Lesson Working regime of microbiological laboratory. The rules of the work in the bacteriological laboratory. Microscopy techniques. Immersion microscopy of bacterial smears. Choose the one correct statement: a – at microscopic examination with immersion objective the latter is immersed into the water whose refractive index is close to that of glass; b – to reveal bacteria laboratory assistant should use objective with magnification in 40 times; c-* resolution of light microscope is about 0,2 µm d – light microscopy is used for studying morphology of viruses; e – for immersion microscopy you can use any liquid for covering of stained smear; Which statement is correct? a-*Luminescent microscopy is the ability of some objects to fluoresce upon their exposure to a beam of electrons; b – ordinary light microscopy allows to achieve the higher magnification then immersion microscopy; c –a dark-field microscopy is used to examine the staining preparations; d – electron microscopy is used to study ultrastructure of a bacterial cell and morphology of virus; e – bacterial cell is colored and clear detected in the smear without staining Choose the one correct statement: a – at microscopic examination with immersion objective the latter is immersed into the water, whose refractive index is close to that of glass; b –* to reveal bacteria laboratory assistant should use objective with magnification in 90 times; c- for immersion microscopy you can use any liquid for covering of stained smear; d – light microscopy is used for studying morphology of viruses; e – resolution of light microscope is about 1 µm Which statement is correct? a- ordinary light microscopy allows to achieve the higher magnification then immersion microscopy; b – luminescent microscopy is the ability of some objects to light upon their exposure to a beam of electrons; c –*a dark-field microscopy is used to examine alive microorganisms (native preparations; d – electron microscopy is used to study ultra structure of a bacterial cell and morphology of virus; e – bacterial cell is colored and clear detected in the smear without staining 5. Choose the one correct statement: a –* at microscopic examination with immersion objective the latter is immersed into the oil (cedar, peachy, "immersiol", etc.) whose refractive index is close to that of glass; b – to reveal bacteria laboratory assistant should use objective with magnification in 40 times; c- for immersion microscopy you can use any liquid for covering of stained smear; d – light microscopy is used for studying morphology of viruses; e – resolution of light microscope is about 1 µm Which statement is correct? a –*electron microscopy provides the flat image of object, and scanning electron microscopy shows three dimensional- image; b –a dark-field microscopy is used to examine the staining preparations; c – Luminescent microscopy is carried out with light microscope and special lenses; d – immersion microscopy is very rare used for detection of bacterial morphology; e -bacterial cell is colored and clear detected in the smear without staining 7. Choose the one correct statement: a –to reveal bacteria laboratory assistant should use objective with magnification in 40 times; 1 b – at microscopic examination with immersion objective the latter is immersed into the water whose refractive index is close to that of glass; c- for immersion microscopy you can use any liquid for covering of stained smear; d –* electron microscopy is used for studying morphology of viruses; e – resolution of light microscope is about 1 µm 8.Which statement is correct? a – bacterial cell is colored and clear detected in the smear without staining; b –a dark-field microscopy is used to examine stained preparations; c –*ultra-violet light is used in luminescent microscope for illumination of a smear; d – immersion microscopy is very rare used for detection of bacterial morphology; e - ordinary light microscopy allows to achieve the higher magnification then immersion microscopy The bank of Krok-1 tests 1. Immunofluorescence reaction is widely used for express-diagnostics of many bacterial and viral infections. Choose the type of the microscope needed for this reaction. A. Light microscope. B. Electron microscope. C. *Fluorescence microscope. D. Phase-contrast microscope. E. Dark-field microscope. 2. The essential drawback of the microscopy method of infection diagnostics is insufficient informativeness caused by morphologic similarity of many microorganisms. Which of the following can increase the informativeness? A. Radioimmunoassay. B. Coombs' test. C. ELISA. D. Opsonization reaction. E. *Immunofluorescence testing. Lesson Morphology of bacteria. Preparation of the smear from bacterial culture. The simple method of staining. 1. Select the correct statement about bacterial genetic apparatus: a – a nucleoid consists of double-stranded DNA surrounded by membrane; b - a nucleoid consists of a single strand of DNA; c – a nucleoid consists of linear DNA; d – a nucleoid can be revealed by Gram’s method; e –* a nucleoid consists of double-stranded DNA localized into cytoplasm without any surrounding membrane. 2. Choose the incorrect statement: a - prokaryote are haploid microorganisms; b- *prokaryote contain one gene; c - prokaryote has no mitochondria, chloroplusts and Golgi apparatus; d –prokaryote has nucleoid, haven’t nuclear membrane; 3. Each of the following statements concerning the surface structures of bacteria is correct EXCEPT: (A) Pili mediate the interaction of bacteria with mucosal epithelium (B) Polysaccharide capsules retard phagocytosis (C)*Both gram-negative rods and cocci have lipopolysaccharide ("endotoxin") in their cell wall (D)Bacterial flagella are nonantigenic in humans because they closely resemble human flagella in chemical composition 2 4. Laboratory assistant isolated from pus spherical bacteria arranged in grape-like clusters in the smear. What group do these microorganisms belong to? A – micrococci; B – streptococci; C- *staphylococci; D- sarcina; E- tetracocci; F- diplococci 5. Select the incorrect statements about bacterial genetic apparatus: a –* a nucleoid consists of double-stranded DNA surrounded by membrane; b - a nucleoid consists of a circular DNA; c – a nucleoid contacts with cytoplasmatic membrane, mesosoms and polysomas; d – a nucleoid can not be revealed by Gram’s method; e – a nucleoid consists of double-stranded DNA localized into cytoplasm without any surrounding membrane. 6. Which one of the statements is the MOST accurate comparison of human, bacterial, and fungal cells? (A) Human cells undergo mitosis, whereas neither bacteria nor fungi do (B) *Human and fungal cells have a similar cell wall, in contrast to bacteria, whose cell wall contains peptidoglycan (C) Human and bacterial cells have plasmids, whereas fungal cells do not (D)Human and fungal cells have similar ribosomes, whereas bacterial ribosomes are different 7. Laboratory assistant isolated from nasopharyngeal mucus spherical bacteria arranged in chains in the smear. What group do these microorganisms belong to? A – micrococci; B – *streptococci; C- staphylococci; D- sarcina; E- tetracocci; F- diplococci 8. Choose the some correct statements: a - prokaryote are diploid microorganisms; b - *prokaryote has no mitochondria, chloroplusts and Golgi apparatus ; c –prokaryote has no flagella ; d –prokaryote has nucleoid, haven’t nuclear membrane; e– prokaryote has no mesosoms. 9. Laboratory assistant prepare the smear from urethral discharge collected from patient with acute urethritis. In the stained smear he reveals kidney bean shaped cocci. What group do these microorganisms belong to? A – micrococci; B – streptococci; C- *diplococci D- sarcina; E- tetracocci; F- staphylococci; The bank of Krok-1 tests 3 1. Furuncle pus smear showed spherical microbes arranged in grape-like clusters. Name these microorganisms. A. *Staphilococci. B. Diplococci. C. Micrococci. D. Streptococci. E. Tetracocci. Lesson Structure and chemical composition of bacterial cell. The structure of the cell wall. L-forms bacteria. The complete methods of staining. Gram’s method. 1. Which structure can be found in both gram-negative and gram-positive bacteria? 1. Lipopolysaccharide (LPS) 2. Outer membrane 3. Teichoic acid 4. *Peptidoglycan 5. Lipid A 2) In the Gram stain, the decolorization of gram-negative bacteria by alcohol is MOST closely related to: (A) Proteins encoded by F plasmids (B) * Lipids in the cell wall (C) 70S ribosomes (D)Branched polysaccharides in the capsule 3) Each of the following statements concerning the structure and chemical composition of bacteria is correct EXCEPT: (A)Some gram-positive cocci contain a teichoic acid (B) *Some gram-positive rods contain lipopolysacharides (C)Some gram-negative rods contain lipid A in their cell wall (D)Some gram-negative cocci contain phospholipids in their cell envelope 4) Choose the correct statement from following: A. all bacteria has a nucleus B. some bacteria may have a nucleoid, others bacteria have nucleus C. bacteria include cellulose in their cell wall D. protists are prokaryotic cells E. *bacteria have nucleoid, a circular double-stranded DNA localized into cytoplasm 5. Which of the following structures is unique to gram-negative bacteria? a. Flagella b. Capsules c. *Outer membrane d. Inclusion body e. Ribosome 6. In the Gram stain after decolorization by alcohol the Gram-negative bacteria will be: a. Pink b. *Invisible (uncolored) c. Blue d. Purple e. Violet 4 7. Each of the following statements concerning the structure and chemical composition of bacteria is correct EXCEPT: a. Some gram-negative rods contain lipid A in their cell wall b. Gram-negative cocci contain lipopolisaccharide c. *Some gram-negative cocci contain a layer of teichoic acid external to the peptidoglycan d. Gram-positive cocci contain teichoic acid in their