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ZoonosesLegionellosis

Pseudomonas Groupand Acinetobacter



Dr. Dr. BerekBerek ZsuzsannaZsuzsanna08 November 201108 November 2011

GRAM NEGATIVE RODS

AEROBBordetellaBrucella (Z)Francisella (Z)

PseudomonasAcinetobacterLegionella

FACULTATIV ANAEROBHaemophilusPasteurella (Z)

Family:EnterobacteriaceaeVibrionaceae

CardiobacteriumEikenellaKingellaActinobacillus

ANAEROBBacteroidesPrevotellaPorphyromonasFusobacterium

MICROAEROPHILCampylobacterHelicobacter

ZoonosesZoonoses: Brucella, : Brucella, Francisella, Pasteurella Francisella, Pasteurella andand

PlaguePlague

1. Brucella1. Brucella

Sir David Bruce (1855-1931)

www.lshtm.ac.uk/.../archives/images/bruce1.jpg

Brucellae

Morphology:Gramnegative Coccobacilli

Cultivation:Agar – nutrient rich(Serum, Glycerine)CO221 Days

Resistant!

Description: Brucella spp. Colony Characteristics: - A. Fastidious, usually notvisible at 24h. - B. Grows slowly on most standard laboratory media (e.g. sheepblood, chocolate and trypticase soy agars). Pinpoint, smooth, entire translucent, non-hemolytic at 48h

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Brucellae

Pathogenesis, Infection, Clinical findings

B. melitensis Goat Maltese feverB. abortus Cattle Morbus BangB. suis Swine swine Brucellosis

Anthropozoonosis All is Brucellosis„Febris undulans” RES!(undulating fever)

- 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

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Brucella – spreading

Medmicro

Figure. Acute unilateral scrotal swellingin a 27-year-old man with brucellosis.

www.medscape.com/.../art-iim441224.fig.jpg

Fig.13.36 Brucellosis. Arthritis of theleft knee. This was accompanied byfever, malaise, generalized myalgia anddepression.Fig. 13.37 Orchitis – B. abortus

Brucellosis - diagnosis

DiagnosisCulture: min. 5 daysSerologyAntibody detection

Tube-agglutination (Wright)IgM ChromatographyELISA

Therapy:Doxycyclin, Rifampicin, StreptomycinProphylaxis:expositionTreatment or anihilation of sick animals

WHO – Bioterror Category B!!!

Brucella IgM

www.kit.nl

Brucellosis


PlaguePlague

2. Francisella 2. Francisella tularensistularensis

Tulare Lake; California, USA

http://upload.wikimedia.org/wikipedia/commons/3/34/Pine_Flat_Lake_1.jpg

Francisella tularensis

Morphology:Gramnegative rods

Resistant to COLD!

Cultivation is prohibited!Only 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 tularensis

Clinical findings:TULARAEMIALymphnodes, small Granulomas+ ulceration+ necrosisPrimar complexcutano-, oculo-, tonsilloglandular, (visible!)thoracal, abdominal - (invisible!) formsGeneralisation – Granulomatous lesions!

Diagnosis: SerologyProphylaxiy:Expositionsprophylaxis

Therapy:Streptomycin, Doxycyclin,Ciprofloxacin

A reported case of exposure of a patient toa wild rabbit, whichsubsequently died, suggested that tularemiawas the likely etiology

staff.vbi.vt.edu/.../Ftularensis

Description: Cervical Lymphadenitis in a Patient With Pharyngeal Tularemia; Patient has marked swelling and fluctuant suppuration of several anteriorcervical nodes. Infection was acquired by ingestion of contaminated food orwater. Source: World Health Organization


Description: ChestRadiograph of a Patient WithPulmonary Tularemia

Description: These Francisella tularensis colonies show characteristicopalescence on cysteine heart agarwith sheep blood (cultured at 37 C for72 hours). Note: On cysteine heartagar, F tularensis colonies arecharacteristically opalescent and donot discolor the medium



PlaguePlague

3. Pasteurella3. Pasteurella

Louis Pasteur (1822-1895)

upload.wikimedia.org/.../4/42/Louis_Pasteur.jpg

http://www.pasteur.fr/

medecinepharmacie.univ-fcomte.fr

Pasteurella multocida

Morphology:Gramnegative, small rods

Cultivation:Blood- and chocolate agar

Pathogenesis:Through dog and/or cat bites

Immunocompromised: Sepsis!

