Page 4

Semester 2: Medical Virology and Parasitology

  • General properties of viruses: Structure and Classification including viroids, prions, satellite RNAs and virusoids

    General properties of viruses: Structure and Classification including viroids, prions, satellite RNAs and viroids
    • Definition and Characteristics of Viruses

      Viruses are acellular infectious agents that can only replicate inside the living cells of an organism. They consist of genetic material, either DNA or RNA, surrounded by a protein coat known as a capsid. Some viruses also have an outer lipid envelope. They lack cellular structures, metabolize independently, and require a host for replication.

    • Structure of Viruses

      Viruses can be classified based on their structure, which includes nucleic acid type, shape, and the presence of an envelope. The basic structures include: 1. Non-enveloped (naked) viruses: These viruses lack an outer lipid membrane, e.g., Adenovirus. 2. Enveloped viruses: These have an outer lipid membrane derived from host cell membranes, e.g., Influenza virus. 3. Capsid shapes: Can be helical, icosahedral, or complex.

    • Classification of Viruses

      Viruses are classified based on various criteria, including: 1. Type of nucleic acid: DNA viruses and RNA viruses. 2. Shape and symmetry: Helical, icosahedral or complex. 3. Replication strategy: Lytic or lysogenic cycles. 4. Host range: Animal, plant, or bacterial viruses.

    • Viroids

      Viroids are small, circular RNA molecules that infect plants. They lack a protein coat and are known to cause various plant diseases. Viroids replicate independently of host cell DNA and do not encode proteins. They are transmitted through mechanical injury or vegetative propagation.

    • Prions

      Prions are infectious proteins that cause neurodegenerative diseases in various animals and humans. They lack nucleic acids and induce misfolding of normal host proteins, leading to aggregation and neuronal damage. Examples include Creutzfeldt-Jakob disease and mad cow disease.

    • Satellite RNAs

      Satellite RNAs are sub-viral agents that lack the genes necessary for replication and depend on a helper virus for their life cycle. They can modify the virulence or pathogenicity of the helper virus. Examples include satellite tobacco mosaic virus RNA.

    • Viroids and Virusoids

      Viroids are small circular RNA molecules infecting plants, whereas virusoids are similar but require a helper virus for their replication. Virusoids may encode some proteins and have been observed in association with specific helper viruses.

  • Cultivation of viruses: embryonated eggs, experimental animals and cell cultures

    Cultivation of viruses: embryonated eggs, experimental animals and cell cultures
    Overview of Virus Cultivation
    Virus cultivation is a critical aspect of virology research, essential for studying viral pathogenesis, vaccine development, and antiviral drug testing. Various methods exist for cultivating viruses, including the use of embryonated eggs, experimental animals, and cell cultures.
    Embryonated Eggs
    Embryonated chicken eggs are a traditional method for viral cultivation. They provide a controlled environment for viral replication. Commonly used viruses include influenza and certain vaccines, where the virus is injected into the allantoic cavity or amniotic sac.
    Experimental Animals
    Animals such as mice, ferrets, and guinea pigs are used to cultivate and study viruses. This method helps in understanding the pathogenesis and immune response. Ethical considerations are crucial, and appropriate protocols must be followed to minimize suffering.
    Cell Cultures
    Cell culture is a widely used technique in modern virology. Cells obtained from various tissues are grown in vitro in a controlled environment. This method supports high-throughput screening of antiviral compounds and the production of vaccines.
    Comparison of Methods
    Each method of virus cultivation has its advantages and limitations. Embryonated eggs are cost-effective but limited to certain viruses. Experimental animals provide a complete biological system but raise ethical issues. Cell cultures allow for precise control and manipulation but may not fully mimic in vivo environments.
  • Purification and Assay of viruses: Physical and Chemical methods, Electron Microscopy, Protein and Nucleic acids studies

    Purification and Assay of Viruses
    • Introduction to Virus Purification

      Overview of the importance of virus purification in medical virology and research. Discussion of the need for pure viral samples for assays and studies.

    • Physical Methods of Virus Purification

      Description of various physical methods such as ultracentrifugation, chromatography, and filtration used to separate viruses from cellular debris and proteins.

