Microbiology
Microbiology comprises three main fields: bacteriology, virology and immunology providing a short summary on veterinary mycology. The students learn general bacteriology and virology and specific bacteriology, mycology and virology with a special focus on pathogens of veterinary importance. They learn the immune response of the host, its protection against infectious diseases and the most important diagnostic methods. In the practicals students have to learn microbiology laboratory techniques and identification procedures. Characteristics of the most important pathogenic bacteria and viruses are studied as well. Teaching is organised in the form of lectures and small group laboratory practicals.
Language:
Year
Term (semester)
Lectures
Practicals
Credits
Examination
Structure of the subject:
Teaching staff
English, Hungarian
2 nd
, 3 rd
4 th
and 5 th
90 lectures (4 th
semester), 45 lectures (5 th
semester)
30 (4 th
semester), 30 (5 th
semester)
4 th semester: bacteriology and virology lectures, bacteriology practicals
5 th
semester: immunology lectures, immunology and virology practicals
3 (4 th
semester), 6 (5 th
semester) practical note (4 th
semester), final examination (5 th
semester)
Bakonyi, T., Fodor, L., Hornyák, Á., Makrai, L. and Tuboly, T.
(lectures), Bakonyi, T., Cságola, A., Forgách, P., Hornyák, Á.,
Makrai, L., Tapaszti, Zs. (practicals)
1.1. Requirements

Lectures serve as the basis of the subject, active participation at the lectures is precondition of a successful examination.

Attending the practicals is compulsory, absence should be remedied. Remedy can happen on the same week by joining an other group or attending the extra practical on the last week of the semester.

Students without knowing the theoretical subject of the practical are regarded absent.

The 4th semester is closed with a practical note. Attending all practicals and showing satisfactory knowledge (oral and written tests) on bacteriology are conditions of gaining the practical note.

Practicals on virology can be commenced only after passing a virology test.

Students can sit to the examination after passing both subjects, attending all practicals and having at least a satisfactory practical note at the end of the 4th semester.
1.1.1. Rules of the practicals

Having a white coat on is compulsory.

Eating, drinking and smoking at the practicals is forbidden.

Accidents at the practicals have to be reported to the teacher at the practicals.

The practical note at the end of the 4th semester is based at the oral and written tests made during the semester. A better note can be received at a voluntary examination at the end of the semester.
1.2. Teaching environment

The computer aided lectures are given in one of the lecture theatres of the campus. The practicals are held at the 16-head-practical room of the department, where individual microscopes, computerised summaries help the understanding. The basic laboratory examinations can be carried out by the students themselves and the complicated methods are demonstrated here as well. The demonstration material is prepared in the different laboratories of the department.
1.3. Topics of the microbiology
1.3.1. Bacteriology lectures

Subject of microbiology, its aim and short history (2 lectures)

Characterization of bacteria, their importance in life. Size and sharp of bacteria, structure of the bacterial cell. Microscopy, native and stained preparations. (5 lectures)

Culture of bacteria, media, metabolism of bacteria, autotrophic and heterotrophic bacteria. Carbohydrate metabolism of bacteria (4 lectures)

Nitrogen metabolism of bacteria, detection of some metabolites and enzymes.
Bacterial activity in practice (2 lectures)

Growth and multiplication of bacteria. Environmental effects on multiplication.

Resistance of bacteria against physical and chemical effects, disinfection, antibiotics
(7 lectures)

Genetics of bacteria (6 lectures)

Pathogenicity and infection (3 lectures)

Systematic of bacteria, identification, specific bacteriology (16 lectures)
1.3.2. Virology lectures

Place and significance of viruses in the biosphere.

Propagation of viruses.

Morphology of viruses.

General characteristics of viral nucleic acid, methods of nucleic acid investigation.
Viral proteins, their role and methods of their investigation.

Virus multiplication.

Mutation of viruses and its role in virus evolution.

Viral interactions.

Viral oncogenicity.

Cytopathic effects, plaque formation.

General features of viral infection.

Laboratory diagnosis of viral infections.

Detection and titration of antiviral antibodies.

Antiviral chemotherapy, interferons, interferon inducers and immune sera in therapy.

Immunization against viral diseases, viral vaccines.

DNA viruses.

RNA viruses.

Bacteriophages.

Viroids, prions.
1.3.3. Immunology lectures


Natural and acquired immunity

Antigens

The tissues and cells of the immune system

Structure and function of immunoglobulins

Monoclonal antibodies

T cell receptors

Genetic background of immunity

Major Histocompatibility Complex.