cell wall 8. Choose the correct statement from following: a. All bacteria have lipoprotein in their cell wall b. Unique structure for any bacterial cell wall is teichoic acid c. *Peptidoglycane is localized into the cell wall of Gram(+) and Gram(-) bacteria d. Peptidoglycane is present into cell wall of Gram(+) bacteria only e. Lipopolysaccharide is a component of the inner layer of Gram(-) bacteria 9.In the Gram stain after decolorization by alcohol the Gram-positive bacteria will be: A) Pink B) Invisible (uncolored) C) Blue D) Purple E) *Violet 10. Which of the following structures is unique to gram-positive bacteria? A) lipopolysaccharide B) peptidoglycane C) *teichoic acid D) protein E) lipid A 11. Choose the correct statement from following: A. *bacteria have 80S ribosomes B. a few bacteria may have a nucleoid, others bacteria have nucleus C. bacteria include peptidoglycane in their cell wall D. all bacteria has a nucleus E. bacterial cytoplasm is surrounded by capsule 12. Staining by Gram is used to reveal: a. capsule b. cytoplasmic membrane c. DNA d. *cell wall structure e. flagella The bank of Krok-1 tests 1. Students have been proposed to stain a mixture of bacteria by Gram's method and explain the mechanism of staining at a microbiology practical class.Name the morphological structures of bacteria that cause Gram-negative or Gram-positive staining of bacteria. A. CPM (cytoplasm membrane). B. *Cell wall. C. Capsule. D. .Flagella. E. Cytoplasm. Lesson 5 Acid-fast bacteria. Bacterial endospores. Methods of their staining with Aujesko`s and ZielhNeelsen`s techniques. 1. Acid-fast bacteria may be revealed with staining by: A) *Ziehl-Neelsen B) Gram C) Burry D) Neisser E) Simple staining 2. Acid-fast bacteria are poorly stained by Gram because their cell wall includes: A) a lot amount of peptidoglycane B) phosphor compounds C) lipoproteins D) *netral lipids, waxes, fat acids E) a lot amount of water 3. Bacterial spores: a. Are formed in response to phosphate deprivation b. Generally can be destroyed by boiling c. Are devoid of chromosomal DNA d. *Have calcium dipicolinate e. Are sensitive to chemicals and antibiotics 4. Which medical important bacteria form spores? a) *clostridia b) spirochetes c) cocci d) vibrio e) enterobacteria 5. Medical important acid-fast bacteria belong to: A)* mycobacteria causing tuberculosis and leprosy B) bacteria causing enteric disorders C) rods causing purulent infections D) sporeforming bacteria E) anaerobic bacteria 6. Due to Ziehl-Neelsen technique the counterstaining is made with: a) hot fuchsine b) *1% methylene blue c) crystal violet d) iodine solution e) safranin 7. Bacteria form spores into the: a) host organism b) environment over some nutrients c) nutrient media d) *environment under unfavorable conditions 8. Bacterial spore may be stained using special technique as: A) *by Aujesky B) by Gram 6 C) by Burry D) by Neisser E) by Romanovsky-Giemsa 9. Into the host bacterial spore germinate after: a) 3-4 days b) one day c) *3-5 hours d) 30 minutes 10. Bacterial spore may be destroyed by: a) boiling b) treatment with antibiotics c) treatment with ethanol d) *autoclaving e) ultra violet radiation 11. Bacterial cell forms spore to: a) *survive under unfavorable conditions b) multiply at environment c) infect the host d) retard phagocytosis Lesson Capsule of bacteria. Flagella. Intracellular inclusions. Methods of their revealing 2. All of the following statements are true concerning the bacterial capsule EXCEPT: 1. Is composed of polysaccharide polymers 2. Is composed of poly D-glutamic acid in Bacillus anthracis 3. *Is composed of lipopolysaccharides 4. Confers upon bacteria resistance to phagocytosis 5. Can be visualized by light microscopy with appropriate staining 3. Choose the correct microscopy technique to reveal motility of microorganism: 1. Staining by Gram 2. Staining by Ziehl-Neelsen 3. Negative staining with Indian ink 4. *Wet mount technique 5. Staining by Neisser 3) Volutin granules may be stained by: a) Gram b) *Neisser c) Ziehl-Neelsen d) Simple staining with fuchsine e) Romanovsky-Giemsa 4) The most part of pathogenic gram-negative bacteria have fimbria. Choose the correct statement about pili (fimbria) from the next: a) they are essential components of bacterial cell b) *they are responsible for bacterial adhesion c) they are organelle visible under light microscopy d) they may be revealed with special method of staining 7 5) Which statement about bacterial flagella is correct? a) *they consist of protein b) they are visible under light microscopy c) they are necessary for surviving bacteria into the moist environment d) they are essential bacterial structure e) they have no antigenic properties 6) Which method is used to reveal bacterial capsule from following? a) by Gram b) *negative staining with Indian ink c) by Neisser d) by Ziehl-Neelsen 7) Which medical important germ forms inclusion body revealed by staining with alkali methylene blue? a) Staphylococcus aureus b) Vibrio cholera c) Escherichia coli d) *Corynebacterium diphtheria e) Clostridium tetani 8) Motile bacteria are divided into creeping and swimming. Swimming bacteria can move due to: a) contractions of the cytoplasmic cylinder b) *rotation of the flagella c) contractions of the fimbria d) pseudopodia formation Lesson Morphology of spirochetes, rickettsia, chlamydia, mycoplasma and actinomycetes., features of their structure and methods of staining 1. Choose the correct statement about Spirochetes: a. *the body of Spirochete consist of an axial filament and cytoplasm wound spirally around the filament; b. they are gram-positive bacteria; c. they produce spore: d. they are acid-fast and poorly stained by Gram: e. spirochetes are capsulated, non-motile bacteria 2. Choose the correct staining technique for Treponema pallidum revealing in the tissue: a. by Gram b.* by Morozov (silver impregnation); c. by Ziehl-Neelsen; d. by Neisser; e. by Loeffler 3. Choose the correct statement about Rickettsia: a. *they are stained with Gram (gram-negative); b. they form spores into the environment; c. rickettsia are facultative parasites; d. their cell wall include a lot amount of peptidoglycane; e. they are motile 4. Which from following is true about Chlamydia? a. they are not pathogenic for human b. they can produce antibiotics 8 c.* they are small cocci-shaped unmotile, asporogenes bacteria d. they are capsulated e. Chlamydia are stained with Gram as gram-positive bacteria 5. Choose the correct statement about Spirochetes: a. they contain teichoic acids in the cell wall; b. they are readily stained with Gram; d. *they are poorly stained and may be revealed with silver impregnation; c. they produce spores; e. they may be branched 6. Choose the correct staining method for Borrelia spp. observation in the blood smear: a. by Neisser; b. by Gram c. by Ziehl-Neelsen; d. *by Romanovsky-Giemsa e. by Loeffler 7. Choose the correct statement about Rickettsia: a. they form spores into the environment; b. they belong to smallest bacteria; c.* rickettsia are obligate intracellular parasites; d. they are capsulated ; e. they are motile 8. Which from following is true about Chlamydia? a. they can produce antibiotics b.*Chlamydia exists into environment in dormant form named elementary body; c. they may be revealed with wet mount tecnique d. Chlamydia are stained with Gram as gram-positive bacteria e. they are capsulated 9. Choose the correct statement about Spirochetes: a. they produce spores; b. they are readily stained with Gram; d. they are capsulated; c. *they are corkscrew shaped bacteria e. they may be branched 10. Choose the correct method to observe motile Leptospira into the urine: a.* wet mount technique; b. by Gram c. by Ziehl-Neelsen; d. by Romanovsky-Giemsa e. by Morozov (silver impregnation); 11. Choose the correct statement about Rickettsia: a. *their dormant cell may be revealed with Ziehl-Neelsen technique b. rickettsia are facultative parasites; c. they form spores into the environment;; d. they are not pathogenic for humans; e. they possess distinct stable morphology 12. Which from following is true about Chlamydia? 9 a. they are not pathogenic for human b. *they may be revealed into host cell with Romanovsky-Giemsa technique c. Chlamydia form spore outside the host cell d. they are motile and capsulated germs; e. they are poorly stained with Gram 13. Treponema pallidum causes in humans: a. relapsing fever; b. typhoid fever; c. generalized spirochetosis; d. *syphilis; e. Q-fever Lesson Physiology of the microorganisms. Nutrition of bacteria. Isolation of a bacterial pure culture (first stage) 1. Unknown bacterial strain can utilize inorganic source of carbon. Due to classification it belongs to: a. auxotroph b. prototroph c. chemotroph d. *autotroph e. heterotroph 2. Glucose may enter the bacterial cell in modified form against gradient concentration. Such type of bacterial nutrition is named: a. facilitated diffusion b. active transport d. *translocation groups c. ion pumps e. passive diffusion 3. Laboratory assistant obtains clinical material collected from patient with intestine infection. He should detect and identify the pathogen in sample. Microscopy investigation was not enough to identify causative agents. What should he do to diagnose disease? a. make a conclusion based on clinical features b.