    • Chemical Methods of Virus Purification

      Overview of chemical methods including the use of detergents, solvents, and precipitation agents to isolate viruses from complex mixtures.

    • Assay Techniques for Virus Measurement

      Discussion of different assay techniques such as plaque assays, hemagglutination assays, and quantitative PCR to measure viral concentration and infectivity.

    • Electron Microscopy in Viral Studies

      Explanation of the role of electron microscopy in visualizing viruses and assessing purity. Techniques involved in sample preparation and imaging.

    • Protein Analysis in Viruses

      Overview of methods for analyzing viral proteins including SDS-PAGE and Western blotting. Importance of protein characterization in understanding virus structure and function.

    • Nucleic Acid Studies in Viruses

      Discussion on the extraction and analysis of viral nucleic acids. Techniques such as RT-PCR and sequencing used for studying viral genomes.

  • Infectivity Assays: Plaque and end-point

    Infectivity Assays: Plaque and End-Point
    Introduction to Infectivity Assays
    Infectivity assays are crucial for measuring the ability of viruses to infect host cells. They help in understanding viral pathogenesis and in evaluating antiviral strategies.
    Plaque Assay
    Plaque assays involve infecting a monolayer of host cells with the virus and overlaying with a semi-solid medium. Each plaque formed represents a single infectious unit, allowing for quantification of viral titer.
    End-Point Dilution Assay
    End-point dilution assays determine the dilution of a virus required to infect a certain percentage of cells. It employs statistical methods to calculate viral concentration based on the presence or absence of infection.
    Comparison of Plaque and End-Point Assays
    Both assays serve similar purposes but differ in methodology. Plaque assays provide visual quantification while end-point dilution assays focus on statistical estimation of viral infectivity.
    Applications in Virology Research
    These assays are essential for determining viral loads, evaluating vaccine efficacy, and studying viral biology within the context of medical virology.
  • Virus Entry, Host Defenses Against Viral Infections, Epidemiology, pathogenic mechanisms

    Medical Virology and Parasitology
    • Virus Entry

      Viruses enter host cells through various mechanisms including receptor-mediated endocytosis and fusion with the host cell membrane. The interaction between viral proteins and host cell receptors plays a critical role in determining host susceptibility to infection.
    • Host Defenses Against Viral Infections

      The immune system employs various strategies to combat viral infections, including innate immune responses such as interferon production and adaptive responses involving the activation of T cells and antibody production. The effectiveness of these defenses can be influenced by factors such as the type of virus and the host's overall health.
    • Epidemiology

      The study of viral epidemiology involves understanding how viruses spread within populations. This includes factors such as transmission routes, population dynamics, and virulence. Surveillance of viral outbreaks and vaccination strategies are key components in controlling viral diseases.
    • Pathogenic Mechanisms

      Viruses employ various pathogenic mechanisms to exploit host cellular machinery. These mechanisms may include direct cell damage, evasion of immune responses, and induction of inflammation. Understanding these pathways is crucial for developing therapeutic strategies.
  • Pathogenesis, laboratory diagnosis, treatment for DNA Viruses (Pox, Herpes, Adeno, Papova, Hepadna) and RNA Viruses (Picorna, Orthomyxo, Paramyxo, Rhabdo, Rota, HIV and other Hepatitis viruses, Arbo, Dengue virus, Ebola virus, Emerging and reemerging viral infections

    Pathogenesis, laboratory diagnosis, treatment for DNA and RNA Viruses
    • DNA Viruses

      Poxviruses cause disease through direct cell lysis and immune response evasion.
      Diagnosis via clinical presentation, PCR, and electron microscopy.
      Supportive care; antiviral therapies like cidofovir may be used in severe cases.
      Establish latency in host cells; can reactivate causing recurrent disease.
      Diagnosis through serology, PCR, and culture.
      Acyclovir and other antivirals are used to manage infections.
      Cause lytic infections; can lead to respiratory, gastrointestinal, and conjunctival diseases.
      PCR and serotyping for diagnosis.
      Supportive treatment; no specific antiviral agents are widely available.
      Includes human papillomavirus which can cause warts and cancers.
      Diagnosis through Pap smear and HPV DNA tests.
      Vaccines available; treatments for lesions include cryotherapy and surgery.
      Infection leads to chronic hepatitis; can cause cirrhosis and liver cancer.
      Hepatitis B surface antigen assays, viral DNA detection.
      Antivirals like tenofovir and entecavir; liver transplant in severe cases.
    • RNA Viruses

