embryonated eggs, and tissue culture. For animal inoculation, mice are commonly used and viruses can be introduced via different routes. Embryonated eggs provide a suitable environment for virus growth and isolation. Tissue culture involves culturing cells or tissue fragments, and cell lines provide indefinite growth. Detection of virus growth involves monitoring for cytopathic effects, hemadsorption, interference, and other methods.
Virology is the study of viruses and their relationship with hosts. Viruses are acellular organisms that can only replicate inside host cells. They have nucleic acid genomes and use host cell machinery to assemble new viral particles. Viruses come in a variety of shapes and sizes, and some have envelopes derived from host cell membranes. They enter host cells, express their genes, replicate their genomes, assemble new viral particles, and exit host cells to infect new targets. Viruses are cultivated using various methods including cell cultures, embryonated eggs, and animal models to study viral replication and pathogenesis.
Viruses cannot be grown on culture media like bacteria, but require living host cells. They can be cultivated in vivo in whole organisms or in vitro in cell cultures. For bacteriophages, a bacterial lawn is used where plaques indicate viral growth. Common methods of culturing animal viruses involve embryonated eggs, live animals, and cell cultures. Cell cultures are now most widely used and include primary cells, diploid cells, and continuous cell lines that can be maintained indefinitely. Organ cultures also allow culturing while preserving cell morphology but are less commonly used now. Virus cultivation is important for diagnosing diseases, producing vaccines and research.
Viruses can only be grown within living host cells. The document discusses three main methods for cultivating viruses: inoculation into animals, embryonated eggs, and tissue culture. It provides details on each method, including commonly used animal and egg types, inoculation sites, advantages and limitations. Tissue culture involves growing viruses in cultured cells, including primary cultures that can only grow briefly, diploid cell strains for limited passages, and continuous cell lines that can divide indefinitely.
This document discusses Leptospira, a pathogenic spirochete bacteria that causes leptospirosis. It is a zoonotic disease transmitted through contact with infected animal urine. Leptospira has a thin, coiled morphology and grows slowly in culture. It can penetrate skin or mucous membranes, causing a mild flu-like illness or the potentially fatal Weil's disease characterized by jaundice and kidney damage. Diagnosis involves culture, serology, or ELISA testing of blood, urine or CSF. Treatment is with penicillin or doxycycline. Prevention involves controlling rodent populations, vaccinating livestock, and taking protective measures during high risk activities like farming.
Staphylococci are Gram positive cocci that commonly cause localized suppurative lesions. Staphylococcus aureus is an important pathogenic species that can cause a variety of infections like food poisoning, toxic shock syndrome, and nosocomial infections. S. aureus has developed resistance to many antibiotics like penicillin through production of beta-lactamases. Methicillin resistant S. aureus strains are a major concern as they are resistant even to methicillin and related antibiotics.
Obligate intracellular, unable to self-replicate.
Once inside living cells, viruses induce the host cell to synthesize virus particles.
The genome is either DNA or RNA (single or double stranded).
Viruses do not have a system to produce ATP.
Viruses range in size from 25 to 270 nm.
Viral tropism!!
The classification of viruses is based on nucleic acid type, size and shape of virion, and presence or absence of an envelope.
Viral Structure
I . Virion is the entire viral particle.
2. Capsid is the protein coat that encloses the genetic material.
3. Capsomer is the protein subunit that makes up the capsid.
4. Nucleocapsid is composed of the capsid and genetic material.
5. The envelope is the outer coating composed of a phospholipid bilayer, which is composed of viral-encoded glycoproteins and sometimes viral encoded matrix proteins. The envelope is derived from a host cell's membrane.
Some viruses use the plasma membrane, whereas others use endoplasmic reticulum, Golgi, or nuclear membranes. Naked nucleocapsids are viruses with no envelopes.
1. The document discusses the structure and classification of several important animal viruses: rhabdovirus, influenza virus, paramyxovirus, hepatitis B virus, and retroviruses.
2. It provides information on their genetic material, morphology from electron micrographs, and proteins. Rhabdovirus is bullet-shaped with a helical nucleocapsid. Influenza virus is spherical with segmented negative-sense RNA.
3. Paramyxovirus is spherical and polymorphic with a helical nucleocapsid. Hepatitis B virus is spherical and enveloped with circular DNA. Retroviruses have spherical cores with two copies of positive-sense RNA and the enzyme reverse transcript
This document discusses the Reoviridae family of viruses. It provides information on Rotavirus, including that it is the most common cause of severe diarrhea in infants and children worldwide, with about 500,000 child deaths annually. Rotavirus has a double-stranded RNA genome and infects the intestinal tract. It spreads easily between young children and causes symptoms like vomiting, diarrhea, and dehydration. While there is no specific treatment, prevention through vaccination is available.