Therapy: surgery, Penicillines

aapredbook.aappublications.org

Pasteurella multocida

Fig. 10.55 Animal bite. Infected wound of fingerfollowing bite of domesticcat. P. multocida wasisolated from the wound.

aapredbook.aappublications.org

Pasteurella multocidacellulitissecondary tomultiple catbites about theface of a one-year-old child


PlaguePlague

4. Pest 4. Pest –– Yersinia pestisYersinia pestis

1863 - 1943

Yersinia pestis Genus: Enterobacteriaceae!

Morphology: Gramnegative rods – bipolar staining

www.lonlygunmen.deGiemsa staining www.mja.com.au

www.idph.state.il.us, www2.cnrs.fr, ww.knowledgenews.net,

Yersinia pestis

NO CULTURE! FORBIDDEN!Only in special LaboratoriesWHO – Bioterrorcategory A!!!

http://www.idph.state.il.us/

http://www.knowledgenews.net/

Pest - XIV. Jh.

Yersinia pestis

VIRULENCE FACTORS

Capsule – Protein!V Antigen (Protein) AntiphagocyticW Antigen = Endotoxin

Extracellular Substances- Plasminogen – Activator – Protein (Pla)

spreading, fibrinolytic-Toxin (kills mice)

Yersinia pestis

Pathogenesis, Infection:Source of infection:Rats (and other rodents)→ Elimination of Rats!!!

Transmission:direct contact,Rat-flea-bites

Penetration: skin

Yersinia pestis

Figure 29-4 Pathogenesis of Y. pestis in plague patients.

Medmicro

Clinical findings:1)Bubonic plague(swollen lymph nodes)

2) Septic form → haemorrhagic inflammation

3) Lung plague = Pneumonia ← direct aerogentransmission from human to human (drop-infection → primary lungplague)!

Yersinia pestis

Fig. 13.55 Plague. Enlarged tender inguinal lymphnodes in a Vietnamesechild with bubonic plague.Fig. 13.56 Advanced stage of inguinal lymphadenitis in bubonoc plague. The nodes have undergone suppuration and the lesion has drainedspontaneously.By courtesy of Dr. J.R. Cantey

Bubonic form

Necrosis of finger tips of septicemicplague.

www.imcworldwide.org

Cutaneous Hemorrhages in Plague. Source www.cdc.gov

Septic form

www.imcworldwide.orgLung plague

Yersinia pestis

DiagnosisClinical pictureDirect detection – microscopic (bipolar!)Serology – tube-agglutination, IF

Therapy:Doxycyclin, Streptomycin

Biological Weapons – Bioterrorism

Biological Weapon: Organism, Toxin, VirusAim:• to kill and/or anihilate individuals and/orwhole 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 – droplets, airwaysHigh letalityTherapy? (too late)„Out of the blue”High number of cases

Biological Weapons – Bioterrorism

LegionellosisLegionella pneumophila

LegionellosisLegionella pneumophila

Morphology:Gram negative rods

Aerob

www.lf3.cuni.cz

Legionella

www.waterscan.co.yu

Legionella

Legionella pneumophila

Flagellum, Fimbriae

mcb.berkeley.edu

Legionella


Cultivation:Special Media!BCYE BCYE (yeast extract, Activ charcoal)(pH=9; Temperature 35°C; 3 days)

Bloodagar

BCYE

LegionellaCulture

www.lf3.cuni.cz

BCYE

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LegionellaCulture

medecinepharmacie.univ-fcomte.fr

Legionella

http://medecinepharmacie.univ-fcomte.fr/bacterio_web/exa_microscopiques/examicro_divers.htm


Pathogenesis-1:Source of infection, habitat:ubiquitous(air-conditions, water-showers,Humidity (floor), Biofilms)

transmission:aerogen - dropinfection!

www.ratsteachmicro.com

www.eco-khlamx.com


Pathogenesis-2:Facultative intracellular!