T and B cell differentiation

Cytokines, lymphokines, interferons

The complement system

Humoral and cellular immune response

Transplantation immunology

Immunotolerance, natural autoimmunity

Immunity to bacteria, parasites and fungi

Immunity to viruses and tumors

Immune response of the fetus and newborn animals.

Active and passive immunization

Autoimmune diseases, immune deficiencies.

Hypersensitivities
1.3.4. Bacteriology practicals
1.
Media, sterilization, bacterial cultures, anaerobic culture methods, pure cultures, microscopy, examination of native bacteria, staining of bacteria.
2.
Bacillus anthracis and aerobic spore-forming bacteria. Clostridium.
3.
Streptococcus, Enterococcus, Anaerobic cocci, Lactobacillus, Staphylococcus,
Micrococcus.
4.
Listeria, Erysipelothrix, Corynebacterium, Renibacterium.
5.
Mycobacteria, Nocardia, Actinomyces, Dermatophilus, Streptomyces.
6.
General characterization of enterobacteria, Escherichia, Klebsiella, Enterobacter.
7.
Salmonella, Shigella, Citrobacter, Proteus, Yersinia, Gram-negative anaerobic bacteria
8.
Haemophilus, Actinobacillus, Pasteurella, Taylorella, Identification of an unknown
.bacterium I.
9.
Brucella, Aeromonas, Pseudomonas, Francisella, Bordetella, Moraxella. Identification of an unknown bacterium II.
10.
Vibrio, Campylobacter, Helicobacter, Arcobacter, Spirochaetes. Antibiotic susceptibility tests.
11.
Mycoplasma, Rickettsia, Chlamydia. Identification of an unknown bacterium III.
12.
Fungi.
13.
Extra practical.
1.3.5. Virology practicals
1.
Virus isolation from diagnostic samples (inoculation of embryonated eggs, and cell cultures)
2.
Virus-induced cytopathic effects and plaque-formation on cell cultures
3.
Direct diagnosis of viral infections using molecular methods I (virus nucleic acid purification, polymerase chain reaction)
4.
Direct diagnosis of viral infections using molecular methods II (agarose gel electrophoresis), indirect diagnosis of viral infections I (virus neutralisation)

5.
Indirect diagnosis of viral infections II (evaluation of virus neutralisation, haemagglutination inhibition)
1.3.6. Immunology practicals
1.
Agglutination and precipitation tests
2.
Complement fixation tests, and neutralization assays
3.
Immunoelectrophoresis, Western blot
4.
ELISA
5.
IF, IP, RIA
6.
Cellular tests (MIT, LST, RCF, CT)
4. Recommended literature

Carter-Chengappa-Roberts: Essentials of Veterinary Microbiology. 5th Ed. Williams-
Wilkins Baltimore 1995. ISBN 0-683-01473-0

Hirsh, D.C., Zee, Y.C.: Veterinary Microbiology. Blackwell Oxford 1999

Murphy, F. A. – Gibbs, E. P. J. – Horzinek, M. C. – Studdert, M. J.: Veterinary
Virology. 3. kiadás San Diego – London: Academic Press, 1999

Quinn, P.J. – Carter, M.E. – Markey, B. – Carter, G.R.: Clinical veterinary microbiology. London: Wolfe, 1994.

Quinn, P.J., Markey, B.K., Carter, M.E., Donnelly, W.J.C., Leonard, F.C.: Veterinary
Microbiology and Microbial Disease. Blackwell. Oxford 2002. ISBN 0-632-05525-1

Tizard, J.: Veterinary immunology. Philadelphia: London: W.B. Saunders, 1992
1.5. Examination

Students can sit to an examination both after the autumn and spring semesters.

Exam days are to be discussed with the teachers.

The minimum numbers of examinees is 8, the maximum is 14 per day including the
English and the Hungarian class.

The examination starts at 7.30, the last report is at 8.30.

The exam is divided into three parts (bacteriology, virology, immunology), students receive 2 questions from each part.

The result is defined after completing the whole exam, it is communicated then.

The examination is successful if at least satisfactory level of knowledge (Note 2) is shown in the case of each question.

Fixed examination date can be postponed only once preferably by e-mail to T. Tuboly or by phone, personally at T. Tuboly.

If a student wants to postpone the exam for a second time or she/he does not appears at the examination she/he will receive a note 1.

