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Sem 2: Microbiology > Week 8 > Flashcards

Week 8 Flashcards

1
Q

Whats a break down of crop losses due to microbial pathogens?

A

Wheat- 12.6%
Rice - 12.2%
Maize - 11.2%
Potato - 20.1%
Soybean - 10.1%

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2
Q

How much loss in global food production is impacted by plant pests and diseases?

A

20-40%

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3
Q

What are saprophytes?

A

Grow in soil on dead organic matter

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4
Q

What are epiphytes?

A

Grow on external plant surfaces

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5
Q

What are pathogens?

A

Grow in internal plant surfaces causing disease

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6
Q

What are biotrophs?

A

Keep the host alive

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7
Q

What are necrotrophs?

A

Kills the host as a way of life

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8
Q

What are hemibiotrophs?

A

Switch from biotroph to necortrophs

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9
Q

What are the main types of bacterial disease in plants?

A

Fruit rot
Leaf blight
Wilt
Stalk rot

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10
Q

What are the genera of bacteria plant pathogens?

A

Pseudomonas, Xanthomonas, Erwinia, and Agrobacterium

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11
Q

What relationship with being a pathogen do the main genera of pathogenic bacteria have?

A

These are facultative pathogens, i.e.:
Exist as saprophytes or epiphytes
Migrate into plant tissues, replication and cause disease

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12
Q

How do you identify a disease-causing agent using Koch’s postulates?

A

Naturally occuring disease
Isolate and grow in pure culture
Inoculate healthy plant
Development of original disease
Re-isolate bacterium

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13
Q

How many species per genus are there of pathogenic plants?

A

Typically ~10 for each genus, e.g. Pseudomonas syringae

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14
Q

How many pathovar are there per species?

A

Up to 150 for some species esp. Xanthomonas and Pseudomonas e.g. Pseudomonas syringae pv. tomato

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15
Q

How many races/strains are there by pathovar of pathogenic bacteria?

A

Typically ~10 for some pathovars
e.g. Pseudomonas syringae pv. tomato DC3000

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16
Q

What is an example of a Gram-negative necrotrophic plant pathogen?

A

Erwinia carotovora
Host range - Wide
Disease - Soft rots

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17
Q

What is an example of a Gram-negative biotrophic plant pathogen?

A

Pathogen - Host range - disease
Erwinia amylovora - Rosacaea - Fireblight
Ralstonia solanacearum - Wide - Wilts
Xanthomonas campestris pv. vesicatoria - Pepper, tomato - Leaf spot

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18
Q

What is pathogenicity?

A

The ability to infect a host

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19
Q

What is virulence?

A

The severity of the infection

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20
Q

How do you measure bacterial virulence?

A

Inoculate
Allow disease to develop
Take samples
Grind in bugger
Make seriel dilution
Plate, incubate and count colonies

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21
Q

What are the typical ways of inoculating leaf pathogens?

A

Syringe infiltration, spraying or dipping

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22
Q

What are potential methods for measuring virulence?

A

One approach is to make mutants by marker exchange to disrupt chromosomal copies of the genes.
Virulence may be assessed by measuring growth in the host.

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23
Q

What is an example of the gene and its impact on virulence?

A

avrRPM1 gene from Pseudomonas syringae pv. maculicola is required for full virulence on Arabidopsis
Bacteria with functional avrRPM1 - 10^5 to 10^6 CFU
Bacteria with non-functional avrRPM1 - 10^3 CFU
Both 5 days after innoculation

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24
Q

What is the two step process of type II secretion system?

A

1 - SecYEG or Tat system transport proteins into the periplasm
2 - Dodecameric secretin GspD transports proteins across the outer membrane

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25
Q

What are the 4 components of the type II secretion system?

A

cytoplasmic ATPase (GspE)
IM platform (four membrane proteins - GspC, GspF, GspL, GspM)
periplasmic pseudopilus (GspG)
OM complex (GspD) aka ‘secretin’

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26
Q

What does the type II secretion system (T2SS) consist of?

