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Institute for Microbiology, Medical Faculty of Masaryk University and St. Anna Faculty Hospital in Brno Miroslav Votava Agents of urinary tract infections Lecture for 3rd-year students 4th November, 2011 Importance of central nervous system infections – revision • CNS infections – relatively rare, but can have a very serious course • Incidence bacterial meningitis: 2/100.000/year viral meningitis: 10/100.000/year • Lethality bacterial meningitis, non-treated: >70 % treated: ~10 % Etiology of acute meningitis – revision I Always distinguish purulent meningitis (nearly always of bacterial origin) from aseptic one (usually of viral origin) Anamnesis Clinical disease Laboratory – above all the examination of CSF cytology (appearance and number of cells) biochemistry (proteins and glucose) microbiology (microscopy, antigens, culture) Etiology of acute meningitis – revision II Cytology and biochemistry of CSF marker norm cells 0-6/μl proteins glucose 20-50 mg/100 ml 40-80 mg/100 ml purulent meningitis aseptic meningitis ↑↑↑ (>1000) ↑↑(100-500) ↑↑ (>100) ↑ (50-100) ↓ (<30) ~ (30-40) Etiology of acute meningitis – revision III Etiology of purulent meningitis by the age in % age 0-1 m. 1-4 y. 5-29 30-59 ≥60 GBS Haem. Neiss. Str. other infl. b men. pneu. 50 70(↓) 45 40 50 Etiology of acute meningitis – revision IV Etiology of purulent meningitis by the age in % age 0-1 m. 1-4 y. GBS Haem. Neiss. Str. List. other infl. b men. pneu. mono. 50 33 70(↓) 15 10 10 5-29 45 25 20 30-59 10 40 33 25 50 ≥60 15 Etiology of acute meningitis – revision V Importance of purulent meningitis according to etiology (lethality and sequelae) impor- Group Haem. Neiss. Str. List. other tance B str. infl. b men. pneu. mono. letha† † (†) † lity seque+++ + + + lae Etiology of acute meningitis – revision VI The most common agents of aseptic meningitis: VIRUSES mumps virus (but CNS infection is clinically silent) enteroviruses: echoviruses (30 serotypes) coxsackieviruses (23 + 6 serotypes) tick-borne encephalitis virus (TBEV) rarely HSV and VZV and other neuroviruses rarely some bacteria leptospirae, borreliae, Mycobacterium tuberculosis Overview of Central-European neuroviruses – revision TBEV (tick-borne enc. v.) other arboviruses enteroviruses: polio LCMV coxsackie /morbilli v./ echo /EBV/ mumps v. /polyomaviruses JC & BK/ HSV, VZV, CMV /HIV/ rabies v. /prions/ Arboviruses in Central Europe – revision I Genus or family : arbovirus Flavivirus: TBEV WNV (West Nile v.) Orbivirus: Tribeč Bunyaviridae: Ťahyňa Batai (Čalovo) Uukuniemi Alfavirus: Sindbis Coltivirus: Eyach Disease Antibodies only + + + + ? ? + + Arboviruses in Central Europe – revision II Arboviruses isolated in Czech Republic, probably nonpathogenic for humans: Bunyaviridae: Lednice Sedlec Other European pathogenic arboviruses, which may be imported: dengue v. (flavivirus, Greece) CCHFV (nairovirus, Ukraine, Bulgaria) Toscana v. (phlebovirus, Italy) Bhanja v. (bunyavirus, Slovakia) chikungunya v. (alphavirus, Italy) Etiology of chronic meningitis – revision • Bacteria: Mycobacterium tuberculosis (meningitis basilaris) Treponema pallidum • Moulds and yeasts: aspergilli Cryptococcus neoformans Etiology of encephalitis – revision Encephalitis – only acute, of viral origin: • • • • tick-borne encephalitis v. HSV enteroviruses mumps v. Etiology of acute brain abscess – revision Acute brain abscesses are only of bacterial origin: • mixed anaerobic and aerobic flora • staphylococci (both S. aureus and coagulase negative staphylococci) • group A and D streptococci Etiology of chronic brain abscess – revision bacteria: Mycobacterium tuberculosis Nocardia asteroides mycotic organisms: Cryptococcus neoformans (yeast) parasites: Cysticercus cellulosae (tissue form of pork tapeworm Taenia solium) Microbiological examination of CNS infections – addendum I 1. PURULENT MENINGITIS Cerebrospinal fluid (CSF) microscopy: Gram; sometimes Ziehl-Neelsen; Heidenhein; Indian ink; wet mount detection of antigens: N. meningitidis A, C, Y, W135, B (Ag as E.coli K1); S. pneumoniae; GBS; H. influenzae b; Cryptococcus neoformans culture: blood agar (BA) with staph. line; chocolate agar in 5% CO2; BA anaerobically; Endo or MacConkey; if required media for mycobacteria, cryptococci, amoebae etc. Blood for haemoculture notably positive in meningococcal meningitis Microbiological examination of CNS infections – addendum II 2. OTHER NEUROINFECTIONS Viral neuroinfections (asept. meningitis & enceph.): CSF for the direct virus detection (PCR, tissue culture) blood for the antibody detection (IgM, 4fold