MYCORRHIZA

• A mycorrhiza (pl mycorrhizae, mycorrhizas) is a symbiotic association

between a fungus & the roots of a vascular plant.

• The fungus colonizes the host plants' roots, either intracellularly as in

arbuscular mycorrhizal fungi (AMF), or extracellularly as in
ectomycorrhizal fungi.

• They are an important component of soil life & soil chemistry.

• Mycorrhizae form a mutualistic relationship with the roots of most

plant species.
MYCORRHIZA
• Mycorrhiza = fungus root.

• They invade the roots & connect

them, one to the other.

• Then send out their hyphae, as

much as 200 times further into the
soil than the roots they colonize.

• Water & nutrients e.g. P -

transmits back to the roots.

• The plant provides glucose to the

fungus.
Benefits of Mycorrhizae
a. Sugar-Water and mineral exchange

• This mutualistic association provides the fungus with relatively

constant & direct access to carbohydrates e.g. glucose & sucrose
supplied by the plant.

• The carbohydrates are translocated from their source (usually leaves)

to root tissue & on to fungal partners.

• In return, the plant gains the benefits of the mycelium's higher

absorptive capacity for water & mineral nutrients.
a. Sugar-Water and mineral exchange…
• Comparatively large surface area of mycelium:root ratio.

• There is improved plant mineral absorption capabilities - 10 to 1000

times.

• Mycorrhizal fungi release powerful chemicals into the soil that

dissolve hard to capture nutrients such as phosphorous, iron & other
"tightly bound" soil nutrients.
a. Sugar-Water and mineral exchange…

• Plant roots alone may be incapable of taking up phosphate ions

that are demineralized e.g. in soils with a basic pH.

• Mycelium of the mycorrhizal fungus can, access these

phosphorus sources, and make them available to the plants they
colonize.

• Mechanisms of increased absorption are both physical &

chemical.

• Mycorrhizal mycelia are much smaller in diameter than the

smallest root.
a. Sugar-Water and mineral exchange…

• They can explore a greater volume of soil, providing a larger

surface area for absorption.

• Cell membrane chemistry of fungi is different from that of

plants.

• Mycorrhizae are especially beneficial for the plant partner in

nutrient-poor soils.
b. Disease and drought resistance
• Mycorrhizal plants are often more resistant to diseases, such as
those caused by microbial soil-borne pathogens.

• Mycorrhizal roots have a mantle (a tight, interwoven socklike

covering of dense filaments) that acts as a physical barrier
against the invasion of root diseases.

• Mycorrhizal fungi attack pathogens entering the root zone.

• E.g. excretions of specific antibiotics produced by mycorrhizal

fungi immobilize & kill disease organisms.
b. Disease and drought resistance…
• Antagonistic function to soilborne disease pathogen.

• Suppression of pathogen growth.

• Induced resistance or systematic resistance in plant.

• Increased resistance or tolerance of mycorrhizal plants to

soilborne diseases.

• Also more resistant to the effects of drought.

c. Colonization of barren soil
• Plants grown in sterile soils & growth media often perform
poorly without the addition of spores or hyphae of mycorrhizal
fungi to colonize plant roots & aid in the uptake of soil mineral
nutrients.

• The absence of mycorrhizal fungi can also slow plant growth in

early succession or on degraded landscapes.

• The introduction of alien mycorrhizal plants to nutrient-deficient

ecosystems puts indigenous non-mycorrhizal plants at a
competitive disadvantage.
d. Resistance to toxicity

• Fungi have a protective role for plants rooted in soils with high
metal concentrations e.g. acidic & contaminated soils.

• Strains of Suillus luteus have varying tolerance of zinc.

• Zinc-tolerant strains of Suillus bovinus confer resistance to

plants of Pinus sylvestris.

• Mechanism - binding of the metal to extramatricial mycelium of

the fungus, without affecting exchange of beneficial
substances.
e. Improved soil structure
 Induced Resistance in plants

• Induced resistance is resistance that is dependent on factors

present only after the host is challenged by the pathogen.

• Plants respond to attacks by insects & diseases by mobilizing

compounds that inhibit plant diseases or reduce feeding by
insects.

• Often, plants in which resistance is induced by one pathogen or

insect will also be resistant to some other pathogens or insects,
but not necessarily the entire spectrum of potential pests.

• It is also possible that mobilizing resistance to one pathogen

could increase susceptibility to another pathogen.
 Systemic resistance

• Refers to resistance that occurs at sites in the host distant

from the point of initial interaction with a potential pathogen.

