TRACING THE CRIMINAL Part six: More gram-negative cocci and bacilli Institute for microbiology shows Survey of topics Clinical characteristics – G– cocci Story One n Johny was very childish, he had no experience with women even in 20 years of age. His friends made fun of him. Once they made a plan: they gave him lots of spirits and paid a prostitute for him. Johny had a feeling, that he is finally a man… only before pus started to drop from his penis… Certainly, you know, what is the criminal n Of course, it is Neiseria gonorrhoeae, (gonococcus) causing gonorrhoea. n Gonorrhoea is urethritis, in females also cervicitis; asymptomatically or symptomatically gonococci are found in pharynx and rectum, too. n In females, it is not a colpitis (vaginal inflamation), so it is not recommended to perform vaginal swabs in gonorrea. Cervical and urethral ones are prefered. Story two n Lucy learned four weeks for the examination of physiology. She did not leave the house at all and only sat at textbooks. At the exam she had intention, that she is not able to say a word, but finally she passed with „E“. n In the evening, she visited a dancing party with friends. The party was full of smoke and they danced during all night. Next day, Lucy was not well, she started to have fever and rash. (continuing) n In this moment, Lucy was hospitalized at infection clinic. In ambulance she failed unconcious and doctors constated metabolic failure. After ten hours of attempts to keep Lucy‘s vital functions, that had no effect, Lucy died. n Such a course of infection may be caused by a dangerous criminal. Some of his strains are present in throat of healthy persons… And the criminal is… Why the infecion comes sometimes, and sometimes does not n The invasive infection is only present, when the strain is highly virulent (specific clones of the microorganism) and the host organism is ready to get infected n Meningococcus is transmitted by a narow contact. Invasive infection is more likely when mucous membranes are dammaged, e. g. by smoking or previous viral infection. n Infection is often present after too big physical activity after long inactivity period How they come into the tissue Meningococcal meningitis is woldwide very important infection Story three n Annie was crying and touching her ear. Her mother measured her temperature, and it was elevated. n At general practicioner‘s, Annie was examined and diagnosis of otitis media was set n As her tympanon was already broken, the pus was taken for examination n Amoxicilin was used for threatment imediatelly. Later, a susceptible pathogen was found. And this pathogen was • Moraxella (sooner Branhamella) catarrhalis • This organism is often present in small amounts even in healthy persons. • In pure culture causes sinusitis, otitis media, pharyngitis etc. • Her original name Branhamella was derived from Sarah Branham, one of first women scientists. She was one of brave women to show man that not only men can be good in science Clinical characteristics – „other G– bacilli“ Story four n There was a big movement in the hospital that day: three more patients, all of them seniors, became ill, and again it was the same – breathing problems and fever n After a thoroughful examination, the laboratory foud the pathogen not only in patient secretions, but also in hot water pipes of the hospital. The pipes had to be rebuilt and only after that the infections were stopped finally. Legionaire‘s disease n It is a disease caused by Legionella pneumophila n Legionaire‘s disease is a variant of disease that is quite severe; another variant, Pontiac fever, is rather mild n The bacterium ofther have reservoir in water instalation, air condition, etc. n When building new hospital departments (but also senior houses, hotels, spa…) measures to prevent legionelosis should be taken, mostly at planing water pipe system (no blind branches). Legionaire‘s disease Legionella and temperature Some ways of water disinfection „Other Gram-negative bacteria“ n It is not a real „group“ or „familly“. Nevertheless, these are quite rare bacteria, usually not growing on Endo agar, some of them growing on blood agar, and causing various diseases n Besides Legionella, we should mention at least three genera: Bordetella, Brucella and Francisella Genus Bordetella n B. pertussis and B. parapertussis cause whooping cough n B. bronchiseptica causes various pathologies in humans and animals. n Whooping cough is very rare due to vaccination n Pernasal swab is used when necessary Bordetella was isolated in 1906 by Jules Bordet et Octave Gengou Bordetella bronchiseptica: cat infection Genus Brucella n It is a causative agent of zoonoses n Brucella abortus is a bovine pathogen. It infects commonly bovine placenta, causing abortions in cattle. In human it causes so named Bang disease (fever, problems with various organs etc.) n Other brucellae are Brucella suis from hogs, Brucella mellitensis from sheep and goats and Brucella canis from dogs Genus Francisella n Most important species – F. tularensis n Causes tularemia – „hare plague“ n Gamekeepers, but even more cooks preparing game are in risk of infections n The organism may infect wounds, but also it is possible to inhalate it, thus leading to pneumoniae From report about epidemiological situation (IX/2008) – I (abbreviated) n Tularemia – A21: Woman, born 1970 from Valtice, gardener. 