Snímka 1

Pseudomonas, Bacillus, Listeria

k ma k 1 m na a n a m 1 a a

Pseudomonas a y f a a ( m) a f n mm n y n nv nm n , k n an n a Of many ff n y f Pseudomonas, n a m f n a nf n n man a Pseudomonas aeruginosa, an a nf n n , n ( n m n a), a f y af y Pseudomonas aeruginosa an a n a a n n y a x a a n am na m R an a n f m an a a n a a n f m n n an n am na an , m n , fa

Bacillus a v y v n m an , an n f a n an a n f n a a a n Bacillus f m a n ma a y n a a an a n n Bacillus n f v n (n n a a ) , an a a a n Bacillus a m a y n f an B. anthracis a an ax; an B. cereus a f n n

Listeria a n f a a a a a an n a a a a n mamma Listeria an f n n , an a v a n am na n An ma an a Listeria a n f n n n k m a , n k v a , f n n an a [1 ] an a ,[13] a z n a z m k, f ma f m m k, an f Pa za n an ff n k n k Listeria; v , n am na n may af k n an f a ka n

Snímka

• Hospital infections • Antrax a bioterrorism • Microscopy - Gram - P. aeruginosa, Bacillus anthracis, Bacillus cereus, Listeria monocytogenes • Wirtz Conklin- B. anthracis, • Cultivation- P. aeruginosa -blood agar and Endo agar, - Bacillus anthracis, Bacillus cereus, Listeria – blood agar • Biochemical properties - P. aeruginosa • ATB therapy of Ps. aeruginosa

Snímka 3

Hospital infections • nf n a n a x a n n a m m a a n m a nf n a a nf n ( A ) • v m n f nf n af a m n

Hospital-acquired infections, also known as healthcare-associated infections (HAI), are nosocomially acquired infections that are typically not present or might be incubating at the time of admission. These infections are usually acquired after hospitalization and manifest 48 hours after admission to the hospital.

For the last few decades, hospitals have taken the hospital-acquired infections seriously. Several hospitals have established infection tracking and surveillance systems in place, along with prevention strategies to reduce the rate of hospital-acquired infections. The impact of hospital-acquired infections is seen not just at an individual patient level, but also at the community level as they have been linked to multidrug-resistant infections. Identifying patients with risk factors for hospital-acquired infections and multidrug-resistant infections is very important in the prevention and minimization of these infections.

an f CW’ a m mm n an m f a n a a fa •A a an m f m a CW a a n kn n a a a a A a nf n ( A ) a n m a nf n•Pa n an an m a n , CW an y f y y an z an •G v a O a f an y n

an m n f a n n a a a nv nm n m x an an n a a a k n n n am na nv nm n R k fa f v m n f CD a kn n n n an , a a an , n n v n n a an nf amma y ( SA D ), an m m R k fa f SS n a n fa a a , a , y, n na a , n za n, x n nf n , an a v fa a a n f , k n an , a n , an m a y ax

nf n an av n f an m y an m a y an av a a f nan a m a n a y a n a ay n f nf a n , n n a a f a n M an (MDR) a n a n f an a f nf n n a , a a y n n n v a n nf n MDR an m a a a an n a n n f ay (LOS), m a y n a , an n a f a S m a n a n n y n z n a a a m an m a a a y a y a n n na an n a n, ax a, an nk C a n a a an f n v n nv nm n

MDR a n a an a a n an f m ff n a ff n m an m f a n MDR an m a f n f AP f n av n an n a 9 ay an m an k fa f MDR nf n O k fa f MDR n n f k an a n f a za n m mm n y f n m a nf n a na y a nf n , n an f nf n [ n n n af n a n na (G ) a ], an n m a n m n a, f n y a a n f a a n z n a ynx a n na a v , n m a nf n f a n na a a a a n f an a f m y an m m m a y n a an n m a a n na nf n an a m a A a z a n a n a n a n m a nf n S m a n a a a k an y n n, y, an n m m mm n y m a m k y an nf n O k fa a n a ay , f n n a , fa f a a k a an , an v f an

