ROLE OF MICROORGANISMS IN DISEASE CONTROL

By EZEUMEH EUCHARIA N

1.0 Introduction

The role of microorganisms in diseases control can be reviewed in two

perspectives. One is their roles in crop and plant production as microbial biological
control agents (MBCA) against plant diseases. On the hand microbes are
Incorporated in animal feeds and human diets to also enhance yield and confer
protection from evading pathogens respectively. In plants, MBCAs are applied to
the crops where they act via different modes of action. In animals they are included
in feeds here they ward off other pathogens thereby increasing the animal survival
and production yield. Similarly in humans, they are used hugely as probiotics ( still
a fresh area for more researches) for the purpose of checking off unwanted foreign
organisms ( pathogens) and radicals. They are also incorporated in foods, usually
sporadic or controlled fermented foods as starter cultures for their nutritive effects
( single cell protein). For proper elucidation, let us x-ray these roles played by
microorganisms in respective areas.

1.1 Microorganisms as Microbial Biological Control Agents

Biological control of plant diseases is the suppression of populations of plant

pathogens by living organisms (Heimpel and Mills, 2017). The beneficial
microorganisms are selected for highly effective biovars which are then cultivated
en mass ( biomass cultivation). The application of these selected highly
antagonistic biovars to the crops at regulated patterns during the planting season is
referred to as augmentative biological control ( Eilenberg et al, 2001; Heimpel and
Mills, 2017;van Lenteren et.al., 2018)

1.2 Criteria for Selection of MBCA

Must be:

 Highly effective, that is lethal dose ( LD 50) must be high at small

concentration or the signalling compound to elicit stimuli must be in small
amount.
 Grown artificially using the appropriate nutrients in g laboratory.
  Antibiotic metabolites must not be cumulatively toxic to the plant or crop to
which it is added.
 The signalling compound produced by MBCA must be easily degradable by
both Microbial and abiotic means and not accumulate in the environment

Commercial MBCAs are available in the market produced by biocontrol

companies.In some cases, antimicrobial metabolites produced by selected
microbial organisms are included in the product, and some products even contain
only antimicrobial metabolites without living cells of the antagonist (Glare et al.,
2012).

1.3 Mechanism of actions of MBCA

As highlighted earlier, these act via a range of mode of actions. And these include

 Interaction with plants by inducing resistance to pathogens or priming

plants without any direct interaction with the targeted pathogen in plant
tissues (Pieterse et.al, 2014)
 Some MBCAs act via nutrient competition or other mechanisms modulating
the growth conditions for the pathogen (indirect interaction).
 Antagonists act through hyperparasitism and antibiosis. This is a classical
direct interference with the pathogens.

Such interactions are highly regulated cascades of metabolic events. Compounds

involved such as signaling compounds, enzymes and other interfering metabolites
are produced in situ at low concentrations during interaction. The potential of
microorganisms to produce such a compound in vitro does not necessarily correlate
with their in situ antagonism.

a. Interactions with plant to enhance resistance: Constitutive mechanisms such as

cuticles are complemented by inducible resistance mechanisms. Induced plant
defense mechanisms are triggered by stimuli recognized by specific recognition
receptors. Resistance inducing stimuli produced by microorganisms are called
MAMPs. This involve the production of reactive oxygen species, phytoalexins,
phenolic compounds, or pathogenesis-related proteins or the formation of physical
barriers like modifications of cell walls and cuticles by the induced plant (Wiesel et
al., 2014). This is energy dependent on the part of the induced plant. In priming of
defense however, plants are allowed to react to stimuli later in a fast and robust
manner with lower energy costs (Mauch-Mani et al., 2017). The most studied
MAMPs are the bacterial proteins flagellin and elongation factor EF-Tu. For both
proteins the receptor systems are known and reactions are induced by
subnanomolar concentrations. Other examples for MAMPs with known receptors
are glucan, chitin, xylan, e.g., produced by Phytophthora megasperma and
Trichoderma viride. Boller and Felix (2009). There are yet other MAMPs with
unknown receptors. Few examples are proteinaceous MAMPs from bacteria, e.g.,
superoxide dismutase and 23-amino-acid peptide, or from oomycetes, e.g., pep13
trans-glutaminase, sterol-binding elicitins, and cellulose-binding lectins; lipophylic
MAMPs, e.g., ergosterol and arachidonic acid e.t.c.

