Volume 04 Issue 01-2024
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International Journal of Advance Scientific Research
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VOLUME
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A
BSTRACT
This research paper is concerned with determining the biocontrol potential of entomopathogenic
nematodes against the Comstock worm in laboratory conditions, presents an optimal standard research
design for the use of virulent species of beneficial nematodes against the pest, and provides information
on the stages of experimental work organization. This scientific information is important in bringing safe
biological control technology against Comstock worm to the field workers in our country.
K
EYWORDS
Comstock worm, e ntomopathogenic nematodes, h eterorhabditis spp, steinernema spp, white trap,
suspension.
I
NTRODUCTION
In Uzbekistan, the integrated control system is
widely used in the fight against pests of
agricultural crops. Its essence is to protect crops
by using biological control methods, which are
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Research Article
THE IMPORTANCE OF USING ENTOMOPATHOGENIC
NEMATODES AGAINST THE MEALYBUG (PSEUDOCOCCUS
COMSTOCKI KUW.) PEST
Submission Date:
January 01,
2024,
Accepted Date:
January 03, 2024,
Published Date:
January 05, 2024
Crossref doi:
https://doi.org/10.37547/ijasr-04-01-05
Kurbonova N.S
Research Institute Of Plant Quarantine And Protection, Uzbekistan
Dusmatova D.
Research Institute Of Plant Quarantine And Protection, Uzbekistan
Karimova R.M
Research Institute Of Plant Quarantine And Protection, Uzbekistan
Volume 04 Issue 01-2024
31
International Journal of Advance Scientific Research
(ISSN
–
2750-1396)
VOLUME
04
ISSUE
01
Pages:
30-35
SJIF
I
MPACT
FACTOR
(2021:
5.478
)
(2022:
5.636
)
(2023:
6.741
)
OCLC
–
1368736135
harmless to the environment, people and animals,
rationally using agrotechnical, mechanical, and
physical control methods, and using chemical
methods as an exception due to necessity.
The advantages of the biological control method
are that the cultivation of agricultural products as
far as possible free of chemical preparations, that
is, pesticides, serves not only human health, but
also the cleanliness of the environment and the
preservation of many types of useful living
creatures living in nature.
For this purpose, in the Republic of Uzbekistan,
the first stage experiments of the use of
entomopathogenic nematodes (EPN), which are
considered useful and low-cost and are being put
into production, were conducted in laboratory
conditions.
EPNs were first identified by Steiner in 1923
under the name Aplectana kraussei (now
Steinernema kraussei). Today, 96 Steinernema, 1
Neosteinernema, 21 Heterorhabdites species
have been identified. They are classified mainly in
the closely related families Heterorhabditidae
and Steinemematidae.
In order to use EPNs against the Comstock worm,
first of all, the step of extracting EPNs from soil
samples was carried out. For this, soil samples
were taken from the greenhouse where cultural
and decorative flowers are grown belonging to
the household in Binokor Kurgan, Nyi Hayat
District. Soil samples taken from a depth of 30 cm
were placed in 500 ml plastic bottles and covered
with lids.
The obtained soil sample was brought to the
laboratory, emptied into trays and cleaned of
unnecessary waste. Soil moisture should be 60-
70% to extract EPNs from the soil. Distilled water
was sprinkled using a water sprayer in order to
create the required moisture in the soil.
Moistened soil was put back into pots, and 15 wax
moth worms were placed on it. Jars were covered
with lids and labels were attached. The label
indicated the address of the soil sample brought
and the date of the experiment. The finished
samples were left in a thermostat at a
temperature of 20 0 C for four days.
After the specified day, the soil samples placed in
a thermostat for the purpose of extracting EPNs
were examined in order to determine the level of
infection of the wax moth with EPNs. Soil samples
in 500 ml plasma jars were poured into a tray,
wax moth worms infected with EPNs were
isolated and placed in petri dishes filled with
water. Live wax moth larvae that had not been
infected with EPNs were left in the soil, re-potted
and placed back into the thermostat.