Pa n na f m v a En n ––m mm n y, a n ´ k n, m m m an m m m an Ex n –– n a f n n am na nv nm n , a n m nn , a , a n m n ma n ym a Role of microbes in hospital infections • S a y a m f n am a n n n , n a n a a y, n MRSA m n n ta h lococcus aureus • G ( f A ) na y a nf n , a y nf n , n nf n, G • O n a n Pseudomonas aeruginosa ( a nv nm n – f , f , a , , m , a n v , nf n n P n a a n an a y, 3 n a , m n m m ) an n n f m n n G – cineto acter s . tenotro homonas malto hilia Bur holderia ce acia n n nv nm n (L n a n m a ma a n) • PC n a v a y n a n f a ma a , n n a • n nf n a n , , C, CM

a y v y a n a n a a nf n n a z a n n y m n m f am v an am n a v a a a n f maj y f n m a nf n Am n m S a y a , E a , P m na a n a an En ak a n

n a m a y n m a a n n n y f nf n, nv v n a n an a n ' n y n n n S a an n n a v n fa a a a a a n f v n a man f a n a a a n an a a za n f m a a n a ma a an m a y n an a f a a y a n

A a y n a nf n • Overuse of ATB - resistence and multiresistence (selection pressure of ATB and transmission in hospital environment), toxic side effects, economic burden, deterioration of physiological microflora • Racional indication - preventive in spread of HI • Prophylaxis - oriented to anaerobic infection - perioperative preparation for GIT and UGT surgery, in instrumental examination of patients with bacteriuria • ATB surveys - monitoring of ATB susceptibility • Restrictive policy - time restricted contraindication of some ATB • Rotation of atb - periodical changes of used - decreasing of selection pressure • Combination of atb - agains possible resistent mutnants, broader antimicrobial spectrum

R an an m a a n m n n a va y f a n , a a y a a n m a nf n A a n n f n a n an , m y an m a y n n m a nf n a n a n f a a n m a nf n a a y y a n an m a a m n a y n f nf n, n a n , a m a a , an f an a a a n S n f a a an m a a y a n an n f k y m a a f nf n, n n a y a n , a a f an m a ava a f a n nf n , nam y m f a v y an n y ( n n a v y v kn n an m an m ), a ma k n f an a y an af y W n an m a an av m a va y f a n v n m n a n n n nf n , C n f D a C n a m n m za n f y a , m , an a an m a an a a m y a n a a a an an y v n a a y an a y f an an m n n n n n v a n a a f an n n v an am Pseudomonas aeruginosa • a f v n a m

• an n a n

• ma y a n m a a n

- a f v n a m, mm n y f n n an a , a y n fa f an an a na y n fa f an ma M fa xam nk, a a , m f , v n a an n a a fav

- an n a n, m an n a x m ak n f n n a an nf n a a n m m n f n y m, m a nj y an m a n f n ma f a n n v Ca n ( C ) an n n a n a m mm n y aff Cy a a m na n n a n n a y nf n

ma y a n m a a n A n CDC, v a n n f P. aeruginosa nf n n S a av a a n ( 1 a ), an a m f m mm n y a n m a a n a n n f 1 1 n f a a a nf n

Pseudomonas aeruginosa – virulence factors

Oportunistic: factors of virulence: Pilli - adherence

Alginat(mucoid exopolysaccharide) capsule – antiphagocytosis, attachment , Endotoxin – LPS - sepsis

Exotoxin A – most important, blocs proteosynthesis of eukaryotic cells

Alkalic protease – destruction of tissue

Phospholipase C – destruction of lipids and lecithin, destruction of tissue

Pyocyanin and Pyoverdine – leads to blue green pigmentation http://www.google.sk/images?hl=sk&biw=1024&bih=585&q=pseudomonas%20aeruginosa%20pictures&um=1&ie=UTF-8&source=og&sa=N&tab=wi

