C hair M icrobiology, V irology, and I mmunology

1. Chair Microbiology, Virology, and Immunology Human Parasitology

2. METHODS OF EXAMINATION IN MICROBIOLOGY BACTERIOSCOPIC BACTERIOLOGICAL SEROLOGICAL BIOLOGICAL ALLERGIC EXPRESS-DIAGNOSIS

3. BACTERISCOPIC METHOD Neisseria meningitidis

4. BACTERISCOPIC METHOD Neisseria meningitidis

5. BACTERISCOPIC METHOD Neisseria gonorrhoeae

6. BACTERISCOPIC METHOD

7. BACTERISCOPIC METHOD Yersinia pestis

8. BACTERISCOPIC METHOD Bacillus anthracis

9. BACTERISCOPIC METHOD Mycobacterium tuberculosis

10. BACTERISCOPIC METHOD Streptococcus pneumoniae

11. BACTERISCOPIC METHOD Clostridium pneumoniae

12. Bacteriological method Isolation of pure culture

13. Bacteriological method Neisseria meningitidis

14. Liquor

15. Serologic method Agglutination test

16. Serologic method Agglutination test

17. Serologic method Agglutination test

18. Serologic method IHAT

19. Serologic method Ring precipitation test

20. Serologic method Immunodiffusion (Ouchterlony) test

21. Serologic method Mancini’s test

22. Serologic method CFT

23. Serologic method ELISA

24. Serologic method ELISA

25. Biological method TBS in rabbit TBC in guinea pig

26. Allergic method Mantoux’s test

27. Express-diagnosis IFT

28. Clinical microbiology is the field of medical microbiology, which study microbial diseases in somatic departments of all specialties. There are general tasks, which provide the help to doctors in diagnosis, treatment and prophylaxis of purulent complications, before clinical microbiology

29. Features of purulent processes in non-infectious clinics: • - polietiology; • - non-specific clinical signs. • There are more then 2000 causative agents of purulent diseases. • The more frequent of them there are: • - genera Staphylococcus, Streptococcus, Bacteroides, Prevotella; • families Enterobacteriaceae (Proteus, Klebsiella, Escherichia, Serratia, Citrobacter, Hafnia etc.), Pseudomonadaceae, Neisseriaceae (Acinetobacter, Moraxella, Branchamella). • In urological and gynecological clinics - Mycoplasmas, Chlamydia.

30. Most of conditionally pathogenic bacteria belong to normal human flora so it’s hard to determine their etiologic role (etiological significance). So, they can present normal microflora of tested fluids and tissues and contaminate them from environment. That’s why for correct interpretation of examination results it’s necessary to know composition of normal microflora of tested samples. In that cases if tested samples are sterile (blood, synovial and pleural fluids, liquor, exudates) all microbes which are present in them may be causative agents of diseases.

31. Bacteria Frequently Present as Normal Flora Occasionally Causing Overt Disease

34. Pure culture of conditionally-pathogenic microbes may be causative agents of disease according to the such signs: -  microbes are present in tested material from pathologic focus in the amount of 104-105colony-forming units (CFU) in 1 mlor 1 g; -  repeated isolation from the same material the same culture; -  increasing in the patient’s serum antibodies to the autostrains or microbial culture, which can be causative agent.

35. There are necessary roles before collection of tested material: -  to take material before antibacterial therapy beginning or after some time after antibiotic inoculation which is necessary for its excretion from the organism (as a rule 8-10 hours); - to take material from infectious focus or examine proper discharges; - hold on to the strict aseptic for the purpose to prevent contamination of the specimen by microflora of environment;

36. -  material is taken into the sterile boxes; clinical specimen with anaerobic bacteria must be protected from atmosphere oxygen action; - the collection of an adequate specimen is useless if the time between collection and culturing allows the disease-producing organism to die (in another cases it’s necessary to use the refrigerator or special transport media); - isolation of viruses, Rickettsia, Chlamidia is made in specialize laboratories; -  to clinical specimen a proper document is added, which has data , which has data for correct microbiological examination.

