Week 3 - Enterococcus, Listeria, Erysipelothryx Flashcards
1
Q
Enterococcus used to be ________ until 1984, when the advent of sequencing such as ____ _____ gene)
A
Streptococcus, 16S rRNA
2
Q
Enterococcus is gram - _______.
A
positive
3
Q
Enterococcus occurs in _____ or ____ _____ like Streptococcus
A
pairs, short chains
4
Q
Enterococcus is Catalase _______ like _______.
A
negative, Streptococcus
5
Q
Enterococcus is Lancefield Group __ antigen (antibody based serogrouping)
A
D
6
Q
Enterococcus is a ________ ______.
A
facultative, anaerobe
7
Q
Some species of enterococcus are ____-_______.
A
non-capsulate
8
Q
Enterococcus is ____-_______ except some species.
A
non-motile
9
Q
What media types are used for growing Enterococcus?
A
- Blood agar
- Media containing up to 40% bile esculin
- MacConkey agar
- Media containing high salt ( 6.5-10%)
- Kenner-fecal agar media
10
Q
Enterococcus
Blood agar = ?
A
Non-hemolytic = gamma
11
Q
Enterococcus
Media containing up to 40% bile esculin produces?
A
dark colonies (NB: Streptococcus does not grow on bile)
12
Q
Enterococcus
MacConkey agar = ?
A
ferment lactose, producing small dark-red magenta colonies
13
Q
Enterococcus
Media containing high salt = ?
A
Grow on media containing high salt (6.5-10%) concentration
(unlike Streptococcus)
14
Q
Enterococcus
Kenner-fecal agar media
A
Selective media for enterococcus
15
Q
A
Bile Esculin Azide Agar
16
Q
Enterococcus is a highly ______ organism in ____ even if they
are __-____ forming bacteria
A
resistant, nature, non-spore
17
Q
Enterococci are able to grow in?
A
hypotonic, hypertonic, acidic, or alkaline conditions
18
Q
Enterococci are able to withstand?
A
detergents, oxidative stress, desiccation, heavy metals
19
Q
Enterococci are resistant to?
A
multiple antimicrobials = member of ESKAPE
20
Q
Enterococci are normal commensals of?
A
Mammals, birds, reptiles, fishes, insects.
21
Q
Enterococci live in what parts of the body?
A
Colon and bile tract, Oral cavity, Urethra, Vulva/vagina in humans and animals
22
Q
Enterococci is the ________ ____ and ____ microbiota of animals and humans. Thus, millions of them are _______
with feces daily to the environment
A
leading, gut, fecal, excreted
23
Q
Name the body structures of Enterococci used for adhesion, colonization, and biofilm formation.
A
- collagen binding proteins
- endocarditis specific antigens (pili)
- surface proteins of enterococci
- Enterococcal polysaccharide on surface = cell wall carbohydrates serve as a capsular
- Aggregation substance = binds to host cells or bacteria-to-bacteria (conjugation)
- Trafficker of AMR genes by transferring them horizontally to enterococci spp.
- Acquired broad-spectrum antimicrobial resistance (AMR).
24
Q
What enzymes do Enterococci use?
A
Gelatinase, Hyaluronidase
25
Gelatinase digests ?
Gelatine, elastin, collagen, haemglobin, and other bioactive peptides.
26
Hyaluronidase destroys blood _____ and ________ of the _______ tissue/_______ for spreading of _______ to the ______ tissue
vessels, mucopolysaccharides, connective, cartilage, bacteria, deeper
27
What toxins or secreted substances do Enterococci use?
1. . Cytolysin/hemolysin (also called bacteriocin or enterocin)
2. Sex pheromones
28
Cytolysin/hemolysin (also called bacteriocin or enterocin) kills by _____-forming on cell ______ of ___ and _____ blood cells
pore, envelope, red, white
29
Cytolysin/hemolysin (also called bacteriocin or enterocin) kills Gram-_______ bacteria competitors = to _____ its territory that contributes to ____ control
negative, defend, niche
30
Sex pheromones ________ expression of ______ substances which results in ________; thus, it is a means of acquiring and accumulating ______.
stimulate, aggregation, conjugation, plasmids
31
E. faecalis, E. faecium, E. durans occupy which host animals?
Multi-host species
32
E. faecalis, E. faecium, E. durans occupy which habitats?
intestinal tract, soil, water, food/feed
33
E. faecalis, E. faecium, E. durans cause which diseases?
1. Septicemia in poultry
2. mastitis in cows
3. endocarditis in cattle & lambs
4. urinary tract infection in pets
5. chronic liver diseases (humans)
34
Overall, 65-95% of the diseases are by __________?