a n n a aff k n m a, nva m a y an y m nf n an a a n z n n m n a n am na a y m n , n z n x na n y a a n ; a nf n af a n ; na y a nf n n n f a a n n ; m n n af n y n a af y a a a n m f v n fa av v a n va y f n a man f a n nf n an n a n x a f n n av an may n a a a f a f On a m a a aff a a, an n a am a n m a n a , a a n f k n m a n a a z y n y ma n , n m na yma an n m Pseudomonas aeruginosa’s nf n f n y aff na y a , a y y m, k n an m a, a n ama aff an a n m f v n fa av m an n va y f n a man f a n a m an – f a m v m y m an n nva n an a a m an m n a a v an y a n nj m a fa En x n – an a , k an D C ( m na n ava a a a n) Ex x n A m a n yn y na va n n a n fa R n yn a n an a nv nm n f n nf n k f m – f m a a f m a y an mak a a an m Py yan n an Py v n – a n m n a n an f y m n, n x a an m M an – a a ama an am n y m fy n nzym mak n f an n m f an n m na

Pseudomonas aeruginosa microscopy • is a Gram-negative rod measuring 0.5 to 0.8 µm y 1 3 µm • Almost all strains are motile - single polar flagellum.

• It is identified on the basis of its Gram morphology, • inability to ferment lactose, • a positive oxidase reaction, • its fruity odor, • its ability to grow at 42°C.

Pseudomonas aeruginosa - cultivation P. aeruginosa strains produce soluble pigments: - fluorescent pigment pyoverdin - blue pigment pyocyanin. Pyocyanin (from "pyocyaneus") refers to "blue pus", which is a characteristic of suppurative infections caused by Pseudomonas aeruginosa.

- ability to grow at 42°C - fruity odour

ENDO, DCA: inability to ferment lactose, H2S negative

A triple sugar iron agar (TSI) tube inoculated with Pseudomonas aeruginosa and incubated at 37°C for 24 hours results in an unchanged slant and butt.

OXIDASE TEST positive

http://www.google.sk/images?hl=sk&biw=1024&bih=585&q=pseudomonas%20aeruginosa%20pictures&um=1&ie=UTF-8&source=og&sa=N&tab=wi

a a P m na a n a a v y m n na m n f n v " n n a ", v n f m n ma n na n n a a y, m m m f f P m na a n a n f a a a a f a n an amm n m fa a a f n n P a n a a may ny y a a a f m a y a y a ma , ny C n a am ny y On y a a f a a an a , m , f a an an va a a an An y , f n y a n f m a y an na y a n , a a m a a an , a n f a na m m an m n a m ay a n n za n an v n D xy a – a a a , En a a P a n a n f m n a , S n a v S a n m n , f n m n y v n an m n y yan n Py yan n (f m " y yan ") f " ", a a a f a v nf n a y P m na a n a A a n a a ( S ) n a Pseudomonas aeruginosa an n a a 3 °C f n an n an an an P m na a n a x a v x a a m a a n a a ay f p f y h x , an nzym m m a n n x a n n f an an m a n a n y m x a nzym , a n m an x z

x a f n a a n , - hy -p-ph y hy h ( KOVÁCS g ), a an a f a n n f y m c

W n a n x z y y m c, an f m a a k m n , ph b . On f n x a v P m na a n aan n x a n a v E a an m n a f a a n n K vá x a a n an a y

ATB susceptibility of Pseudomonas • Therapy - Frustrating – immunocompromised defence of patient, typical ATB rezistence induction of ATB inactivating enzymes resistence transmission via plasmids Isolation without signs of infection is not indication for ATB therapeutic intervention • Combination of ATB – beta lactams + aminoglycosides, Meronem, Imipenem, Sulperason, Cefoperason • Preventive approaches - Interruption of contamination of steril materials and cross contamination. Decontamination of fits and and tubes and catheters and environment - nosocomial strains in intensive care units. Broad spectrum ATB use - very carefully - suppression of normal flora and overgrowth of resistent bacteria and mutants

Pseudomonas aeruginosa an v fa an an , n m a n y n mm n a za n f an am n an a y an a a m m y n PA nf n’ a m n a n n a P a n an a n, am n y k G n am n an Am ka n, m n m, Q n n , Az nam an C a n a a Listeria • The genus Listeria consists of 10 species, with • Listeria monocytogenes and • Listeria ivanovii the only recognized pathogens.