* inoculate material onto solid medium to obtain pure culture c. inoculate material onto liquid medium to detect growth characteristics of a germ d. infect laboratory animal to observe development of the same disease e. repeat microscopy investigation of another sample collected from the same patient 4. Select the correct statement: a- parasites grow and multiply without any lesion into the host; b – the hetero (organo) trophs utilize inorganic compounds for synthesis of all carbon-containing compounds c – prototroph can not grow into minimal media d – *all medical important bacteria belong to chemoorganotrophs: e – medical important bacteria belong to photoheterotrophs 5. Unknown bacterial strain uses organic compounds as source of carbon. Due to classification it belongs to: a. phototroph b. prototroph c. chemotroph 10 d. autotroph e. *heterotroph 6. Some gases can pass through cytoplasm membrane due to concentration gradient. Such type of transport is named as: a. facilitated diffusion b. active transport d. translocation groups c. ion pumps e. *passive diffusion 7.Select the correct statement: A – saprophytes can not be cultivated at ordinary media; b –* the hetero (organo) trophs utilize organic compounds for synthesis of all carbon-containing compounds c – auxotroph can grow into minimal media d – all bacteria belong to chemoorganotrophs: e – medical important bacteria belong to photoheterotrophs 8.Unknown bacterial strain can obtain energy by splitting organic compounds. Due to classification it belongs to: a. phototroph b. prototroph c. *chemotroph d. autotroph e. heterotroph 9. . Most part of nutrients enters the bacterial cell through special proteins localized into cytoplasm membrane against concentration gradient. Such type of transport is named as: a. facilitated diffusion b.* active transport d. translocation groups c. ion pumps e. passive diffusion 10.Select the correct statement: a –* the prototrophs utilize a glucose and ammonium salts as a unique source for synthesis; b – the autotrophs utilize organic compounds for synthesis of all carbon-containing compounds c – auxotroph can grow into minimal media d – all bacteria belong to chemoorganotrophs: e - saprophytes parasitize into human tissue 11.Unknown bacterial strain can not grow onto the simplest media and requires group B vitamins for successful cultivation. Due to classification it belongs to: a. phototroph b. prototroph c. chemotroph d. *auxotroph e. heterotroph 12.Bacterial cells possess so named holo-phytos type of nutrition. It means that: a. Bacteria absorb some nutrients in unsolved state b. Bacteria use sun light as source of energy c. Bacteria have specific organoid for nutrition 11 d. e. *Bacteria absorb nutrient in dissolved state by the whole surface Bacteria can take any nutrient by endocytosis into cytoplasm 13. For isolation of pure culture of aerobic bacteria tested materials should be: a –* inoculated onto agar media; b – injected into laboratory animal; c – injected into chicken embryo; d – inoculated into cell cultures e – inoculated into the liquid media. 14. Select the correct statement: a – *the autotrophs utilize СО2 for synthesis of all carbon-containing compounds; b – the autotrophs utilize organic compounds for synthesis of all carbon-containing compounds c – auxotroph can grow into minimal media d – all bacteria belong to chemotrophs: e - saprophytes parasitize on animal and vegetative substances Lesson Physiology of the microorganisms. Multiplication of bacteria. Isolation of a bacterial pure culture (second stage) 1. Generally bacteria are multiplied by (choose the most accurate answer from following): a) Sporulation b) Budding c) *Binary division d) Fragmentation of bacterial cell e) Mitosis 2. Bacterial cells do not multiply but start to increase their mass and synthesize new components during growth period of the batch culture named as: a) Log phase b) *Lag phase c) Stationary phase d) Death phase 3. In the closed system (batch culture) bacterial cells grow and divide at the maximal rate possible given their genetic potential during period named as: a) Lag phase b) *Exponential phase c) Stationary phase d) Death phase 4. In which period the total number of viable microorganisms remains constant in the batch culture? a) Death phase b) Lag phase c) *Stationary phase d) Log phase 5. In which period total number of viable cells starts to decrease with logarithmic rate in the closed system? a) Lag phase b) *Death phase c) Stationary phase d) Exponential phase 12 4. Medical important bacteria can grow at optimal temperature as following: a. *370C b. 250C c. 450C d. 180C 5. Bacteria causing infections in animals and humans may be cultivated on the media with pH ranging as next: a) From 3 to 6 b) From 5 to 7 c) *From 6 to 8 d) From 8 to 10 6. Which nutrient medium is the most convenient to isolate pure culture from the clinical sample? a) Semisolid medium b) *Solid medium c) Liquid medium d) Minimal medium 7. Which medium supports the growth of fastidious microorganisms with requirements in additional growth factors? a) Simple medium b) *Special (enriched) medium c) Selective medium d) Conservation medium 8. Which medium should be used to isolate and distinguish bacteria with hemolytic properties? a) Meat-peptone gelatin b) *Blood agar c) Hiss media d) Meat-peptone broth e) Serum agar 9. Which medium should be used to isolate and distinguish bacteria based on their ability to split lactose? a) Meat-peptone agar b) Meat-peptone broth c) Blood agar d) Serum agar e) *Endo agar or Ploskirev one Lesson Physiology of the microorganisms. Bacterial enzymes. Biochemical activity of bacteria. Isolation of a bacterial pure culture (third stage) 1. Laboratory assistant has isolated pure culture of the pathogenic microorganism. He should detect ability of isolated culture to split carbohydrates. Which media from following should be used with such purpose? a) Blood MPA b) MPA with sucrose c) *Hiss` media d) MacConky agar e) Endo agar 13 2. Laboratory assistant has isolated pure culture of the pathogenic microorganism. He should detect proteolytic activity of isolated culture. Which medium from following should be used with such purpose? a) MPA with sucrose b) Hiss` media c) *Gelatin medium d) MacConky agar e) Endo agar 3. Biochemical activity of bacteria play important role in their metabolism. Which group of enzymes inconstantly synthesize of bacterial cell? a) *Adaptive enzymes b) Constituent enzymes c) Isomerizes d) Enzymes of respiratory chain 4. Biochemical activity of bacteria play important role in their metabolism. Which group of enzymes constantly synthesize of bacterial cell? a) Sugar splitting enzymes b) Adaptive enzymes c) *Constituent enzymes d) Hemolysin 5. Which characteristics of isolated germ are determined during third period of the bacteriological investigation for its identification? a) Morphology b) Features of colony c) Growth into the liquid medium d) *Biochemical properties e) Tinctorial properties 6. Laboratory assistant should detect ability of bacterial culture to form hydrogen sulfide. Which medium and indicator may be used? a) Hiss media b) *Meat peptone broth and indicator paper with lead acetate c) Meat peptone broth and indicator paper with lithmus d) Meat peptone broth and indicator paper with oxalic acid 7. Laboratory assistant should detect ability of bacterial culture to form indol. Which medium and indicator may be used? a) Hiss media b) Meat peptone broth and indicator paper with lead acetate c) Meat peptone broth and indicator paper with lithmus d) *Meat peptone broth and indicator paper with oxalic acid 8. During second stage of bacteriological investigation separated bacterial colonies have been obtained on the solid media. To obtain pure culture from separated colony laboratory assistant should use next method of cultivation: a) *Stroke culture b) Liquid culture c) Carpet culture d) Pour plate culture 14 9. During the third stage of bacteriological investigation laboratory assistant prepare special media to detect bacterial enzymes. Why is it important to detect biochemical activity of isolated strain? a) It is necessary to propagate bacterial strain b) *It may be useful for identification of bacterial strain c) Saccharolytic activity is in a base of bacterial pathogenecity d) All medical important bacteria have the same proteolytic activity 10. Detection of the biochemical activity may be done with either special media or commercial kits for definite group of bacteria. Why do many of laboratory personal prefer the commercial kits? a) They are more available b) They do not require pure culture of bacterial strain c) They are less accurate but simpler in interpretation d) *They include the maximal important tests for identification Lesson Physiology of the microorganisms. Bacterial respiration. Cultivation of anaeaerobs. Pure culture isolation of an anaerobic micrororganism 1. To isolate pure culture of an obligate anaerobe from collected material you should choose the most corresponding medium from the list: a) Serum agar b) Yolk agar c) MPA d) *Robertson’s cooked meat broth or Kitt-Tarozzi medium e) MPB with glucose 2. Surgeon has suspected anaerobic wound infection. To isolate causative germs collected wound discharge should be cultivated into the such medium as : a – Ploskirev agar b –* Kitt-Tarozzi medium c – meat peptone broth d – Endo agar e – meat peptone agar 3) Laboratory assistant has to isolate Clostridia tetani, anaerobic sporeforming bacterium, from wound exudation. What will he do before inoculation of material into appropriate culture medium? a. he filtrates the material through cellulose filter b. *he heats the sample to kill asporogenous bacteria at 800C for 20 min c. wound discharge should be frozen before inoculation to inhibit facultative anaerobes d. he will not do anything before investigation e. he heats the sample to kill vegetative forms bacteria at 600C for 5 min 4) Anaerobic bacteria may be propagated into the special media. Choose from following main requirements to such media: a. *they should have low rH and be free from oxygen b. they should have high rH and be free from oxygen c. they should contain minimal nutrients and be free from oxygen d. they may have any rH and nutrients 5) According to type of respiration bacteria are divided into aerobes and anaerobes. Which group of bacteria can grow as at presence oxygen and at its absence? a. obligate anaerobes b. obligate aerobes c. *facultative anaerobes 15 d. capnophilic bacteria 6) It is possible to cultivate obligate anaerobes on the special medium under absence of air. The most reliable method to create anaerobic condition will be next: a. usage of thermostat b. *usage of anaerostat c. by inoculation into solid media (pour agar culture) d. usage of chemostat 7) Recently it was estimated anaerobic bacteria can obtain energy by biological oxidation (respiration). Their end acceptors of electrons will inorganic compounds containing carbon, nitrogen or sulfur. Which chemical is used as end electron acceptor by obligate anaerobes? a. nitrate b. *sulphate c. carbonate d. nitrite e. sulphite 8) Contact with oxygen may cause death of anaerobic bacteria. Which from following statements can explain this phenomenon? a. anaerobic enzymes are inhibited by oxygen b. oxygen provokes activation of anaerobic autolysins c.