Gram-Negative Cocci Gram-Negative Bacilli
Gram-negative Cocci Gram-negative bacilli
Miklos Fuzi Neisseria Pyogenic Cocci GRAM -
Aerobic: Oxidase + Neisseria N. gonorrhoeae N. meningitidis others (N. sicca, N. subflava, N. flavescens and apathogenic spp.) Moraxella M. catarrhalis
Anaerobic: Veillonella spp.
Veillonellae vietsciences.free.fr N. gonorrhoeae and N. meningitidis
Morphology Gram-negative Diplococci
www.waterscan.co.yu/images Gram-negative Diplococci
path.upmc.edu N. gonorrhoeae and N. meningitidis
Oxidase + Culture: Special demands Nutrient rich medium (Chocolate agar and
5-10% CO2)
Resistance: Sensitive bacteria: Dry, Heat, Disinfectants, Antibiotics
path.upmc.edu http://www.mfi.ku.dk/ppaulev/chapter33/images/33-3.jpg N. gonorrhoeae = Gonococcus
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
Pili textbookofbacteriology.net N. gonorrhoeae = Gonococcus
Gonococcus-Lymphocyte Interaction
neisseria.org/images/ng-lym2.jpg N. gonorrhoeae = Gonococcus
Source of infection Sick humans
Transmission - Direct (sexual) Contact
Clinical findings Gonorrhea = Tripper Ophthalmoblenorrhea neonatorum
NO IMMUNITY! (Antigenic variations!) Pathogenesis
Medmicro Gonorrhea – acute Urethritis
www.stdservices.on.net www.boltonlgb.co.uk Gonorrhea – acute Urethritis Gonorrhea – acute Cervicitis
www.boltonlgb.co.uk Gonorrhea – acute Cervicitis Gonorrhea – acute Conjuctivitis Blenorrhea neonatorum
www.mc3.edu Corneal ulcers due to gonococcus are very destructive and have a tendency to perforate the cornea. www.slackbooks.com Gonorrhea – Chronic and disseminated Form
Endometritis, Salpingitis, Prostatitis purulent 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
Microscopic www2.mf.uni-lj.si, www.uni-ulm.de , Direct detection – phagocytosed diplococci pathmicro.med.sc.edu Gram staining Methylenblue staining, Direct Immunofluorescent (DIF) 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 Gonorrhea Therapy: 3. Generation Cephalosporin (Ceftriaxone) or Spectinomycin (Aminoglycoside)
Prophylaxis: GO - Exposition (safe sex) - Source of infection: find and treat! - Early Diagnosis and treatment www.tiscali.co.uk
Ophthalmia neonatorum: Application of 1% silver-nitrate in conjunctival sack
NO VACCINE! (Antigenic variants!) N. meningitidis = Meningococcus
scanning EM textbookofbacteriology.net N. meningitidis = Meningococcus
Antigens and Virulence factors: Capsule – Polysaccharide, antiphagocytic Multiple serotypes (A, B, C, W135, Y!) Capsule change Pili/Fimbriae IgA-Proteases! Outer Membrane Proteins (OMP) LOS (Mimicry, sialisation: Serum resistant, very toxic)
Meningococcus
zdsys.chgb.org.cn N. meningitidis = Meningococcus
Source of infection human – carriers (sick, healthy)
Transmission, Portal of entry - Direct, drop-infection - Nose, throat
Clinical finding Pharyngitis Meningitis cerebrospinalis epidemica Sepsis = Waterhouse-Friderichsen Syndrome Fig. 10.56 Acute meningococcaemia. Note the variable 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 bulbar 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 septicaemia, caused by Neisseria meningitidis . (Courtesy of Wellcome Trust Photographic Library) srs.dl.ac.uk ufl. © Urban © Urban & Fischer 2003 – A Roche LexikonMedizin, 5. Waterhouse- Friderichsen Syndrome: schwere nekrotisierende Hautläsionen bei Meningokokkensepsis mit Verbrauchskoagulopathie (R. E. Rieger, Univ.-Kinderklinik Marburg). www.gesundheit.de The patient with Waterhouse-Friderichsen syndrome has sepsis with DIC and marked purpura. medlib.med.utah.edu pathy.fujita-hu.ac.jp
Purulent meningitis with hemorrhage in the frontal lobe (gross findings). 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 punction Blood carriers: throat Meningitis Diagnosis
Detection Microscopic examination (Liquor, blood culture) Culture Liquor, Blood, Throat Direct detection of antigen (Liquor) – Latex-agglutination Direct detection of bact. DNA Real-time PCR from blood, CSF Diagnosis N. meningitidis
Culture: Bloodagar, Chocolate agar
Identification: glu+, mal+
MIC (E-test) Meningococcus meningitis
Therapy: Penicillin and/or Ceftriaxone; cefotaxime NO Beta-lactamase production