This document discusses virus isolation and cultivation. It explains that viruses require living cells to replicate and the primary purposes of cultivation are to isolate viruses from clinical samples, conduct research, and produce vaccines. Viruses can be cultivated in experimental animals, embryonated eggs, or tissue culture. Tissue culture is now most commonly used and involves growing viruses in primary cells, diploid cell strains, or continuous cell lines. The document describes different tissue culture methods and how viral growth can be detected using cytopathic effects, hemadsorption, interference, transformation, and microscopy.
Largest viruses that infect vertebrates
Can be seen under light microscope
Poxvirus diseases are characterized by skin lesions – localized or generalized
Important diseases caused by poxviruses are-
Smallpox
Monkeypox
Cowpox
Tanapox
Molluscum contagiosum
The document defines various terminologies related to microbial pathogenicity and infection. It discusses terms like saprophytes, parasites, commensals, pathogens, opportunistic pathogens, infection, colonization, infestation, and more. It also classifies infections based on factors like source, clinical manifestation, epidemiological patterns, and more. Finally, it covers various mechanisms of microbial pathogenicity like adhesion, invasiveness, toxins, inhibition of phagocytosis, and more.
The document discusses the history and development of medical microbiology from its earliest concepts to modern times. It describes key contributions from Antonie van Leeuwenhoek, who first observed microorganisms under a microscope; Louis Pasteur, considered the father of microbiology; Joseph Lister, the father of modern surgery; and Robert Koch, the father of bacteriology. It was during the 1800s that microbiology emerged as a scientific discipline, aided by advances like staining techniques, pure culture isolation, and Koch's postulates for identifying microbes that cause disease. The early 1900s marked the discovery of viruses and antibiotics like penicillin. Overall, the document provides a comprehensive overview of the scientific milestones that established microbiology
Viruses can be cultivated within suitable host cells like bacteria to allow for viral replication. Bacteriophages are commonly grown by adding them to a culture of bacteria in a growth medium. The bacteriophages will multiply within the bacteria, destroying cells and releasing new viral particles. Viruses are also cultivated in tissue culture systems using cell lines grown in suspension. The virus infects the host cells, forcing them to produce new virus copies until the cells are lysed. Viral titers can be determined using plaque assays, where viral plaques are counted, or animal inoculation studies.
This presentation contains 45 slides on general virology comprises of topics on viral classification, transmission, pathogenesis, viral cytopathic effect, stages of viral infections, antiviral drugs and viral vaccines. It also have a slide noting an outline of laboratory diagnosis of viral infection. This power point presentation was designed for medical students, nurses and academicians teaching virology and microbiology in medical universities, schools or colleges.
Fungi are a separate kingdom from plants and animals that play an important role in decomposition. Dermatomycoses are fungal infections of the skin, hair, or nails caused by dermatophytes from three genera: Trichophyton, Microsporum, and Epidermophyton. Dermatophytes are transmitted through direct or indirect contact with infected skin or hair. Diagnosis involves microscopic examination of skin or hair samples in potassium hydroxide or culturing samples on growth media. Treatment depends on the severity but may include topical imidazole creams for mild cases or oral antifungals like griseofulvin for more severe or persistent infections.
Louis Pasteur and Robert Koch were two of the founders of bacteriology. Pasteur developed the process of pasteurization to prevent contamination and disproved spontaneous generation. He also discovered vaccines for anthrax, cholera, and rabies. Koch isolated pure bacterial cultures and invented techniques like the hanging drop method. He discovered the specific bacteria that cause anthrax, tuberculosis, and cholera and proposed Koch's postulates for identifying the microorganisms that cause diseases. Both scientists greatly advanced the germ theory of disease.
Sporulation is the process of spores formation by bacteria during unfavorable condition
Germination is the process of spores return to give vegetative cells under favorable condition.
Bacteriophage typing involves identifying viruses that infect bacteria up to the strain level. It is used to control infections and for epidemiological purposes. Strains are differentiated based on phenotypic and genotypic differences, known as 'typing'. Various typing methods exist, including phenotypic methods like biotyping, phage typing, serotyping, and genotypic methods like plasmid profiling, ribotyping, and pulse field gel electrophoresis. Each method has advantages and limitations in terms of typeability, reproducibility, discriminatory power, and ease of use. Molecular typing techniques are increasingly used for outbreak detection and epidemiological investigations.
Cultivation of viruses uhf copy - copyheena thakur
This document discusses viruses and methods for cultivating viruses. It describes viruses as obligate intracellular parasites that can only multiply inside living host cells. The three main methods for cultivating viruses discussed are inoculation of viruses into animals, embryonated eggs, and tissue culture. For animal inoculation, mice are commonly used and viruses can be introduced via different routes. Embryonated eggs provide a suitable environment for virus growth and isolation. Tissue culture involves culturing cells or tissue fragments, and cell lines provide indefinite growth. Detection of virus growth involves monitoring for cytopathic effects, hemadsorption, interference, and other methods.