• It is generally the result of necrosis brought about by the

inducing inoculation, and is usually detected by challenge
inoculation at a different time and location on the plant.

• Note: Systemic resistance is a form of induced resistance which

can be localized or systemic.
 Characteristics of induced resistance (localized or systemic)

(a) They are time-dependent - resistance is established only after

certain metabolic changes occur in the host during a specific
interval following the inducing inoculation.

(b) They are temperature dependent - systemic resistance is

more sensitive to this factor than localized resistance.

(c) They are affected by light to different degrees - but light is

essential only in certain cases of systemic resistance.
 Characteristics of induced resistance (localized or systemic)

(d) In both cases, resistance is persistent once the machinery for

induction of the plant response has been turned on.

(e) Both types are nonspecific in terms of the organisms that

induce the response and of the range of pathogens against
which the plant is protected.
Types of mycorrhiza
1. Endomycorrhiza - Their hyphae penetrate the cell wall &
invaginate the cell membrane.

• Are variable & have been further classified as arbuscular,

ericoid, arbutoid, monotropoid & orchid mycorrhizae.

i. Arbuscular mycorrhizas - mycorrhizas whose hyphae enter

into the plant cells, producing structures that are either
balloon-like (vesicles) or dichotomously-branching
invaginations (arbuscules).
i. Arbuscular mycorrhizas…

• The fungal hyphae do not in fact penetrate the protoplast (i.e.

the interior of the cell), but invaginate the cell membrane.

• The structure of the arbuscules greatly increases the contact

surface area between the hypha & the cell cytoplasm to
facilitate the transfer of nutrients between them.

• Formed only by fungi in the division Glomeromycota.

i. Arbuscular mycorrhizas…

• Are found in 85% of all plant families; occur in many crop spp.

• Their hyphae produce the glycoprotein glomalin - one of the

major stores of carbon in the soil.

• They have been asexual for millions of years & individuals can
contain many genetically different nuclei - heterokaryosis
ii. Ectomycorrhiza

• Hyphae of ectomycorrhizal fungi do not penetrate individual cells

within the root.

• Ectomycorrhizas, are formed between the roots of around 10% of

plant families, mostly woody plants e.g. eucalyptus, oak, pine & rose
families and fungi belonging to the Basidiomycota, Ascomycota, and
Zygomycota.

• Ectomycorrhizas consist of a hyphal sheath, or mantle, covering the

root tip and a net of hyphae surrounding the plant cells within the
root cortex.
iii. Ericoid mycorrhiza
• Have a simple intraradical (grow in cells) phase, consisting of dense
coils of hyphae in the outermost layer of root cells.

• There is no periradical phase & the extraradical phase consists of

sparse hyphae that don't extend far into the surrounding soil.

• They might form sporocarps but their reproductive biology is little

understood.

• Have considerable saprotrophic capabilities, which would enable

plants to receive nutrients from not-yet-decomposed materials via the
decomposing actions of their ericoid partners.
Microorganisms as disease causing agents

• Pathogen - An infectious disease-causing agent.

• A pathogen is a living micro-organism (bacterial, fungal, viral or

parasitic) that can cause disease to its host.

• The major infectious disease causing agents are bacteria, fungi,

viruses, rickettsia & parasites.
a. BACTERIA

• Are single-celled (unicellular) living organisms with tiny flagella

(a tail like appendage that they use to swim).

• Invisible to the naked eye & can only be seen with a microscope.

• Generally have cell walls & may appear in one of several shapes.
a. BACTERIA

• Bacillus (rodlike), coccus (spherical or ovoid), spiral

(corkscrew/curved).

• Reproduce by binary fission - dividing themselves into equal

cells (daughter cells).

• Live in & reproduce in warm, moist environments in the body &

other areas where they grow quickly, causing an infection.
a. BACTERIA
• Many bacterial diseases generally produce inflammation,
swelling & pain from nerve irritation & fever caused by
increased body temperature by the body fighting the disease.

• Bacterial infections can usually be treated with antibiotics.

• Some examples of bacterial disease are:

Cholera - caused by the ingestion of contaminated water &

food containing the Vibrio cholerae.
a. BACTERIA
• Syphilis - spread by sexual intercourse of an infected person to
the sexual partner or from an infected mother to her fetus
containing the disease agent Treponema pallidium.

• Other bacterial diseases:

• Dysentery, Leprosy, Plague, Scarlet Fever etc.

