Gram-negative Cocci Gram-negative bacilli
Miklos Fuzi
Neisseria
Pyogen ic Cocci GRAM -
Aerobic: Oxidase +Neisseria N. gonorrhoeae
N. meningitidisothers (N. sicca, N. subflava, N. flavescens and apathogenic spp.)
Moraxella M. catarrhalis
Anaerobic: Veillonella spp.
Veillonellae vietsciences.free.fr
N. gonorrhoeae and N. meningitidis
Morpholog yGram-negativeDiplococci
www.waterscan.co.yu/images
path.upmc.edu
Gram-negative Diplococci
N. gonorrhoeae and N. meningitidis
Oxidase +
path.upmc.edu
Culture:Special demandsNutrient rich medium(Chocolate agar and5-10% CO2)
Resistance:Sensitive bacteria:Dry, Heat, Disinfectants, Antibiotics
http://www.mfi.ku.dk/ppaulev/chapter33/images/33-3.jpg
Antigens and Virulence factors:Pili/Fimbriae (Antigenic variations!)IgA-Proteases!outer membrane proteins (OMP)
(Antigenic variations!)LOS (Mimicry!- sialiation in serum prevents
immune response)Cell-wall Peptidoglycan (Toxic)Inhibits apoptosis in macrophages (prevention
of immune response)
N. gonorrhoeae = Gonococcus
N. gonorrhoeae = Gonococcus
N. gonorrhoeae = Gonococcuste
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Pili
neisseria.org/images/ng-lym2.jpg
Gonococcus-Lymphocyte Interaction
N. gonorrhoeae = Gonococcus
N. gonorrhoeae = Gonococcus
Source of infectionSick humans
Transmission- Direct (sexual) Contact
Clinical findingsGonorrhea = TripperOphthalmoblenorrhea neonatorum
NO IMMUNITY!(Antigenic variations!)
Medmicro
Pathogenesis
Gonorrhea – acute Urethritis
www.stdservices.on.net ww
w.b
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nlgb
.co.
uk
Gonorrhea – acute Urethritis
www.boltonlgb.co.uk
Gonorrhea – acute Cervicitis
Gonorrhea – acute Cervicitis
Gonorrhea – acute ConjuctivitisBlenorrhea neonatorum
www.mc3.edu
www.slackbooks.com
Corneal ulcers due to gonococcus are very destructive and have a tendency to perforate the cornea.
Gonorrhea – Chronic and disseminated Form
Endometritis, Salpingitis, Prostatitispurulent Arthritis, Vasculitis
Important! anorectal GO and Pharyngitis(„alternative Genitals”)
Fig. 8.33 Gonococcal arthritis. Dactylitis secondary to gonococcal bacteriaemia. By courtesy of Dr. S.E. Thompson
Fig. 8.33 Gonococcal septic arthritis. Arthritis due to N. gonorrhoeae in a 24-year-old woman, showing marked erythema and swelling of the right ankle and leg. By courtesy of Dr. T.F. Sellers Jr.
Gonorrhea – Diagnosis – acute Disease
MicroscopicDirect detection – phagocytosed diplococciGram stainingMethylenblue staining, Direct Immunofluorescent (DIF)
www2.mf.uni-lj.si,www.uni-ulm.de,
pathmicro.med.sc.edu
GO – Gram staining – presumptive Diagnosis only!
GO – Gram staining – only presumptive Diagnosis!
www.med.uni-giessen.de
Gonorrhea – Diagnosis
Culture :„bedside” Thayer-Martin medium and chocolate agar, 5% CO2
Identification: ox+, glu+, mal-
Antigen detection (direct):Latex-agglutination
Detection of bact. DNA: PCR
www2.mf.uni-lj.si,www.uni-ulm.de,
pathmicro.med.sc.edu
Therapy:3. Generation Cephalosporin (Ceftriaxone) or Spectinomycin (Aminoglycoside)
Prophylaxis:GO- Exposition (safe sex)- Source of infection: find and treat! - Early Diagnosis and treatment
Ophthalmia neonatorum : Application of 1% silver-nitrate in conjunctival sack
NO VACCINE! (Antigenic variants!)
Gonorrhea
www.tiscali.co.uk
N. meningitidis = Meningococcus
scanning EM
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N. meningitidis = Meningococcus
Antigens and Virulence factors:Capsule – Polysaccharide, antiphagocytic
Multiple serotypes (A, B, C, W135, Y!)Capsule change
Pili/FimbriaeIgA-Proteases!Outer Membrane Protein s (OMP)LOS (Mimicry, sialisation: Serum resistant,
very toxic)
zdsys.chgb.org.cn
Meningococcus
Source of infectionhuman – carriers (sick, healthy)
Transmission, Portal of entry- Direct, drop-infection- Nose, throat
Clinical findingPharyngitisMeningitis cerebrospinalis epidemicaSepsis = Waterhouse-Friderichsen Syndrome
N. meningitidis = Meningococcus
Fig. 10.56 Acute meningococcaemia. Note the variabl e size of the lesions and their peripheral distribution. Some of the lesions are obviously purpuric, others macular or papular.
