Microbial control
Definition
When microbial organisms or their products (toxins) are employed by man for the
control of insects animals, and plants in a particular area, it is referred to as microbial control.
Obviously, this is an extension of biological control where instead of the microbes (parasites and
predators), microbes (pathopgens) are employed.
Traits desirable in pathogens
Four qualities go to make a microorganism suitable for the control of insect pests they are
(i) a suitable strain; (ii) virulence; (iii) toxins and (iv) persistence (long shelf-life). Even within
the same species one strain of microorganism may be more effective as a control agent than the
other. It is fortunate for us that microorganisms occur in many strains so as to allow a suitable
selection. Strains are more common in bacteria and fungi than in viruses and protozoa. Virulence
is the power of the pathogens to invade and produce diseases in their hosts. The faster it can
spread, the more virulent it will be. Virulence can be artificially enhanced by (a) passing the
pathogens successively through susceptible hosts, (b) introducing chemicals that may aid in
increasing the invading power of the pathogens, (c) associating it with other microorganisms
that may render it more capable of invading tissue and (d) changing nutritional factors and
culturing conditions. Toxin-producing pathogens are effective in two ways: by their own viru
lence and by the poisonous nature of the toxins they produce. Toxins could themselves be
utilized as insecticides and, therefore, toxin-producing pathogens which are largely bacteria and
fungi are attracting the attection of the entomologists to a greater extent. Persistence is the
quality which enables a microorganism to retain its viability and virulence on storage. The spores
of bacteria, fungi, protozoa and polyhedral inclusions of viruses are known to possess a long
shelf-life.
Some descriptive terms
Some of the common terms used in description of the microorganisms are as follows.
Entomophilic/entomogenous pathogens. Pathogens that are found in or are harmful to insects.
Epizootics. Pathogens infecting large populations of insects.
Epizootiology. Science that deals with infectious diseases as they occur in groups of animals;
this has the same meaning as epidemiology used with reference to human diseases.
Enzootics. These are pathogens infecting small population of insects.
Portal entry. This is the point at which the pathogens enter the body of the host.
Septicemia this is the condition when the pathogen multiplies in the blood and tissue throughout
the body cavity.
Morbidity. Diseased condition, sickness.
Epizootic disease. Diseased of high morbidity and the one that is only occasionally present in a
recognizable form.
Enzootic disease. A disease that has a low incidence but is constantly present in a population.
T oxemia. Dissemination of toxins in blood.
V. Principal group of pathogens
The microorganisms that have been found suitable for insect control are: bacteria,
viruses, rickettsiae, protozoa, nematodes and fungi. These can be placed in two groups: ingested
microbes (bacteria, viruses, rickettsiae and protozoa) which enter insect body along with food
(and are likened to stomach insecticides) and penetrating microbes (nematodes and fungi) that
enter by penetrating the integument (and are likened to contact insecticides). The sections that
follow describe these pathogens and the way they kill their insect hosts.
Bacteria. Entomogenous bacteria are classified as sporeformers and non-sporeformers which are
further subdivided into obligate and facultative. The latter, under sporeformers, could be crystal
forming (crystalliferous) or non-crystalliferous. The examples of these different groups are listed
in Table 21.1
Sporeformers. The sporeforming or sporulating bacteria form endospores (Fig. 21.1) which are
highly resistant to environmental changes (Temperature, humidity, etc.) and allow them to
persist in a dormant condition outside the intended host to develop into a bacterial cell on being
ingested. The sporeforming bacteria are therefore, more promising in insect control than the non-
sporeforming ones. Amongst the sporeformers, again, it is the crystalliferous ones which are
better than the non-crystalliferous ones because of the toxic nature of the crystals they produce.
The crystal. The protein crystal or the parasporal body is formed only when sporulation is
to take place and not before in the vegetative cell. They young bacterial (vegetative) cells are,
therefore, non-toxic to the larvae which are susceptible to sporulated cultures. Crystals of
different species of the bacillus differ in their shape and size, but those found in the varieties of
B. thuringiensis are regular diamond-shaped bodies. One cell gives rise to only one crystal, the
crystal toxins are referred to as endotoxins to distinguish them from the exotoxins that the
bacterial cell produces in addition.
Mode of action. On being ingested by the susceptible host, the endospore germinates in
the gut (only if the pH is below 9), producing the bacterial cell. The cells migrate into the
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,
the thick-walled refract-tile spores are formed; the bacillus body is then called sporangium,
Besides the endotoxins of the crystal, B. thuringiensis also produces a few more toxins-at
least two kinds of exotoxios and some toxic enzymes.
Symptoms of bacterial infection. The main symptoms produced by B. thuringiensis in its
insect host are stoppage of feeding. Regurgitation and diarrhea due to gut paralysis followed by
general body paralysis.
Commercial products. Some of the commercial names of the products containing B.