      Causes diseases like poliomyelitis; infects gastrointestinal tract.
      Isolation from clinical specimens and PCR.
      Supportive care; vaccines available for poliovirus.
      Influenza viruses cause respiratory infections with seasonal pandemics.
      RT-PCR and rapid antigen tests.
      Antiviral medications like oseltamivir; vaccination is key for prevention.
      Includes measles, mumps; causes respiratory symptoms.
      Serology and PCR are used for diagnosis.
      Supportive care, vaccination for prevention.
      Rabies virus causes encephalitis; transmitted through bites.
      PCR and direct fluorescent antibody tests.
      Post-exposure prophylaxis with rabies vaccine.
      Leading cause of severe diarrhea in children.
      Enzyme immunoassays for detection in stool.
      Supportive care; oral rehydration therapy is crucial.
      HIV attacks CD4+ T-cells; hepatitis viruses lead to liver diseases.
      HIV-DNA, antibody tests; hepatitis serologies.
      Antiretroviral therapy for HIV; antiviral therapy for hepatitis.
      Transmitted by arthropods; causes febrile illnesses.
      Serologic tests and PCR.
      Symptomatic treatment; vaccines available for some.
      Causes dengue fever, can lead to severe dengue.
      Dengue NS1 antigen, serology, PCR.
      Supportive care; no specific antiviral treatment.
      Causes severe viral hemorrhagic fever; transmitted through bodily fluids.
      PCR and ELISA for antigen detection.
      Supportive care; investigational vaccines available.
      Include novel viruses entering populations; often zoonotic.
      Virus-specific serologies and genomic sequencing.
      Varies widely; emphasis on prevention and outbreak control.
  • Bacterial viruses - X174, M13, MU, T4, lambda, Pi: Structural organization, life cycle and phage production, Lysogenic cycle - typing and application in bacterial genetics

    Bacterial viruses - X174, M13, MU, T4, lambda, Pi: Structural organization, life cycle, and phage production, Lysogenic cycle - typing and application in bacterial genetics
    • Structural Organization

      Bacterial viruses, or bacteriophages, exhibit diverse structural organizations. Key structural features include capsid shapes, nucleic acid types, and tail structures. X174 is an icosahedral virus with single-stranded DNA, while T4 is a complex virus with a head-tail structure and double-stranded DNA. Lambda has a linear double-stranded DNA genome and a complex tail structure, enabling efficient attachment to host bacteria.

    • Life Cycle

      The life cycle of bacteriophages generally includes two main pathways: the lytic cycle and the lysogenic cycle. The lytic cycle involves attachment, penetration of the host cell, replication of viral components, assembly of new virions, and lysis of the host cell, releasing new phages. In contrast, the lysogenic cycle allows the viral genome to integrate into the host genome, remaining dormant until activated.

    • Phage Production

      Phage production varies among different bacteriophages. In the lytic cycle, phages increase in number rapidly, often leading to host cell death. The efficiency of phage production depends on factors such as the multiplicity of infection and environmental conditions. Bacterial strains can exhibit different susceptibilities to phage infection, influencing production rates.

    • Lysogenic Cycle

      In the lysogenic cycle, temperate bacteriophages, such as lambda and Mu, integrate their DNA into the bacterial chromosome as a prophage. This integration can regulate bacterial gene expression and contribute to horizontal gene transfer. Under specific conditions, the prophage can be excised from the host genome, re-entering the lytic cycle.

    • Typing and Application in Bacterial Genetics

      Bacteriophages serve as fundamental tools in bacterial genetics and molecular biology. Techniques such as phage typing help identify bacterial species and strains based on their susceptibility to specific bacteriophages. Bacteriophages are utilized in genetic engineering to introduce or modify genes within bacteria, facilitating studies in gene function and regulation.