If a student fails at the exam she/he cannot take an other within 5 days after the unsuccessful one.
 If more information is needed, please ask Assoc. Prof. Tamás Tuboly.
1.6. Questions at the examination
1.6.1. Bacteriology
1.
Position and significance of bacteria in the biosphere. Eukaryotes and prokaryotes. The size and shape of bacteria, their examination by light, dark-field, phase contrast, fluorescence and electron microscope.

2.
The structure of bacterial cell, cell wall, cytoplasm, cytoplasmic membrane, nuclear material.
3.
Capsule, flagella, fimbria and spore of bacteria.
4.
Examination of native and stained bacteria. Simple and differential staining methods.
5.
Bacterial metabolism. Autotrophic and heterotrophic bacteria. Vitamin and additive requirements of bacteria. Bacterial pigments.
6.
Utilization of bacterial activity.
7.
Nitrogen metabolism of bacteria. Detection of the most important enzymes and metabolites of the nitrogen metabolism.
8.
Carbohydrate metabolism of heterotrophic bacteria, aerobic and anaerobic bacteria.
Detection of enzymes and metabolites of the carbohydrate metabolism.
9.
Culture of bacteria, media, pure cultures. Anaerobic cultures.
10.
Growth and multiplication of bacteria. Growth phases in cultures, methods of counting bacterial cells.
11.
Environmental effects on the growth of bacteria (water, temperature, pH, osmosis). The resistance of bacteria against physical effects.
12.
Disinfection and disinfectants.
13.
Sterilization methods.
14.
The concept and the aim of chemotherapy. Sulphonamides. Types of bacterial resistance to antibacterials. Examination of antibiotic resistance of bacteria.
15.
Antibiotics.
16.
Structure and function of bacterial genetic material.
17.
Mutations. Extra chromosomal genetic elements of bacteria. The most important plasmids.
18.
Recombination of bacterial genetic material, bacterial transformation, transduction, phage conversion.
19.
Bacterial conjugation. Phenotypic modification of bacteria.
20.
Examination of the genetic material of the bacteria. Gene manipulations, genetic engineering.
21.
Pathogenic and saprophytic microorganisms. Pathogenicity and virulence. Measurement and changing of virulence.
22.
Factors of virulence attached to the bacterial cell surface, extra cellular enzymes.
23.
Toxins of bacteria. Exotoxins and endotoxins.
24.
Bacillus anthracis and the aerobic spore forming bacteria.
25.
Clostridium.
26.
Staphylococcus. Micrococcus.
27.
Streptococcus, Enterococcus, anaerobic cocci, lactobacilli.
28.
Erysipelothrix, Listeria.
29.
Mycobacterium.
30.
Corynebacterium, Nocardia, Actinomyces, Dermatophilus, Renibacterium, Rhodococcus.
31.
The general characteristics of Enterobacteriaceae, Gram negative anaerobic bacteria.
32.
Escherichia coli.
33.
Salmonella, Citrobacter.
34.
Klebsiella, Shigella, Yersinia, Proteus.
35.
Pasteurella, Ornithobacterium rhinotracheale
36.
Haemophilus, Actinobacillus, Taylorella.
37.
Francisella, Bordetella, Moraxella.
38.
Brucella.
39.
Pseudomonas, Aeromonas.
40.
Vibrio, Campylobacter, Helicobacter, Arcobacter.
41.
The general characteristics of Spirochaetes. Borrelia, Treponema, Serpulina.