Viruses can only be grown within living host cells. The document discusses three main methods for cultivating viruses: inoculation into animals, embryonated eggs, and tissue culture. It provides details on each method, including commonly used animal and egg types, inoculation sites, advantages and limitations. Tissue culture involves growing viruses in cultured cells, including primary cultures that can only grow briefly, diploid cell strains for limited passages, and continuous cell lines that can grow indefinitely.
Viruses must infect living cells in order to replicate and be grown in culture. There are several methods for culturing and isolating viruses, including animal inoculation, embryonated eggs, and various types of cell culture. The primary purposes of virus cultivation are to isolate and identify viruses from clinical samples, study viral structure and replication, and produce viruses for vaccines. Viruses can be grown in experimental animals, embryonated eggs, or different types of cell cultures, including primary cell cultures, diploid cell strains, and continuous cell lines. Growth of viruses in cell cultures can be detected by observing cytopathic effects, hemadsorption, or heterologous interference.
This document discusses methods for isolating and cultivating animal and plant viruses. There are three main methods for animal viruses: embryonated chicken eggs, tissue cultures, and laboratory animals. Embryonated chicken eggs are commonly used as they provide several inoculation sites and are economical. Tissue cultures include primary cell cultures from animal tissues, diploid cell strains with normal chromosome numbers, and continuous cell lines with abnormal numbers that can undergo infinite divisions. Laboratory animals like primates and small mammals are also used but their use is now limited. For plant viruses, whole plants, plant tissue cultures, and protoplast cultures are used, with protoplasts being a standard technique. Insect cell cultures can also be used
Viruses can only multiply within living cells and must be grown using one of three cultivation methods: animal inoculation, embryonated egg inoculation, or tissue culture. Animal inoculation involves injecting animals like mice and observing for disease signs. Embryonated eggs are commonly used by inoculating virus into areas like the chorioallantoic membrane or allantoic cavity. Tissue culture uses organ fragments, explants, or cell lines derived from tissues to provide an environment for virus growth.
Viruses can only reproduce inside host cells and are obligate intracellular parasites. There are several approaches to studying viruses including inoculation into animals or embryonated eggs, and culturing viruses in cells. Tissue culture involves growing cells in vitro in growth medium, then inoculating them with viruses. There are different types of cell cultures including primary cultures from tissues that can only grow for a limited time, diploid cell lines that can be subcultured 50 times, and continuous cell lines from cancer cells that can be cultured indefinitely. Viruses infect and replicate within cells, sometimes causing cell lysis and formation of plaques that can be observed. Cytopathic effects indicate viral growth. Tissue culture allows broad virus cultivation
Viruses can be grown through three main methods: inoculation into animals, inoculation into embryonated eggs, and tissue culture. Embryonated eggs are commonly used because they provide an ideal substrate for viral growth and replication. Viruses are inoculated into 7-12 day old chick embryos through the shell and incubated for 2-3 days, then isolated from embryo tissue. While eggs are inexpensive and easy to maintain, different viruses grow in different locations within the egg. Tissue culture involves digesting tissues and mixing individual cells with viral growth medium in flasks, providing a broad method for cultivation but requiring trained technicians. Animal inoculation allows for studying pathogenesis but is expensive and raises welfare issues.
Viral cultivation methods involve growing viruses within suitable host cells in order to obtain sufficient virus particles for research and applications. Viruses can be cultivated using bacterial cultures, plant tissue cultures, whole plants, embryonated eggs, animal tissues, and cell cultures. The type of host system used depends on the type of virus. Bacterial viruses are grown in bacterial cultures by plating phage and bacteria together. Plant viruses are grown in whole plants or plant tissue cultures. Animal viruses can be grown in embryonated eggs, animal tissues, or cell cultures. Cell cultures are the most widely used method and allow viruses to be easily detected and grown in large quantities. Primary cell cultures have limited growth potential while continuous cell lines can be grown
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive function. Exercise causes chemical changes in the brain that may help protect against developing mental illness and improve symptoms for those who already suffer from conditions like anxiety and depression.
Viruses are obligate intracellular parasites which means they can only grow or reproduce inside a host cell.
The primary purpose of virus cultivation:
To isolate and identify viruses in clinical samples.
To do research on the viral structure, replication, genetics, and effects on the host cell.
To prepare viruses for vaccine production.
Isolation of the virus is always considered a gold standard for establishing the viral origin of the disease
topics covered
CULTIVATION OF VIRUSES
Animal inoculation
Embryonated eggs
CAM
Allantoic cavity
Amniotic cavity
Yolk sac
Tissue culture
Organ culture
Explant culture
Cell culture
Primary cell culture
diploid cell culture
Continues cell lines
This document discusses techniques for cultivating viruses. It explains that viruses require living host cells to replicate and describes three main cultivation methods: animal inoculation, embryonated egg inoculation, and tissue culture. Animal inoculation involves infecting animals like mice to isolate and study viruses, but it is expensive and raises welfare issues. Embryonated egg inoculation is commonly used to grow viruses by inoculating eggs at specific sites, and it is cost-effective but each virus grows at different sites. Tissue culture uses cell monolayers and is versatile but requires specialized facilities and technicians. The document provides details on each technique's advantages, disadvantages and applications.