b. Nutrient completion with pathogens:Obligate biotrophic pathogens use

exclusively nutrients from infected living host cells and do not depend on any
exogenous nutrient sources in the environment outside the host plant (Agrios,
2005). The majority of plant pathogens exploit nutrient sources in a much less
specific way by degrading dead organic plant matter. Necrotrophic plant
pathogenic bacteria, fungi and oomycetes kill and subsequently invade tissues of
host plants and utilize the available nutrients as primary colonizers of these killed
tissues. Once necrosis has been induced by the pathogen, non-pathogenic
microorganisms with saprophytic life style can potentially also colonize necrotic
tissues so that a saprophytic competitive substrate colonization between different
populations is common resulting in competition for nutrients and space.Common
for all necrotrophic pathogens during their saprophytic stage is that they depend on
exogenous nutrients available in the environment, This dependency on exogenous
nutrients during significant parts of their life cycle makes non-biotrophic
pathogens vulnerable to nutrient competition (Köhl and Fokkema, 1998).
Consequently, highly competitive microorganisms are potential candidates for
biological control using competition for nutrients and space as mode of action.An
example of use of this efficient mode of action is wound protection of fruits from
pathogen invasion by fast colonizing yeasts . They compete for carbohydrat and
even nitrogen sources. Pichia guilliermondii and pathogenic Penicillium digitatum,
P. expansum, B. cinerea, or Colletotrichum spp. in wounds of different fruits and
Aureobasidium pullulans and P. expansum in apple wounds.
c. Hyperparasitism and antibiosis: Parasitism is the direct competitive interaction
between two organisms in which one organism is gaining nutrients from the other.
If the host is also a parasite, e.g., a plant pathogen, the interaction is defined as
hyperparasitism. This kind of relationship usually occurs among fungi and rarely
reported in bacteria. Main mechanisms of parasitism is the excretion of CWDEs
combined in some cases with excretion of secondary metabolites in close contact
with the host cell leading to openings in the cell wall (cell wall degradation) and
subsequent disorganization of the cytoplasm. Examples include hyoerparasitism of
Cladosporium uredinicola against Puccinia violae, Alternaria alternata against
Puccinia striiformis f. sp. tritici .

Mycoparasitism mechanism in Trichoderma spp and Clonostachys spp: They

produce structures for attachment and infection, and kill their hosts by CWDEs,
often in combination with antimicrobial secondary metabolites (Harman et al.,
2004; Harman, 2006; Mukherjee et al., 2012; Karlsson et al., 2017; Nygren et al.,
2018). These lytic enzymes are not constitutive but their production is triggered by
complex signaling after recognition of the host. Surface compounds such as lectins
from the host cell wall together with surface properties and diffusible host-released
secondary metabolites play important roles in the recognition and signaling
pathways such as MAPK cascades, cAMP pathway and G-protein signaling. These
lead to transcriptional reprogramming and expression of the “molecular weapons”
involved in host attack and lysis.

d. Antibiosis: this involves production of antimicrobial metabolites (secondary

metabolites) belonging to heterogeneous groups of organic, low-molecular weight
compounds by microorganisms that are deleterious to the growth or metabolic
activities of other microorganisms (Thomashow et al., 1997). They are produced
and released to the environment in small quantities by many microorganisms.
Huge numbers of known antibiotics are produced by actinomycetes (8700 different
antibiotics), bacteria (2900) and fungi (4900). Less than 1% of microscopically
counted bacteria can be cultured on usual culture media (Amann et al., 1995). They
are known only through rRNA sequences (Clardy et al., 2006).This means that
majority of antibiotics produced in situ in the environment is still unknown
(Raaijmakers and Mazzola, 2012). Biocontrol bacteria that excrete specific
antibiotic as secondary metabolites include Agrobacterium, Bacillus, Pantoea,
Pseudomonas, Serratia, Stenotrophomonas, Streptomyces, and many other
genera.Many antibiotics are produced only when a microbial population reaches
certain thresholds. This is quorum-sensing phenomenon. Examples of compounds
produced by different bactia which antimicrobial activities are phenazine, DAPG
and pyrrolnitrin, by certain Pseuodomonas spp; lipopeptides eg iturin, surfactin,
and fengycin in certain Bacillus genera. Among fungi, Trichoderma and closely
related Clonostachys (former Gliocladium) produce 6-PAP, gliovirin, gliotoxin,
viridin.