n Listeria n y m f 1 , n n 1 Listeria n y n 9 Af n v a nam an n a a f n 1911, nam a ff a y an Listeria monoc togenes n 19 n J L , n v a z n a n m n f a n k f nf n L. monoc togenes a n nv nm n , a y n an a a a an v v n f many m n An ma , a a y a , an a y L. monoc togenes a a n k an a a n f a a a a v n a an f n n an nv nm n — y an a a an a y n n , an m a , an a fa y m n n a a a L. monoc togenes v v a n m n a va y f f an f n an a a a an m y an n f n an f y a L. monoc togenes a y a n an an n n m n n am na n f f n k m a a, L. monoc togenes an an m y a m a , mak n a a a n a m v n n y f a f On y a f n a a v y n a f f z n a f n am na L. monoc togenes

P m nf L. monoc togenes y a n n am na f P an a m nf y an n n am na f , a n am na f , n n am na fa an n an n a n a y an f n a a f m an m a an m nf n a f m a n am na f n nan y a a an n am na a va y f f , a Ra m a ; R a y a m a a an m a ( fa y a a ka an a n ); Ra v a ; R f a a ; R a y a m k af an a af ; P a a a , n n a an f f a a ; M n ; S f ma n a z m k; an n a z m k an m k Pa za n, k n , an m nf n a n k L. monoc togenes L a m x v y n nan m n, n n , y, an ak n mm n y m ( a an a ) P nan m n a 1 m m k y an a y a , an 1 f a L. monoc togenes nf n a n nan m n S n n’ kn y nan m n a a Af a n n L. monoc togenes, a a k y n v an n m v n am an an nva many a n y, n n a n; M m an n n a n an na ; Ga n na , v , a ( ma an n n ); an am

L. monocytogenes • Granulomatosis infantiseptica - severe form of early-onset listeriosis, • abscesses and granulomas in multiple organs • high mortality rate unless treated promptly

G an ma nfan a an n a n nf n a f a n na a m a y a v y m a y f m n f nf n n a z a a z y x n v f a n a m n y nf a n f v , n, an m a y f n an n n a a m n n n m a

L. Monocytogenes - diagnosis

• The diagnosis is typically made by microscopy and culture of : • Blood • cerebrospinal fluid (CSF) • amniotic fluid.

Sym m f a nf n a n n f an m a nf a L a na y aff a y, y n , n n nan n v a n n n a k P nan m n E y n na n v a mm n m m y an an an a n, an , a m n m n fa an a n a n y a y ma y f , na f (CS ), amn f

L. monocytogenes - microscopy

• significant human pathogen, • facultative intracellular pathogen gram-positive rod • The short rods appear singly, in pairs, or in short chains • Motile „ n v n “ tumbling motion

L a m n y n a fa a v y n a a , am v a v a a m a an , n n refrigeration m a an m m n a m a , m n a tumbling m y a m a A y m a (3 °C) n a n ym za n n a n an f m “rocket tails” a fa a n a a m v m n an a a m m an , ay avoids antibodies L. Monocytogenes - culture

• blood agar - hemolysis • Mueller-Hinton agar • cultures : 1-2 days

Treatment • Ampicillin

L a m n y n an a a y n n m a, a m ak n n an m f m G am v , a a y L a n m a a M n n a a On a a a a ma z n f m y an v a n an n n , ak 1 ay f T An a y a m n f a m a a f k f a n mm n m n L n may n mm n m m a n M n n a f 3 k Am n n a y n f a n , n am n a f n y f yn y

Bacillus

a a a a a fam y f G am v , , a a a a may n M m m f fam y a a a z y f a a S m a a a a a , a fa a v ana M a n a n , a a kn n a a n man A fam y f a an y n , , n a m na y, m na y, n a y a m an m y a y , mm n y a a n n S m a a a n n an an ma

Bacillus anthracis. • Anthrax - fatal disease - Bacillus anthracis. • anthrax spores are • inhaled, • ingested, • contact with the skin lesion