  • Diagnosis of viral infections - conventional serological and molecular methods

    Diagnosis of viral infections - conventional serological and molecular methods
    • Introduction to Viral Infections

      Viral infections can cause a wide range of diseases. Accurate diagnosis is crucial for effective treatment and control. Understanding the methods for diagnosing these infections is essential for healthcare professionals.

    • Conventional Serological Methods

      Serological methods detect the presence of antibodies or antigens related to viral infections. Common techniques include ELISA, Western blotting, and immunofluorescence. These methods are valuable for diagnosing infections such as HIV, hepatitis, and influenza.

    • Limitations of Serological Methods

      Serological tests may have false positives or negatives, particularly in early infections where antibodies may not yet be detectable. Timing of specimen collection is critical for accurate results.

    • Molecular Methods of Diagnosis

      Molecular methods, such as PCR and RT-PCR, detect viral nucleic acids. These techniques are highly sensitive and specific, allowing for early detection of viral infections and helping in managing outbreaks.

    • Comparison of Serological and Molecular Methods

      Molecular methods offer faster and more accurate results compared to serological methods. However, serological methods can provide information about previous infections and immune status.

    • Conclusion

      Both serological and molecular methods are important for diagnosing viral infections. The choice of method depends on various factors, including the type of virus, stage of infection, and clinical context.

  • Antiviral agents and viral vaccines

    Antiviral agents and viral vaccines
    • Introduction to Antiviral Agents

      Antiviral agents are medications specifically designed to treat viral infections. They function by inhibiting the development of the virus or eliminating it from the host. Antivirals differ from antibiotics as they are ineffective against bacterial infections.

    • Mechanisms of Action

      Antiviral agents operate through various mechanisms: 1. Inhibiting viral entry into host cells 2. Interfering with viral replication and transcription 3. Blocking viral enzyme activity 4. Enhancing host immune response.

    • Types of Antiviral Agents

      Common classes of antiviral drugs include: 1. Nucleoside analogs 2. Protease inhibitors 3. Neuraminidase inhibitors 4. Polymerase inhibitors. Each type targets specific viruses, such as HIV, influenza, and herpes.

    • Viral Vaccines

      Vaccines are biological preparations that provide acquired immunity to a particular viral infection. They stimulate the body's immune response without causing the disease.

    • Types of Viral Vaccines

      Viral vaccines can be classified into: 1. Live attenuated vaccines 2. Inactivated or killed vaccines 3. Subunit, recombinant, or conjugate vaccines. Each type varies in terms of efficacy and safety.

    • Immunization Strategies

      Effective immunization strategies may include: 1. Standard immunization schedules 2. Booster doses 3. Vaccination during outbreaks. These strategies aim to increase herd immunity and reduce viral transmission.

    • Future Perspectives

      Research continues on developing novel antiviral agents and vaccines, including mRNA vaccines and broadly neutralizing antibodies. Future advancements may enhance efficacy, coverage, and ease of administration.

  • Introduction to Medical Parasitology: Classification, host-parasite relationships

    Introduction to Medical Parasitology
    • Definition and Scope of Medical Parasitology

      Medical parasitology is the study of parasites that affect human health. It encompasses various types of organisms including protozoa, helminths, and ectoparasites. This field explores the biology of these organisms, their life cycles, and their impact on human diseases.

    • Classification of Parasites

      Parasites are classified based on several criteria: 1. Type of Organism: Protozoa (single-celled) and Helminths (multicellular). 2. Host Relationship: Endoparasites (live inside a host) and Ectoparasites (live on the surface). 3. Life Cycle: Direct and indirect life cycles, based on the number of hosts involved.

    • Host-Parasite Relationships

      The interaction between hosts and parasites can be classified as mutualism, commensalism, or parasitism. In parasitism, the relationship is detrimental to the host, leading to disease. Understanding these interactions is crucial for developing control strategies.

    • Pathogenesis of Parasitic Infections

      Parasitic infections can lead to various pathophysiological effects on the host. These include tissue damage, immune response evasion, and nutrient depletion. The severity of these effects varies depending on the parasite and host factors.