20^th June GP visit for T 38°C, sore throat, enlarged lymphonodes next to right clavicula. The lymphonode was extirpated. First serological examination negative, second (2^nd July) positive n Brucellosis – A23: Man, born 1972, Brno. 10^th May intestinal problems, febrilia, hepatopatia, dg. proven serologically. Brucella abortus positive, CFT 1:8. In anamnesis a travel to Borneo (Kalimantan) in april, trek in a tropical forest. Delayed report. (Reported by Public Health Office for South Moravia) From report about epidemiological situation (IX/2008) – II n Petussis (A37.0): 4 cases reported (Brno-environment, Hodonín), people aged 14 to 17, all vaccinated, one with missing re-vaccintion n Parpetussis (A37.1): 3 cases of disease, coinfection, Brno environment, Hodonín Diagnostics of G– cocci Neisseriae and Moraxellae – characteristics n Microscopy: G – diplococci, coffee bean shape, often intraleucocytar n Culture: tiny, colourless or yellowish (according to the species) colony, growing (species specific) on blood or chocolate agar n Biochemical diagnostic: catalase positive, oxidase positive, differenciation possible n Antigen analysis: very important in Meningococcus, serogroup B is not preventable by vaccination! Observation of gonorrhoea smear n Gonococci (but also meningococci) are Gram-negative diplococci, coffee-bean shaped, mostly intracelullar. Presence of cocci inside leucocytes is their typical property. Culture of Neisseria and Moraxella n There exist differences bethween G– cocci in culture properties. Oral neisseriae and moraxellae grow on blood agar (BA). Meningococci grow only on nutrient-rich variants of BA. Gonococci do not grow on BA at all, they require chocolat agar. Basic biochemical tests n Quick tests with diagnostic strips simplify the diagnostics n Neisseriae are oxidase positive, moraxellae too, but their reaction might be late. n Moraxella is typically positive in INAC test n INAC test is simillar to oxidase test, but the strip should be moistened and one has to wait 5 minutes. The colour is blue-green. Species determination of neisseriae and moraxellae n For detailed identification of neisseriae and moraxellae, biochemical tests are used, in Czechia mostly NEISSERIAtest, in other countries other tests (at those below or on the next slide) n Both pathogenous neisseriae have little biochemical activity: Gonococcus splits glucose only, Meningococcus glucose and maltose. Antibiotic susceptibility testing in neisseriae and related bacteria n Antibiotic susceptibility in pathogenous neisseriae is determined on media, on which they are able to grow, i. e. not on MH agar n First election drug for meningococcus is even today classical penicillin. It is used also for Gonococcus. Other drugs are macrolids, quinolones or cefriaxone. In neisseriae, zones are often large and confluent. A set of antibiotics againts pathogenic neisseriae Somewhere, E-test is used Antigen analysis Antigens detected at CSF antigen analysis n Neisseria meningitidis A n Neisseria meningitidis B teens, children n Neisseria meningitidis C n N. meningitidis Y/W135 n Haemophilus influenzae b children n Streptococcus pneumoniae seniors n Streptococcus agalactiae newborns n In green colour there is the age group, where the infection is the most typical Diagnostics of „other G– bacilli“ „Other gram negative“ characteristic n Microscopy: G – bacilli, often short n Culture: we use mostly special media (BG for Bordetella, BCYE for Legionella etc.) n Biochemical diagnostic: some characteristics might be used n Antigen analysis: sometimes usefull n Indirect methods used, mostly for tularemia Diagnostics of Bordetella, Brucella, Legionella and Francisella n To diagnostics of these bacteria, special media are used, or indirect diagnostics (antibody detection) n Each subtask describes one of the genera mentioned here BCYE medium for Legionella Francisella diagnostics: Reading the agglutination set Titre is counted, i. e. the highest dilution giving still a positive reaction Pictures of G– cocci and „other G– bacilli“ Criminal photos: Gonococcus Gonococci Meningococcus Moraxella catarrhalis Francisella Legionella Bordetella Brucella The End