• Important bacterial genera in plant diseases include:

Xanthomonas, Pseudononas and Erwinia.
 Plague Leprosy

Yersinia Pestis
Mycobacterium lepromatosis

Banana Bacterial Bean Halo Blight Soft Rot of Potato

Wilt

Xanthomonas campestris Pseudononas syringae pv Erwinia carotovora

pv. musacearum phaseolicola subsp. atroseptica
b. FUNGI
 Rice blast disease Dutch elm disease

Fusarium Head Blight Maize Head Smut Tomato early blight

 Apple scab Coffee berry disease

Venturia inaequalis Colletotrichum kahawae

b. FUNGI

• Can cause serious diseases in humans.

• May be fatal if untreated e.g. Aspergilloses, candidoses,

coccidioidomycosis, cryptococcosis, histoplasmosis, mycetomas.

• Persons with immuno-deficiencies are particularly susceptible to

diseases by genera such as:

- Aspergillus, Candida, Cryptoccocus, Histoplasma & Pneumocystis.

b. FUNGI

• Other fungi can attack eyes, nails, hair, and especially skin.

• Dermatophytic & keratinophilic fungi cause local infections such

as ringworm and athlete’s foot.

• Fungal spores are also a cause of allergies.

Candida albicans
Microsporia & Trichophytosis capitis
Athlete's foot Ringworm
c. VIRUSES

• Can only be seen using an electron microscope.

• They are acellular (not cellular).

• Viruses the smallest life forms existing.

• Nucleic acid - either DNA or RNA - contains the virus's genes.

c. VIRUSES…
• Covered by a protein coat; sometimes encased by an envelop
(lipid or fat membrane).

• They can be rod-shaped, sphere-shaped, or multisided.

• Cannot self-reproduction outside a host cell.

• Depend on their host cellular system to reproduce.

c. VIRUSES…

• Unlike parasites, they are not considered to be truly alive.

• Virus DNA enters cells & uses its DNA to make copy of itself,
similar to what the ordinary cell would do.

• Host body cell is tricked into making many copies of the virus
inside the host cell.

• The virus Kills the cell where the viruses then enter (infecting)
other cells to repeat the process.
c. VIRUSES

• Inside the body, viruses produce toxins (poisons) that can cause
rashes, aches and fevers.

• A Virus is very difficult to kill & cannot be killed with antibiotics.

• Some examples of viral diseases in humans are: Aids, Smallpox

Influenza, Yellow Fever etc.

• Potato virus Y, potato virus X and Tobacco mosaic virus are

economically import plant diseases.
Small pox Tobacco mosaic virus
e. PARASITES
• Organisms whose survival depends on other organisms (the
host) to feed, grow & live.

• Live in or on the living tissue of a host organism causing disease

generally without killing the host.

• Parasites can be either single-celled protozoa or multi-celled

parasites (e.g. worms, flukes, and insects).

• The infection from a parasite is often transmitted thro’ contact

with an intermediate vector or from the result of direct contact
with the parasite.
HOST PATHOGEN RELATIONSHIPS…
 HOST PATHOGEN RELATIONSHIPS

• The pathogen must be able to attack the host.

• The environment must favor the development of the pathogen.

• Disease is most severe when the environment favors pathogen

growth & development over the host.
THE DISEASE TRIANGLE: a plant pathological paradigm
 THE DISEASE TRIANGLE: a plant pathological paradigm

• It is a simple model that plant pathologists use to represent the

key factors that lead to the development of disease & in an
indirect way, clues about how to manage a particular disease.

• The existence of a disease caused by a biotic agent requires the

interaction of a susceptible host, a virulent pathogen & an
environment favorable for disease development.
 THE DISEASE TRIANGLE: a plant pathological paradigm

• Plant disease is prevented upon elimination of any one of these

three causal components.

• The interactions of the 3 components of disease have often

been visualized as triangle generally referred to as the
“DISEASE TRIANGLE”.
 HOST PATHOGEN RELATIONSHIPS
• Each side of the triangle represents one of the 3 components.

• The length of each side is proportional to the sum total of the

characteristics of each component that favor disease.

• e.g. if the plants are resistant, the wrong age, or widely spaced,
the host side - and the amount of disease - would be small or
zero.

• If any of the three components is zero, there can be no disease.

 Plant Disease Epidemiology (Development)
• An epidemic - when a pathogen spreads to & affects many
individuals within a population over a relatively short time.