E. faecalis
35
E. faecalis, E. faecium, E. durans cause?
36
E. faecalis affects ______ of all ____, but exceptionally severe in ______ and ______ _____.
birds, ages, embryos, young chicks
37
E. faecalis usually colonizes the _____ first followed by E. ______, and then E. ______.
intestines, faecium, cecorum
38
E. cecorum has emerged as a major cause of _______ disease in adult _____ _____ causing ?
skeletal, broiler chickens
- osteomyelitis,
- femoral head necrosis,
- Spondylitis (i.e. back & neck pain due to inflammation of the vertebrate joints)
- arthritis.
39
Spondylitis in broiler chickens (showing it in humans)
40
Currently, enterococci rank among the top ___ leading cause of ________ infection in humans
3rd, nosocomial
41
In humans in the USA, Enterococcus spp. contributes to:
1. 20% of _______, ______ and
2. 10-16% _____ infection
endocarditis, endodontic, urinary
42
The majority of enterococcal infections are caused by E. _____ or E. _____
• E. faecalis accounts for the majority (65-95%) of ______ Enterococci infections in humans.
• E. faecium constitutes the majority of _______ and ______ resistant isolates of enterococci
faecalis, faecium, nosocomial, vancomycin, ampicillin
43
To a significantly lesser extent, infections are caused by other Enterococci species such as ?
E. durans, E. avium, E. gallinarum, or E. casseliflavus
44
What are the entry and exit transmission routes of Enterococci?
Entry = oral route: ingestion
Exit = feces from gut (their ecological niche) and other body parts via urine or milk (in case of mastitis)
45
Enterococci are excreted everyday in _____ concentrations in ____, usually between ____ and ___ bacteria per ___ wet weight
high, feces, 104, 106, gram
46
Enterococcus is routinely used as an indicator of fecal _______ and ______ quality of ____, ____, _____, ____ and ______ contamination/pollution by fecal
materials
contamination, hygienic, food, milk, meat, water, environmental
47
Collect samples and culture.
48
Treatment of Enterococci is challenging due to its _____
MDR
49
Faecium is more resistant, but faecalis causes disease more. Do AMR test to see which AMR is the most effective. Expensive ones are best (listed here).
50
Relative to the Streptococci, Enterococci are naturally _____to many commonly used antimicrobial agents such as _____ ______
resistant, beta-lactams
51
Enterococci generously give their AMR genes (_____) to other bacteria species by ______ (_____ _____)
plasmid, conjugation, horizontal transfer
52
MDR is more common E. ______ than E. ______
faecium, faecalis
53
How do you treat cases of Enterococcus infection?
In case the isolates are susceptible, use Beta-lactam antimicrobials e.g. amoxicillin, vancomycin OR Carbapenem
54
How do you control cases of Enterococcus infection?
1. effective waste treatment of feces/manure and hygiene
2. Water sanitation to reduce the incidence.
3. Ensuring proper cleaning and disinfection of the facilities can reduce
environmental reservoirs of the bacteria.
4. Preventing immunosuppressive diseases and conditions, because
enterococcosis often occurs secondary to another disease.
55
What is the most clinically important strain of listeria?
Listeria monocytogenes
56
Listeria ivanovii (?)
Less pathogenic
57
Listeria is gram _______
positive
58
Listeria is _____-shaped ______.
rod, bacilli
59
Listeria is a ________ ______.
facultative anaerobe
60
Listeria is ___-____ forming.
non-spore
61
Listeria is catalase _______.
positive
62
Listeria is an ________ pathogen of animals and humans
intracellular
63
Listeria possess ________ for motility and invasion.
flagella
64
Listeria is ________-_______ on blood agar.
beta hemolytic
65
Listeria is resistant to ______ _____ (?) in the media.
high salt, 7-12
66
Listeria grows at ___ temperature (?).
cold, 4 degrees celsius
67
Tumbling motility or “umbrella” shaped colony growth at
20-28C in a semisold motility media (because of its 1-5
flagella)
actin jet motility inside host cell
Umbrella-shaped subsurface listeria colony growth in semi-solid
motility media.
68
L. monocytogenes has ____ serotypes based on its ____ (O) and _______ (H) antigens. Of these, __ serotype are important (virulent) since they cause the vast majority of clinical case = ?