Fig. 10.60 Acute meningococcaemia. Petechia on bulb ar conjunctiva.
Fig. 10.62 Acute meningococcaemia. Gangrene of the extremities following a near-fatal illness with hypotension.
Fig. 10.63 Acute meningococcaemia. Gangrene of both legs in a black man with acute meningococcal infection. Bilateral below knee amputations were later required.
The characteristic skin rash of meningococcal septi caemia, caused by Neisseria meningitidis . (Courtesy of Wellcome Trust Photographic Library) srs.dl.ac.uk
Waterhouse- Friderichsen Syndrome: schwere nekrotisierende Hautläsionen bei Meningokokkensepsis mit Verbrauchskoagulopathie (R. E. Rieger, Univ.-Kinderklinik Marburg).
© U
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Lex
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Med
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www.gesundheit.de
The patient with Waterhouse-Friderichsen syndrome h as sepsis with DIC and marked purpura. medlib.med.utah.edu
Purulent meningitis with hemorrhage in the frontal lobe (gross findings).
path
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Acute hemorrhage in bilateral adrenals caused acute adrenal insufficiency (Waterhouse-Friderichsen syndrome).
pathy.fujita-hu.ac.jp
Meningitis Diagnosis
Samples, specimen:Liquor (cerebrospinal fluid) ! – Lumbar punctionBloodcarriers: throat
DetectionMicroscopic examination(Liquor, blood culture)CultureLiquor, Blood, ThroatDirect detection of antigen(Liquor) – Latex-agglutinationDirect detection of bact. DNAReal-time PCR from blood, CSF
Meningitis Diagnosis
Culture: Bloodagar, Chocolate agar
Identification:glu+, mal+
MIC (E-test)
DiagnosisN. meningitidis
Meningococcus meningitis
Therapy:Penicillin and/orCeftriaxone; cefotaximeNO Beta-lactamase production
Prophylaxis:Active ImmunisationVaccine for:- Risk groups- Traveler(Meningitis belt!)
Chemoprophylaxis:Ciprofloxacin; rifampicin(Contacts)
Meningitis belt
Neisseria meningitidis - B
Europe!
NO VACCINE!
Rifampicin only
www.versapharm.com
Haemophilus
GRAM-NEGATIVE COCCOBACILLI
Genus SpeciesHaemophilus H. influenzae
H. parainfluenzaeH. aegyptius H. ducreyi
Bordetella B. pertussis B. parapertussis
P: Pathogen
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzae
Morphology:Gram - Coccobacillus, ca. 1 µm
Cultivation:Growth factors !(chocolate, X= Haem, V= NAD, Satellite-phenomenon; Satellitism
www.waterscan.co.yu/images
phil.cdc.gov Blood agar plate culture showing Haemophilus influe nzae satelliting around Staphylococcus aureus.
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzae
Antigens and Virulence factors:Capsule – Polysaccharide
Types: a, b, c, d, e, f (HiB!)
IgA-Protease!Surface antigens:
Outer Membrane Proteine (OMP)LPS
Haemophilus influenzae Type b (Hib)
www.soundmedicine.iu.edu
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzae
Clinical findings:Meningitis , Sepsis
Cellulitis
Upper respiratory tract:Epiglottitis !, Nasopharyngitis, Sinusitis, Otitis media
Lower respiratory tract:Bronchitis, Pneumonia,
An infant with severe vasculitis with disseminated intravascular coagulation (DIC) with gangrene of the hand secondary to Haemophilus influenzae type b septicemia - prior to the availability of the Hib vaccine. -Image provided by: Visual Red Book on CD-ROM--(2000 Red Book: 25th Edition, Report of the Committee on Infectious Diseases)
www.ecbt.org
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzae
Sepsis
Periorbital cellulitis. © Neal Halsy, MD www.cispimmunize.org
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzae
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzae
Clinical findings:Meningitis , Sepsis
Cellulitis
Upper respiratory tract:Epiglottitis! , Nasopharyngitis, Sinusitis, Otitis media
Lower respiratory tract:Bronchitis, Pneumonia,
HiB-epiglottitis
Haemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeHaemophilus influenzaeDiagnosis:Samples, specimen
�LIQUOR! (CSF)� Site of infection (Nose, throat, Sputum etc.)