A

Type II Secretion System (T2SS) consists of 12-15 protein, called general secretion pathway (Gsp) proteins in enterotoxigenic Escherichia coli.

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27
Q

What are other names of the general secretion pathway called?

A

In other bacteria GspC and GspD interact to insert the substrate protein into the secretin channel, through which it is pushed in piston-like as the pseudopilus extends.

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28
Q

What is the Tat system?

A

Twin-arginine translocation system

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29
Q

What is the function difference of the SecYEG and Tat transports?

A

SecYEG transports unfolded proteins into the PM. Tat transports fully folded proteins into the periplasm

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30
Q

What are the proteins secreted by T2SS?

A

In free-living bacteria T2SS secretes enzymes that degrade compounds in the environment (including proteases, lipases, chitinases)

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31
Q

What is an example of mutating (ATPase) impacting virulence of a bacteria?

A

T2SS is required for virulence of some animal/human pathogens, e.g. Yersinia enterocolitica yts1E (ATPase-) mutants show reduced infection in mouse

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32
Q

What are examples of cell wall degrading enzymes used by necrotrophs that go through a T2SS seen in Erwinia species?

A

Pectate lyase, endo- and exo- enzymes (at least 9 enzymes)
Pectin acetylesterase
Pectin methylesterase (2)
Polygalacturonase
Cellulase (2)
Endoxylanase
Protease (at least 4)

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33
Q

What are T3SS?

A

Multi-protein nano-machine, structurally related to bacterial flagella
Transfers effector proteins from the bacterium into the host cytoplasm

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34
Q

What the components of T3SS in salmonella?

A

Hexameric ATPase (InvC)
Sorting platform (SpaO, OrgA,OrgB)
Secretin (InvG)
Needle (PrgI)

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35
Q

What molecules get secreted through the T3SS system?

A

Effectors
Translocator proteins - forms pores in cellular membranes
Class 1 and 2 chaperones which binding and folding proteins

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36
Q

What is an example of TS33 being required for virulence?

A

Shigella flexneri T3SS effector IpaB induces apoptosis in macrophages
ipaB mutant has 0% cytotoxity compared to ~70% cytotoxicity in wildtype Shigella flexneri

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37
Q

What are examples of effectors used in TS33 against plants?

A

Several Hrp proteins form a pilus through which effectors (including Avrs) are translocated

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38
Q

How have TS33 delivery been seen?

A

Seen with immuno-gold labelling of effector AvrPto

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39
Q

What types of enzymes are commonly used as effectors?

A

Proteases and Ubiquitine ligase shows that they have a function of degredation

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40
Q

What is an overview of TALE T3 effectors?

A

AvrBs3, PthA, AvrXa7 and related proteins from Xanthomonas species make up the TALE family
TALE: transcription activator-like effectors
Exhibit sequence-specific DNA binding
Act as transcriptional activators in the plant cell nucleus

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41
Q

How many repeats are there in each Tales?

A

Each 33-35 amino acid repeat binds a single nucleotide. Specificity is determined by a two amino acid sequence. HD binds C, NN binds G, NG binds T, NI binds A (H,D,N,G,I are amino acids)

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42
Q

What is an example of an effector and the gene it targets?

A

Xcv AvrBs3 targets Bs3 gene in pepper which is a resitance gene

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43
Q

What is the function of FLS2 and back protein BAK1?

A

Detects the presence of flagella
Greats a signalling cascade which triggers immunity-related transcription
PAMP-triggered immunity

44
Q

What are examples of type 3 effectors that inhibits FLS2?

A

AvrPto
AvrPtoB
AvrPphB

45
Q

What is the function of AvrPtoB?

A

AvrPtoB causes degradation of FLS2 by tagging it with ubiquitine

46
Q

What causes Crown gall?