NB: Microorganisms producing antimicrobial metabolites with the potential to

interfere with antibiotics in human and veterinary medicine must be excluded from
use as MBCAs (Anonymous, 2013a).

1.4.1 Mechanism of actions:

Some antibiotics, especially lipopeptides support the mobility of bacteria, most

likely via changing the viscosity of the colonized surfaces. Surface-active
antibiotics allow bacteria to move to nutrient rich locations and also change the
water dynamics on leaf surfaces which indirectly affects pathogen development.
Other groups of antibiotics influence the nutritional status of plants. For example,
DAPG-producing Pseudomonas upregulates the nitrogen fixation by plant growth-
promoting Azospirillum brasilense, and redox-active antibiotics support
mobilization of limiting nutrients such as manganese and iron.

1.5 Microorganism as Agent of Disease Control in Humans and Animals

Definition of common terminology

 A pathogen is a micro-organism that has the potential to cause disease.

 An infection is the invasion and multiplication of pathogenic microbes in an
individual or population.
 Disease is when the infection causes damage to the individual’s vital
functions or systems.
 Probiotics: culture of live microroganisms which are beneficial to health.
 An infection does not always result in disease

The immune system: An infection can be seen as a battle between the invading
pathogens and the host. Our bodies are equipped to fight off invading microbes
that may cause disease. These are called our natural defences.
The surfaces of the body – the skin, digestive system, and the lining of the nose –
are covered by a community of microbes called the normal body flora. They help
protect the host from becoming infected with more harmful micro-organisms by
acting as a physical barrier. The normal body flora ( commensals) colonise these
linings which reduces the area available for pathogens to attach to and become
established. It also means that the harmful microbes have to compete with the
normal body flora for nutrients. The average human gut contains around one kilo
of these good bacteria (probiotics) which is equivalent to one bag of sugar!

1.6 Health Benefits of Probiotics

 Improved digestion of lactose and reduction of intestinal bloating, flatulence

and discomfort
 Prevention of traveller's diarrhoea
 Prevention of the potential outgrowth of spores of Clostridium botulinum in
the GI-tract, the associated toxin production and a possible cause of sudden
infant death syndrome (SID)
 Enhancing the immune system, improving resistance to infection and
improving well-being
 Protection against certain types of cancer
 Lowering serum cholesterol levels and reducing the incidence of coronary
heart disease
 Prevention or helping treat peptic ulcer disease
 Treating intractable diarrhoea following antibiotic therapy
 Reducing allergic inflammation

1.6.1 Mechanism of action of probiotics

The growth of lactobacillus and other beneficiary micororganisms in the GI- tract
would displace other putrefactive bacteria, reduce the concentrations of toxins in
the gut and thus improve health. This utilize competitive mechanism; for available
nutrients and space as discussed in hyperparasitism above. Also production of
certain bacteriocin that are antagonistic to other pathogenic bacteria.

In human infants, the GI tracts consist of Bifidobacterium accounting for 95% of

the total Flora. The predominance of these bacteria is due to selective agents in
meconium (the sterile fluid in the GI-tract of human neonates), human colostrum
and human milk. These selective factors are known as 'bifidus growth factors'. The
factor was found to consist of several large oligosaccharides ( secondary
metabolite) and are interestingly low in ruminant's milk.

Again several metabolic compounds produced by lactic acid bacteria including

organic acids, fatty acids, hydrogen peroxide, and diacetyl have antimicrobial
activities. Of these, nisin, which is produced by some Lactobacillus lactis subsp.
lactis strains is at present the only purified bacteriocin approved for use in products
intended for human consumption. The gut bacteria can also affect your blood
sugar, brain health and heart health by reducing cholesterol levels, blood pressure
and inflammation particularly L. plantarum and L. reuteri. They do this by binding
with cholesterol in the intestines to stop it from being absorbed. They also help
produce certain bile acids, which help metabolize fat and cholesterol in your
body.Certain probiotics can also produce short-chain fatty acids, which are
compounds that can help prevent cholesterol from being formed by the liver.