    • Diagnosis and Treatment of Parasitic Diseases

      Diagnosis of parasitic infections often involves laboratory techniques such as microscopy, serology, and molecular methods. Treatment may include antiparasitic medications, which target specific life stages of the parasites. Preventive measures are essential for controlling transmission.

  • Epidemiology, life cycle, pathogenic mechanisms, laboratory diagnosis and treatment for Protozoa causing human infections

    Epidemiology, life cycle, pathogenic mechanisms, laboratory diagnosis and treatment for Protozoa causing human infections
    • Epidemiology

      Protozoan infections are prevalent in tropical and subtropical regions, with transmission occurring through vectors, contaminated water, and food. Major diseases include malaria (caused by Plasmodium spp.), amoebiasis (Entamoeba histolytica), and giardiasis (Giardia lamblia). The epidemiology of these infections is influenced by factors such as climate, sanitation, and human behaviors.

    • Life Cycle

      Protozoa exhibit complex life cycles, often including both sexual and asexual stages. For instance, Plasmodium undergoes a life cycle involving mosquito vectors and human hosts, alternating between different forms like sporozoites, merozoites, and gametocytes. Understanding these life cycles is crucial for developing control strategies.

    • Pathogenic Mechanisms

      Protozoa can cause disease through various mechanisms, including direct tissue damage, immune evasion, and toxin production. For example, Entamoeba histolytica invades the intestinal epithelium, leading to ulceration, while Giardia lamblia disrupts intestinal absorption, resulting in diarrhea.

    • Laboratory Diagnosis

      Diagnosis of protozoan infections typically includes microscopic examination of stool, blood smears, or tissue biopsies. Specific staining techniques and molecular methods such as PCR can enhance detection sensitivity and specificity. Serological tests may also be utilized, especially for diseases like leishmaniasis.

    • Treatment

      Treatment of protozoan infections varies depending on the pathogen. Antimalarial drugs such as chloroquine and artemisinin-based therapies are effective against malaria. Metronidazole is commonly used for amoebiasis and giardiasis. Resistance to these treatments is emerging, highlighting the need for ongoing research and development of new therapies.

  • Classification, life cycle, pathogenicity, laboratory diagnosis and treatment for parasites including Helminthes (Cestodes, Trematodes, Nematodes) and other parasites

    Classification, life cycle, pathogenicity, laboratory diagnosis, and treatment for parasites including Helminthes and other parasites
    • Classification of Parasites

      Parasites can be classified into different groups based on various criteria. Major classes include: 1. Protozoa: Single-celled organisms. 2. Helminthes: Multicellular organisms; further divided into: a. Cestodes (tapeworms) b. Trematodes (flukes) c. Nematodes (roundworms) 3. Ectoparasites: External parasites like lice and fleas.

    • Life Cycle of Helminthes

      Helminthes exhibit complex life cycles, often involving multiple hosts. Life stages may include: 1. Eggs 2. Larvae (various stages) 3. Adult forms Each species may have specific requirements for host transition and environmental conditions.

    • Pathogenicity of Helminthes and Other Parasites

      Pathogenicity refers to the ability of parasites to cause disease. Factors influencing pathogenicity include: 1. Adherence and invasion of host tissues. 2. Production of metabolic by-products and toxins. 3. Eliciting immune responses from the host.

    • Laboratory Diagnosis of Parasites

      Diagnosis often involves: 1. Microscopic examination of stool or tissue samples. 2. Serological tests for antibodies. 3. Molecular techniques like PCR. Each helminthic infection may require specific diagnostic approaches.

    • Treatment of Parasitic Infections

      Treatment strategies vary by type of parasite and can include: 1. Anthelminthic drugs for helminthes (e.g., praziquantel for cestodes, albendazole for nematodes). 2. Supportive care for symptomatic relief. 3. Preventive measures such as hygiene and sanitation practices.

  • Cultivation of parasites

    Cultivation of parasites
    • Introduction to Parasite Cultivation

      Cultivation of parasites involves growing and maintaining parasites in controlled laboratory environments. This practice is crucial for studying their biology, pathogenicity, and developing treatments.

    • Types of Parasites

      Parasites can be classified into several categories such as protozoa, helminths, and ectoparasites. Each type requires specific cultivation techniques and conditions.