* bacterial cell does not have specific enzyme capable of splitting hydrogen peroxide have been formed in small amount from oxygen during bacterial respiration d. Oxygen inhibit bacterial propagation by specific action onto the saccharolytic enzymes 9). Medical important anaerobic spore forming bacteria belong to next genus: a. Bacillus b.* Clostridia c. Fusobacterium d. Streptococcus e. Escherihia 10). During the first stage of anaerobic pure culture isolation clinical sample is inoculated into the: a. sugar agar b. *Robertson’s cooked meat broth or Kitt-Tarozzi medium c. serum agar d. sugar broth c. meat peptone broth Lesson Bacterial genetics. Variation of bacteria. Brief review of the gene engineering. Transformation. Transduction. Conjugation. 1. Each of the following events involves recombination of chromosomal DNA EXCEPT: (A) Transduction of a chromosomal gene (B) Transposition of a mobile genetic element (C) Integration of a temperate bacteriophage (D)*Conjugation, eg, transfer of an R (resistance) factor 2. Which of the following events is MOST likely to be due to bacterial conjugation? (A) A strain of Corynebacterium diphtheriae produces a toxin encoded by a prophage (B) *A strain of Pseudomonas aeruginosa produces β-lactamase encoded by a plasmid similar to a plasmid 16 of another gram-negative organism (C)An encapsulated strain of Streptococcus pneumoniae acquires the gene for capsule formation from an extract of DNA from another encapsulated strain (D)A gene encoding resistance to gentamicin in the Escherichia coli chromosome appears in the genome of a virulent bacteriophage that has infected E. coli 3. Each of the following statements concerning the mechanisms of resistance to antimicrobial drugs is correct EXCEPT: (A) R factors are plasmids that carry the genes for enzymes which modify one or more drugs (B) Resistance to some drugs is due to a chromosomal mutation that alters the receptor for the drug (C) Resistance to some drugs is due to transposon genes that code for enzymes which inactivate the drugs (D)*Resistance genes from bacterial chromosome are rarely transferred by transduction 4. During an outbreak of gastrointestinal disease caused by an Escherichia coli strain sensitive to ampicillin, tetracycline, and chloramphenicol, a stool sample from one patient yields E. coli with the same serotype resistant to the three antibiotics. This phenomenon is the most closely associated with the next process: (A) Transduction (B) *Conjugation (C) DNA transformation (D) Transposition 5. A mutant cell line lacking a functional thymidine kinase gene was exposed to a preparation of DNA from normal cells; under appropriate growth conditions, a colony of cells was isolated that makes thymidine kinase This phenomenon is the most closely associated with the next process: (A) Transduction (B) Conjugation (C) *DNA transformation (D) Transposition 6. Corynebacteria diphtheria strain without a tox-gene was cultivated into the media with temperate βphage. After several passages toxigenic strain of C.diphtheria was isolated. It arose due to following type of recombination (choose from the list one correct answer): (A) *Transduction (B) Conjugation (C) DNA transformation (D) Transposition 7. Bacterial genome may include next DNA molecules EXCEPT: (A) Chromosomal DNA (B) IS-elements (C) plasmids (D) transposons (E) *phage DNA 8. Mutations may arise under influence of mutagens onto the bacterial genome. Choose physical mutagen from the list: (A) desiccation (B) sun light (C)* ultraviolet radiation (D) pH variation 9. Mutations may arise under influence of mutagens onto the bacterial genome. Choose chemical mutagen from the list: 17 (A)* 5-bromuracil (B) sulfuric acid (C) nitrates (D) thymidine 10. In genetic engineering transferring of necessary genetic material from eukaryotic cell to prokaryotic cell may be done with vectors. Choose from the list the most reliable and often used vectors: (A) temperate bacteriophages (B) *plasmids (C) DNA fragments (D) adenoviruses 11. From patient with diarrhea pure culture of Shigella sonnei was isolated. It grew on the solid medium with formation different types of colonies. This type of bacterial variation is known as: (A) mutation (B) pleomorphism (C) heteromorphism (D) *dissociation 12. In 1928 F. Griffith established that from mice infecting with small doses of a non-pathogenic decapsulated type II Streptococcus pneumoniae culture together with pathogenic capsulated type III culture killed by healing, virulent capsulated S. pneumoniae will be isolated. Ability to produce capsule by previously non-capsulated strain arise due to next type of genetic recombination as: (A) Transposition (B) Transduction (C)* Transformation (D) Conjugation Lesson Influence of the physical and chemical factors on microorganisms. Sterilization techniques 1. Each of the following statements concerning the killing of bacteria is correct EXCEPT: (A) Lysozyme in tears can hydrolyze bacterial cell walls (B) Silver nitrate can inactivate bacterial enzymes (C) Detergents can disrupt bacterial cell membranes (D) *Ultraviolet light can degrade bacterial capsules 2. Each of the following statements concerning the killing of bacteria is correct EXCEPT: (A) A 70% solution of ethanol kills more effectively than absolute (100%) ethanol (B) An autoclave uses steam under pressure to reach the killing temperature of 121 °C (C) The pasteurization of milk kills pathogens but allows many organisms and spores to survive (D) *Iodine kills by causing the formation of thymine dimers in bacterial DNA 3. Find the false statement concerning sterilization and disinfection from following: (A) Chlorine is an oxidizing agent that effectively kills bacteria and viruses (B) *Boiling water effectively kills spores (C) Filtration is used to sterilize heat labile fluids (D) Chlorhexidene is a routine hand and skin disinfectant (E) Steam under pressure is the most reliable method of sterilization 4. Choose the correct method for sterilization of the plastic items: (A) boiling into the sodium carbonate for 30 minutes (B) *gas sterilization with ethylene oxide (C) autoclaving 18 (D) sterilization into the hot air oven at 160°C 5. After smear preparation student have to sterilize the charged loop. Which from following methods will be the most reliable? (A) *flaming until it becomes red hot (B) treatment with disinfectant solution (C) autoclaving (D) boiling 6. During the work with pathological material (suspension of feces from patient with diarrhea) laboratory assistant pour it off occasionally onto the table. He has to eliminate material from contaminated area next way: (A) wash off the residual suspension and dry surface (B) switch on the bactericidal ultraviolet lamp and treat contaminated surface with UV rays (C) *treat the contaminated surface with chloramines’ solution for 5-10 min (D) treat the place of accident with boiling water 7. Choose incorrect statement about desinfectants: (A) *iodine solutions disrupt bacterial membranes (B) they decrease number of pathogenic bacteria onto the treated surface (C) phenolic derivates inhibit bacterial enzymes and denaturate proteins (D) chlorine oxidizes microbial enzymes 8. Choose correct statement about desinfectants: (A) *they are used to decrease the number of microorganisms on the treated objects (B) they are effective to kill spores in the environment (C) they are enough reliable to treat surgical instruments (D) desinfectants are applied on the wound surface to prevent suppurative complication The bank of Krok-1 tests 1. With the purpose of acceleration sugar-containing media were sterilized by live, nonpressurized steam in one day: during 30 minutes in the morning, in the evening, and in the afternoon. In what way should the media have been sterilized? A. During 1 hour. B. During 15 min., three days in succession. C. During 30 min., two days in succession. D. *During 30 min., three days in succession. E. During 15 sec. twice a day. 2. After practical activity in a laboratory students must put in order and disinfect their working place. Choose the chemical agent, which should be used. A. Ether. B. Chloric acid. C. Formalin. D. Chloroform. E. *Chloramine 3. In a laboratory, researches on tetanus diagnostics were carried out. What method of sterilization should be used to destroy the isolated cultures of tetanus pathogens? A. *Autoclaving (steam under pressure). B. Boiling. C. Tyndallization. D. Dry heat. 19 E. Pasteurization 4. Control of instrument sterilization in autoclave was carried out with the help of a biologic method. What bacteria are the most rational to use as test-microorganisms? A. *Spore forming. B. Capsule forming. C. Acid-fast. D. Pathogenic. E. Thermophilic. Lesson Chemotherapeutic agents 1. Which statement is MOST accurate regarding the aminoglycosides? (A) It is bacteriostatic (B) *It inhibits protein synthesis (C) It prevents formation of folic acid (D) It inhibits DNA synthesis 2. Each of the following statements regarding the selective action of antibiotics on bacteria is correct EXCEPT: (A) Chloramphenicol affects the large subunit of the bacterial ribosome, which is different from the large subunit of the human ribosome (B) *Isoniazid affects the DNA polymerase of bacteria but not that of human cells (C)Sulfonamides affect folic acid synthesis in bacteria, a pathway that does not occur in human cells (D)Penicillins affect bacteria rather than human cells because bacteria have a cell wall, whereas human cells do not 3. Each of the following statements concerning the mechanism of action of antimicrobial drugs is correct EXCEPT: (A)Vancomycin acts by inhibiting peptidoglycan synthesis (B) Quinolones, such as ciprofloxacin, act by inhibiting the DNA gyrase of bacteria (C)*Erythromycin is a bactericidal drug that disrupts cell membranes by a detergent like action (D)Aminoglycosides such as streptomycin are bactericidal drugs that inhibit protein synthesis 4. Each of the following statements concerning the resistance of bacteria to antimicrobial drugs is correct EXCEPT: (A) Resistance to chloramphenicol is known to be due to an enzyme that acetylates the drug (B) Resistance to penicillin is known to be due to reduced affinity of transpeptidases (C) Resistance to penicillin is known to be due to cleavage by β-lactamase (D) *Resistance to tetracycline is known to be due to an enzyme that hydrolyzes the ester linkage 5. Each of the following statements concerning the mechanism of action of antibacterial drugs is correct EXCEPT: (A) Cephalosporins are bactericidal drugs that inhibit the transpeptidase reaction and prevent cell wall synthesis (B) *Tetracyclines are bacteriostatic drugs that inhibit cell wall synthesis by binding to peptide chains in peptidoglycan (C) Aminoglycosides are bacteriostatic drugs that inhibit protein synthesis by activating ribonuclease, which degrades mRNA (D) Gramicidin is a bactericidal drugs that disrupts cells membranes by a detergent like action 6. Chemical modification of benzylpenicillin (penicillin G) has resulted in several beneficial changes in the clinical use of this drug. Which one of the following is NOT one of those beneficial changes? 20 (A) Lowered frequency of anaphylaxis (B) *Increased activity against gram-negative rods (C) Increased resistance to stomach acid (D) Reduced cleavage by penicillinase 7. Each of the following statements concerning resistance to antibiotics is correct EXCEPT: (A) Resistance to aminoglycosides can be due to phosphorylating enzymes encoded by R plasmids (B) *Resistance to sulfonamides can be due to enzymes that hydrolyze the β-lactam ring structure (C) Resistance to penicillins can be due to alterations in binding proteins in the cell membrane (D) Resistance to cephalosporins can be due to cleavage of the β-lactam ring 8. Each of the following statements concerning penicillin is correct EXCEPT: (A) β-lactam ring is a feature of penicillin molecule (B) The structure of penicillin resembles that of a dipeptide of alanine, which is a component of peptidoglycan (C) *Penicillin is a bacteriostatic drug because autolytic enzymes are not activated (D) Penicillin inhibits transpeptidases, which are required for cross-linking peptidoglycan 9. Each of the following statements concerning the mechanisms of resistance to antimicrobial drugs is correct EXCEPT: (E) R factors are plasmids that carry the genes for enzymes which modify one or more drugs (F) Resistance to some drugs is due to a chromosomal mutation that alters the receptor for the drug (G)Resistance to some drugs is due to transposon genes that code for enzymes which inactivate the drugs (H)*Resistance genes from bacterial chromosome are rarely transferred by conjugation 10. Each of the following statements concerning the quinolones is correct EXCEPT: (A) *The drug is bacteriostatic (B) The drug inhibits bacterial DNA-gyrase (C) It is artificially synthesized drug (D) It has broad spectrum of antibacterial activity 11. All of the following inhibit cell wall synthesis EXCEPT: (A) *Chloramphenicol (B) Penicillin (C) Cephalosporin (D) Bacitracin (E) Vancomycin 12. Isolated culture from wound discharge should be tested in sensitivity to antibiotics. Choose the appropriate method from following: a. *Disk diffusion method b. Cultivation on the special media c. Staining by Gram d. Phagotyping of isolated culture e. Agglutination test The bank of Krok-1 tests 1. The staphylococcus, isolated from the nasopharynx of a nurse during hospital infection, differs from the strain, isolated from patients, by containing R-plasmid. What feature can the hospital strain acquire due to the conjugation with the strain isolated from the nurse? A. *Antibiotics resistance. B. Production of enterotoxin. 21 C. Production of epidermolitic toxin. D.Coagulase production. E. Rough colonies formation. Lesson Infection. Factors of a bacterial pathogenicity, their detection. Biological method of a laboratory diagnostics 1. Exotoxins are responsible for: a. Fever b. Inflammatory reactions c. General intoxication d. Decreasing of the blood pressure e. *Specific action depending on the type of toxin 2. Each of the following statements concerning endotoxins is correct EXCEPT: (A)They are less potent (ie, less active on a weight basis) than exotoxins (B) They are more stable on heating than exotoxins (C)*They bind to specific cell receptors, whereas exotoxins do not (D)They are part of the bacterial cell wall, whereas exotoxins are not 3. Which one of the following BEST describes the mode of action of endotoxin? (A) Degrades lecithin in cell membranes (B) Inactivates elongation factor 2 (C) Blocks release of acetylcholine (D)*Causes the release of biologically active mediators, responsible for shock reaction 4. The effects of endotoxin include each of the following EXCEPT: (A)*Opsonization (B) Fever (C)Activation of the coagulation cascade (D)Hypotension 5. Each of the following statements concerning exotoxins is correct EXCEPT: (A) When treated chemically, some exotoxins lose their toxicity and can be used as im-munogens in vaccines (B) Some exotoxins are capable of causing disease in purified form, free of any bacteria (C) Some exotoxins act in the gastrointestinal tract to cause diarrhea (D) *Some exotoxins contain lipopolysaccharides as the toxic component 6. Each of the following statements concerning endotoxins is correct EXCEPT: (A) The toxicity of endotoxins is due to the lipid portion of the molecule (B) *Endotoxins are found in most gram-positive bacteria (C) Endotoxins are located in the cell wall (D) The antigenicity of somatic (O) antigen is due to repeating oligosaccharides 7. Each of the following statements concerning exotoxins is correct EXCEPT: (A) Exotoxins are polypeptides (B) Exotoxins are more easily inactivated by heat than are endotoxins (C) *Exotoxins are less toxic than the same amount of endotoxins (D) Exotoxins can be converted to toxoids 8. Some bacterial enzymes belong to invasion factors and play important role in the pathogen spreading from the portal of entry. Choose from the list culture media for detection bacterial hemolysins: 22 (A) (B) (C) (D) MPB MPA *Blood agar Salt yolk agar 9. The first stage of any infection is adhesion of the pathogen to host cells. All from following are adhesion factors EXCEPT: (A) Peptidoglycan (B) Pili (C) Flagella (D) *Lipopolysaccharide in the cell envelope (E) Capsule 10. Some bacteria can cause generalized infection due to ability to split host tissue and spread from the site of entry. Choose from the list invasion factors of a pathogen: (A) Endotoxins (B) *Virulent enzymes (C) Adhesins (D) Exotoxins (E) Antiphagocytic factors 11.Which virulent factor from following capable of splitting connective tissue of a host? (A) Plasmocoagulase (B) *Hyaluronidase (C) IgA protease (D) Fibrinolysine (E) Hemolysine 12. Which virulent factor from following capable of splitting connective tissue of a host? (A)Plasmocoagulase (B) IgA protease (C)* Collagenase (D) Lecitinase (E) Deoxyribonuclease The bank of Krok-1 tests 1. Bacteriological examination of the feces of a 38-year-old woman, who had been ill with typhoid fever 1.5 year ago, revealed Salmonella typhi. Name the form of the infectious process. A. *Bacteria carrying. B. Secondary infection. C. Reinfection. D. Superinfection. E. Recidivation. 2. A child, who is recovering after measles, has developed pneumonia caused by opportunists. What is the most probable form of this infection? A. Reinfection. B. *Secondary infection. C. Superinfection. D. Persistence infection. E. Mixed infection. 23 3. A patient recovered completely after Sonne shigellosis and was repeatedly infected by this causative agent. How is such form of infection called? A. Superinfection. B. Recidivation. C. *Reinfection. D. Persistent infection. E. Chronic infection. 4. A patient has acute gonorrhea diagnosed. It is known, that the patient was ill with gonorrhea before, but was cured completely. Name the form of this infection. A. Secondary infection. B. Superinfection. C. Recidivation. D. *Reinfection. E. Autoinfection. 