A

Agrobacterium tumefaciens

47
Q

Why does the Crown gall occur?

A

Type 4 secretion system inserts T-DNA found in the Ti Plasmid into wounded cells and moves into nucleus, integrating randomly into a nucleus. This induces uncontrolled cell division –> crown gall

48
Q

Outside of uncontrolled cell division what else does Agrobacterium do to a plant cell?

A

Produce unusual amino acids - Octopine and Nopaline (Opines) which is an energy source only the bacteria can use

49
Q

What is the structure of the T-DNA?

A

LB: left border repeat; RB right border repeat: (C/T)GGCAGGATATA(T/A)C(A/(A/G)TTGTAA(A/T)T
aux: auxin biosynthesis genes
cyt: isopentyl transferase (involved in cytokinin biosynthesis)
tm1: tumour size regulation
ocs: octopine synthase

50
Q

What is the mechanism for the detecting and insertion of T-DNA into a cell?

A

VirA and VirG sense chemicals from wounded plant cell
Expression of virulence proteins
ss-T-DNA forms a complex VirD2 which is transported through a pore by a VirB protein into plant cell
The T-DNA integrates into the host DNA

51
Q

What are exopolysaccharides?

A

Extracellular polysaccharide (EPS)
Form a capsule around most bacterial cells
Used to hides cell surface molecules

52
Q

How did they test for exopolysaccharide required for virulence?

A

Exopolysaccharide required for virulence of some animal pathogens, e.g. Klebsiella pneumoniae K antigen (capsular polysaccharide):
Bacteria mixed with polymorphonuclear leukocytes (PMNL)
Bacteria which lacked capsular polysaccharide were quickily killed off (within 30 mins), wildtype increased in number over 90 mins

53
Q

How did they test for exopolysaccharide required for virulence in plant pathogens?

A

Exopolysaccharide (= extracellular polysaccharide, EPS) is required for virulence of some plant pathogens, e.g.
Xanthomonas campestris pv. campestris (Xcc):
Capsule minus bacteria died off after 3 days compared to capsule plus which increased in number over 3 days

54
Q

What is a function of EPS in Xanthomonas capestris?

A

EPS suppresses Ca2+ signalling and thus the expression of plant defence genes like PR1.

55
Q

What is the virulence function of animal pathogen, cholera toxin?

A

The B subunit of the toxin binds to GM1 gangliosides on the surface of gut epithelial cells.
The A subunit is internalised, binds to a G protein and activates adenylate cyclase.
Converts AMP to cAMP
Increased intracellular cyclic AMP levels over activate protein kinase A.
PKA phosphorylates cystic fibrosis transmembrane conductance regulator (CFTR) causing loss of Cl-, Na+, K+, and HCO3− and H2O from the cell

56
Q

What are examples of Toxins from necrotic plant pathogenic bacteria?

A

Toxin, Pathovar, structure, target/action, symptoms
Syringomycin, syringae, Cyclic lipodepsipeptide, Pores in PM, Necrosis
Syringopeptin, syringae, Cyclic lipodepsipeptide, Pores in PM, Necrosi

57
Q

What are examples of Toxins from chlorosis plant pathogenic bacteria?

A

Toxin, Pathovar, structure, target/action, symptoms
Tagetitoxin, tagetis, Hemithioketal, Inhibitor of chloroplastic RNA Pol, Chlorosis
Tabtoxin, tabaci and others, B-lactam dipeptide, Inhibitor of glutamine synthase, chorosis

58
Q

What is the virulence function of coronatine?

A

Mimics of the plant hormone jasmonic acid
Suppresses stomatal closure (a consequence of PTI), by forcing them open to allow for bacteria to enter leaf

59
Q

What is the structure of coronatine?

A

Polyketide and cyclised amino acid

60
Q

What bacteria can use coronatine?

A

Glycinea impacts soybeans and others

61
Q

What is mycetism?