titre rise): in susp. infections with TBEV, HSV 1 and 2, VZV, CMV, mumps virus Borreliosis and neurosyphilis: blood for the antibody detection Brain abscess (both acute and chronic): punctate or excission for bacteriology (incl. tbc), mycology (cryptococci) & histology (cysticerci) Urinary tract infections (UTIs) Frequency of UTIs: The 2nd most common infections (after respiratory ones) In adults: the most common infections in a general practitioner’s office Afflicting mainly females (because of shorter urethra) Examples of UTIs The most common UTI: cystitis develops ascendently caused by intestinal microflora main symptoms: dysuria (difficult urination with sharp and burning pain) pollakisuria (urgent need to urinate accompanied by urination of a small amount of urine only) Other UTIs: mainly pyelonephritis (more serious) origin: ascendent or hematogenous urethritis – will be dealt with as STD Etiology of UTIs Proportional representation of microbes differs in • non-complicated UTIs • infections accompanying structural abnormalities (prostatic hypertrophia, urinary stones, strictures, pregnancy, congenital defects, permanent catheters) • infections accompanying functional disorders (vesicoureteral reflux, neurological disorders, diabetes mellitus) Etiology of non-complicated UTIs circa 80 % Escherichia coli circa 10 % enterococci (Enterococcus faecalis) circa 5 % Proteus mirabilis rest: other enterobacteriae (Klebsiella pneumoniae, Kl. oxytoca, Ent. cloacae, C. freundii etc.) Streptococcus agalactiae coagulase neg. staphylococci (S. epidermidis, S. saprophyticus, S. haemolyticus etc.) yeasts (mainly Candida albicans) Etiology of complicated UTIs circa 80 %: the rest: Escherichia coli Klebsiella pneumoniae Proteus mirabilis Pseudomonas aeruginosa enterococci other enterobacteriae acinetobacters other G-neg. non-fermenting rods candidae Lege artis taking a urine sample 1. Only after a thorough cleaning of genital, incl. external orificium of urethra by means of soap and water 2. Take the middle stream of urine only 3. Use a sterile vessel 4. Pour urine into a sterile tube & stopper it promptly 5. If not possible to process it within 2 hours, place the specimen into 4 °C for 18 hours at most Semi-quantitative examination of the urine sample – I We are interested • not only in the kind of microbe present in the urine sample, but especially in • the amount of the microbe Why are we interested in the number of microbes in 1 ml of urine? Because • high numbers only stand for the UTI • low numbers mean usually contamination acquired during urination Semi-quantitative examination of the urine sample – II Therefore, the urine is inoculated on culture media by means of calibrated loop, usually taking exactly 1 μl of urine In this case 1 colony means 103 CFU/ml 10 colonies mean 104 CFU/ml 100 colonies mean 105 CFU/ml (CFU = colony-forming unit = 1 bacterial/yeast cell) Significant concentrations of bacteria in urine Type of specimen, symptoms Type of microbe Significant number (CFU/ml) Primary urine pathogen 103 Dubious urine pathogen 105 Middle stream, no symptoms Any 5 10 Suprapubic punction Any 1 10 Middle stream, symptoms present Media used in microbiological examination of urine • Blood agar inoculated by means of calibrated loop • Chromogenic medium oriented on the most frequent urinary pathogens; their colonies are of different colour inoculated by means of calibrated loop • According to requirements further media e.g. chromogenic medium for yeasts or a medium for MRSA Primary urine pathogens Escherichia coli & most of other enteric bacteria enterococci (mostly Enterococcus faecalis) Streptococcus agalactiae staphylococci (mostly coagulase negative: S. epidermidis, S. saprophyticus, S. haemolyticus etc.; but also S. aureus) yeasts (in the main Candida albicans) Pseudomonas aeruginosa & some other Gramnegative non-fermenting rods ... Homework 5 – solution Jacques-Louis David (17481825): Death of Marat (1783) What is the connection between this painting and medicine? • Jacques-Louis David, had a facial tumor • Jean Paul Marat, murdered by Charlotte Corday in 1793, was initially a physician • He was run through when taking a bath for treatment his skin disorder (probably dermatitis herpetiformis Dühring) Homework 5 Successful homework solvers: Roye Mauthner Mavis Araba Koufie At least partially successful solver: Karin Sharafi Congratulations! Homework 6 Who is the author of this painting and what is its name? Answer and questions The solution of the homework and possible questions please mail to the address [email protected] Thank you for your attention