5. A 2-year-old boy was not given APDT vaccine according to schedule. On his pharyngeal tonsils there were revealed microorganisms, which by morphological and biochemical properties are identical to Corynebacterium diphtheriae. But precipitation test in gel with antitoxic serum has been negative. What form of infectious process can this agent cause without urgent treatment? A. *Asymptomatic bacteria carrying. B. Easy nontoxic form of disease. C. Toxic complicated form of disease. D. Chronic disease with the damage of internal organs. E. Persistent infection. Lesson Species immunity. Detection of the nonspecific resistance factors 2. Host defense mechanisms which protect the respiratory from infection include all of the following EXCEPT: a. Viscous layer of mucus b. Lysozyme and lactoferrin c. *Low pH d. Ciliated epithelial cells e. Secretory IgA 3. a. b. c. d. e. Immunity may be natural or acquired. Which of the following best describes acquired immunity? Increase in C-reactive protein (CRP) Presence of natural killer (NK) cells Complement cascade *Maternal transfer of antibody with breast milk and colostrum Inflammatory response 4. a. b. c. d. e. Antigens can best be processed for presentation by: *Macrophages Kupffer cells B cells Young erythrocytes Suppressor T cells 5. a. b. Which type of immunity will be form after infection? Artificial, acquired, active *Natural, acquired, active 24 c. d. e. Artificial, acquired, passive Natural, acquired, passive Innate, non-specific 6. Choose from the list the correct statement about mechanisms of complement activation: a. *An immune complex activates complement by classical pathway b. Classical pathway of complement activation starts with activation of C3 and C4 c. Bacterial polysaccharides, endotoxins, properdin are capable to activate complement system by alternate pathway d. Complement system is self activated e. Complement belongs to specific defence 6. During cascade mechanism of complement activation C3 is cleaved to form C3a and C3b by C3 convertase. C3b is involved in all of the following EXCEPT: (A) *Altering vascular permeability (B) Promoting phagocytosis (C) Forming alternative-pathway C3 convertase (D) Forming C5 convertase 7. Activated proteins C3a and C5a from complement system can cause (A)Bacterial lysis (B) *Vascular permeability (C)Phagocytosis of IgE-coated bacteria (D)Aggregation of C4 and C2 8. Complement fixation refers to (A)The ingestion of C3b-coated bacteria by macrophages (B) The destruction of complement in serum by heating at 56°C for 30 minutes (C)*The binding of complement components by antigen-antibody complexes (D)The interaction of C3b with mast cells 9. The classic complement pathway is initiated by interaction of Cl with (A)Antigen (B) Factor B (C)*Antigen-IgG complexes (D)Bacterial lipopolysaccharides 10. Natural killer cells are (A)B cells that can kill without complement (B) Cytotoxic T cells (C)Increased by immunization (D)*Able to kill virus-infected cells without prior sensitization 11. Lactoferrin is a humoral factor of non-specific resistance. Which from the next statements is the true about lactoferrin? a. it modifies milk sugars b. *it binds iron ions and competes with bacterial iron-bindings proteins c. lactoferrin may activate complement system d. it belongs to antiviral substances of the host e. it belongs to cellular mediators The bank of Krok-1 tests 25 1. A 1.5-year-old boy who hasn't been inoculated according to the schedule contacted with a patient with measles. For emergency specific prevention the child was given donor γ-globulin. What kind of immunity developed in this case? A. Natural. B. *Passive. C. Antitoxic. D. Postvaccinal immunity. E. Local. 2. After BCG vaccination of infants immunity to tuberculosis is preserved until there are live bacteria of vaccine strain in the body. Name this kind of immunity. A. Type specific. B. Humoral. C. *Non-sterile (infectious). D. Innate. E. Crossed. Lesson Acquired (specific) immunity. Antigens and antibodies. Serological method of the laboratory diagnostics of infectious diseases. Agglutination test, its modifications 1. The identification of bacteria by serologic tests is based on the presence of specific antigens. Which one of the following bacterial components is LEAST likely to contain useful antigens? (A) Capsule (B) Flagella (C) Cell wall (D)*Ribosomes 2. Bacterial surface structures that show antigenic diversity include each of the following EXCEPT: (A) Pili (B) Capsules (C) Flagella (D) *Peptidoglycan 3. All bacterial antigens from following may be detected with agglutination test, EXCEPT: (A) O-antigen (B) H-antigen (C) Surface protective antigen (A-protein) (D) *Exotoxins 4. Laboratory assistant has to identify E.coli based on its antigenic structure. Which from following is taken to detect O-antigen of the isolated strain with agglutination test? (A) Group O-diagnosticums of the E.coli (B) *Group specific O-sera (C) Type specific H-sera (D) Type specific K-sera 5. Serological tests are used to diagnose infection diseases by antibody detection in the patient sera. What is antibody titer of examined serum? (A) The highest amount of the tested serum that is agglutinated with specific diagnosticum (B) *The highest dilution of the tested serum that is agglutinated with specific cellular antigen (C) The lowest dilution of the tested serum that gives positive result in agglutination test (D) The average amount of the tested serum that agglutinates specific antigen 26 6. In laboratory practice some modification of the agglutination test are used. Which from the list is used as antigen in the indirect hemagglutination test? (A) Hemolytic serum (B) Sheep erythrocytes (C) *Sheep erythrocytes covered with specific either bacterial or viral antigen (D) Sheep erythrocytes covered with antiimmunoglobulin antibody 7. Molecule of a complete antigen consists from some parts. What is epitopes of antigen? (A) *Specific part that reacts with antibody paratope (B) Carrier of antigen determinants (C) Specific parts of antigen molecule located inside the polypeptide chain (D) Non-specific parts of antigen molecule with the same structure for different antigens 8. Antigen binds to specific antibody by special sites on the molecule surface. What is valence of antigen? (A) The number of possible variants of antigen dimensional structure (B) *The number of the different epitopes on the antigen molecule (C) The number of the same epitopes on the molecule surface (D) The number of a protein molecule configuration 9. Student has to detect somatic antigens of the tested strain of Salmonella. Which components should be taken for presumptive agglutination test? (A) Diagnostic serum, tested culture (B) *Diagnostic serum, tested culture, isotonic solution (C) Diagnosticum, serum and isotonic solution (D) Diagnosticum, tested culture, isotonic solution (E) Diagnostic serum, sterile water, and tested culture The bank of Krok-1 tests 1. Production of antibodies is one of the most important parts of immune response of an organism. Which type of cells actually secretes antibodies? A. T-lymphocytes. B. *Plasma cells. C. T-helpers. D. Macrophages. E. Microphages. 2. Scheduled vaccination against measles has been carried out in a kindergarten. What method can be used to check up the development of postvaccinal immunity? A. Bacteriological. B. Virologic. C. *Serologic. D.Bacterioscopic. E. Allergic. 3. After penetration into an organism bacteria are ingested by macrophages. What is the function of microphages in immunocompetent cells cooperation at the first stage of immune response? A. Activation of NK-cells. B. T-killers activation. C. *Processing and presentation of antigen to T-helpers. D. Antibodies production. E. Processing and presentation of antigen to T-killers. 27 4. A patient referred to a doctor on the second week of illness, which resembled of typhoid-paratyphoid disease by clinicoepidemiologic findings. The doctor decided to confirm the diagnosis by detecting specific antibodies. What preparation should be used for this purpose? A. Adsorbed monoreceptor serums. B. Diagnostic serums. C. Labeled serums. D. Monoclonal antibodies. E. *Diagnosticums. 5. Serologic diagnostics of infectious diseases is based on specific antigen-antibody interaction. Name the serological reaction, which consists in microorganisms clumping under the influence of specific antibodies in the presence of electrolyte. A. Hemadsorption. B. Precipitation. C. Complement fixation test. D. *Agglutination. E. Neutralization. 6. Serologic diagnostics of infectious diseases is based on specific antigen-antibody interaction. Name the serological reaction,during which highly refined antigens are adsorbed on erythrocytes. A. Neutralization. B. Precipitation. C. Complement fixation test. D. Hemadsorption. E. *Indirect hemagglutination. 7. A causative agent has been isolated from a patient with acute gastroenteritis. It should be identified by antigen structure. Which serological reaction should be used? A. Precipitation. B. Complement fixation test. C. Neutralization. D. *Agglutination. E. Opsonization 8. Effective diagnostics of enteric bacteria carrying is based on detecting antibodies to certain bacteria antigens during indirect hemagglutination test. Name the standard preparation which should be used. A. Sheep erythrocytes and hemolytic serum. B. Antibodies against immunoglobulins of basic classes. C. Monoclonal antibodies. D. Monoreceptor diagnostic serum. E. *Erythrocytic diagnosticum (erythrocyte adsorbed by bacteria antigens). Lesson Antibodies. Serological method of the laboratory diagnostics of infectious diseases. Precipitation tests, its modifications 1. The effects of antibody on bacteria include each of the following EXCEPT: (A) Lysis of gram-negative bacteria in conjunction with complement (B) Enhancement of phagocytosis (C)*Inhibition of bacterial metabolism (D)Inhibition of adherence of bacteria to mucosal surfaces 2. Choose correct statement about important function of antibody in a host defense from the next: (A) Activation of lysozyme that degrades the cell wall 28 (B) Acceleration of proteolysis of exotoxins (C)*Facilitation of phagocytosis (D)Inhibition of bacterial protein synthesis 3. The MAIN host defense against bacterial exotoxins is (A) Activated macrophages secreting proteases (B) *IgG and IgM neutralizing antibodies (C)Helper T cells (D)Modulation of host cell receptors in response to the toxin 4. The fragment required for an IgG molecule to bind complement is (A) Fab (B) *Fc (C) Variable part of a L-chain (D) Variable part of a H-chain (E) Constant parts of the L-chains 5. The antibody-binding site is formed primarily by (A) The constant regions of H and L chains (B) *The hypervariable regions of H and L chains (C) The hypervariable regions of H chains (D)The variable regions of H chains (E) Tke variable regions of L chains 6. The class of immunoglobulin present in highest concentration in the blood of a human is (A) *IgG (B) IgM (C) IgA (D)IgD (E) IgE 7. Which one of the following properties of antibodies is NOT dependent on the structure of the heavychain constant region? (A) Ability to cross the placenta (B) Isotype (class) (C)Ability to fix complement (D)*Ability to bind antigen 8. Each of the following statements concerning the variable regions of heavy chains and the variable regions of light chains in a given antibody molecule is correct EXCEPT: (A) *They have the same amino acid sequence (B) They define the specificity for antigen (C) They are encoded on different chromosomes (D)They contain the hypervariable regions 9. A primary immune response in an adult human requires approximately how much time to produce detectable antibody levels in the blood? (A) 12 hours (B) 3 days (C) *1 week (D) 3 weeks 10. Your patient became ill 10 days ago with a viral disease. Laboratory examination reveals that the patient's antibodies against this virus have a high ratio of IgM to IgG. What is your conclusion? 