Microbial control of insect-pests

Satish Kumar
 History
• Aristotle ( 384-322BC) was the first to mention
that bees suffered from diseases
• Bassi in 1835 demonstrated the infectious nature
of Beauveria bassiana in respect of Bombyx mori
• Le Conte ( 1873) for the first time advocated the
use of disease as means of insect control
• Krassiilstschik (1888) mass produced spores of
Anisoplia austriacu and applied them in field
tests.
• The term microbial control was coined only in
1949 by Steinhaus
• Between 1920 and 1960 many species of
bacteria, virus and nematodes were tried out
• Bacillus thuringiensis proved effective for
control of Lepidopterous insects
• Among the viruses, NPV and among
protozoans micros-porodians and among
nematodes neoaplectanids have been more
commonly tested and found promising
• When microbial organisms or their products
are employed by man for control of insects,
animals and plants in a particular area, it is
referred to as microbial control
 Traits desirable in pathogens
• Suitable strain
• Virulence
• Toxins
• persistance
 Terms used in microbial control
• Entomophilic/ entomogenus pathogens
• Epizootics
• Enzootics
• Portal entry
• Septicemia
• Morbidity
• Toxemia
• Virulence
• Infectious disease
• Contagious disease
 Principal group of pathogens
• Bacteria
• Viruses
• Rickettsiae
• Protozoa
• Nematodes
• fungi
 Bacteria
Sporeformers Non-sporeformers
• Obligate Facultative • Potential facultative
• Pseudomonas , Serratia
• Septica
• Crystaliferous NCF • Aerobactor
• B. popillae • Cloaca
• Clostridium • Proteus
 Bacillus
• Thuringiensis
• Var. galleriae
• Var. sotto
• Var. thuringiensis
• Var. subtoxicus
 Sporeformers
• The sporulating bacteria form endospores which
are highly resistant to environmental changes and
allow them to persist in dormant conditions
• On ingestion they develop in to bacterial cell
• Among the sporeformers, crystalliferous are
better than non-crystalliferous because of toxic
nature of crystals they produce
• One of the best studied species crystalliferous
bacteria is Bacillus thuringiensis
• The first commercial product containing this
bacteria was available in USA in 1958
• This bacteria is effective against caterpillars of
many Lepidoptera
• Var. thuringiensis is more toxic to Pieris
brassicae and Bombyx mori
• It produces a toxic proteinaceous crystal in its
cell
 The crystal
• Protien crystal or the parasporal body is formed
when the sporulation is to take place and not
before the vegetative cell
• Young bacterial cells are non-toxic to larvae
• Crystals of different species of bacillus size and
shape
• Crystals of B. thuringiensis are regular diamond
shaped bodies a bipyramid with prominent
surface
• The crystals are quite stable in water but are
broken up by alkaline treatments
 Mode of action
• On being ingested by the susceptible host the endospore
germinate in the gut producing the bacterial cell
• Cells migrate in to haemocoel where they multiply rapidly
and invade and destroy certain tissues and soon fill much of
the haemocoel .
• This stage of infection is termed as septicemia
• Prior to the death of the host thick walled refractile spores
are formed
• After death of the host spores are released in to the soil
• Clostridium does not invade haemocoel but multiply in gut
and host cadaver gets mummified instead of disintegrating
 Role of crystal
• The crystal enters the midgut epithelium quickly and
induces an intense secondary activity in the cells which
get separated from basement membrane
• This leads to gut paralysis
• Blood pH changes due to change in permeability of the
midgut epithelium
• This finally leads to paralysis of the body
• Blockade of the nerve conduction due to impairment
of the axonal membrane permeability, precipitation of
the blood protiens and inhibition of enzymes
 Symptoms of bacterial infection
• Stoppage of feeding
• Regurgitation
• Diarrhoea
• Gut paralysis
• General body paralysis
• The proteolytic enzyme working at high pH
dissolve the crystals releasing endotoxins
 Non sporeformers
• They are heterogeneous group of bacteria
which infect host only under extra ordinary
circumstances
• They occur in gut they do not multiply there
readily nor invade haemocoel
• These are not very promising
 viruses
• Inclusion virsuses
• Non inclusion viruses
IV PV
• Polyhedrosis viruses ( PV) • Nuclear polyhedrosis
• Granulosis viruses (GV) viruses (NPV)
• Cytoplasmic polyhedrosis
viruses (CPV)
 Nuclear polyhedroses viruses
• They are rod shaped 20-50um broad and 200-
400um long
• They are enclosed in two proteinaceous
membranes which to gather with virus
particles constitute polyhedral body
• There could be one or more virus particle in a
polyhedron
 Mode of action
• On ingestion membrane of the polyhedra are
dissolved by the alkaline gut juices setting the
virus particles free
• These particles then penetrate gut epithelium
and come in contact with blood cells, tracheal
matrix or epidermal cells
• Inside cells, they enter the nuclei, attach
themselves with chromatin and multiplya ring of
minute virus particles showing strong Brownain
movement is formed on the nuclear periphery
• Membranes are reformed round these
particles to give rise to polyhedra
• The new polyhedra shed their membranes
and attach to more chromatin and multiply
• Nuclear and cell membrane rupture, releasing
the polyhedra in to the body cavity to invade
other cells.
• In advance stage of infection tissues like silk
glands, ganglia, imaginal buds may also get
infected
 Symptoms