A

Poisoning by mushroom

62
Q

What is mycotoxicoses?

A

Poisoning by fungi (on other food stuffs)

63
Q

What is an overview of Mycetism?

A

Mycetism, results from the ingestion of a mushroom that contains preformed toxic metabolites
Examples include ibotenic acid from Amanita muscaria and amatoxins from Amanita phalloides (Death cap)

64
Q

What is an overview of Amanita muscaria and its toxins?

A

A. muscaria produces ibotenic acid. This is an unstable thermolabile amino acid which can be decarboxylated to muscimol.
ibotenic acid activates the glutamate receptor
Muscimol activates the gaba receptor
Both affect neurotransmission signalling

65
Q

What are the symptoms of Amanita muscaria and its toxins?

A

Symptoms similar to acute alcohol intoxication. Victim staggers, becomes delirious and loses consciousness.
Death results in only 1% of cases, but when it does occur it is from respiratory failure.
Muscimol has insecticidal properties; the common name for this fungus is the fly agaric because it was used to kill flies.

66
Q

What is Amanita phalloides and its toxins?

A

Amanita phalloides produces amatoxins
There are at least 9 known amatoxins
Among the most deadly poisons in nature
α-Amanitin is a potent inhibitor of RNA polymerase II.
It causes kidney failure, liver failure, coma and death.
One mushroom or less may be fatal.

67
Q

What is an overview of the treatment of mushroom poisoning?

A

Treatment needs to be quick, even before the mushroom has been identified.
Evacuation of the gastrointestinal tract is performed by inducing vomiting, stomach lavage/ and or enemas.
Carbon may be given to absorb toxins.
Kidneys and liver function are monitored

68
Q

What is an overview of mycotoxicoses?

A

This is the intoxication following ingestion of fungi producing toxins (mycotoxins) when growing on foodstuffs.
Associated with a specific food.
Seasonal, non-transmissible and unresponsive to drug and antibiotic treatment.
>100 species of filamentous fungi are known to produce mycotoxins

69
Q

What is an example of mycotoxicoses?

A

In 1960 more than 100,000 Turkeys died within a few months in a 100 mile radius around London. The new disease was called Turkey X
It was caused by aflatoxins produced by Aspergillus flavus, present in the peanut meal used in turkey feed.
Aflatoxins are the most dangerous mycotoxins, mainly effecting animals

70
Q

Where have Aflatoxins been found?

A

The feed had been prepared with contaminated peanut meal from which a mycotoxin was extracted.
Most reports of aflatoxins refer to stored foodstuffs such as mouldy peanuts, cottonseed, coconuts and corn.
They have been found in dried fermented fish and identified in the milk of animals eating contaminated feed.

71
Q

What is an overview of Aflatoxins?

A

There are four principle aflatoxins B1, B2 G1 and G2. They are produced by a range of fungi including Aspergillus flavus, Aspergillus parasiticus, Aspergillus ostianus, plus other species of Aspergillus and Penicillium.

72
Q

How does Aflatoxins effect people?

A

Children are more sensitive than adults and development may be impaired.
The characteristic effect of high-level aflatoxin poisoning is liver damage – necrosis, cirrhosis and carcinoma.
Aflatoxins intercalate into DNA, causing DNA damage (alkylation) and chronic exposure increases risk of cancer, especially of the liver and gall bladder.

73
Q

What is an overview of pathogenic fungi?

A

Approximately 300 species of fungi cause human disease.
Most are soil saprophytes.
They are facultative parasites, infecting hosts when the opportunity arises.
Combined, they kill more than either malaria or TB

74
Q

What are examples of fungal

A

Disease, location, estimated life threatening infections, mortality rates
Cryptococcosis (Cryptococcus neoformans), worldwide, >1,000,000, 20-70%, opportunistix invasive mycoses
Penicillious (Penicillium marneffei), Southeast Asia, >8,000, 2-75%, endemic dimorphic mycoses

75
Q

How have fungi diseases been recognised as a problem?