Detection:Microscopic, Culture, Capsule Ag detection (Latex-agglutination)Real-time PCR from blood, CSF
Therapy:1. Ampicillin + III. gen. Cephalosporins2. Ampicillin + Aminoglycosides
Prophylaxis:Active Immunisation - HiB Conjugate-Vaccine
(Polysaccharide + Protein)
Lipopolysaccharid Extract - Vaccine
ibs-isb.nrc-cnrc.gc.ca
www.kmhk.kmu.edu.tw
Haemophilus ducreyiHaemophilus ducreyiHaemophilus ducreyiHaemophilus ducreyi
Causing: Ulcus molle = Chancroid == soft Chancre
Haemophilus aegyptiusHaemophilus aegyptiusHaemophilus aegyptiusHaemophilus aegyptius
Causing: Brasilian Purpuric Fever
Haemophilus parainfluenzaeHaemophilus parainfluenzaeHaemophilus parainfluenzaeHaemophilus parainfluenzae
Pharyngitis, Endocarditis, Conjunctivitis
Ulcus molle
Ulcus molle
medinfo.ufl.edu
www.smu.eduChancroid in female
Bordetella
Bordetella pertussis
Morpholog y:Gram-negative Coccobacillus, ca. 1 µm
www.waterscan.co.yu/images
Bordetella pertussis
Culture:Special MediumBordet – Gengou
www.szu.cznobelprize.org
Antigens and Virulence factors:CapsuleFimbriae, filamentous HaemagglutininOuter Membrane Proteine (OMP)LPSPertactin
Extracellular Toxins:Pertussis ToxinAdenylate-cyclase ToxinTracheal cytotoxinDermatonecrotic Toxin
Bordetella pertussis
FIGURE 31-2 Virulence factors of B pertussis .
Medmicro
Pertussis toxin
www.med.sc.edu:85
Pathogenesis, Infection:Source: sick – in prodromal and catarrhal Stadium
Portal of entry: Respiratory tract
Transmission: drop-infection → sensitive! 55°C; 30’
Bordetella pertussis
FIGURE 31-1 Pathogenesis of whooping cough.
Medmicro
www.my-pharm.ac.jp
FIGURE 31-3 Binding of pertussis toxin to cell memb ranes.
Medmicro
FIGURE 31-4 Synergy between pertussis toxin and the filamentous hemagglutinin in binding to ciliated respiratory epithelial cells.
Medmicro
Bordetella pertussis
Clinical finding:Whooping caugh / Pertussis(Peribronchial inflammation, Intersticiale Pneumonia)
4-Phases:Prodromal, Catarrhal, Paroxysmal, Convalescent
Colonization of tracheal epithelial cells by B. pertussis web.umr.edu/~microbio
www.gesundes-kind.de,www.vaccineinformation.org
Pertussis –paroxysmal Phase
www.thecrookstoncollection.com aapredbook.aappublications.org
www.med.sc.edu
Pertussis - Diagnosis
Lymphocytosis
Bordetella pertussis
Diagnosis
Cultivation:Bordet – GengouDirect caugh!Charcoal MediumSerology:IgM, IgA, IgG Detection of DNAPCR
medinfo.ufl.edu
Bordetella pertussis
Therapy:Macrolides
Prophylaxis:Active Immunisation – acellular Vaccine DaPT
ToxoidFH/PilusPertactin
DPT = DiPerTe – killed B. pertussis
Pertussis in the USA – 2012 (CDC)
< 1 year 4516 1-6 years 7312 7-10 years 8349 11-19 12484 20+ years 8890 Unknown 329 Total 41880
Brucella
BrucellaeBrucellaeBrucellaeBrucellae
Morpholog y:Gram-negative coccobacilli
Cultivation :Agar – nutrient rich(Serum, Glycerine)Atmosphere: CO2Incubation: days-weeks
Description: Brucella spp. Colony Characteristics: - A. Fastidious, usuall y not visible at 24h. - B. Grows slowly on most standard l aboratory media (e.g. sheep blood, chocolate and trypticase soy agars). Pinpoin t, smooth, entire translucent, non-hemolytic at 48h
staf
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BrucellaeBrucellaeBrucellaeBrucellae
Pathogenesis, Infection, Clinical findings
B. melitensis Goat Maltese feverB. abortus Cattle Morbus BangB. suis Pig Swine Brucellosis
Anthropozoonosis All is Brucellosis„Febris undulans” RES!(undulating fever: „wavelike”)
- from sick animals (meat, milk)- through direct contact or contaminated food- invasion through skin lesions or conjunctiva or GI tract
mucosa
Brucella – source of infection
Medmicro
Brucella –
portals of entry
Figure 28-1 Portals of entry for Brucella species.