42.
Leptospira.
43.
Rickettsieae, Chlamydia.
44.
Mycoplasma.
45.
Characterisation of fungi, their morphology, culture and resistance.
46.
Zygomycota, Ascomycota, Basidiomycota, Deuteromycota, imperfect and dimorph fungi.
1.6.2. Virology
1.
Place and significance of viruses in the biosphere, evolution of viruses.
2.
Propagation of viruses I. (Inoculation of embryonated eggs, experimental infection of laboratory animals).
3. Propagation of viruses II. (Production of cell cultures.)
4. Concentration and purification of viruses.
5. Morphology of viruses.
6. General characteristics and purification of viral nucleic acid. Methods of nucleic acid investigation I. (size, morphological characteristics, restriction enzyme analysis, bacterial cloning, Southern blot).
7. Methods of nucleic acid investigation II. (heteroduplex technique, nucleic acid hybridization, DNA microarray, sequencing, polymerase chain reaction).
8. Viral proteins, their role and methods of their investigation (polyacrylamide-gel electrophoresis, Western blot, immunoperoxidase staining, monoclonal antibodies). Viral lipids and carbohydrates.
9. General features of virus multiplication (adsorption, penetration, decapsidation, maturation, virus release).
10. Transcription, translation and nucleic acid replication of DNA viruses.
11. Transcription, translation and nucleic acid replication of RNA viruses.
12. Mutation of viruses and its role in virus evolution.
13. Viral interactions (recombination, complementation, phenotypic mixing, interference, virus exaltation).
14. Viral oncogenicity.
15. Cytopathic effects, plaque formation.
16. General features of viral infection (infection routes, factors influencing the virus - host interactions).
17. Types of viral infections (acute, latent, tolerated, persisting and slow infections).
18. Laboratory diagnosis of viral infections (collection and transport of samples).
19. Direct demonstration of virus infections I (virus isolation and identification).
20. Direct demonstration of virus infections II (detection of viral antigens and viral nucleic acid).
21. Titration of viruses (infective titer, hemagglutinating titer).
22. Detection and titration of antiviral antibodies (serumneutralization, hemagglutination inhibition).
23. Antiviral medicines, interferons, interferon inducers and immune sera in therapy.
24. Immunization against viral diseases, types of viral vaccines.
25. Papillomaviridae, Polyomaviridae, Adenoviridae.
26. Herpesviridae.
27. Poxviridae.
28. Asfarviridae, Hepadnaviridae
29. Parvoviridae, Circoviridae.
30. Reoviridae, Birnaviridae.
31. Picornaviridae, Caliciviridae, Astroviridae.

32. Togaviridae, Flaviviridae.
33. Nidovirales (Coronaviridae, Arteriviridae.)
34. Orthomyxoviridae.
35. Paramyxoviridae.
36. Rhabdoviridae, Filoviridae.
37. Bunyaviridae, Arenaviridae, Bornaviridae.
38. Retroviridae.
39. Bacteriophages.
40.
Viroids, prions.
1.6.3. Immunology
A1. Essential features of antigens. Factors that influence immunogenicity.
A2. Immunoglobulin supergene family. Characterization of immunoglobulins (methods).
A3. IgG and IgM, structure and function.
A4. IgA, IgE and IgD, structure and function.
A5. Monoclonal antibodies, features, hybridoma. Genetically engineered monoclonal antibodies
A6. T cell receptors, the TCR/CD3 complex.
A7. Theories of antigen recognition. Antibody and TCR diversity.
A8. Genetic basis of antibody and TCR formation.
A9. MHC I, general characteristics, process of antigen presentation by MHC I.
A10. MHC II, general characteristics, process of antigen presentation by MHC II.
A11. Cytokines, lymphokines and interleukins in general
A12. Interferons.
A13. Tumour necrosis factor α and β.
A14. Humoral immune response.
A15. Cellular immune response.
A16. Primary and secondary immune response.
A17. Regulation of the immune response by the antigens and the antibodies.
A18. Regulation of the immune response by anti-idiotypes, T cells, suppressor cells, neural system, and others.
A19. Immunotolerance in general, B and T cell tolerance.
A20. Autoimmunity in general, central and peripheral tolerance.
A21. The complement system in general, complement proteins, consequences of complement activation, regulation of the complement system.
A22. Complement activation, classic pathway.
A23. Complement activation, alternative and lectin activated pathways
A24. Types of vaccines.
A25. Foetal, newborn and maternal immunity. Colostrum.
A26. Inflammation
B28. Tissue and organ transplantation.
B29. Active and passive immunization.
B30. Routes of vaccination, limitations of vaccination.
B31. Type I and Type II hypersensitivity.
B32. Type III and Type IV hypersensitivity.
B33. Organ specific autoimmune diseases.
B34. Systemic autoimmune diseases.
B35. Immunoglobulin isotype, sub-isotype, allotype, idiotype.
B36. Interleukin-1, –2, -3, -4, -5 and –6.

B37. Maturation of B and T cells.
B38. Natural killer cells. T independent antigens.
B39. Use of monoclonal antibodies in diagnostics and therapy.
B40. Non-specific immunity. Physical, chemical factors.
B41. Primary lymphoid organs.
B42. Secondary lymphoid organs
B43. Resistance and immunity to viruses.
B44. Resistance and immunity to bacteria
B45. Immunity and resistance to parasites
B46. Resistance and immunity to tumours.
B47. Primary immune deficiencies.
B48. Secondary immune deficiencies.
B49. General principles of serology. Titration of antibodies
B50. Agglutination tests. Precipitation tests.
B51. Passive agglutination, antiglobulin test, ABR-test.
B52. Immune-electrophoresis. Complement fixation test.
B53. Radioimmunoassay. Immunofluorescent test. ELISA.
B54. Cellular tests, tuberculin test.