• Insect stops feeding and become sluggish and

pale in colour
• May swell slightly and then become limp and
flaccid
• Integument becomes fragile, rupture easily to
emit blood with disintegrated tissues and
polyhedra.
• Prior to death insect climbs to higher positions
and dead larvae usually hang by their pro legs
 Host range

• Diptera
• Hymenoptera
• lepidoptera
 Cytoplasmic polyhedroses viruses
• They attack columnar cells of midgut epithelium
• Infection is restricted to midgut but may spread to
foregut and hindgut
• Polyhedra develop in the cytoplasm and enlarge to
loose polyhedral shape and become spherical
• Small size of larvae, disproportionately large heads
• Loss of appetite
• Hind gut may be completely evarted due to persistent
efforts on the part of larvae to void the polyhedra
which block the gut lumen
 Granuloses viruses
• They inhibit fat body, epidermis, blood cells
and tracheal matrix
• They develop in cytoplasm
• Inclusion bodies are ellipsoidal rather than
polyhedral
• They infect tissues eventually disintegrate and
bogy fluid becomes filled with inclusions
 Rickettsiae
• Group of microorganisms which are regarded
as intermediate form between virus and
bacteria
 Nematodes
Mermithidae Tylenchida
• Hydromermis • Tripius
• Mermis • Heterotylenchus
• Agamermis
• Rhabdita
• Neoaplectana
• Portal entry of nematodes is through the
integument
• Insecticidal property of nematodes was
discovered by Dutky and Hough (1955) in the
codling moth larva
• Theses are free living soil stage which picks up
bacteria or other micro-organisms on their
cuticle and enter their host along with them
 Mode of action
• Neoplectana carpocaopsae has symbiotic association
with bacterium Achromebacter nematophilus
• Infective stage ( 3rd instar) carries the bacterium in the
lumen of the intestine and releases it through its anus
into the body of the host
• Infected juveniles of the nematode are ingested with
food by the host
• They moult in gut penetrate the foregut and invade the
haemocoel
• Bacterium produces antibiotic that inhibits growth of
other micro-organisms
 Fungi
Phycomycetes Ascomycetes
• Coelomomyces • Cordyceps
• Entomophthora
• Massospora
 Fungi
Basidiomycetes Deuteromycetes
• Septobasidium • Beauveria
• Metarrhizium
• Aspergillus
• Spicara
• Hirssutela
• Isara
• Paecilomyces
 Mode of action
• Most common portal entry of fungi is through integument
through invasion via respiratory or alimentary tract
• Infective unit is spore
• Upon germination spore give out small swellings called
appressoria
• Apperessorium attaches itself to cuticle and sends out an
infection peg
• Hypae penetrate the layers of integument by enzymatic
dissolution of chitin and protein ramify in cuticle and then
reach the haemocoel and internal organs
• Conidiophores are produced which erupt through cuticle
• Death of host is by obliteration of the tissues and by toxins
produced by the fungus
 Symptoms
• Loss of appetite and attempt to climb higher up
• Decreased irritability
• General or partial paralysis
• Dicoloured patches on integument
• Increased acidity in blood
• Fungi need a high degree of atmospheric
humidity to germinate and this is the limiting
factor in their use as microbial control agent in
the field
 Host range
• Lepidoptera
• Homoptera
• Hymenoptera
• Coleoptera
• Diptera
 Fungal toxins
• Aflatoxin cause serious change in the
metabolic activity of the vertebrates and
insects
• Destraxin A and destraxin B cause paralysis
and death
Thank you