A

In 2022 the WHO released the first list of fungal pathogens of particular concern eg critical group includes

76
Q

How have fungi diseases been recognised as a problem?

A

In 2022 the WHO released the first list of fungal pathogens of particular concern eg critical group includes Cryptococcus neoformans

77
Q

What is an overview of dimorphism in fungi?

A

Dimorphism, is where fungi can exist in tissues as either single cells (yeast) or a mycelial form.
Dimorphic fungi which are pathogenic in animals are more likely to exist outside the host in a mycelial form.
Conditions inside the host cause the conversion of the mycelium to the yeast form. eg Elevated temperatures, High CO2 and low O2, Levels of nutrients

78
Q

What are the types of fungi infections?

A

Superficial
Cutaneous
Subcutaneous
Systemic

79
Q

What is cutaneous infection?

A

Fungi which cause cutaneous damage are called dermatophytes

80
Q

What are examples of a cutaneous infection?

A

Tinea corporis, ringworm. Lesions are red, raised, itchy, and often circular (hence ‘ringworm’).
Tinea pedis, athletes foot. The most common type. Typically found in the toe webbing.

81
Q

What causes most cases of tineo pedis?

A

Trichophyton rubrum,
Trichophyton mentagrophytes
Epidermophytom floccosum

82
Q

What is an overview of subcutaneous infection?

A

The fungi gain entry to the subcutaneous tissue by wounds, typically scratches caused by thorns.
These diseases tend to be prevalent in rural and tropical regions.
Localised and form abscesses called mycetomas.
Examples include Sporothrix schenckii, which occurs in farmers and gardeners
Tissue damage is variable, but can be very disfiguring.

83
Q

What is an example of subcutaneous infection?

A

Sporothrix schenckii
There have also been outbreaks among building workers exposed to heavily infected timbers.

84
Q

What is an example of systemic infection?

A

The fungus spreads from one internal organ to another often via the bloodstream
Frequently systemic mycosis originates in the lungs by the inhalation of spores. If a vital organ such as the brain becomes involved it frequently leads to death.

85
Q

How are systemic infections diagnosed?

A

This is based on clinical observation and laboratory investigation.
Specimens should be taken from as many sites as possible, including urine and sputum. A lung biopsy may need to be performed for pulmonary mycotic infections.

86
Q

How easy is it to grow pathogenic fungi on artificial culture?

A

Most pathogenic fungi are easy to grow in artificial culture. The agar medium most commonly used is Sabouraud’s glucose agar and 4% malt extract agar

87
Q

What types of agar can be used to grow pathogenic fungi?

A

For dimorphic fungi agar such as brain-heart infusion agar and blood agar are effective for yeast phase growth.

88
Q

What is India Ink?

A

A variety of treatment and staining techniques can be used.
Add a drop of CSF to a drop of India Ink

89
Q

What would Cryptococcus neoformans look like using India Ink?

A

Capsule is seen as a clear halo around the yeast
Round yeast with mucopolysaccharide capsule

90
Q

What is an overview of detection using antibodies?

A

Serological (antibody-based) methods are used to detect fungal antigens
Types of test include:
immunodiffusion
enzyme-linked immunosorbent assay (ELISA)

91
Q

How can PCR be used to identify fungi pathogen?

A

Organism-specific primers are employed.
Care must be taken in the interpretation of results, as the sensitivity of PCR may lead to false positive results.

92
Q

What is an overview of Aspergillus fumigatus?

A

Causes aspergillosis
Very effective pathogen in immunocompromised patients and patients with lung disease (including cystic fibrosis, tuberculosis, asthma).

93
Q

What is an overview of Aspergillus fumigatus infection?