• Epidemiology - Study of epidemics & the factors that influence

them.

• A plant disease develops when the host plant is susceptible;

there is a virulent pathogen & the environmental conditions
optimum for pathogen growth, reproduction & spread.

• The interaction of the 3 components can be expanded to

include time & humans.
 1. HOST FACTORS
i) Levels of genetic resistance or susceptibility of the host
• Host plants carrying race-specific (vertical) resistance do not
allow a pathogen to become established in them.

• Host plants carrying partial (Horizontal) resistance (i.e. will

probably become infected but the rate at which the disease &
the epidemic will develop depends on the level of resistance &
the environmental conditions in the presence of a virulent
pathogen) will favour the development of disease epidemics.
 ii) Degree of genetic uniformity of host plants
• When genetically uniform host plants are grown over large
areas there is a greater likelihood that a new pathogen race will
appear that can attack their genome & result in an epidermic.

• e.g. southern corn leaf bight of corn carrying Texas male sterile
cytoplasm.

• Epidemics generally occur in vegetatively propagated crops &

the lowest rates in cross pollinated crop.
 ii) Age
• Pythium damping-off and downy mildew diseases occur at the
early stages during growth.

• Host plants become resistant during the adult period.

• Stem rusts attack crops at the blooming stage, not before or

after.

• Botrytis attacks hosts at the adult period when host plants

become senescing.
 PATHOGEN FACTORS

i. Levels of virulence
Virulent pathogens capable rapidly produce large amounts of
inoculum & thereby disease than pathogens of lesser virulence.

ii) Amount of inoculum

The greater the no. of pathogen propagules (bacteria, fungal
spores and sclerotia, nematode eggs, virus infected plants, etc)
within or near fields of host plants, the more inoculum reaches
the hosts & at an earlier time, thereby increasing the chances of
an epidemic greatly.
 iii) Type of pathogen reproduction

a) TYPE I - Some plant pathogens have short reproduction cycles

& therefore are polycyclic i.e. they can produce many
generations in a single growing season e.g. rusts, mildews, leaf
spots.
- They are responsible for most of the sudden catastrophic
plant disease epidemics.

Wheat leaf rust (Puccinia spp.) Powdery mildew of cucurbits

(Erysiphe & Sphaerotheca spp.)
 b) TYPE II
a) Some soil fungi e.g. Fusarium, Verticillium & most nematodes
usually have one to a few (up to four) reproductive cycles per
growing season.
Fusarium and Verticillium spp. reproduce inside the plant.
 Nematodes have one or a few reproductive cycles.
 Rhizoctonia and Sclerotium do not produce spores.

• Such pathogens with smaller no. of offspring & especially the

conditions of their dispersal limit their potential to cause sudden
& widespread epidemics in a single season.
 c) TYPE III - Monocyclic pathogens
• Several pathogens e.g. smuts & bunts require an entire year to
complete a life cycle.

• Can only cause one series of infections per year.

• In monocyclic diseases, the inoculum builds up from one year to

the next & an epidemic usually develops over several years.

• Such pathogens produce inoculum each year & cause a series of

infections that lead to long-term epidemics e.g. Cedar-apple
rust.
 iv) Ecology of the pathogen
• Most fungi & all parasitic higher plants, produce their inoculum
on the surface of the aerial parts of the host.

• Spores & seeds can be dispersed with ease over a range of

distances & can cause widespread epidemics.

• Vascular fungi, bacteria, viruses, protozoa & mollicutes

reproduce inside the plant.

• Spread of the pathogen is rare or impossible without the help of

vectors.
 iv) Ecology of the pathogen
• Such pathogens can only cause epidemics when vectors are
plentiful & active.

• Soilborne fungi, bacteria & nematodes produce their inoculum

on infected plant parts or the soil.

• The inoculum disperses slowly & presents little danger for

sudden or widespread epidemics.
 iv) Mode of Spread

a) Airborne
• Spores of plant pathogenic fungi causing rusts, mildews & leaf
spots are released into the air.

• Spores are dispersed by air breezes or strong winds over

distances from a few centimeters up to several kilometers.

• Such pathogens are responsible for the most frequent & most
widespread epidemics.
 b) Vector dependency
• Most viruses are transmitted by aphids, whiteflies & other
insects.

• Mollicutes & fastidious bacteria are transmitted by leafhoppers,

plant hoppers & psyllids.