VIRAL VACCINES
Since viruses are intracellular parasites they will grow only within other living cells.
Methods of viral vaccine production:
Cultivation of virus using free living animals
Fertile eggs
Tissue cultures
Viruses can only replicate inside host cells. Three common substrates used for cultivating viruses are embryonated eggs, tissue culture, and animal inoculation. Embryonated eggs are economical and support growth of many viruses but not all. Tissue culture uses cells or organs grown in vitro in nutrient media and allows detection of viruses through cytopathic effects, metabolic changes, or plaque formation. Animal inoculation is ideal when the natural host is impractical, using lab animals like mice. Effective diagnosis requires proper collection, transport, and storage of specimens at early stages of illness for virus isolation in host systems and subsequent identification tests.
This document discusses three methods for cultivating viruses: laboratory animals, embryonated eggs, and tissue culture. Laboratory animals like monkeys were historically used but pose risks, while embryonated eggs are economical and sterile but some viruses don't grow in them. Tissue culture became important when Enders grew polio virus in human cells, allowing cultivation of many viruses. The three tissue culture types are organ culture, explant culture, and cell culture.
Viruses can be cultivated through several methods, including inoculation in animals, embryonated eggs, and tissue/cell culture. Inoculation in animals allows study of viral replication and immune responses but is expensive. Embryonated eggs are widely used as they are inexpensive and viruses can replicate, though not all human viruses grow well. Tissue/cell culture is now preferred, using primary cultures, continuous cell lines, or explant cultures. Growth is detected through cytopathic effects, staining, or metabolic changes in infected cells.
The document discusses the cultivation of viruses using embryonated eggs. It notes that embryonated eggs are a simpler technique than animal inoculation for growing viruses, as eggs do not have an immune response and contain suitable cells for virus growth. Viruses can be inoculated into different areas of embryonated eggs, including the chorioallantoic membrane, allantoic cavity, amniotic sac, and yolk sac. The allantoic cavity is most commonly used due to its simplicity and ability to produce large quantities of viruses like influenza. Embryonated eggs remain an important method for growing stocks of viruses for research and vaccine production.
Basic Information on Virus & Hepatitis Virusssuserc16f64
Viruses can infect all life forms and replicate only inside living cells. They have either DNA or RNA as their genetic material and a protein coat called a capsid that may be surrounded by an envelope. Viruses are cultivated using experimental animals, embryonated eggs, or tissue culture methods in order to study them and develop vaccines. Hepatitis viruses are a group that target the liver and can cause inflammation and damage.
This document discusses methods for cultivating and isolating viruses, including animal inoculation, embryonated hen egg inoculation, and primary cell culture. Viruses can be grown in animal tissues, eggs, or cell cultures. Eggs provide routes of inoculation like the allantoic cavity, amniotic cavity, and chorioallantoic membrane. Primary cell cultures are made from chicken embryos. Cell lines like vero and HeLa support viral growth and replication. Detection of viral growth involves monitoring for cytopathic effects, interference, or detecting viral antigens.
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Know the difference between Endodontics and Orthodontics.Gokuldas Hospital
Your smile is beautiful.
Let’s be honest. Maintaining that beautiful smile is not an easy task. It is more than brushing and flossing. Sometimes, you might encounter dental issues that need special dental care. These issues can range anywhere from misalignment of the jaw to pain in the root of teeth.
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdfOsvaldo Bernardo Muchanga
GASTROINTESTINAL INFECTIONS AND GASTRITIS
Osvaldo Bernardo Muchanga
Gastrointestinal Infections
GASTROINTESTINAL INFECTIONS result from the ingestion of pathogens that cause infections at the level of this tract, generally being transmitted by food, water and hands contaminated by microorganisms such as E. coli, Salmonella, Shigella, Vibrio cholerae, Campylobacter, Staphylococcus, Rotavirus among others that are generally contained in feces, thus configuring a FECAL-ORAL type of transmission.
Among the factors that lead to the occurrence of gastrointestinal infections are the hygienic and sanitary deficiencies that characterize our markets and other places where raw or cooked food is sold, poor environmental sanitation in communities, deficiencies in water treatment (or in the process of its plumbing), risky hygienic-sanitary habits (not washing hands after major and/or minor needs), among others.
These are generally consequences (signs and symptoms) resulting from gastrointestinal infections: diarrhea, vomiting, fever and malaise, among others.
The treatment consists of replacing lost liquids and electrolytes (drinking drinking water and other recommended liquids, including consumption of juicy fruits such as papayas, apples, pears, among others that contain water in their composition).
To prevent this, it is necessary to promote health education, improve the hygienic-sanitary conditions of markets and communities in general as a way of promoting, preserving and prolonging PUBLIC HEALTH.
Gastritis and Gastric Health
Gastric Health is one of the most relevant concerns in human health, with gastrointestinal infections being among the main illnesses that affect humans.