An ax an nf an n a y fa a a a y a m Bacillus anthracis. a n an ax a n a , n , m n n a k n n n a An ax mm n y aff a m an ma , a y a an ; an a a na y aff man a n nf n n man nv v k n, a n na a n man an m nf an ax y an n f m nf an ma y a n n an ax f m nf an ma ( k ) P a an m nf a n na an ax y a n n k m a f m nf an ma Bacillus anthracis – virulence factors Exotoxin – 3 components:

- Edema Factor (EF)

- Lethal Factor (LF)

- Protective Antigen (PA)

v n fa f B. anthracis n a n m f x x n an a Ex x n a m n , a a , n n m x ma f 3 m n n E ma a (E ),L a a (L ), P v An n (PA) fa a yn a y (f x ff ) n v v , n f na v m y B. anthracis a n f x n n n f L an E a n n a a x n n n f L an PA a an a v a x n, an n n f PA an E a an ma x n PA n L an E av a an ma ff a n f a y f D am a n y a a m an an a y n a mm n n an , v n f any an , y a n v a a n a W n v n a , a a a k y a a a n n an an af y m y

Bacillus anthracis – virulence factors

ama n na f B. anthracis v a n a n m f a a n a x x n a a a v an n, ma fa , an a fa a a a m a a n a v y m v W n ma fa n v an n, ma fa m a va an a f f m an n W n a fa n v an n, a a x n a a k ma a an f mm n y m a an na v f m a , v y n v a a a f a f n n f an a n a

Types of anthrax:

Cutaneous anthrax

Inhalation anthrax TREATMENT: penicillin ciprofloxacin

An nf n v n man af x B. anthracis n f a n n y n n n f x C an an ax n a a n k n a n ak n k n, a n a k n M an 9 f a an ax a v f m an x C an x an av a m a y a a n

nf n an a f m n a n n a n a f m man fa n n a a n an ax n m a y f m f an ax nf n a y v n a k af x , an ak m n W a m n , n y a 1 – 1 f a n n a a n an ax v v v , a v a m n , a f a n v v

L mm n y, an ax nf n an n v y m f m a n n am na f k n k an a n m a nf n a y v f m 1 ay af x W a m n , m an a f f a n a n na an ax v , a m n , f a n v v

An ax nf n n a a y a an f n n n z n a n a a , a n m a y a An a m n a ava a an f n a m n a y, y a n’ a v ama n y x n nf n an a y a n n, an an a a a y n nk f y an n a a a’ a O an k f xa n a n n n nf n a y B. anthracis. P n n an a a an ma , k a , a av n a an ax a m n a ava a , an ax va n a ava a man an an ma ff n a a n B. anthracis man va n a n ava a n a a n, y a v n k n y an ma an ma a a a k f nf n C n y a m n man an ax va n n a a ( SA) an v (R a) va A n y an ax va n a an n a a n y ( y ma, n a n, n , f v ) an a x ma y a , , k an , 1 , 1 m n , ann a ma n a n mm n y http://www.youtube.com/watch?v=i g1Cz2tdVjY

An ax an a a a a n n n S a n 1 An ax a a y a a y m y n n n a n n an ax a a f an ax nf n an a an a a n, a v y v n ( x n), a y a y a a , m , an a n y n n 19 , n S n W Wa , a a a a n G na y m a y n f m y D a m n v nm n a nv a n f a y f a a n a a k n an ax a n a a n a n n am na n f mm a a a y an ax

Bacillus cereus

- opportunistic pathogen

- relatively low capacities for virulence

- Gastroenteritis

- ocular infections

- intravenous catheter-related sepsis

https://www.sciencedirect.com/science/article/pii/S0014483519307493

W B. cereus a a ma n y f n n , n n a n y a a f an n a y fa a n n a n na a nf n a n y f B. cereus, n na n n n na , n ma y a a n f v x nzym Am n x n a f m y n , n a , an m n n x n, an a

a m a n n m na n a m f n a n n a f n n an v y nf n , an ax k v n m n a, f m nan , an va a n n a n v y m nf n , a a y n mm n n v a , n av n a , an n na n n m a a a m a an n nf n n a a n a a n f n , a y n n ava a v a a a n P ma y an nf n m m k n a a an n n n a ma av a n m n ym m f a a y f n n m m f C m f n n f n n n f a y a a, a m na am , an a n 1 af n m n f n am na f a a may a m any a a, v m n ( m ) a y Sym m f n m n an m y f f n n a a z y na a an v m n n af n m n f n am na f O a na y, a m na am an a a may a D a n f ym m n a y an ym m f y f f n n a a a y S a y a f n n x a n