Medm
icro
Brucella – spreading
Medmicro
Figure. Acute unilateral scrotal swelling in a 27-year-old man with brucellosis.
www.medscape.com/.../art-iim441224.fig.jpg
Fig.13.36 Brucellosis. Arthritis of the left knee. This was accompanied by fever, malaise, generalized myalgia and depression.Fig. 13.37 Orchitis – B. abortus
DiagnosisCulture: min. 5 daysSerologyAntibody detection
Tube-agglutination (Wright)IgM ChromatographyELISA
Direct detection of DNA : PCRTherapy:Doxycyclin, Rifampicin, Streptomycin
Prophylaxis:Avoid expositionTreatment or annihilation of sick animals
WHO – Bioterror Category B!!!
Brucella IgM
www.kit.nl
BrucellosisBrucellosisBrucellosisBrucellosis
Francisella
Francisella tularensisFrancisella tularensisFrancisella tularensisFrancisella tularensis
Morphology:Gram-negative rods
Survives in wet and cold environment.
Cultivation is prohibited!Only in special LaboratoriesWHO – Bioterror category A!!!
Pathogenesis, Infection- From sick animals- through direct contact or inhalation or per os or via ectoparasites
NO HUMAN-TO-HUMAN TRANSMISSION
Francisella tularensisFrancisella tularensisFrancisella tularensisFrancisella tularensis
Clinical findings:TULAREMIALymphnodes, small granulomas+ ulceration+ necrosisAssociated with diverse symptomscutano-, oculo-, tonsilloglandular, (visible!)thoracal, abdominal - (invisible!) formsGeneralisation – Granulomatous lesions!
Diagnosis: SerologyDetection of DNA: PCRTherapy:Streptomycin, Doxycyclin,Ciprofloxacin
A reported case of exposure of a patient to a wild rabbit, which subsequently died, suggested that tularemia was the likely etiology
staff.vbi.vt.edu/.../Ftularensis
Description: Cervical Lymphadenitis in a Patient With Pharyngeal Tularemia ; Patient has marked swelling and fluctuant suppurati on of several anterior cervical nodes. Infection was acquired by ingestion of contaminated food or water. Source: World Health Organization
staff.vbi.vt.edu/.../Ftularensis
Description: Chest Radiograph of a Patient With Pulmonary Tularemia
Description: These Francisella tularensis colonies show characteristic opalescence on cysteine heart agar with sheep blood (cultured at 37 C for 72 hours). Note: On cysteine heart agar, F tularensis colonies are characteristically opalescent and do not discolor the medium
staff.vbi.vt.edu/.../Ftularensis
Yersinia
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis Genus: Enterobacteriaceae!
Morphology: Gram-negative rods – bipolar staining
www.lonlygunmen.deGiemsa staining www.mja.com.au
www.idph.state.il.us, www2.cnrs.fr, ww.knowledgenews.net,
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis
NO CULTURE! FORBIDDEN!Only in special LaboratoriesWHO – Bioterrorcategory A!!!
Plague: 14th century
Plague in medieval Europe
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis
VIRULENCE FACTORS
Capsule – Protein!V Antigen (Protein) AntiphagocyticW Antigen = Endotoxin
Extracellular Substances- Plasminogen – Activator – Protein (Pla)
spreading, fibrinolytic-Toxin (kills mice)
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis
Pathogenesis, Infection:Source of infection:Rats (and other rodents)→ Elimination of Rats!!!
Transmission:direct contact,Rat-flea-bites
Penetration: skin
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis
Figure 29-4 Pathogenesis of Y. pestis in plague patients.
Medmicro
Clinical findings :1)Bubon ic plague(swollen lymph nodes)
2) Septic form → haemorrhagic inflammation
3) Pulmonary form = Pneumonia ← direct aerogen transmission from human to human (airborne infection → primary pulmonary plague)!
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis
Fig. 13.55 Plague. Enlarged tender inguinal lymphnodes in a Vietnamese child with bubonic plague.Fig. 13.56 Advanced stage of inguinal lymphadenitis in bubonoc plague. The nodes have undergone suppuration and the lesion has drained spontaneously.By courtesy of Dr. J.R. Cantey
Bubonic form
Necrosis of finger tips of septicemic plague.
www.imcworldwide.org
Cutaneous Hemorrhages in Plague. Source www.cdc.gov
Septic form
www.imcworldwide.orgPulmonary plague
Yersinia pestisYersinia pestisYersinia pestisYersinia pestis
DiagnosisClinical pictureDirect detection – microscopic (bipolar!)Real-time PCRSerology – tube-agglutination, IF
Therapy:Doxycyclin, Streptomycin
Biological Weapons – Bioterrorism
Biological Weapon: Microbe, ToxinAim:• to kill individuals and/or whole population • economic damage
Biological war (military conflicts)Bioterrorisms (ideology!)Biological crime (personal)
Categories: A, B, CMost dangerous: AB. anthracis, C. botulinum, F. tularensis, Y. pestis
Easy to cultureEasy to spread/transmit – airborneHigh rate of mortalityTherapy? (too late)High number of cases
Biological Weapons – Bioterrorism
THE ENDKorfu, 2006