Among gastric problems, we have GASTRITIS AND GASTRIC ULCERS as the main public health problems. Gastritis and gastric ulcers normally result from inflammation and corrosion of the walls of the stomach (gastric mucosa) and are generally associated (caused) by the bacterium Helicobacter pylor, which, according to the literature, this bacterium settles on these walls (of the stomach) and starts to release urease that ends up altering the normal pH of the stomach (acid), which leads to inflammation and corrosion of the mucous membranes and consequent gastritis or ulcers, respectively.
In addition to bacterial infections, gastritis and gastric ulcers are associated with several factors, with emphasis on prolonged fasting, chemical substances including drugs, alcohol, foods with strong seasonings including chilli, which ends up causing inflammation of the stomach walls and/or corrosion. of the same, resulting in the appearance of wounds and consequent gastritis or ulcers, respectively.
Among patients with gastritis and/or ulcers, one of the dilemmas is associated with the foods to consume in order to minimize the sensation of pain and discomfort.
Pictorial and detailed description of patellar instability with sign and symptoms and how to diagnose , what investigations you should go with and how to approach with treatment options . I have presented this slide in my 2nd year junior residency in orthopedics at LLRM medical college Meerut and got good reviews for it
After getting it read you will definitely understand the topic.
How to Control Your Asthma Tips by gokuldas hospital.Gokuldas Hospital
Respiratory issues like asthma are the most sensitive issue that is affecting millions worldwide. It hampers the daily activities leaving the body tired and breathless.
The key to a good grip on asthma is proper knowledge and management strategies. Understanding the patient-specific symptoms and carving out an effective treatment likewise is the best way to keep asthma under control.
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chemistry investigatory project
The development of nanogold-based cancer therapy could revolutionize oncology by providing a more targeted, less invasive treatment option. This project contributes to the growing body of research aimed at harnessing nanotechnology for medical applications, paving the way for future clinical trials and potential commercial applications.
Cancer remains one of the leading causes of death worldwide, prompting the need for innovative treatment methods. Nanotechnology offers promising new approaches, including the use of gold nanoparticles (nanogold) for targeted cancer therapy. Nanogold particles possess unique physical and chemical properties that make them suitable for drug delivery, imaging, and photothermal therapy.
Computer in pharmaceutical research and development-Mpharm(Pharmaceutics)MuskanShingari
Statistics- Statistics is the science of collecting, organizing, presenting, analyzing and interpreting numerical data to assist in making more effective decisions.
A statistics is a measure which is used to estimate the population parameter
Parameters-It is used to describe the properties of an entire population.
Examples-Measures of central tendency Dispersion, Variance, Standard Deviation (SD), Absolute Error, Mean Absolute Error (MAE), Eigen Value
“Psychiatry and the Humanities”: An Innovative Course at the University of Mo...Université de Montréal
“Psychiatry and the Humanities”: An Innovative Course at the University of Montreal Expanding the medical model to embrace the humanities. Link: https://www.psychiatrictimes.com/view/-psychiatry-and-the-humanities-an-innovative-course-at-the-university-of-montreal
Breast cancer: Post menopausal endocrine therapyDr. Sumit KUMAR
Breast cancer in postmenopausal women with hormone receptor-positive (HR+) status is a common and complex condition that necessitates a multifaceted approach to management. HR+ breast cancer means that the cancer cells grow in response to hormones such as estrogen and progesterone. This subtype is prevalent among postmenopausal women and typically exhibits a more indolent course compared to other forms of breast cancer, which allows for a variety of treatment options.
Diagnosis and Staging
The diagnosis of HR+ breast cancer begins with clinical evaluation, imaging, and biopsy. Imaging modalities such as mammography, ultrasound, and MRI help in assessing the extent of the disease. Histopathological examination and immunohistochemical staining of the biopsy sample confirm the diagnosis and hormone receptor status by identifying the presence of estrogen receptors (ER) and progesterone receptors (PR) on the tumor cells.
Staging involves determining the size of the tumor (T), the involvement of regional lymph nodes (N), and the presence of distant metastasis (M). The American Joint Committee on Cancer (AJCC) staging system is commonly used. Accurate staging is critical as it guides treatment decisions.
Treatment Options
Endocrine Therapy
Endocrine therapy is the cornerstone of treatment for HR+ breast cancer in postmenopausal women. The primary goal is to reduce the levels of estrogen or block its effects on cancer cells. Commonly used agents include:
Selective Estrogen Receptor Modulators (SERMs): Tamoxifen is a SERM that binds to estrogen receptors, blocking estrogen from stimulating breast cancer cells. It is effective but may have side effects such as increased risk of endometrial cancer and thromboembolic events.
Aromatase Inhibitors (AIs): These drugs, including anastrozole, letrozole, and exemestane, lower estrogen levels by inhibiting the aromatase enzyme, which converts androgens to estrogen in peripheral tissues. AIs are generally preferred in postmenopausal women due to their efficacy and safety profile compared to tamoxifen.