Prophylaxis: Active Immunisation Vaccine 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 Species Haemophilus H. influenzae H. parainfluenzae H. aegyptius H. ducreyi
Bordetella B. pertussis B. parapertussis
P: Pathogen www.waterscan.co.yu/images m µ
HaemophilusHaemophilus influenzae influenzae Morphology: -Gram Coccobacillus, 1 ca. Cultivation: factorsGrowth ! (chocolate, NAD, Haem, V= X= Satellite-phenomenon; Satellitism phil.cdc.gov Blood agar plate culture showing Haemophilus influenzae satelliting around Staphylococcus aureus. , c, d, e, f f (HiB!) e, , c, d, b – Polysaccharide antigens: Types: a, Types: -Protease! Outer Membrane Proteine (OMP) ProteineMembrane Outer LPS Surface Antigens and Virulence factors: Virulence Antigens and Capsule IgA Haemophilus influenzae influenzae Haemophilus Haemophilus Haemophilus influenzae Type b (Hib)
www.soundmedicine.iu.edu Sepsis !, Nasopharyngitis, Sinusitis,Otitismedia , Cellulitis Lower respiratory tract: HaemophilusHaemophilus influenzae influenzae Clinical findings: Meningitis Upper respiratory tract: Epiglottitis Bronchitis,Pneumonia, Haemophilus influenzae
Sepsis
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- www.ecbt.org -(2000 Red Book: 25th Edition, Report of the Committee on Infectious Diseases) Haemophilus influenzae
Periorbital cellulitis. © Neal Halsy, MD www.cispimmunize.org Otitismedia , Nasopharyngitis, Sinusitis, , Sepsis
HaemophilusHaemophilus influenzae influenzae Clinical findings: Meningitis Cellulitis Upper respiratory tract: Epiglottitis! Lower respiratory tract: Bronchitis,Pneumonia, HiB-epiglottitis
) (Polysaccharide + Protein + (Polysaccharide HiB Conjugate-Vaccine HiB Site of infection (Nose, throat, Sputum etc.) Sputum throat, (Nose, infection of Site LIQUOR! (CSF) LIQUOR! ‹ ‹
HaemophilusHaemophilus influenzae influenzae Detection: Culture, Microscopic, detection Ag Capsule (Latex-agglutination) Diagnosis: specimen Samples, CSF blood, PCR from Real-time Therapy: 1.Cephalosporins gen. III. Ampicillin + Aminoglycosides Ampicillin + 2. Prophylaxis: - Immunisation Active Lipopolysaccharid Extract - Vaccine
ibs-isb.nrc-cnrc.gc.ca
www.kmhk.kmu.edu.tw Causing: Ulcus molle = Chancroid = = soft Chancre Pharyngitis,Endocarditis, Conjunctivitis
Haemophilus ducreyi ducreyi Haemophilus Haemophilus aegyptius aegyptius Haemophilus Haemophilus Causing: Brasilian Purpuric Fever Haemophilus parainfluenzae parainfluenzae Haemophilus Haemophilus Ulcus molle Ulcus molle medinfo.ufl.edu Chancroid in female www.smu.edu Bordetella Bordetella pertussis Morphology: Gram-negative Coccobacillus, ca. 1 µm
www.waterscan.co.yu/images Bordetella pertussis
Culture: Special Medium Bordet – Gengou
nobelprize.org www.szu.cz Bordetella pertussis
Antigens and Virulence factors: Capsule Fimbriae, filamentous Haemagglutinin Outer Membrane Proteine (OMP) LPS Pertactin Extracellular Toxins: Pertussis Toxin Adenylate-cyclase Toxin Tracheal cytotoxin Dermatonecrotic Toxin FIGURE 31-2 Virulence factors of B pertussis .
Medmicro Pertussis toxin
www.med.sc.edu:85 Bordetella pertussis
Pathogenesis, Infection: Source: sick – in prodromal and catarrhal Stadium
Portal of entry: Respiratory tract
Transmission: drop-infection → sensitive! 55°C; 30’ FIGURE 31-1 Pathogenesis of whooping cough.
Medmicro www.my-pharm.ac.jp FIGURE 31-3 Binding of pertussis toxin to cell membranes.
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 Pertussis – paroxysmal Phase
www.gesundes-kind.de , www.vaccineinformation.org www.med.sc.edu Pertussis - Diagnosis
Lymphocytosis
www.thecrookstoncollection.com aapredbook.aappublications.org Bordetella pertussis Diagnosis
Cultivation: Bordet – Gengou Direct caugh! Charcoal Medium Serology: IgM, IgA, IgG Detection of DNA
PCR medinfo.ufl.edu Bordetella pertussis
Therapy: Macrolides
Prophylaxis: Active Immunisation – acellular Vaccine DaPT Toxoid FH/Pilus Pertactin
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 2 : y: -negative coccobacilli BrucellaeBrucellae Morpholog Gram Cultivation Agar – nutrient rich (Serum, Glycerine) Atmosphere: CO Incubation:days-weeks staff.vbi.vt.edu/pathport/pathinfo_images/Bru...