14, somatic, flagella, 3
1. 1/2a = the most frequently isolated from food
2. 1/2b
3. 4b = causes the majority of human epidemics
Ruminants are very susceptible to listeria, humans are second
69
Isolation of Listeria on a culture media has two key issues:
1. It require enrichment media as it is fastidious bacteria
2. It requires prolonged inubationtime for the recovery of stressed Listeria cells.
70
How do you create an enriching media for Listeria?
1. Mullen-Hinton agar with 5% sheep blood as enrichment
2. Beta-hemolytic colonies on blood agar
3. Ferments glucose, maltose, lactose …produces acid but no
gas.
71
Describe how you recover stressed Listeria cells?
1. Incubation for weeks at 4°C on agar plates until the formation of visible colonies
2. This method of isolation usually does not allow for the isolation of injured or weak Listeria cells
72
FDA and ISO developed broth media that gives you, in 48 hrs, results.
Animal fecal sample or discharge from aborted animals or from meat samples or anything that you suspect has listeriosis -> culture in broth -> incoulate -> add supplements after 4 hours (acriflavin, etc). to kill other organisms in case if they are growing in the broth. -> incubate for 48 hours at 30 degrees -> do gram stain to observe gram positive rod and perform catalase test to find bubbles
73
Listeria is resistant to ____ salt (?%) in the media
high, 7-12%,
74
Listeria survive and multiply on ______ objects. For example?
inanimate
• E.g. silage through the transition processes from one host to another
75
Listeria adapt to _____ temperature (-0.4-45 degrees C),
pH (4.4-9.6). Can multiply at refrigerator Tm (4C)
changing
76
Listeria grow as _______ in soil and _______ vegetation in the environment.
saprophytes, decaying
77
Listeria is not ______ rather a real pathogen
commensal
78
Listeria is isolated from ?
sewage, water, feed, food
79
Listeria is abundant in ____ temperate.
What countries/regions of the world?
At what temperatures?
cold
EU, North America
Can grow at low temperatures < 4°C
80
Describe Listeria's host range.
It has diverse host ranges and isolated from:
Humans
42 species of mammals
22 species of birds
fish
insects
81
Listeria is most common in what species?
It is more common in ruminants (mainly sheep)
It causes seasonal outbreak in ruminants
It is sporadic in pig and horse
82
_______ is a well recognized food borne pathogen in humans.
Listeria
83
Describe the point of entry and exit of Listeria.
• Entry: It enters the host by ingestion (oral)
1. Livestock: highly associated with feed/feeding of Silage (decaying plants) with high iron content is a risk factor for acquiring Listeria in ruminants
As you increase iron concentration in silage, it promotes listeria contamination
2. Humans: Consumption of contaminated raw vegetables, meat, and milk
Exit: It exits from the host via feces, vaginal discharge (abortion), milk (mastitis)
Detected in feces, milk, slurry, soil, feed, water, food
84
What body structures does Listeria possess?
1. 1-5 Flagella - for motility, adhesion & invasion
2. Internalin (A, B & C) - for host cell adhesion & invasion
3. invasion-associated protein (iap) - for host cell adhesion
& invasion
85
Listeria form _______.
Biofilms
86
Listeria is an _______ pathogen and multiply in _____ cells, _______, & ________.
intracellular, tissue, monocytes, macrophages
87
Listeria hide from?
the immune patrolling of the host cells,
antibodies & antimicrobials
88
What enzymes does Listeria use?
1. super-oxide dismutase from listeria
2. Phospholipase
89
Super-oxide dismutase protects listeria against _____ ______ from the host _______ cells
free radicals, phagocyte
90
Phospholipase = _____-forming _____ on endocytosis membrane of the host to free itself from ______ (vacuoles) in the _____ of the host cell
Pore, lysis, lysosome, cytoplasm
91
What toxins or secreted substances does Listeria use?
1. Beta-hemolysin
2. listeriolysin O
3. Bacteriocins (listeriolysin S) –
4. Actin-polymerizing protein (actA)
92
Beta-hemolysin and listeriolysin O = _____-forming _____ on endocytosis membrane of the host to free itself from _____ (vacuoles) in the _____ of the host cell
Pore, lysis, lysosome, cytoplasm
93
Bacteriocins (listeriolysin S) – ____ competing gut microbiota
kill
94
Actin-polymerizing protein (actA) instructs ____ cell for deposition of its ___ filaments on the end of Listeria. Then Listeria uses the host ___ filaments for _____ to the nearby cells. Such host ____-based zipper mechanism for
propelling and transmission from cell to cell are
used by?
host, actin, actin, propelling, actin, Listeria, Rickettsia, and Yersinia pseudotuberculosis
95
Listeria enters the 4 known barriers of the body systems
1. Blood-brain barrier
2. Placental barrier
3. Intestinal barrier
4. Cell membrane lipid bilayer barrier (intracellular).
96
1. Attachment and adhesion
2. Enter deep into cytoplasm
3. Multiply here and then jump to adjacent cells.
4. When they come out of the first cell, it is already dead (kill and jump).
97
Listeriosis is characterized by ?