• Some fungi, bacteria & nematodes are disseminated primarily by

beetles.
 c) Wind-blown rain
• Such pathogens which are blown by wind or water splash are
almost annually responsible for severe but localized epidemics
within a field, a country or a valley.

d) Seedborne
• Such pathogens are often placed in the midst of susceptible
plants, but their ability to cause epidemics depends on the
effectiveness of their subsequent transmission to new plants.
 e) Soilborne
• Pathogens present in & spreading thro’ the soil, because of the
physical restrictions imposed by the soil, are generally unable to
cause sudden or widespread epidemics but often cause local,
slow-spreading diseases of considerable severity.
 3. ENVIRONMENTAL FACTORS
i) Moisture
• Abundant, prolonged or repeated high moisture, is the
dominant factor in the development of most epidemics of
diseases caused by fungi, bacteria & nematodes.

• Moisture promotes new succulent & susceptible growth.

• Increases sporulation of fungi & multiplication of bacteria.

 3. ENVIRONMENTAL FACTORS

i) Moisture…
• Facilitates spore release & oozing of bacteria to the host
surface.

• Enables spores to germinate.

• Enables zoospores, bacteria & nematodes to move.

 ii) Temperature

• Epidemics are sometimes favored by temp. higher or lower

than the optimum for the plant - reduce plant's level of partial
resistance.

• At certain levels, temp. may reduce or eliminate the race-

specific resistance of host plants.

• Plants growing at such temp. become "stressed" & predisposed

to disease, provided the pathogen remains vigorous.
 ii) Temperature
• Low temp. reduces the amount of inoculum of oomycete fungi,
bacteria & nematodes that survives cold winters.

• High temp. reduces the inocuIum of viruses & mollicutes that

survive hot temperatures.
ii) Temperature
 ii) Temperature…
• Low temp. reduces the no. of vectors that survive cold temps.

• Low temp. during growing season reduces the activity of vectors.

• Favourable temp. - a polycyclic pathogen can complete an

infection cycle within a very short time (usually days).

• Polycyclic pathogens can produce many infection cycles within a

growing season.
 4) EFFECT OF HUMAN CULTURAL ACTIVITIES
a) Site selection and preparation
Low-lying & poorly drained and aerated sites if near other
infected fields favour appearance & development of epidemics.

b) Selection of propagative material

• Infected propagative material increases the amount of initial
inoculum within the crop & greatly increases devt of epidemics.

• Use of pathogen free or treated propagative material reduces

the chances of epidemics.
c) Cultural practices
d) Disease control measures
 e) Introduction of new pathogens
• These pathogens are introduced thro’ human activity &
movement.

• Such pathogens may result in epidemics since they lack

natural enemies.
CONTROL OF MICRORGANISMS BY PHYSICAL & CHEMICAL AGENTS

Sterilization - A treatment that kills or removes all living cells,

including viruses & spores, from a substance or object.
o The means used to sterilize materials is called a sterilant.
oCould be a chemical or physical agent e.g. ethylene gas or high
temp.

Disinfection - A treatment that reduces the total No. of microbes on

an object or surface, but does not necessarily remove or kill all of
the microbes.
CONTROL OF MICRORGANISMS BY PHYSICAL & CHEMICAL AGENTS
Antisepsis - the procedure which inhibits the pathogenic effects of
MOs. or kill MOs. in their vegetative state on living tissue or skin.
o The chemical agent used is called an antiseptic.

Sanitation - Reduction of the microbial population to levels

considered safe by public health standards.
CONTROL OF MICRORGANISMS BY PHYSICAL & CHEMICAL AGENTS
-cidal - A suffix meaning that “the agent kills.”
e.g. a bacteriocidal agent kills bacteria.

-static - A suffix that means “the agent inhibits growth.”

e.g. a fungistatic agent inhibits the growth of fungi, but
doesn’t necessarily kill it.
 Pattern of Microbial Death

• Mos. are not killed instantly when exposed to a lethal agent.

• Population death decreases by a constant fraction at constant

intervals (exponential killing).

• A microorganism is considered dead when it is unable to grow

in conditions that would normally support its growth.
Conditions influencing the effectiveness of an antimicrobial agent

i.Population size - larger populations take longer to kill than

smaller populations.

ii. Population composition - microorganisms differ markedly in

their sensitivity to various agents.
• Endospores are more resistant than the vegetative form.
• Younger cells are destroyed quicker than older cells.
• Mycobacterium tuberculosis is more resistant than most
bacteria.
Conditions influencing the effectiveness of an antimicrobial agent

iii. Concentration of the antimicrobial agent - higher

concentrations are generally more efficient, but the relationship
is not linear.