Selective Estrogen Receptor Downregulators (SERDs): Fulvestrant is a SERD that degrades estrogen receptors and is used in cases where resistance to other endocrine therapies develops.
Combination Therapies
Combining endocrine therapy with other treatments enhances efficacy. Examples include:
Endocrine Therapy with CDK4/6 Inhibitors: Palbociclib, ribociclib, and abemaciclib are CDK4/6 inhibitors that, when combined with endocrine therapy, significantly improve progression-free survival in advanced HR+ breast cancer.
Endocrine Therapy with mTOR Inhibitors: Everolimus, an mTOR inhibitor, can be added to endocrine therapy for patients who have developed resistance to aromatase inhibitors.
Chemotherapy
Chemotherapy is generally reserved for patients with high-risk features, such as large tumor size, high-grade histology, or extensive lymph node involvement. Regimens often include anthracyclines and taxanes.
The biomechanics of running involves the study of the mechanical principles underlying running movements. It includes the analysis of the running gait cycle, which consists of the stance phase (foot contact to push-off) and the swing phase (foot lift-off to next contact). Key aspects include kinematics (joint angles and movements, stride length and frequency) and kinetics (forces involved in running, including ground reaction and muscle forces). Understanding these factors helps in improving running performance, optimizing technique, and preventing injuries.
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1. VIRUSES
intracellular
❖ Viruses are obligate
parasites.
❖ They multiply only inside the living
host cells. Animals ,plants ,humans ,
bacteria, fungus ,protozoa and algae
are the natural hosts of viruses.
2. ❖Viruses are host specific and grow only in
selective hosts. Virologists use only a suitable
hostsystemfor cultivation of a virus.
❖Viruses cannot grow in artificial media .They
cannot grown in non-living culture or on agar plates
alone , they must require living cells to support their
replication.
❖There is no universal cell that will support all
viruses.
3. Main purpose of virus cultivation
❖ To isolate and identify viruses in
clinical samples.
❖ To prepare viruses for vaccine
production.
❖ To do research on viral structure
, replication , genetics and
effects on host cells.
4. ❖ Reed and colleagues (1900) used human
volunteers for their pioneering work on yellow fever . Due to
the serious risk involved , human volunteers are used only
when no other method is available and when virus is relatively
harmless.
❖ Monkeys were used for the isolation of the poliovirus by
Landsteiner and Popper (1909). However, due to
their cost and risk to handlers, monkeys find only limited
applications in virology.
❖ The use of white mice, pioneered by Theiler (1903)
extended thescope of animal inoculation greatly.
5. ❖ Good Pasture in 1931 first used the embryonated
hen’s egg for the cultivation of virus and this method is
further developed by Burnet.
❖The first application of tissue culture in virology was by
Steinhardt and colleagues (1913), who maintained
the vaccinia virus in fragments in rabbit cornea.
❖Maitland (1928) used chopped tissue in nutrient
media for cultivation of vaccinia viruses.
❖The turning point which made tissue culture the most
important method for cultivation of virus was the
demonstration by Enders, Weller and Robins (1949),
that poliovirus till then considered a strictly neurotropic
virus,could be grown in tissue culture of non-neural origin.
6. METHODS FOR CULTIVATION OF VIRUSES
Inoculation of virus
into animals.
Inoculation of virus
into embryonated eggs.
Tissue culture.
7. ❖Viruses which are not cultivated in
embryonated egg and tissue culture are
cultivated in laboratory animals. e.g: mice,
guinea pig , hamester , rabbits and
primates are used.
❖The selected animals should be healthy
and free from any communicable diseases.
❖Suckling mice (less than 48 hours old) are
most commonly used.
1). Inoculation of Virus in
Animals
8. Different ways of inoculation in mice are:
1) Intracerebral.
2) Subcutaneous.
3) Intraperitoneal.
4) Intranasal.
9. 1). Intracerebral
➢It occurring or
introduced
within
or
administered into the
cerebrum. It means
when a diseased blood
vessel within the brain
bursts allowing blood
to leak inside the brain.
13. Advantages and disadvantages
of animal inoculation :
Advantages :
❖ Production of antibodies can be identified.
❖Diagnosis , pathogenesis and clinical
symptoms are determined.
❖ Primary isolation of certain viruses.
❖Mice provide a reliable model for
studying viral replication.
❖Used for the study of immune
responses,epidemology and oncogenesis.
14. Disadvantages :
❖Expensive and difficulties in maintaince
of animals.
❖Difficulty in choosing of animals for
particular virus.
❖Some human viruses cannot be grown
in animals or can be grown but do not
cause diseases.
❖Mice do not provide models for vaccine
development.
15. 2). Inoculation of virus into
embryonated egg
The process of cultivation of viruses in
embryonated eggs depend upon the
type of egg being used.
Egg provide a suitable means for :
i. The primary isolation and
identification of viruses.
ii. The production of vaccines.
iii. The maintaince of stock culture.