Febrile gastroenteritis, septicemia/shock, placentitis, brain stem & cranial nerve dysfunction
98
Swallow contaminated food and water, listeria establish themselves in digestive tract, then pili of epithelial cells are eroded and majority of them come out to the environment with diarrhea to infect another host but also some go to the deep tissue to lymph nodes. They can stay inside the MQ and MQ and neutrophils, monocytes will stay there (?) and use a ?bot?, so when MQ, neutrophil circulate around the body to lymph node -> go with monocytes to spleen or liver -> blood stream to brain, placenta -> death
Shooting their enzymes with toxins; fighting wherever they go.
write side effects on separate cards
99
The two major clinical manifestations of listeriosis are: ?
neural form vs. visceral form
100
Neural form of listeriosis in silage-fed sheep shows?
microabscess in the brain
Ataxia and ‘circling disease’
Unilateral facial paralysis
head tilting to one-side
ear dropping (one ear)
tongue protrusion
salivation
101
Visceral form of listeriosis is linked with ?
________
________
_____ and ______ damage
___________ tropism (________)
_________
_______ (bone + bone marrow)
_______
Gastroenteritis,Septicemia,Liver,spleen,Fetoplacental,abortion,Myocarditis,Osteomyelitis,mastitis
102
____________ is the third leading cause of death from food-borne illnesses in the United States, with approximately 260 deaths annually
L. monocytogenes
103
Collect fecal sample, blood, milk, urogenital in case of abortion -> use FDA or ISO enrichment media -> gram stain, etc.
104
How do you treat Listeria infections?
Treatment : Listeria is a Gram-positive intracellular pathogen (more responsive than ESKAPE)
Ampicillin (200 mg/kg per day, IV)
Chlortetracycline (10 mg/kg per day for 5 days, IV)
Penicillin (44,000 U/kg per day for 7 days, IM)
Trimethoprim-sulphmethoxazole (10 mg/kg per day, IV)
105
How do you control Listeria outbreaks?
Its control is difficult as ubiquitously present
Little is known as to its risk factor (except silage) to control
Control Listeria growth in the feed mainly in silage
Vaccination with live attenuated listeria vaccine (in Norway)
and commercial killed vaccine in some countries
106
Which strain of Erysipelothrix is medically the most important?
Erysipelothrix rhusiopathiae
107
Erysipelothrix rhusiopathiae is gram ______.
positive
108
Erysipelothrix rhusiopathiae is shaped like ?
Causes?
Long filament (rod)…. (causes emboli)
109
Erysipelothrix rhusiopathiae is a _______ ________.
Facultative anaerobe
110
Erysipelothrix rhusiopathiae is Catalase ________
Negative
111
Erysipelothrix rhusiopathiae is oxidase ________
negative
112
Erysipelothrix rhusiopathiae is ____-motile
non
113
Erysipelothrix rhusiopathiae is _____-spore forming.
non
114
Erysipelothrix rhusiopathiae
115
Erysipelothrix has ____ serotypes based on _____ antigen on the cell wall
28, peptidoglycan,
116
The pig is susceptible to at least ___ serotypes of Erysipelothrix.
15
117
Field cases of swine erysipelas are predominantly caused by three E.
rhusiopathiae serotypes:
1. 1a
2. 1b, or
3. 2
118
In swine, ?% of isolates are classified into serotype 1 or 2
75–80
119
What culture media can be used for Erysipelothrix?
• Blood agar (-hemolytic after 24h)
• Heart infusion + supplemented with sodium azide & crystal violet
• Nutrient broth + 1% glucose
• Triple sugar iron
120
Describe how to achieve nutrient rich media for Erysipelothrix
It require enrichment media
1. Alpha-hemolytic colonies on blood agar
2. Heart infusion agar with sodium azide & crytal violet for plating pig
sample & incubated for 48h
3. Ferments glucose, lactose producing acid but not maltose and mannitol
121
Hydrogen sulfide H2S is produced by 95% of strains of Erysipelothrix species on triple sugar iron (TSI) agar
H2S smells like fart
122
Erysipelothrix is unique & probably the only Gram-positive bacillus/rod bacteria that produce H2S (gas)
123
124
125
What habitat do Erysipelothrix occupy?