• Most often, the more concentrated a chemical agent, the more

rapidly microbes are destroyed.

• Sometimes, the agent is more effective at lower concentrations:

70% ethanol is more effective than 95% ethanol, because its
activity is more enhanced by the presence of water.
Conditions influencing the effectiveness of an antimicrobial agent

IV. Duration of exposure - The longer the population is exposed

to the antimicrobial agent, the more organisms are killed.
• To achieve sterilization, exposure should reduce population
to 10 -6 or less.

v. Temperature - An increase in temp. at which the chemical acts

often enhances its activity.
• Higher temperatures will often (but not always) increase the
effectiveness of killing.
Conditions influencing the effectiveness of an antimicrobial agent

vi. Local environment - environmental factors e.g. pH, viscosity, &

concentration of OM can profoundly influence the effectiveness
of a particular antimicrobial agent.

• The environment will either protect or aid in destruction of a

pathogen.

• OM can protect microbes against heating & chemical

disinfectants.

Biofilms – OM in a surface biofilm will protect the mos.

Biofilm and mos are usually hard to remove.

Conditions influencing the effectiveness of an antimicrobial agent

• Presence of too much OM can increase the risk of pathogens

by protecting the pathogen.

• Drinking Water - the more OM, the more chlorine needed.

• Syringes and medical & dental equipment - Should be cleaned

then sterilized.
Physical Methods in Control of microrganisms
A. Heat
i. Moist heat
• Boiling water is effective against vegetative cells & spores.

• Autoclaving (steam under pressure) is effective against

vegetative cells and most bacterial endospores.

• Mechanism of killing is a combinantion of protein/nucleic acid

denaturation & membrane disruption.

• Effectiveness is heavily dependent on type of cells present &

environmental conditions (type of medium or substrate).
i. Moist heat …
Methods of moist heat:
• Boiling at 100°C
Effective against most vegetative cells
Ineffective against spores
Unsuitable for heat sensitive chemicals & many foods.

• Autoclaving/pressure canning
Temp. above 100°C achieved by steam pressure
Most procedures use 121.1 °C
Achieved at approx. 15 psi pressure
15 - 30 min autoclave time to ensure sterilization.
i. Moist heat …
• Bacterial spores are more difficult to kill than vegetative cells.

• Sterilization in autoclave in biomedical or clinical lab must be

periodically validated by testing with spores of Clostridium or
Bacillus stearothermophilus.
 Pasteurization
• A process involving brief exposure to temp. below the boiling
point of water.

• It reduces the total microbial population.

• This method is often used for heat-sensitive materials.

• Pasteurization is used to reduce microbial Nos. in milk & other

beverages while retaining flavor & food quality of the beverage.
 Pasteurization…

• It retards spoilage but does not sterilize.

• Flash pasteurization = high-temp. short term pasteurization.

• Involves quick heating to about 72°C for 15 sec, then rapid

cooling.
 Ultrahigh-temperature (UHT) sterilization

• Milk & similar products heated to 140 - 150°C for 1 - 3 sec.

• Very quickly sterilizes the milk while keeping its flavor & quality.

• Used to produce the packaged “shelf milk” that does not

require refrigeration.
ii. Dry heat
• Can be used to sterilize moisture-sensitive materials such as
powders, oils & similar items.

• However, it is less efficient than moist heat.

• It usually requires higher temp. (160 to 170 °C) & longer

exposure times (2 to 3 hrs).
 Incineration

• Burner flames

• Electric loop incinerators

• Air incinerators - used with fermenters; generally operated at

500 °C.
 Oven sterilization

- Used for dry glassware & heat-resistant metal equipment.

- Typically 2 hr at 160°C is required to kill bacterial spores by dry

heat.

- This does not include the time for the glass to reach the

required temp. (penetration time) & the cooling time.

b. Low temperatures

i. Freezing
• Freezing at -20 °C does not necessarily destroy microbes.

• It inhibits growth by slowing metabolism & removing available

liquid water.

• Can be used for long-term storage of microbes.

• “Ultracold” lab. freezer typically -80°C.

• Freezing inhibits all microbial growth - however many bacteria &

other microbes may survive freezing temperatures.
ii. Refrigeration

• Slows microbial growth by lowering rates of metabolism except

for special types of microbes (e.g. psycrophiles).