White trap method was used to isolate nematodes
from wax moth worms infected with EPNs. For
this, a jar lid was placed on a petri dish and a filter
paper was placed on it. With the help of tweezers,
the wax moth worms were arranged in a circular
pattern and distilled water equal to half of the
petri dish was added for evaporation. It was
covered and placed in a thermostat at a
temperature of 20 0 C. After each experiment, a
label was attached and the date of the experiment
was recorded.
Volume 04 Issue 01-2024
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International Journal of Advance Scientific Research
(ISSN
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VOLUME
04
ISSUE
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Pages:
30-35
SJIF
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(2021:
5.478
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(2022:
5.636
)
(2023:
6.741
)
OCLC
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1368736135
Figure 1. White trap method entomopathogenic nematodes incubation to do
Seven days later, the experiment placed in the "White Trap" was removed from the thermostat and studied
under a microscope to determine the level of infestation of wax moth worms with EPNs, and it was found
that EPNs of various stages were developing. About 30-40 ml of distilled water was added to the isolated
EPNs to form a suspension ready for use against Comstock worm.
Figure 2. Separately received entomopathogenic nematodes isolates
In order to use the prepared suspension against
Comstock worm in laboratory conditions, filter
paper was placed in 3 petri dishes, and 10
Comstock worm larvae were placed in each petri
dish. For the first experiment, Heterorhabditis
spp was pipetted from a suspension of 2 ml of
EPN and inoculated onto Comstock worm larvae.
For the second experiment, Steinernema spp was
withdrawn from the suspension of EPN using a 2
ml pipette and inoculated onto Comstock larvae.
As the third experiment was taken as a control
option, only distilled water was sprayed. The date
Volume 04 Issue 01-2024
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International Journal of Advance Scientific Research
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VOLUME
04
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Pages:
30-35
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of the experiment and the name of the EPN used
for the lesion were recorded on top of all three
experiments. It was left for 2 days in an
environment with a room temperature of 28 0 C.
Figure 3. Local entomopathogenic nematodes using Comstock worm laboratory conditions
damage
After the specified period, the experiment was
controlled. Heterorhabditis spp . When examining
the first experiment conducted on the basis of
EPN suspension, it was found that 5 out of 10
larvae of Comstock worms of different ages were
dead. The dead larvae were 1st, 2nd, and 3rd
instar larvae, which were black in appearance and
had broken side growths. The remaining large,
mature worms were observed to be barely
moving.
For the second experiment, a suspension
prepared on the basis of Steinernema spp EPN
was used, and this experiment was also studied in
turn. 3 out of 10 Comstock worms at different
stages were dead, and they were young larvae. 2
larvae of the 3rd age and 5 worms in the mature
imago state were moving in the market.
A third control was taken and when the normal
distilled water spray experiment was examined,
each of 10 larvae of different ages were alive, and
some imago-stage Comstock worms were found
to have laid eggs.
To continue the experiment, 1 ml of the
suspensions prepared on the basis of the EPNs
corresponding to the experimental options 1 and
2 were withdrawn and sprayed with a pipette.
Distilled water was used in 3 control experiments.
Each experiment was covered and left for another
2 days in a room with a temperature of 28 0 C.
When the first experiment based on the
suspension of Heterorhabditis spp EPN was
examined after the specified period, it was found
that all Comstock worms were killed.
Volume 04 Issue 01-2024
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International Journal of Advance Scientific Research
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VOLUME
04
ISSUE
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Pages:
30-35
SJIF
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(2021:
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(2022:
5.636
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6.741
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OCLC
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1368736135
When examining the second experiment
conducted on the basis of the suspension of
Steinernema spp EPN, it was observed that 2 of
the adult Comstock worms were moving and the
rest of the worms were dead.
A control experiment using plain distilled water
showed that Comstock worms of all stages were
alive and could be seen moving freely.
A laboratory experiment to determine the
effectiveness of EPNs against the Comstock worm
can be concluded that Heterorhabditis spp EPNs
are the most effective species. To get a general
and clear conclusion, two more repetitions of the
laboratory experiment are required. Research
processes aimed at identifying the most effective
EPNs against the Comstock worm are being
carried out in stages.