    • Cultivation Techniques

      Common methods for cultivating parasites include in vitro cultures, animal models, and cell cultures. Each method has advantages and limitations depending on the parasite.

    • Nutritional Requirements

      Parasites have specific nutritional needs that must be met for successful cultivation. These may include amino acids, vitamins, and other growth factors.

    • Contamination Prevention

      Preventing contamination is essential during cultivation. Techniques include maintaining sterile conditions, using antibiotics, and regularly monitoring cultures.

    • Applications of Cultivated Parasites

      Cultivated parasites are used in research to understand their biology, test drugs, and develop vaccines. They also play a role in diagnostics.

    • Ethical Considerations

      Cultivating parasites raises ethical issues, particularly in the use of animal models. Guidelines must be followed to ensure humane treatment of all organisms involved.

  • Diagnosis of parasitic infections: Serological and molecular diagnosis

    Diagnosis of parasitic infections: Serological and molecular diagnosis
    • Introduction to Parasitic Infections

      Parasitic infections are caused by organisms that live on or in a host organism, often causing harm to the host. Accurate diagnosis is crucial for effective treatment.

    • Serological Diagnosis of Parasitic Infections

      Serological tests detect antibodies or antigens related to parasitic infections. Common tests include ELISA and Western blot. These tests vary in specificity and sensitivity depending on the parasite.

    • Molecular Diagnosis of Parasitic Infections

      Molecular techniques, such as PCR, allow for the detection of parasite DNA or RNA. These methods are highly sensitive and specific, enabling early diagnosis and monitoring.

    • Comparison of Serological and Molecular Techniques

      Serological methods are generally quicker and less expensive than molecular techniques but may provide false positives or negatives. Molecular methods, while more accurate, can be more costly and time-consuming.

    • Clinical Applications and Limitations

      Both diagnostic methods have unique applications in clinical settings. While serological methods are useful for population studies, molecular approaches provide insights in complex cases.

    • Future Trends in Parasitic Infection Diagnosis

      Advancements in technology may enhance the accuracy and speed of diagnosis. Next-generation sequencing and point-of-care tests are promising developments in the field.

  • Anti-protozoan drugs

    Anti-protozoan drugs
    • Introduction to Anti-protozoan Drugs

      Anti-protozoan drugs are medications used to treat infections caused by protozoa, which are single-celled organisms. These infections can lead to various diseases, including malaria, amoebiasis, and leishmaniasis. Understanding the mechanisms of action, types, and clinical applications of these drugs is essential in medical microbiology.

    • Types of Anti-protozoan Drugs

      1. Antimalarials: These drugs are specifically used to treat malaria. Common examples include chloroquine, quinine, and artemisinin derivatives. They work by targeting the life cycle stages of the malaria parasite.

    • Amebicides

      Amebicides are used to treat infections caused by amoebae, particularly Entamoeba histolytica. Metronidazole and iodoquinol are widely used agents that target the intestinal and extraintestinal forms of the parasite.

    • Antileishmanial Drugs

      These drugs are formulated to treat leishmaniasis, caused by Leishmania spp. Treatments include antimonials, amphotericin B, and miltefosine, each acting through different mechanisms on the intracellular parasites.

    • Mechanisms of Action

      Anti-protozoan drugs typically function by inhibiting DNA synthesis, disrupting metabolic pathways, or interfering with the parasite's ability to reproduce. For example, antimalarials may inhibit heme detoxification in the parasite, leading to cell death.

    • Resistance to Anti-protozoan Drugs

      Resistance against anti-protozoan drugs poses a significant challenge, particularly in malaria treatment. Factors contributing to resistance include overuse, incomplete treatment courses, and genetic mutations in protozoan populations.

    • Current Research and Development

      The ongoing research focuses on discovering new anti-protozoan agents, understanding resistance mechanisms, and developing combination therapies to enhance efficacy and reduce resistance emergence. This area is crucial as global travel and changing climates can influence the spread of protozoan infections.

Medical Virology and Parasitology

M.Sc. Microbiology

Medical Virology and Parasitology

2

Periyar University, Salem

Core V

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