Habitat – Tonsils
- lymphoid tissue of intestinal tracts in carrier animals
- Detected in the slurry of cattle herds or sewages from abattoirs
126
Describe the non-human animal host range of Erysipelothrix?
Mammals (pig, horses, cattle), birds (turkey), reptiles, amphibians, fish
Pigs including wild boars (wildlife)
Detected in horses with vegetative endocarditis
Turkey with swollen snood
127
Describe the human host range of Erysipelothrix.
Humans due to occupational zoonotic disease
• people who have contact with pigs, poultry, fish (vets, abattoir workers, butchers,
farmers)
It causes a purple colored hardened finger swelling in humans
It may disseminate from skin to cause endocarditis, pneumonia and meningitis.
128
Describe the points of entry and exit of Erysipelothrix
Entry: oral route
enter the host orally to colonize tonsils
Flies may transmit it
• Exit: fecal route (urine, saliva, nasal
secretion)
exit from the host mainly with feces to spread
to the environment (effluent) and new host
129
Describe the virulence factors of Erysipelothrix
No well known virulence factors except the following surface structures and enzymes
130
Describe the structural surfaces of Erysipelothrix
1. Capsule (lipopolysaccharide, slime or glycocalyx) – for resistance to phagocytosis
2. Surface protective proteins (spA, B and C antigens) – for biofilm formation,
adhesion, and to elicit immune response.
These surface proteins can be used as a protective vaccine.
131
What enzymes does Erysipelothrix posess?
1. Neuraminidase (sialidase)
2. Hyaluronidase
3. Superoxide dismutase
132
Neuraminidase (sialidase) – for bacterial _______, ____, ____ blood vessels leading to ?
attachment, invasion, destroy
hemorrhage and thrombosis
This can lead to bacterial emboli and infarction in the heart valves (endocarditis), spleen, liver, lung, kidney, and joints (polyarthritis)
133
Hyaluronidase – for spreading via destroying ______ ___ and ______ between adjacent cells
hyaluronic acid, polysaccharides
134
Superoxide dismutase – for protection from _______ killing by ______.
intracellular, macrophages
135
Erysipelothrix is the problem of ___, ________, and _____ …. it is also?
pigs, ruminants, turkey, zoonotic/humans
136
Diamond skin disease - dark/bluish in color due to lack of O2 to skin.
Go into circulation -> attack vital organs
137
Erysipelas diamond shaped skin form
rhomboid urticarial purple/black and hard skin lesion mainly around belly, inside the thighs, throat,
neck, ears
138
Erysipelas
pneumonia
139
Erysipelas cardiovascular form
• Purple/black discoloration of the skin of belly with bluish (cyanotic) of the extremities due to
• septicemia, emboli, infarcts, & vegetative endocarditis that blocks heart valve orifice.
Erysipelas ranked 2nd next to Sterpt. suis in causing endocarditis in pig industry
140
Erysipelas joint form
Polyarthritis (swelling joints, lameness and stiffness) mainly elbow, hip, hock, stifle, and knee joints
141
Erysipelas uterine form
Abortion mainly due to fever, but the bacteria is isolated from fetuses (i.e. congenital/vertical infections
indicating it can pass the placental layers)
142
Describe the clinical manifestation of Erysipelothrix
143
What are the clinical signs of Erysipelothrix
• Clinical signs (lace-like or diamond
shape skin lesion) that is purplish
144
How can you diagnose Erysipelothrix via bacteriology?
1. Gram positive - Long filamentous (Rod)
2. Catalase negative
3. Alpha-hemolytic
4. H2S in TSI agar
145
How can you diagnose Erysipelothrix via serology?
serotyping) - 28 serotypes
146
How do you treat Erysipelothrix infections?
Penicillin (50,000U/kg
per day for 3 days, IM)
ampicillin
ceftiofur
147
How do you control Erysipelothrix infections?
• Vaccinate pigs > 3 months age with formalin-killed
vaccine bi-annually
but vaccine fails due to several serotypes i.e. 28
short duration of immunity i.e. protection lasts less
than 6 months
• Therefore, the feasible control methods are:
1. Remove clinically sick or test positive animals from the
herd immediately (but no reliable test?)
2. Good hygiene (biosecurity): all-in all-out policy followed by
disinfection of the house before restocking