R
EFERENCES
1.
Rubén Blanco-Pérez, Francisco Ángel Bueno-
Pallero , Ignacio Vicente- Díez , Vicente
Santiago Marco -Mancebón , Ignacio Pérez-
Moreno, Raquel Campos-Herrera, Scavenging
behavior and interspecific competition
decrease
offspring
fitness
of
the
entomopathogenic nematode Steinernema
feltiae , Journal of Invertebrate Pathology,
Volume 164, 2019, Pages 5-15, ISSN 0022-
2011,
https://doi.org/10.1016/j.jip.2019.04.002 .
2.
Randy Gaugler , Ramon Georgis , Culture
method and efficacy of entomopathogenic
nematodes ( Rhabditida : Steinernematidae
and Heterorhabditidae ), Biological Control,
Volume 1, Issue 4, 1991, Pages 269-274, ISSN
1049-9644, https://doi. org/10.1016/1049-
9644(91)90077-D .
3.
Kaya HK, Gaugler R. 1993. Entomopathogenic
nematodes. Annual Review of Entomology,
38: 181
–
206.
4.
Poinar GO 1990. Taxonomy and biology of
Steinernematidae and Heterorhabditidae . In:
Gaugler R., Kaya HK (Eds.) Entomopathogenic
Nematodes in Biological Control, Boca Raton
(FL): CRC Press, pp. 23
–
60.
5.
Hominick WM 2002. Biogeography. In:
Gaugler
R.
(Ed.)
Entomopathogenic
Nematology.
Wallingford,
UK:
CABI
Publishing, pp. 115-143.
6.
Hominick WM, Briscoe BR 1990. Occurrence
of entomopathogenic nematodes ( Rhabditida
: Steinernematidae and Heterorhabditidae ) in
British soil. Parasitology, 100: 295
–
302.
7.
Griffin CT, Moore JF, Downes MJ 1991.
Occurrence of insect-parasitic nematodes (
Steinernematidae , Heterorhabditidae ) in the
Republic of Ireland. Nematologica , 37: 92
–
100.
8.
Stock SP, Pryor BM, Kaya HK 1999.
Distribution of entomopathogenic nematodes
( Steinernematidae and Heterorhabditidae )
in natural habitats in California, USA.
Biodiversity and Conservation, 8: 535
–
549.
9.
Grewal PS, Ehlers RU., Shapiro- Ilan DI 2005.
Nematodes as Biological Control Agents.
Wallingford: CABI Publishing.
10.
Koppenhofer AM, Fuzy EM 2006. Nematodes
for white grub control: Effects of soil type and
soil moisture on infectivity and persistence.
Volume 04 Issue 01-2024
35
International Journal of Advance Scientific Research
(ISSN
–
2750-1396)
VOLUME
04
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01
Pages:
30-35
SJIF
I
MPACT
FACTOR
(2021:
5.478
)
(2022:
5.636
)
(2023:
6.741
)
OCLC
–
1368736135
USGA Turfgrass Environmental Research
Online, 5: 1
–
10.
11.
Lacey LA, Georgis R. 2012. Entomopathogenic
nematodes for control of insect pests above
and below ground with comments on
commercial
production.
Journal
of
Nematology, 44(2): 218
–
225.
12.
Kurbonova NS (2023): Can entomopathogenic
nematodes be the best measure for growing
eco-friendly agricultural products? American
Journal of Applied Science and Technology.
ISSN-2771-2745. Sjifimpact Factor (2023:
7.063). Issue: Vol. 3 No. 07: Volume 03 Issue
07
|
Pages:
23-31
Crossref
DOI:
https://doi.org/10.37547/ajast/Volume03
Issue07-06 .
13.
Kurbonova NS (2023): First report on local
entomopathogenic nematode Steinernema
feltiae in Uzbekistan. International Journal of
Advance Scientific Research, 3(07), 225
–
235.
https://doi.org/10.37547/ijasr-03-07-38 .