37. Examination of the blood Bacteriemia, septicemia, septicopyemia

38. Bacteremia — bacteria in the blood — frequently is accompanied by the onset of chills and fever, an increase in pulse rate, and a drop in blood pressure. Even in infections in which bacteremia is a major aspect of the disease, the organisms in the bloodstream are not always constantly present in sufficient numbers to be grown from a single blood specimen. Patients with such infections may have to provide several blood specimens before the causative agent can be isolated. When an intermittent bacteremia is suspected, it is routine to obtain three 10 to 20-ml blood samples over a 24 hour period to maximize chances for isolation of the organism

39. Collection of a Blood Specimen In taking a blood specimen for culture, one should be aware that although blood is normally sterile, the skin that must be penetrated is not sterile. Routinely, the skin should be cleansed first with 70% to 95% alcohol to remove dirt, lipids, and fatty acids. The site then should be scrubbed with a circular, concentric motion (working out from the starting point) using a sterile gauze pad soaked in an iodophor. The iodine should be allowed to remain on the skin for at least 1 minute before it is removed by wiping with a sterile gauze pad soaked with 70% to 95% alcohol. It must be emphasized, however, that all this will be useless if the person drawing the blood palpates the vein after the cleaning process, thereby contaminating the very site that had been cleaned. After cleansing the penetration site, the blood can be withdrawn using either a sterile needle and syringe or a commercially available, evacuated blood collection tube.

40. Media Inoculated With Blood Specimens Blood always should be inoculated into the appropriate medium at the bedside partially evacuated, commercially available, blood culture bottles, which contain 30 to 100 ml of a rich, liquid medium such as brain-heart infusion or trypticase soy broth is routinely used. If possible, 10 to 20 ml of blood should be taken from the patient and inoculated into an approximately10 fold excess of the blood culture medium when possible, two such bottles should be inoculated. One is vented to permit the growth of aerobic bacteria (by inserting a sterile, cotton plugged needle through the rubber stopper until the bottle has filled with air), and the other is not vented to allow the growth of anaerobic organisms. Special media for aerobic and anaerobic culture are available. Some commercially available bottles are provided with a venipuncture set, which allows the blood to be injected into the medium.

41. Identification of Blood Isolates Blood cultures are incubated at 36°C and observed daily for at least 1 week for evidence of turgidity or hemolysis. Gram's stains, streak plates, and antibiotic susceptibility tests should be carried out as quickly as possible after the observation of visible growth in the original broth culture. In the absence of obvious growth in 1 or 2 days, blind subcultures on chocolate blood agar plates may speed the appearance of obligately aerobic organisms. Commercially available penicillinase can be added to blood cultures from patients who have received penicillin therapy. Penicillinase preparations should be checked for sterility to eliminate them as a potential source of contamination. Resins incorporated in special blood culture media can neutralize a broad spectrum of antibiotics.

42. Once a Gram's stain has rendered some information concerning the type of organism involved, special supplementary or differential media should be inoculated. MacConkey or eosin-methylene blue plates should be streaked if gram negative rods are present, and prereduced media should be inoculated if obligate anaerobes such as Bacteroides or Fusobacterium are suspected. The finding of organisms that constitute the normal flora or are frequent inhabitants of the skin (eg, diphtheroids, Staphylococcus epidermidis. Bacillus sp.) usually is viewed with suspicion, unless the frequency of isolation or the clinical setting indicates they did not arrive as contamination during the collection of the blood

43. Pathogenesis of toxic shok

44. If microbes are absent after 10 days of incubation it means, that inoculation results are negative and blood is sterile.

45. URINE Most urinary tract infections are initiated by organisms that gain entrance to the bladder by ascending through the urethra, and they are more common in women than in men In men, however, a chronic infection of the prostate gland also can be the source of a bladder infection (cystitis) or a kidney infection (pyelonephritis). In both sexes, most urinary tract infections are caused by normal flora enteric organisms, among which can be species of Escherichia, Klebsiella, Fnterobacter, Proteus, Pseudomonas, and Enterococcus.

46. Examination of urina Etiology of urethritis

47. Major causes of urinary tract infections

48. Pathogenesis of genital tract infections

49. Major causes of vaginitis Major causes of cervicites

50. Collection of Urine Specimens Urethral catheterization can yield samples with minimal contamination, but the danger of introducing organisms from the urethra into the bladder provides some risk to this procedure. Moreover, microbial flora in the urethra, particularly in men, can contaminate the specimen, leading the microbiologist to an erroneous conclusion. Therefore, catheterization is not performed routinely for the collection of urine samples. Instead, voided samples are obtained after careful cleansing of the external genitalia. However, the following considerations must be adhered strictly if bacteriologic reports on voided urine samples are to be meaningful.