B. Anthracis B. cereus – Gram stain

G+ huge rod , long chains of rods with centrally located spores: Bacillus anthracis: In smear from tissue - not spores. Bacillus cereus: G+ rod without capsule, motile

Bacillus anthracis v y a , G am v , f m n (1 1 μm n x 3 μm n n ) a an n a n v y m a a , av am a z an m y A a a a n (“ x a ” a ) a a an a a B. anthracis v a v Bacillus cereus a G am v a fa a v y ana , m , f m n , a a m C

• B. anthracis on blood agar- aerobe, facult. anaerobe rod, small adherent colonies lining in paralel way - growing in picture of caput medusae, colonies with wave edges. • colonies are rough, • Older colonies growing in aerobic environment contain spores – dry, wrinkled look. • Growth on medium with PNC - chain of pearls

• B. cereus on blood agar – • capsules are not present, • gray colonies, • Beta-haemolysis B. anthracis B. cereus

B. anthracis a n n fa , fa a v y ana a m an m a y n m a a y m a m m m a f 3 –3 °C Af v n n a n a 3 –3 °C n a a , –3 mm n am , a , a , , a an y “f a ” ( n a ) a a an O a na y n may av f n n ( a n ) a “M a a a a an ” Ca m a Ca a f m n n a na a a n a n n m n a – a n x a m , n f na n m ( f na m k ) B. cereus W n n n a n n n a a a 3 °C, B. cereus n a ay an a a ma fa C ny m a a an n nf a n f a m n f m f n a n a n, a B. cereus a m n m y Z n f a m y n an nf m ny m y

Sporulation

S m a a av a y n a a f n an ma n n n n a nfav a An n an x m y an man y m a a ( m n ' n ' m an ' n ' an ' ' f ' man ,') n a f m n a many xam f n f m n a a m mm n a Clostridium an Bacillus a a n ma y , ma an y ma W n n n n nv nm n a ( a , a a n, m a nf n, a n), f m a a n f ma n f n f ma n a a n S n f n v v n f an m f , a av m z any a a

Af n f a nv nm n a v a v , n f an n v a v a a m na n

P f En • C a n a n a m m m , a f m n n f n yn z n a a a , an an n y n a n y m a n y y A n m f n nzym a f m ( , n a yn a ) a an f n a y a a an n a n n f a am n f a m na • S Wa nn m ay n n nn m m an a a n a n n ma y an an m a f m na n v a v • C x x k ay f nv C x y an x m y n v y zym , an a y ay a n m na n • C a a m f a k a n k n m m a y f ay nf n a v an an a a m a a n • Ex m x m a n m m an n a n n a y a

W z C nk n

• Differencial staining form spores • Spore forming bacteria – red • Spores – green 1. Prepare fixed smear: susspension dried on the air, heat fixed 2. colorization: 5% malachit green - heating untill evaporation 3times (3-6 minutes). 3. Rinse under water flow 4. add 0,5% water solution of carbol fuchsin - counterstaining (60 sec.) 5. Rinse with water, dry and read

Bacillus anthracis an Bacillus cereus n v a n f a f m a n a n , y an n a y v a z n m a f an a a a y a a a ( a a n n a a ) af – f n a n a 3 °C 3 °C a n a m na , a an n v a v S a n n n m a a f m n a m n En a a n y W z C nk n m ma a n f a n n f n an a n a a n f n n a n a nk af an n a f n Ascoli termoprecipitation reaction • D n f an n x a y a y f m a ma a y an n a a

ASCOL 'S ERMOPREC P A O ES f a n f an ax ( a an a ) a n a n , n n am v n f m an a a k y n nv a am a n n a n , an f an n x a f m ay v an ax an m n a na a a y A n f a a a a j n n f n m n D a n may a y A ’ m n y m n a n f an ax an n n x a a m n • Bacillus cereus infections

• Bacillus anthracis

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