Similarly, High blood pressure is another risk factor for heart disease, and it may
be lowered by certain probiotics.One study of 36 smokers found that taking
Lactobacilli plantarum for 6 weeks significantly reduced blood pressure. Also,
when combining different strains of bacteria. However, not all probiotics are
effective for improving high blood pressure. The ability to reduce high blood
pressure stem from the ability to adhere to the ileum and human cell lines both HT-
29 and Caco-2 cell lines. By this, together with other Cascades of reactions
induced mobilization of second line of immune defense and stabilization by the
body; the macrophages recruited by the body from the liver and heart engages in
clearing and mopping up of pathogens and toxins, thereby normalizing the blood
pressure.

1.7 Conclusion

Role of microorganisms in disease control in both plants and animals are well
documented. But care should be taken in assaying the antimicrobic activities of the
strains to ensure safety of the host. More studies should also be taken up to assess
other Microbial biocontrol that hitherto are unaccounted for.
References

Agrios, G. N. (2005). Plant Pathology, 5th Edn. Amsterdam: Elsevier Academic

Press.Google Scholar

Ajouz, S., Nicot, P. C., and Bardin, M. (2010). Adaptation to pyrrolnitrin in

Botrytis cinerea and cost of resistance. Plant Pathol. 59, 556–566. doi:
10.1111/j.1365-3059.2009.02230.xCrossRef Full Text | Google Scholar

Ajouz, S., Walker, A. S., Fabre, F., Leroux, P., Nicot, P. C., and Bardin, M. (2011).
Variability of Botrytis cinerea sensitivity to pyrrolnitrin, an antibiotic produced by
biological control agents. Biocontrol 56, 353–363. doi: 10.1007/s10526-010-9333-
7CrossRef Full Text | Google Scholar
Amann, R. I., Ludwig, W., and Schleifer, K. H. (1995). Phylogenetic identification
and in situ detection of individual microbial cells without cultivation. Microbiol.
Rev. 59, 143–169.PubMed Abstract | Google Scholar

Anonymous (2011). Commission Regulation (EU) No 546/2011 of 10 June 2011

implementing Regulation (EC) No 1107/2009 of the European Parliament and of
the Council as regards uniform principles for evaluation and authorisation of plant
protection products. Off. J. Eur. Union L 155, 127–175.Google Scholar

Badosa, E., Montesinos, L., Camó, C., Ruz, L., Cabrefiga, J., Francés, J., et al.
(2017). Control of fire blight infections with synthetic peptides that elicit plant
defense responses. J. Plant Pathol. 99, 65–73. doi: 10.4454/jpp.v99i0.3915
CrossRef Full Text | Google Scholar

Bakker, R. A. H. M., Raaijmakers, J. M., and Schippers, B. (1993). “Role of iron

in the suppression of bacterial plant pathogens by fluorescent pseudomonads,” in
Iron Chelation in Plants and Soil Microorganisms, eds L. L. Barton and B. C.
Hemming (San Diego: Academic Press), 269–278.

GBadosa, E., Montesinos, L., Camó, C., Ruz, L., Cabrefiga, J., Francés, J., et al.
(2017). Control of fire blight infections with synthetic peptides that elicit plant
defense responses. J. Plant Pathol. 99, 65–73. doi: 10.4454/jpp.v99i0.3915

CrossRef Full Text | Google Scholar

Bakker, R. A. H. M., Raaijmakers, J. M., and Schippers, B. (1993). “Role of iron

in the suppression of bacterial plant pathogens by fluorescent pseudomonads,” in
Iron Chelation in Plants and Soil Microorganisms, eds L. L. Barton and B. C.
Hemming (San Diego: Academic Press), 269–278.Google Scholar
Bardin, M., Ajouz, S., Comby, M., Lopez-Ferber, M., Graillot, B., Siegwart, M., et
al. (2015). Is the efficacy of biological control against plant diseases likely to be
more durable than that of chemical pesticides? Front. Plant Sci. 6:566. doi:
10.3389/fpls.2015.00566.PubMed Abstract | CrossRef Full Text | Google
Scholaroogle Scholar

Bardin, M., Ajouz, S., Comby, M., Lopez-Ferber, M., Graillot, B., Siegwart, M., et
al. (2015). Is the efficacy of biological control against plant diseases likely to be
more durable than that of chemical pesticides? Front. Plant Sci. 6:566. doi:
10.3389/fpls.2015.00566PubMed Abstract | CrossRef Full Text | Google Scholar