29 (A) *It is unlikely that the patient has encountered this organism previously (B) The patient is predisposed to IgE-mediated hypersensitivity reactions (C) The patient has encountered this organism previosly (D)It is likely that the patient has an autoimmune disease 11. Idiotypic determinants are located within (A) *Hypervariable regions of heavy and light chains (B) Constant regions of light chains (C) Constant regions of heavy chains (D) The hinge region 12. An antibody directed against the idiotypic determinants of a human IgG antibody would react with (A) The Fc part of the IgG (B) *An IgM antibody produced by the same plasma cell that produced the IgG (C) All human kappa chains (D) All human gamma chains 13. Which one of the following statements concerning immunoglobulin allotypes is CORRECT? (A) Allotypes are found only on heavy chains (B) Allotypes are determined by class I MHC genes (C)Allotypes are confined to the variable regions (D)*Allotypes are due to genetic polymorphism within a species 14. Which one of the immunoglobulins BEST fits the following description: It is found in plasma as a dimer with a J chain. As it passes through mucosal cells, it acquires a secretory piece that protects it from degradation by proteases. (A) IgM (B) IgG (C) *IgA (D) IgD (E) IgE The bank of Krok-1 tests 1. Serologic diagnostics of infectious diseases is based on specific antigen-antibody interaction. How is the antigen sedimentation from solution reaction under the influence of immune serum in the presence of electrolyte called? A. Neutralization. B. Agglutination. C. Complement fixation. D. Hemadsorption. E. *Precipitation. 2. Bloodstains are revealed on the clothes of a person accused of a murder. What test can prove that the blood is human? A. Neutralization. B. Immunofluorescence test. C. Complement fixation test. D. Agglutination. E. *Precipitation. Lesson Serological tests. Complement fixation test 30 1. In setting up a complement fixation test for antibody, the reactants should be added in what sequence? (Ag = antigen; Ab = antibody; C = complement; EA = antibody-coated indicator erythrocytes.) (A)Ag + EA + C/wait/ + patient's serum (B) C + patient's serum + EA/wait/ + Ag (C)Ag + patient's serum + EA/wait/ + C (D)*Ag + patient's setum + C/wait/ + EA 2. You suspect syphilis at patient with typical lesion on the glans penis. Complement fixation test is used to detect specific antibody in patient serum. How is tested serum prepared before serological test? A) It should be kept at +40C into the fridge for 2 hours B) *It should be heated at 560C for 30 min to destroy own complement C) It should be used without any delay D) It should be heated at 600C for 90 min to destroy own complement 3. To indicate whether compliment has fixed or no, laboratory assistant adds to mixture #1 (tested serum+Ag+complement ) hemolytic system. From which components does hemolytic system consist of? A) Human erythrocytes and tested serum B) *Sheep erythrocytes and immune serum from rabbit immunized with sheep erythrocytes C) Sheep erythrocytes and immune serum from volunteer D) Rabbit erythrocytes and human serum 4. Complement fixation test is made with two stages and two phases. First stage consists of the tested mixture (phase 1 ) and indicator system (phase 2) preparation. Which components are mixed in tested mixture? A) Hemolytic serum and erythrocytes B) *Tested serum, diagnosticum (Ag), complement C) Tested serum, complement, erythrocytes D) Hemolytic serum, erythrocytes, tested serum/ antigen E) Antigen, complement, hemolytic serum, erythrocytes 5. Complement fixation test is widely used to detect complement-fixing antibody in the patient serum and to confirm clinical diagnosis with serological testing. From which serum from following complement is taken? A) Human B) Rabbit C) *Guinea pig D) Mouse E) Rat 6. Complement fixation test is made to confirm ricketsiosis, viral diseases, chlamydiosis, some fungal diseases, etc. Which from following is titer of tested serum? A) The lowest dilution of patient serum that demonstrate negative result B) The lowest dilution of patient serum that demonstrate positive result C) The highest dilution of patient serum that demonstrate negative result D) *The highest dilution of patient serum that demonstrate positive result E) The average dilution of patient serum that demonstrate positive result 7. Which result is read as positive at complement fixation test? A) *Erythrocytes in the indicator system are not hemolysed B) Erythrocytes in the hemolytic system are lysed C) Erythrocytes in the indicator system are partly destroyed D) The most part of erythrocytes is lysed 8. Which result is read as negative at complement fixation test? 31 A) B) C) D) *Erythrocytes in the hemolytic system are completely lysed Erythrocytes in the indicator system are not hemolysed Erythrocytes in the indicator system are partly destroyed The most part of erythrocytes is not lysed 9. Laboratory assistant takes for complement fixation fresh patient serum, ricketsial antigen, titred complement and indicator hemolytic system. He mixes tested serum, antigen and complement, and after 40 minutes` exposure at thermostat this mixture he adds to it hemolytic system. Mixture is thermostated again at same exposure. After exposure in the tested mixture hemolysis will arise. Assistant estimates result as negative. Which mistake from following was made and lead to false-negative result? A) *He did not heat tested serum to destroy own complement that lead to complement excess B) He did not cool tested serum and “cool antibody” did not react in CFT C) He did test by incorrect way D) The exposure was too short for reaction 10. Laboratory assistant have to prepare hemolytic system for complement fixation test. He titrated hemolytic serum, washed sheep erythrocytes with isotonic solution and prepared 3% suspension of them. Then he mixed serum and erythrocytes and incubated into the thermostat to check it before test. After 30min. incubation erythrocytes will be lysed. Which mistake ahs made by laboratory assistant? A) He did not wash erythrocytes thoroughly B) It is not necessary to wash erythrocytes C) The concentration of red cells is too small for testing D) *The hemolytic serum should be heated at 560C for 30 min before testing E) The hemolytic serum should be heated at 560C for 30 min before titration The bank of Krok-1 tests 1. A patient with a suspicion of the chronic form of gonorrhea was admitted to a hospital. Choose a twosystem serological reaction for detecting specific antibodies in the serum. A. *Complement fixation test. B. Neutralization. C. Agglutination. D. Radioimmunoassay. E. Enzyme immunoassay. 2. Hemolytic serum against sheep erythrocytes is necessary for work in the laboratory, where serologic diagnostics of infectious diseases is carried out. Name the test, during which the serum is used. A. Viral hemagglutination test. B. Indirect hemagglutination test. C. Test for the diagnostics of newborns' diseases. D. Passive hemagglutination. E. *Complement fixation test. Lesson Serological tests with labelled antibody or antigen, their application in the laboratory diagnostics 1. In the determination of serum insulin levels by radioimmunoassay, which one of the following is NOT needed? (A)Isotope-labeled insulin (B) Anti-insulin antibody made in goats (C)Anti-goat gamma globulin made in rabbits (D)*Isotope-labeled anti-insulin antibody made in goats 2. High specific microscopic examination with fluorescent antibody allows diagnosing some 32 infectious disease during 4-6 hours after sample collection. Which type of antibody is used in direct immunofluorescent test? A) Antibody to specific antigen of a causative agent B) *Fluorescent-labelled antibody to specific germ antigen C) Antihuman immunoglobulin antibody D) Fluorescent-labelled antihuman immunoglobulin antibody E) Fluorescent-labelled antirabbit immunoglobulin antibody 3. Enzyme-linked assay is the modern test to detect either specific antibody in the tested serum or specific antigen of a pathogen in the collected sample. Which reagent is linked with enzyme at indirect immunosorbent assay? A) Tested serum antibody B) Known antigen absorbed in the well C) Human immunoglobulin added for competition with tested sample antibody D) *Antihuman immunoglobulin 4. Which from following microscopic test is useful to identify causative virus in the collected sample? a) Electron microscopy b) *Immune electron microscopy c) Light microscopy