In water: ProtozoonIn humans: Leukocytes, Inflammatory reaction, Proteases,Phospholipases

www.lifespan.org

FIGURE 40-7 ElectronmicrographshowingL. pneumophila

serogroup 1 in theprocess of dividing (arrows) within a vesicle of an amoeba(Hartmanellaveriformis) cell.

Legionella andamoeba

Pathogenic cycle of Legionella

microbewiki.kenyon.edu

textbookofbacteriology.net/Legionella.jpeg


Clinical findings:Legionellosis1) Pneumonia → Legionair’s disease2) Pontiac-Fever (non-pneumonia)

Diagnosis:detection – Biopsy! BAL: direct IFCultivationDirect antigen detection – Urine!Antibody detection – Serology; ELISA

Prophylaxis: avoiding of Legionella containing Aerosole

Therapy:Erythromycin, Tetracyclin, Rifampicin, FQ

Fig. 2.29 Legionnaires’ disease. Chest radiograph showing extensiveconsolidation affecting parts of all lobes of the lungs.

Fig. 2.28 Legionella pneumophila. Specimen from bronchial biopsy taken throughfibreoptic bronchoscope in a patient with fulminant Legionnaires’s disease. The organismcan be isolated on selective culture media or by guinea pig inoculation. By courtesy of Dr. S. Fischer-Hoch

Diagnosis

Fig. 2.30 Legionnaires’ disease. Autospy specimen showingconsolidation of upper and lower lobes of right lung.

Pseudomonas GroupPseudomonas Group

Pseudomonas P. aeruginosa P

Burkholderia B. mallei PB. pseudomallei PB. cepacia

Stenotrophomonas S. maltophilia(Xanthomonas)

P = obligate pathogen

Pseudomonas aeruginosa

Morphology and culture:Gramnegative rods 1-2 μm, no demands; Biofilmbuilding!

Pigments:1. Pyocyanin2. Fluorescein

Haemolysis (β)


VERY RESISTANT !

- to HeatLightDry

Desinfectants

Antibiotics


www.fiu.edu

ANTIGENS AND VIRULENCE FACTORS:Adhaesion and Colonisation- "0", "H", Pili/Fimbriae- Capsule = Glycocalyx- Alginate slime = mucoid Exopolysaccharid BiofilmbuildingInvasion, Penetration- Extracellular Proteases, Exoenzymes (many!)- Cytotoxin = Leukocidin and Haemolysins- PigmentsDissemination- Exotoxin A – inhibit Proteinsynthesis (EF2 Diphtheria)- Exotoxin/Exoenzym S – at burns; detecting in blood- Endotoxin

LD50 – at burnt wounds 30Normal skin 108


Pseudomonas aeruginosa – Structure

Medmicro

No single factor is decisive for virulence.

www.ratsteachmicro.com

Pseudomonas aeruginosaToxin-A – mode of action

Pseudomonas aeruginosa – elmi


Pathogenesis, Infection:

Habitat (normally):Soil, water (swimming-pools, Pools!), sewege waterHuman GI tractRespiratory tract: animals

Source of infection:sick, carrierscontaminated enviromentSolutions (humid Milieu!) Kunststoffe

transmission: direct, indirect Contact

Clinical findings:Nosokomial infections

- Meningitis, Pneumonia (Respirator!) - Sepsis- Burns! skin, Wounds- Urogenital Infections (Katheters), - GI tract (!), newborn/babies- Otitis media, externa- EYE + Contact lenses

Cystic Fibrosis (mucoid strains)


Fig. 10.2 Pseudomonas folliculitis. Papulopustuler rash over thebuttocks and thights following use of a spa pool.

Fig. 13-6 Ecthyma gangrenosum. Necrotic round lesion on the buttock of a childwith Pseudomonas septicaemia associated with immunodeficiency.