16. After incubation , the egg is broken and virus is
isolated from tissue of egg.
For inoculation , eggs are first prepared for cultivation ,
the sheel surface are first prepared for cultivation , the
shell surface is first disinfected with iodine and
penetrated with a small sterile drill.
Viruses are inoculated into chick embryo of 7-12
days old.
18. ▪ Virus growth and multiplication in
the egg embryo is indicated by the
death of the embryo , by embryo cell
damage , or by the formation of
typical pocks or lesions on the egg
membrane.
▪ Viruses can be cultivated in
various pats of egg like :
1). Chorioallantoic membrane (CAM)
2). Allantoic cavity
3) Amniotic sac
4) Yolk sac Pock
19. 1). Chorioallantoic
membrane (CAM)
▪ Inoculation is
growing poxvirus.
▪ After incubation
mainly for
and visible
lesions called pocks are
observed , which is grey white
area in transparent CAM.
▪ Herpes simplex virus is also
grown.
▪ Single virus gives single pocks.
▪ This method is suitable for
plaque studies.
20. Plaque : A clear area in a lawn of
host cells that results from the lysis of
host cells by viruses.
21. 2). Allantoic cavity:
❖Inoculation is mainly done for
production of vaccine of
influenza virus , yellow fever ,
rabies.
❖Most of avaian viruses can be
isolated using this method.
❖Allantoic inoculation is a quick
and easy method that yields
large amounts (8-15ml) of
virus-infected egg fluids.
22. 3). Amniotic sac :
▪ Inoculation is mainly done
for primary isolation of
influenza virus and the
mumps virus.
▪ Growth and replication of
virus in egg embryo can be
detected by
haemagglutination assay.
▪ The virus is introduced
directly into the amniotic fluid
that bathes the developing
embryo.
23. 4). Yolk sac inoculation:
❖ It is
method
also a simplest
for growth and
multiplication of virus.
cultivation
viruses
of
and
❖ It is inoculated for
some
some
bacteria ( Chlamydia ,
interference
Rickettsiae ).
❖Immune
mechanism can be
detected in most of avian
viruses.
24. Advantages of inoculation into
embryonated egg
❖ Widely used method for the isolation of virus and
growth.
❖ Cost effective and maintenance is much easier.
❖ The embryonated eggs are readily available.
❖ They are free from contaminating bacteria and
many latent viruses.
❖ Ideal substrate for the viral growth and replication.
❖ less labor is needed.
❖ Widely used method to grow virus for some
vaccine production.
❖ Defense mechanisms are not involved in
embryonated eggs.
25. Disadvantage of inoculation into
embryonated egg
• The site of inoculation for varies with
different virus . That is , each virus
have different sites for growth and
replication.
26. 3). Tissue culture
❖Cultivation of bits of tissues and organs in
vitro had been used by physiologists and
surgeons for the study of morphogenesis
and wound healing.
❖Before the advent of cell culture , animal viruses
could be propagated only on whole animals or
embryonated chicken eggs.
27. ✓ Cell cultures have replaced
embryonated eggs as preferred
type of growth medium for
many viruses.
✓Cell culture consists of cells
grown in culture media in the
laboratory.
28. There are three types of tissue culture:
1) Organculture.
2) Explantculture.
3) Cell culture.
29. 1). Organ culture
Example: Tracheal ring organ culture is employed for the isolation of
coronavirus, a respiratory pathogen.
Organ culture is useful for the isolation of some viruses which appear
to be highly specialized parasites of certain organs.
Small bits of organs can be maintained in vitro for days and weeks,
preserving their original architecture and function. Formalin is used for
the preservation.
30.
31. 2). Explant culture
The explants taken from laboratory animals such as mice , rabbit , guineapigs
, hamester and man can be grown in petri dishes.
An explant is aseptically transferred into a sterile petri dish by using a fine
tipped forceps and then a coverslip is placed over that explant.
Enough volume of medium is poured into the petri dish , which is then
incubated at 37 ⁰C untill cell growth.
Example : Adenoid tissue explant culture were used for the isolation of
adenoviruses.
A small portion of tissue excised from animal’s body is explant.
33. 3). Cell culture
The essiential constitutents of the growth medium are physiologic amounts of
essiential amino acids and vitamins , salts , glucose and a buffering system
generally consisting of bicarbonate in equilibrium with atmosphere containing
about 5% carbon dioxide.
Tissues are dissociated into the components of cells by the
action of proteolytic enzymes such as trypsin and
mechanical shaking.
This is the type of culture routinely employed for growing
viruses.
34. Antibiotics are added to prevent the bacterial
contaminants and phenol red as indicator.
Such media will enable most cell types to multiply with
a division time of 24-48 hours.
The cell suspension is dispensed in bottles , tubes or
petri dishes.
The cell adhere to the glass surface and on incubation ,
divide to form a confluent monolayer sheet of cells
covering the surface within about a week.
This is supplemented with up to 5% calf or fetal calf
serum.