Thuringiensis and var. thuringiensis are Bakthane L-69®, Parasporin® and Thuricide® in the USA
and Sporeine and Biospore2808 in Europe. All commercial products declare their bacterial
concentration in terms of billions of spores per gram powder. Both spray and dust formulations
of a bacterial insecticide could be used. Dust formulations generally contain 2.5-7.5 billion
spores per gram and wettable powder, some 25-100 billion spores per gram powder.
Host range. Facultative [Link] has been found to be generally effective against
a wide range of lepidopterous larvae. belonging to Coleoptera, Hymenoptera, and Lepidoptera,
while the obligate B. popilliae is good for coleopterans of the family Scarabaeidae, particularly
the destructive Japanese beetle Popillia japonica. The refractile spores filling the haemocoel
appear white under the integument and so the disease has been named the milky disease.
The milky disease bacillus, B. popilliae. Was isolated and described by Dutky in 1937. It
was one of the first bacterial organisms to be extensively tried out in fields in the USA during
1942-50 with great success against the Japanese beetle, Popillia japonica which plagued some
parts of that country.
Viruses: Entomogenous viruses fall in two categories: inclusion viruses (IV) producing
inclusion bodies in the host cell and non-inclusion virus (NIV) which do not do so. The IV are
further subdivided into polyhedrosis viruses (PV) or polyhedroses which produce polyhedral
(i.e., many-sided) bodies and granular bodies and granulosis viruses (GV) which produce
granular bodies. Polyhedroses could inhabit the nucleus and are called nuclear polyhedrosis
viruses (NPV) or the cytoplasm which are called cytoplasmic polyhedrosis viruses (CPV). The
summarized classification of these viruses is shown diagrammatically.
The inclusion viruses are more promising as insect control agents than non-inclusion
viruses, the portal entry of all the viruses, as mentioned earlier, is by ingestion, less commonly
sub-cutaneously, and doubtfully through spiracles.
Nuclear polyhedroses. The NPV (Fig. 21.3) are rod-shaped, 20-50 m broad and, 200-400
m long. They are enclosed in two proteinaceous membranes which together with the virus
particle constitute the polyhedral body or polyhedron (Fig. 21.4). There could be one or more (up
to 100) virus particles in a polyhedron (Fig. 21.5). A polyhedron is usually hexagonal but could
be of other shapes. There is some evidence that the virus rod itself is divisible into 6-8 sub-units
and also that it bears a protrusion at one end by which it attaches itself to the host cell. The virus
rod here contains DNA; no RNA has been found.
One ingestion, the membranes of the polyhedra are dissolved by the alkaline gut juices
setting the virus particles free. The virus Particles then penetrate the gut epithelium and come to
infect the blood cells (haemocytes), tracheal matric or epidermal cells. Inside these cells, they
enter the nuclei, attach themselves to the chromatin and multiply (i.e., break up into their sub-
units). Soon a ring of minute virus particles showing a strong Brownian movement is formed on
the nuclear periphery which later on fill the entire nucleus. Membranes are re-formed round
these particles to give rise to polyhedral. The new polyhedral shed their membranes, attach
themselves again to more chromatin and multiply. As multiplication goes on, chromatin is
consumed. Eventually, the nuclear and cell membranes rupture releasing the polyhedral into the
body cavity cell membranes rupture releasing the polyhedral into the body cavity to invade other
cells.
Symptoms. Symptoms of NVP infection appear only at a late stage when the insect is near death.
The incubation period is 1-3 weeks. The insect stops feeding and becomes sluggish and pale in
colour. It may swell slightly, then become limp and flaccid. The integument becomes fragile,
ruptures easily to emit blood with disintegrated tissue and polyhedral (a turbid fluid). Prior to
death, the insect climbs to higher positions and the dead larvae usually hanglby their prologs and
dry down to dark brown or black cadavers
Host range. Nuclear polyhedrosis is primarily a lepidopterous disease, occurring also in
some Diptera and Hymenoptera.
Cytoplasmic polyhedroses. The CPV attacks the columnar cells of the midgut
epithelium. The infection is restricted to the midgut but at best may spread to the foregut and
hindgut. The polyhedral develop in the cytoplasm where they enlarge to lose their polyhedral
shape and become spherical. They contain RNA rather than DNA.
Granuloses. The GV largely inhabit the fat body, epidermis, frequently blood cells, and
the tracheal matrix. Three is a controversy over the site of their development. Some believe that
they develop in the nucleus, while others think that they develop in the cytoplasm. The inclusion
bodies called capsules are ellipsoidal rather than polyhedral. They are fairly large-sized (300:500
µm) and can be seen in a light microscope. The capsules contain usually one and occasionally
two rod-shaped virus particles. Except for their size and shape, the properties of the capsules are
the same as those of the nuclear polyhedroses. The infected tissues eventually disintegrate and
the body fluid becomes filled with inclusions.