Description: Brucella spp. Colony Characteristics: - A. Fastidious, usually not visible at 24h. - B. Grows slowly on most standard laboratory media (e.g. sheep blood, chocolate and trypticase soy agars). Pinpoint, smooth, entire translucent, non-hemolytic at 48h „Febris undulans” „Febris undulans” RES! (undulating „wavelike”) fever: through lesions skin or conjunctiva ortract GI direct contact or contaminated food
BrucellaeBrucellae sick animalsmilk) (meat, mucosa from through invasion Pathogenesis, Infection, Clinical findings Clinical Infection, Pathogenesis, melitensisB. abortusB. suisB. GoatAnthropozoonosis Cattle Pig Maltese fever All is Brucellosis Morbus Bang Swine Brucellosis - - - Brucella – source of infection
Medmicro Brucella – portals of entry
Medmicro Figure 28-1 Portals of entry for Brucella species. Brucella for entry of Portals 28-1 Figure 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 Brucellosis Diagnosis Culture: min. 5 days Serology Antibody detection Tube-agglutination (Wright) IgM Chromatography ELISA Direct detection of DNA: PCR Brucella IgM Therapy: Doxycyclin, Rifampicin, Streptomycin www.kit.nl Prophylaxis: Avoid exposition Treatment or annihilation of sick animals WHO – Bioterror Category B!!! Francisella -HUMAN -HUMAN TRANSMISSION or per os or ectoparasites via -TO direct contact sick sick animals -negative rods -negative inhalation
Francisella tularensis tularensis Francisella Francisella From through Morphology: Gram Survives wet in and cold environment. prohibited! is Cultivation special Laboratories in Only –WHO A!!! Bioterror category Infection Pathogenesis, - - or HUMAN NO PCR Serology Therapy: Doxycyclin, Streptomycin, Ciprofloxacin Diagnosis: DNA: Detection of as atous atous lesions! – Granulom
Francisella tularensis tularensis Francisella Francisella findings: Clinical TULAREMIA smallLymphnodes, granulom ulceration + necrosis + symptoms diverse Associated with (visible!) oculo-, cutano-,tonsilloglandular, thoracal, abdominal - (invisible!) forms Generalisation 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 staff.vbi.vt.edu/.../Ftularensis
Description: Cervical Lymphadenitis in a Patient With Pharyngeal Tularemia ; Patient has marked swelling and fluctuant suppuration of several anterior cervical nodes. Infection was acquired by ingestion of contaminated food or water. Source: World Health Organization 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
Description: Chest Radiograph of a Patient With Pulmonary Tularemia
staff.vbi.vt.edu/.../Ftularensis Yersinia www.mja.com.au Genus: Enterobacteriaceae! Genus: Giemsa staining Giemsa -negative rods -negative – bipolar staining Gram Morphology:
YersiniaYersinia pestis pestis www.lonlygunmen.de
Yersinia pestis NO CULTURE! FORBIDDEN! Only in special Laboratories WHO – Bioterrorcategory A!!!
www.idph.state.il.us , www2.cnrs.fr, ww.knowledgenews.net , Plague: 14th century Plague in medieval Europe spreading, fibrinolytic spreading, VIRULENCE FACTORS VIRULENCE – Capsule Protein! Antiphagocytic (Protein) Antigen V = Endotoxin Antigen W Substances Extracellular -– Plasminogen – Activator (Pla) Protein (killsmice) -Toxin
YersiniaYersinia pestis pestis Elimination Rats!!! of → skin
YersiniaYersinia pestis pestis Pathogenesis, Infection: Source of infection: Rats (and other rodents) Transmission: directcontact, Rat-flea-bites Penetration: Figure 29-4 of 29-4 FigurePathogenesis patients. plagueinpestis Y. Medmicro
Yersinia Yersiniapestis pestis Yersinia pestis
Clinical findings: 1)Bubonic 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)! Bubonic form
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 Necrosis of finger tips of septicemic plague.
Septic form
Cutaneous Hemorrhages in Plague. Source www.cdc.gov
www.imcworldwide.org Pulmonary plague www.imcworldwide.org Diagnosis Clinical picture Direct detection – microscopic (bipolar!) Real-time PCR Serology – tube-agglutination, IF Therapy: Doxycyclin, Streptomycin YersiniaYersinia pestis pestis Biological Weapons – Bioterrorism
Biological Weapon: Microbe, Toxin Aim: • to kill individuals and/or whole population • economic damage
Biological war (military conflicts) Bioterrorisms (ideology!) Biological crime (personal) Biological Weapons – Bioterrorism
Categories: A, B, C Most dangerous: A B. anthracis, C. botulinum, F. tularensis, Y. pestis
Easy to culture Easy to spread/transmit – airborne High rate of mortality Therapy? (too late) High number of cases Korfu, 2006 THE END