35. Based on their origin , chromosomal
characters and the number of generations
through which they can be maintained , cell
culture are classified into three types :
Some fastidious virus grow only in suchroller
cultures.
Cell culture tubes may be incubated in a
sloped horizontal position , either as
‘stationary culture’ or may be in special ‘roller
drums’ to provide better aeration.
38. 1). Primary cell culture
Examples : Kidney cells of monkey and man , embryo
cells , alveolar cells , macrophages and amniotic cells
are usually grown in primary cell cultures.
It is capable of only limited growth and hence it can
be subcultured once or twice.
The cell culture established directly from cells taken
from animal’s tissue is called primary culture.
39. Primary cell cultures are widely used for the isolation
of animal viruses and cultivation of viruses for vaccine
production.
The culture vessel is incubated at 37⁰C for a few days to
get a primary culture.
A small volume of cell suspension is aseptically
transferred to a culture flask or petri dish containing
nutrient medium , with the help of pipette.
40. Secondary cell cultures are used for the isolation of wide group of animal
viruses and growing fastidious viruses. Some secondary cultures are usedfor
vaccine production.
Examples : Human embryonic kidney cells and skin fibroblast cells.
As secondary cell cultures can be maintained and subcultured for 20-50 times
, they are called semi-continuous cells.
The cell culture established from primary cell culture are called secondary
culture or sub-culture.
41. The fragments of monoculture grow into large
monolayers. These are called secondary culture.
It is then cut into small fragments. 2or 3fragments are
inoculated into a roller drum containing nutrient medium
and the roller drum is incubated at 37⁰Cfor few days.
Monolayer produced as a result of primary cell culture is
detached from the bottom of the culture flask by adding
trypsin or EDTA.
42. ➢ Animal cells capable of indefinite growth are
called continuous cell lines or cell lines.
➢ These are the cells of a single type , usually
derived from cancer cells , that are capable of
continuous serial cultivation indefinitely.
➢ Standard cell lines derived from human
cancers , such as HeLa , HEp – 2 and KB cell
lines have been used in laboratories
throughout the world for many years.
43. ❖These cell lines may be maintained by
serial subcultivation or stored in the cold (
-70⁰C ) for use when necessary.
❖Some cell lines are now permitted to be
used for vaccine manufacture, for example
: Vero cells for rabies vaccine.
44. Advantages of cellculture
❖Relative ease, broad spectrum, cheaper
and sensitivity
Disadvantage of cellculture
❖The process requires trained technicians
with experience in working on a full time
basis.
❖State health laboratories
laboratories do not isolate
and hospital
and identify
viruses in clinical work.
❖Tissue or serum for analysis is sent to
central laboratories to identify virus.
45. Detection of virusgrowth
• The following methods are available to
detect the virus growth in the cell or tissue
cultures.
a). Cytopathic effect.
b). Haemadsorption.
c). Interference.
d) Transformation.
e)Immunofluorescence.
f). Metabolic inhibition.
46. ❖Many viruses cause morphological changes
in cultured cells in which they grow. These
changes can be readily observed by
microscopic examination of the cultures.
❖These changes are known as ‘cytopathic
effects’ (CPE) and the viruses causing CPE
are called ‘cytopathogenic viruses’.
❖The CPE produced by different groups of
viruses are characteristic and help in
presumptive identification of virus isolates.
47. • For example, enteroviruses produce
rapid CPE with crenation of cells and
degeneration entire cell sheet ; measles
virus produce syncytium formation;
herpes virus causes discrete focal
degeneration; adenovirus produce large
granular clumps of grapes.
48.
49. ❖When hemagglutinating viruses (such as
influenza and parainfluenza viruses) grow
in cell cultures, their presence can be
indicated by the addition of guinea pig
erythrocytes to the cultures.
❖If the viruses are multiplying in the cultures,
the erythrocytes will adsorb onto the surface
of cells. This is known as
‘hemadsorption’.
50.
51. 3). Interference
• The growth of first virus will inhibit second
virus infection due
effect. This property
to some inhibitory
of cell cultures is
celled interference. it is useful to detect
the growth of non-cytopathic viruses in cell
cultures
52. 4). Transformation
• If oncogenic viruses are inoculated into
cell cultures, the infected cell grow fast
and produce microtumours in the culture.
This is called transformation. It indicated
the presence of oncogenic viruses in the
culture.
53. 5). Immunofluorescence test
• Some cell from the cell culture are stained
with a fluorescent dye conjugated antiserum
and viewed under an UV microscope.
• Viral antigen present on the cell surface bind
with the antiserum.
• Fluorescence from the cell is the positive
indication for presence of virus in the cell.
This is a widely used method in diagnostic
virology.
55. 6). Metabolic inhibition
❖ In normal cell cultures, the medium
turns acid due to the cellular
metabolism.
❖When viruses grow in cell cultures , the
cell metabolism is inhibited and there is
no acid production.
❖This can be made out by the indicator
(phenol red) incorporated in the
medium.