• Operates at around 4°C which inhibits growth of mesophiles or

thermophiles while psychrophiles will grow.
 c. Filtration
• Used for physically removing microbes & dust particles from
solutions & gases.

• Often used to sterilize heat-sensitive solutions or to provide a

sterilized air flow.

i. Depth filters - thick fibrous or granular filters that remove

microbes by physical screening, entrapment, or adsorption.

ii. Membrane filters - thin filters with defined pore sizes that
remove Mos., primarily by physical screening.

iii. High-efficiency particulate air (HEPA) filters - used in laminar

flow biological safety cabinets to sterilize air.
 d. Radiation
i. Ultraviolet (UV) radiation
• Effective, but its use is limited to surface sterilization.

• UV radiation does not penetrate glass, dirt films, water &

other substances.

• DNA absorbs ultraviolet radiation at 260 nm wavelength.

• Causes damage to DNA in the form of thymine dimer

mutations.

• It is useful for continuous disinfection of work surfaces, e.g. in

biological safety cabinets.
 ii. Ionizing radiation
• X rays, gamma rays, etc. is effective & penetrates the material.

• Powerful sterilizing agent.

• Penetrates & damages both DNA & protein.

• Effective against both vegetative cells & spores.

• Often used for sterilizing disposable plastic labware e.g. petri

dishes; antibiotics, hormones & other heat-sensitive materials.

• Can also be used for sterilization of food.

• Approved but has not been widely adopted by the food industry.
 Chemical Agents in Control
i. Phenolics
• Lab. & hospital disinfectants.
• Phenolics act by denaturing proteins & denaturing
membranes.

ii. Alcohol
• Widely used as disinfectants & antiseptics.
• Do not kill endospores.
• Act by denaturing proteins & dissolving membrane lipids.
• Kills vegetative cells of bacteria & fungi but not spores.
• Used in disinfecting surfaces & thermometers.
• “Ethanol-flaming” technique is used to sterilize glass plate
spreaders or dissecting instruments at the lab bench.
iii. Halogens
• Act as oxidizing agents.

• Oxidize proteins & other cellular components.

• They are widely used antiseptics & disinfectant e.g.

 Iodine - oxidizes cell constituents and iodinates cell

proteins.

 Chlorine - oxidizes cell constituents.

iv. Heavy metals

• Effective but usually toxic.

• They act by combining with proteins & inactivating them.

v. Aldehydes

• Reactive molecules that can be used as chemical sterilants.

• But may irritate the skin.

• They act by combining with proteins & inactivating them.

vi. Quaternary ammonium compounds

• Are cationic detergents of low toxicity.

• Used as disinfectants for food utensils & small instruments.

• Also used as skin antiseptics.

• Act by disrupting biological membranes & denaturing proteins.

vii. Sterilizing gases (e.g. ethylene oxide)
• Used to sterilize heat-sensitive materials.

• Act by combining with proteins & inactivating them.

• Vapor-phase hydrogen peroxide has been used to

decontaminate biological safety cabinets.
 BACTERIA MOTILITY
• Many bacteria can move using a variety of mechanisms:
i. Flagella - used for swimming through water.
• Swimming bacteria frequently move near 10 body lengths per
second & a few as fast as 100.
• Are as fast as fish, on a relative scale.

ii. Bacterial gliding & twitching - move bacteria across surfaces.

• In twitching motility, bacterial use their type IV pili as a grappling
hook, repeatedly extending it, anchoring it & then retracting it
with remarkable force.

iii. Changes of buoyancy - allow vertical motion.

BACTERIA MOTILITY…

• Bacterial species differ in the no. & arrangement of flagella on

their surface:

i. Monotrichous - have a single flagellum

ii.Amphitrichous - a flagellum at each end

iii.Lophotrichous - clusters of flagella at the poles of the cell

iv.Peritrichous - flagella distributed over the entire surface of cell

BACTERIA MOTILITY…

• The bacterial flagella is the best-understood motility structure in

any organism and is made of about 20 proteins.

• Approx. another 30 proteins required for its regulation &

assembly.

• Flagellum is a rotating structure driven by a reversible motor at

the base that uses the electrochemical gradient across the
membrane for power.
 BACTERIA MOTILITY…

• This motor drives the motion of the filament, which acts as a

propeller.

• Many bacteria (e.g. E. coli) have two distinct modes of

movement: Forward movement (swimming) & tumbling.