Fig. 12.46 Bacterial keratitis. Contact lens-associated keratitis due toPseudomonas aeruginosa. By courtesy of Dr. A.N. Carlson

Fig. 12.47 Bacterial keratitis, in this case due to P. aeruginosa. Aninfiltrate is seen with corneal thinning. By courtesy of Mr. P.A. Hunter

Diffuseinfiltrate

Edge of epithelium

Thinned corneal stoma

Fig. 12.48 Bacterial keratitis. A massive inflammatory response inanterior uveitis leads to precipitation of the cells as pus in the anteriorchamber. This is called hypopyon. By courtesy of Mr. S. Harding

Fig. 12.49 Bacterial keratitis. P. aeruginosa eye infection showingcorneal ulceration and hypopyon formation in this rapidly progressiveeye infection.


Clinical findings:Nosokomial infections

- Meningitis, Pneumonia (Respirator!) - Sepsis- Burns! skin, Wounds- Urogenital Infections (Katheters), - GI tract (!), newborn/babies- Otitis media, externa- EYE + Contactlenses

Cystic Fibrosis (mucoid strains)

Diagnosis:Detecting the bacterium, identification, Oxidase +

Therapy and Prophylaxis:Antibiogram!!!Aminoglycosides, Carbenicillin, antipseudomonas Cephalosporines, Fluoroquinolones

Expositionsprophylaxis: CLEAN! Disinfection!Active Immunisation – in cystic fibrosis


A conjugate vaccine in the final clinical phase is Aerugen®, the first and onlyvaccine for the prophylaxis of fatal Pseudomonas aeruginosa infections in cystic fibrosis patients. The polyvalent conjugate vaccine combines 8 prevalentP. aeruginosa serotypes and the bacterial exotoxin A. It is the first conjugatevaccine based on a lipopolysaccharide component.

www.bernabiotech.com

Vaccine

Burkholderia malleiClinical finding: Malleus (glanders) – horse, donkeyHumans: often fatal!Occupational diseaseBioterror category B!

www.vef.hr/vetarhiv/papers/68-5/alani2.jpg

A horse with glandersand with positive mallein test

microbewiki.kenyon.edu/images/3/39/Horse.jpg

http://microbewiki.kenyon.edu/images/3/39/Horse.jpg

Clinical finding: Melioidosis(pseudoglanders)

– Pneumonia, Sepsis

subtropical andtropical area

Burkholderia pseudomallei

B. pseudomallei

www.co.collin.tx.us

B. pseudomallei

www.asm.org

B. pseudomallei onAshdown's agar afterincubation at 37°C in air for 3 days

Burkholderia cepacia,Stenotrophomonas maltophilia

Nosokomial infections

Figure 1. Electron micrographs showing expression of flagella by SMDP92. Stenotrophomonas maltophilia strains can have one (A) to several flagella(B,C). The flagella on these bacteria show a polar disposition. Bars, 0.5 µm

www.cdc.gov/.../web%20images/01-0535-8t.jpg

Figure 8. Ultrastructuralanalysis of Stenotrophomonas maltophilia adheringto plastic. (A) Scanningelectron micrographsshowing the tight adhesion of SMDP92 to the plasticsurface. (B) Structures resembling flagella seem tobe protruding andinterconnecting bacteria(arrowheads) or connectingbacteria to the plastic(arrows). (C) In addition tothe flagellalike filaments(arrowheads), high-powermagnification shows thepresence of thin fibrillarstructures connectingbacteria to the abioticsurface. Bars: A 10 mm, B 1 mm, C 2 mm.

www.cdc.gov/.../web%20images/01-0535-8t.jpg

Stenotrophomonas maltophilia www.uni-ulm.de

www.oregon.govBurkholderia sp.

www.uni-ulm.deB. cepacia

Acinetobacter spp.Acinetobacter spp.

Morphology:Gramnegative dimorphicrods, coccobacilli

www.acinetobacter.org

Acinetobacter spp.Culture:Agar, Bloodagarimportant: Temperature 40-44°CNon-motile – „akinetic”

Pathogenesis and clinical findings:like Pseudomonas aeruginosa

www.acinetobacter.org

Nosokomial infections!Multiresistant!

THE ENDIstanbul, 2006

dr. berek zsuzsanna - semmelweis egyetem | kutató...

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