A

Initial siter of infection is the lungs.
A type of aspergillosis in which a ‘fungus ball’ (compact ball of mycelia) may form in an existing cavity in the lung, particularly old tuberculosis lesions
May cause no symptoms or may cause bleeding (which can be severe or even fatal).
Systemic infection occurs via the bloodstream – the fungus can disseminate to the brain, liver and kidneys. Liver and kidney failure can be rapid and may be fatal.

94
Q

What is an overview of Candida albicans?

A

Most common fungal infection
Most Candida species are commensals – part of the human GI and skin microbiomes in healthy people
There is a concerning emergence of multi-drug resistant Candida strains
Oral thrush is common in new-born babies, resulting from infection during birth

95
Q

What can cause ans Candida albicans infection?

A

Causes candidiasis when the immune system is weakened or the equilibrium of the microbiome is disrupted (e.g. by antibiotics)

96
Q

What is an overview of Candida albicans infection?

A

Survives phagocytosis by forming hyphae and puncturing the cell – also used to disrupt epithelial cells. When get to blood stream in gut the tranform into yeast form and spread arounf the body
It is a very effective pathogen in immunocompromised patients
Easily adapts to different nutrient availabilities
It forms biofilms, particularly on implanted foreign surfaces, helping it survive internally

97
Q

What is an overview of Cryptococcus neoformans?

A

Cryptococcus species are yeasts that cause cryptococcosis
HIV/AIDS sufferers are particularly at risk
The fungus can persist for years in the lung, reactivating only upon weakened immune surveillance

98
Q

How does a Cryptococcus neoformans infection occur?

A

Infection occurs from inhalation of particles from soil or bird droppings – colonises the lungs
Easily disseminates via the blood to the central nervous system (cryptococcal meningitis), and also encephalitis

99
Q

What are examples of diseases caused by Candida albicans?

A

Brain abcesses
Focal absesses (lung and liver)
Endocarditis
Retinitis

100
Q

How does the polysaccharide capsule play a large role in Cryptococcus neoformans infection?

A

It protects from phagocytosis and can even trigger macrophage death
This is suggested to be the way Cryptococcus crosses the blood-brain barrier, although the actual mechanism is not yet confirmed

101
Q

What is an overview of the treatment of fungal infections?

A

The treatment is often difficult, since both the host and the parasite are eukaryotic.
Hard to attack the fungus without damaging the host cells.
There are very few antimycotic compounds which can be used in human treatment.

102
Q

What are an overview of Polyene macrolide?

A

Polyene macrolide antibiotics are produced by Streptomyces spp and include amphotericin B, nystatin and pimaricin. They have in common a large ring structure.
Bind specifically to the sterol in the fungal membrane (ergosterol). They increase the permeability of the plasma membrane –> leakage of the cellular constituents, lysis and cell death

103
Q

What is an overview of Amphotericin B?

A

Amphotericin B is administered intravenously in a bile salt suspension and is active against most fungi.
It is used to treat systemic infections and severe superficial mycosis. At low concentrations this antibiotic can also boost the immune system. Unfortunately in higher concentrations it is toxic to the kidneys

104
Q

What is an overview of Nystatin?

A

Nystatin is too toxic to be administered intravenously, and cannot be absorbed by the digestive tract it is therefore along with pimericin used topically to treat superficial infections such as thrush (Candida albicans).

105
Q

What is an overview of newer antifungal drugs?

A

Of the newer anitifungal drugs the imidazoles and the triazoles are the most effective. They have a broad spectrum and effect the ergosterol in the plasma membrane.

106
Q

What is an overview of Ketoconazole?

A

Ketoconazole can be absorbed through the gastrointestinal tract and inhibits the synthesis of ergosterol. Another advantage is that it is non-toxic to human cells

107
Q

What antifungal is used when faced life-threatening mycosis?

A

Despite these new drugs Amphotericin B still remains the drug of choice when faced with life-threatening mycosis.

Sem 2: Microbiology (12 decks)

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