• Tumbling allows them to reorient & makes their movement a

three-dimensional random walk.
 BACTERIA MOTILITY
• Motile bacteria are attracted or repelled by certain stimuli in
behaviors called taxes.

• Include chemotaxis, phototaxis, energy taxis & magnetotaxis.

• For myxobacteria, individual bacteria move together to form

waves of cells that then differentiate to form fruiting bodies
containing spores.

• The myxobacteria move only when on solid surfaces, unlike E.

coli which is motile in liquid or solid media.
 Bacteria Chemotaxis

• The phenomenon in which bacteria & other single-cell or

multicellular organisms direct their movements according to
certain chemicals in their environment.

• This is important for bacteria to find food (e.g. glucose) by:

 Swimming towards the highest concentration of food molecules.

To flee from poisons (e.g. phenol).

 Bacteria Chemotaxis

• In multicellular organisms, chemotaxis is critical to early (e.g.

movement of sperm towards the egg during fertilization) &
subsequent phases of development (e.g. migration of neurons
or lymphocytes) as well as in normal function.

• Positive Chemotaxis - if movement is in the direction of a higher

concentration of the chemical in question.

• Negative Chemotaxis - if movement is in the opposite direction

of a higher concentration of the chemical in question.
 Chemoattractants and chemorepellents

• Chemoattractants & chemorepellents are inorganic or organic

substances possessing chemotaxis-inducer effect in motile cells.

• Effects of chemoattractants are elicited via described or

hypothetic chemotaxis receptors.

• A chemoattractant is target cell specific & concentration

dependent.
 Chemoattractants and chemorepellents

• Most frequently investigated chemoattractants are formyl

peptides & chemokines.

• Responses to chemorepellents result in axial swimming & they

are considered as a basic motile phenomena in bacteria.

• The most frequently investigated chemorepellents are inorganic

salts, amino acids & some chemokines.
a. Behavior

• The overall movement of a bacterium is the result of alternating

tumble & swim phases.

• If one watches a bacterium swimming in a uniform environment,

its movement will look like a random walk with relatively
straight swims interrupted by random tumbles that reorient the
bacterium.

• Bacteria such as E. coli are unable to choose the direction in

which they swim & are unable to swim in a straight line for more
than a few seconds due to rotational diffusion.
a. Behavior…

• In other words, bacteria "forget" the direction in which they are

going.

• By repeatedly evaluating their course & adjusting if they are

moving in the wrong direction, bacteria can direct their motion
to find favorable locations with high concentrations of
attractants (usually food) and avoid repellents (usually poisons).

• In the presence of a chemical gradient, bacteria will chemotax

or direct their overall motion based on the gradient.
a. Behavior…

• If the bacterium senses that it is moving in the correct direction

(toward attractant/away from repellent), it will keep swimming
in a straight line for a longer time before tumbling.

• If it is moving in the wrong direction, it will tumble sooner & try

a new direction at random.

• Bacteria like E. coli use temporal sensing to decide whether life

is getting better or worse.
a. Behavior…

• This way, it finds the location with the highest concentration of

attractant (usually the source) quite well.

• Even under very high concentrations, it can still distinguish very

small differences in concentration.

• Fleeing from a repellent works with the same efficiency.

a. Behavior…
• This purposeful random swimming is a result of simply choosing
between two methods of random movement - tumbling &
straight swimming.

• Chemotactic responses such as forgetting direction & choosing

movements resemble the decision-making abilities of higher life-
forms with brains that process sensory data.
a. Behavior…
• The helical nature of the individual flagella filament is critical for
this movement to occur.

• The protein that makes up the flagella filament, flagellin, is quite

similar among all flagellated bacteria.

• However, there are bacteria that do not follow this rule.

a. Behavior…
• Many bacteria, such as Vibrio, are monoflagellated and have a
single flagellum at one pole of the cell.

• Their method of chemotaxis is different.

• Others possess a single flagellum that is kept inside the cell wall.

• These bacteria move by spinning the whole cell, which is shaped

like a corkscrew.
b. Signal transduction

• Chemical gradients are sensed thro’ multiple transmembrane

receptors, called methyl-accepting chemotaxis proteins (MCPs).

• MCPs vary in the molecules that they detect.

• These receptors may bind attractants or repellents directly or

indirectly thro’ interaction with proteins.
b. Signal transduction…

• The signals from these receptors are transmitted across the

plasma membrane into the cytosol, where Che proteins are
activated.

• The Che proteins alter the tumbling frequency, and alter the
receptors.
 The end

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