d) Hemabsorption test 5. Viral hepatitis, HIV infection may be diagnosed with ELISA. But to confirm suspected diagnosis immunobloting is used. Why this method is preferred to confirm diagnosis? a) High specific antigen is used in immunoblotting test b) It is impossible to obtain false-positive result at it c) It is more sensitive than ELISA d) *It is made to reveal antibody to some important antigens of causative virus 6. Which one of the following sequences is appropriate for testing a patient for antibody against the AIDS virus with the ELISA procedure? (The assay is carried out in a plastic plate with incubation and a wash step after each addition except the final one.) (A) Patient's serum/enzyme substrate/HIV antigen/enzyme-labeled antibody against HIV (B) *HIV antigen/patients serum/enzyme-labeled antibody against human gamma globulin/ enzyme substrate (C)Enzyme-labeled antibody against human gamma globulin/patient's serum/HIV antigen/enzyme substrate (D)Enzyme-labeled antibody against HIV/HIV antigen/patient's serum/enzyme substrate 7. A causative agent of infectious disease may be detected in the smear from collected sample with specific antibody. Which types of sera are used in indirect immunofluorescent test? A) *Rabbit specific antibody and fluorescent-labelled antirabbit immunoglobulin antibody B) Fluorescent-labelled rabbit antibody to specific germ antigen C) Antibody to specific antigen of a causative agent and fluorescent-labelled antihuman immunoglobulin antibody D) Fluorescent-labelled antihuman immunoglobulin antibody 8. Enzyme-linked assay is the modern test to detect either specific antibody in the tested serum or specific antigen of a pathogen in the collected sample. Which reagent is linked with enzyme at “double antibody sandwich assay”? A) Tested antigen B) Known antibody absorbed in the well C) Human immunoglobulin 33 D) *Specific to unknown antigen antibody, added into the well after antigen The bank of Krok-1 tests 1. Indirect immunofluorescence test has been used to detect toxoplasmas antibodies in the blood serum of a pregnant woman. At first, the fixed smear of toxoplasmas was processed with the investigated serum. With what should the specimen be processed at the following stage? A. *Luminescent serum against human immunoglobulins. B. Fluorescent solution. C. Human normal immunoglobulin. D. Diagnostic serum containing toxoplasmas antibodies. E. Serum containing fluorescent-labeled toxoplasmas antibodies. 2. Causative agent antigens can be revealed in blood of patients with different infectious diseases. What reaction should be used, taking into account that antigenemia level is low? A. *ELISA. B. Agglutination test. C. Indirect hemagglutination test. D. Latex agglutination method. E. Immunoelectrophoresis. 3. A patient is hospitalized with the previous diagnosis of hepatitis B. To diagnose the disease a serological reaction based on interaction of antigen and antibody, chemically linked to peroxidase or alkaline phosphatase, was carried out. Name this reaction. A. Complement fixation test. B. Radioimmunoassay. C. Immunofluorescence test. D. *Enzyme immunoassay. E. Immobilization. Lesson Immunoprophylaxis and immunotherapy of infectious diseases. Vaccines and toxoides 1. All of the following are used for active immunization in humans EXCEPT: a) Toxoids b) Polysaccharides c) *Immunoglobulins d) Inactivated (killed) bacteria e) Inactivated (killed) viruses 2. Killed bacterial vaccine is: a) BCG b) *Pertussis c) B.anthracis non-capsulated strain d) Rabies vaccine e) MMR 3. Each of the following statements concerning viral vaccines is correct EXCEPT: A) In live, attenuated vaccines, the virus has lost its ability to cause disease but has retained its ability to induce neutralizing antibody B) In live, attenuated vaccines, the possibility of reversion to virulence is of concern C) *With inactivated vaccines, IgA mucosal immunity is usually induced D) With inactivated vaccines, protective immunity is due mainly to the production of IgG 34 4. Which one of the following strategies is MOST likely to induce lasting intestinal mucosal immunity to poliovirus? (A) Parenteral (intramuscular) vaccination with inactivated vaccine (B) Oral administration of poliovirus immune globulin (C) Parenteral vaccination with live vaccine (D)*Oral vaccination with live vaccine 5. Each of the following statements concerning rubella vaccine is correct EXCEPT: (A) The vaccine prevents reinfection, thereby limiting die spread of virulent virus (B) *The immunogen in the vaccine is killed rubella virus (C) The vaccine induces antibodies that prevent dissemination of die virus by neutralizing it during the viremic stage (D)The incidence of both childhood rubella and congenital rubella syndrome has decreased significantly since the advent of the vaccine 6. The two most common types of viral vaccines are killed vaccines and live, attenuated vaccines. Regarding these vaccines, which one of the following statements is the MOST accurate? (A) Killed vaccines induce a longer-lasting response than do live, attenuated vaccines (B) Killed vaccines are no longer used because they do not induce secretory IgA (C) Killed vaccines induce a broader range of immune responses than do live, attenuated vaccines (D)*Killed vaccines are safer to give to immunocompromised patients than are live, attenuated vaccines 7. To create active immunity against tetanus and diphtheria children are immunized with corresponding toxoids prepared from germs` exotoxins. Which method from following is used to make toxoid? a) Exotoxin is inactivated by utraviolet radiation b) Bacterial exotoxins are inactivated by heating at 800C for a hour c) *Exotoxin is treated with 0.4% formaldehyde at 38-400C for four weeks d) Exotoxin is treated with phenolic solution for four weeks 8. All from following associated vaccines are used for immunization according national schedule of immunization EXCEPT: a) DPT vaccine b) MMR vaccine c) DT vaccine d) DPT-Polio vaccine e) *TABte vaccine 9. Health care personnel are a high risk group of hepatitis B infection. Which modern vaccine is used to immunize groups that have great possibility to be infected with hepatitis B virus? A) HBsAg prepared from blood of chronic carries B) *Recombinant vaccine including HBsAg that is obtained from recombinant E.coli strain C) Recombinant strain of E.coli, producing HBsAg (vector vaccine) D) Attenuated hepatitis B virus E) antiHBs human immunoglobulin from immunized volunteers 10. Immunization of immune compromised children has some features. Which type of vaccine is strictly forbidden for immunization of this group? A) Recombinant vaccine due to presence additional fractions B) *Alive vaccine due to residual virulence C) Killed vaccine due to negative influence of preservations D) Chemical vaccine due to ability cause side effect The bank of Krok-1 tests 35 1. Numerous vaccine preparations have been used to induce human active immunity. Which preparation is presented by live attenuated bacteria? A. APDT vaccine. B. *BCG vaccine. C. Salk's vaccine. D. TABTe vaccine. E. Hepatitis B vaccine. Lesson Immunoprophylaxis and immunotherapy of infectious diseases. Immune serums and immunoglobulin 1. Monoclonal antibodies are: (F) Less specific than conventional antisera (G) Produced by a single clone of T cells (H) *Secreted by myeloma-thymic lymphocyte hybridoma (I) Antibodies with different specifity (J) Different classes of immunoglobulins 2. Antitoxins are sterilized by: (A) Autoclave (B) Hot air oven (C) *Filtration (D) Steam under low pressure (E) X rays 3. The main advantage of passive immunization over active immunization is that (A) It can be administered orally (B) *It provides antibody more rapidly (C) Antibody persists for a longer period (D)It contains primarily IgM 4. Each of the following statements concerning a hybridoma cell is correct EXCEPT: (A) The spleen cell component provides the ability to form antibody (B) The myeloma cell component provides the ability to grow indefinitely (C) *The antibody produced by a hybridoma cell is IgM, because heavy-chain switching does not occur (D) The antibody produced by a hybridoma cell is homogeneous; ie, it is directed against a single epitope 5. Diagnostic sera are used to identify antigens of a causative agent with serological test. How is diagnostic serum obtained? a) By immunization of volunteers b) *By immunization of rabbits c) By immunization of mice d) By immunization of horses 6. Antitoxins are used for treatment infections caused by bacterial exotoxins (tetanus, diphtheria, botulism, etc.). Which way is used to obtain heterogenous antitoxin? a) Immunization of goats b) *Immunization of horses c) Immunization of mice d) Immunization of guinea pigs 7. Which type of immunity will be created after treatment with specific serum preparations? 36 a) b) c) d) e) *Artificial acquired passive Artificial acquired active Natural acquired active Natural acquired passive Natural inherited passive 8. To prevent tetanus wounded person is immunized with human antitetanus immunoglobulin. How long will passive immunity last? A) Two weeks B) *Two months C) 7-10 days D) Six months 9. To prevent tetanus wounded person is treated with heterogenous antitoxin. How long will passive immunity last? A) Two months B) *At least two weeks C) Six months D) More then three weeks 10. Immune serum preparations are used in medicine to: a) Treat severe infections b) *Treat and immediate prevent severe infection under high risk of disease c) Prevent infections of childhood d) Decrease transmission of dangerous germ from carrier The bank of Krok-1 tests 1. When using treatment antitoxic serums a patient is always given precisely defined doses. In what units is the activity of antitoxic antidiphtheric serum measured? A. *International. B. Flocculation. C. Antigenic. D. Agglutination. E. Neutralization. 37