Bacillus subtilis (Bs) and B. megaterium (Bm), Pseudomonas fluorescens (Pf) as well as Trichoderma harziamum (Th), Trichoderma vierns (Tvr)and T. viride (Tv) were applied alone as well as in combination in pot experiment for controlling M. incognita in Cowpea. Results showed that Bs, Bp and Pf had nematicidal effects against M. incognita parameters viz., J2 in soil and roots, galls and egg-masses compared to untreated check. Bm alone had highly nematicidal activity against J2 in soil, while Bs was the best against J2 in roots, galls and egg-masses. Bs + Bm in combination was effective against M. incognita parameters. The tested species of Trichoderma also reduced the parameters of M. incognita.Th alone was highly effective in reducing the tested nematode parameters, followed by Tvr and Tv treatments. Tv + Tvr highly reduced the numbers of J2 in soil and roots, while Th+Tv+Tvr significantly reduced the galls and egg-masses numbers. All treatments also increased the growth parameters of cowpea viz., shoot length, fresh and dry shoot weights, and leaf numbers.

1. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
IJEN
Application of certain bacterial and fungal species for
controlling Meloidogyne incognita parameters in
cowpea
Wafaa M.A. El-Nagdi1* and H. Abd-El-Khair2
1Nematology Laboratory, National Research Centre, Giza, Egypt, P.O. Box 12622, Cairo, Egypt.
2Plant Pathology Department, National Research Centre, Giza, Egypt, P.O. Box 12622, Cairo, Egypt.
Bacillus subtilis (Bs) and B. megaterium (Bm), Pseudomonas fluorescens (Pf) as well as
Trichoderma harziamum (Th), Trichoderma vierns (Tvr)and T. viride (Tv) were applied alone as
well as in combination in pot experiment for controlling M. incognita in Cowpea. Results showed
that Bs, Bp and Pf had nematicidal effects against M. incognita parameters viz., J2 in soil and
roots, galls and egg-masses compared to untreated check. Bm alone had highly nematicidal
activity against J2 in soil, while Bs was the best against J2 in roots, galls and egg-masses. Bs +
Bm in combination was effective against M. incognita parameters. The tested species of
Trichoderma also reduced the parameters of M. incognita.Th alone was highly effective in
reducing the tested nematode parameters, followed by Tvr and Tv treatments. Tv + Tvr highly
reduced the numbers of J2 in soil and roots, while Th+Tv+Tvr significantly reduced the galls and
egg-masses numbers. All treatments also increased the growth parameters of cowpea viz., shoot
length, fresh and dry shoot weights, and leaf numbers.
Key words: biological control, Bacillus spp., Pseudomonas fluorescens, Trichoderma spp., Meloidogyne incognita,
Cowpea.
INTRODUCTION
Cowpea is an annual herbaceous legume from the
genus Vigna. Cowpea also called black-eyed pea or
southern pea as well as it is used as a protein-rich food
crop (Sawadogo et al., 2009). The nematode attacks
cowpea resulting in reduction in quality and quantity of
yield, where the yield loss ranged from 5% to complete
loss (Sasser and Carter, 1984). The application of
synthetic pesticides is problematic due to their side effects
on the environment, concerns for public health and the
rapid development of resistance. Therefore, applications
of bio-control agents, as alternatives to chemical
insecticides are promising for they are able to efficiently kill
soil pathogens and are considered to be environmentally
sound and harmless to mammals. Biological control
becomes widely used for controlling plant parasitic
nematodes, to decrease the extent of environment
degradation and the effect of the excessive toxic
nematicide. Plant-beneficial rhizobacteria and
Trichoderma spp. have the remarkable ability to suppress
soil-borne plant pathogens, promote plant growth, and
induce systemic plant defenses (Kupferschmied et al.,
2013).Trichoderma harzianum and Pseudomonas
fluorescens, as soil treatment, were significantly effective
against M. incognita and improved the growth of green
gram plants (Haseeb et al., 2005). Application of Bacillus
subtilis also significantly reduced hatching of larvae of M.
javanica root knot nematode , where maximum inhibition
of galls was observed in cowpea.
*Corresponding author: Wafaa M.A. El-Nagdi,
Nematology Laboratory, National Research Centre, Giza,
Egypt, P.O. Box 12622, Cairo, Egypt. E-mail:
wafaaelnagdi@yahoo.com, Co-Author Email:
khairhz@yahoo.com
International Journal of Entomology and Nematology
XXXX-XXXX:ISSN© www.premierpublishers.org..7, 201September,760-070), pp.2Vol. 3(
Research Article

2. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
El-Nagdi and Abd-El-Khair 071
The treatments increased the germination of seeds of
cowpea as well as growth parameters viz., shoot length,
root length, shoot weight and root weight (Dawar et al.,
2008). Siddiqui et al. (2009) showed that
Pseudomonas caused greater inhibitory effect on hatching
and penetration of M. incognita than caused by Bacillus in
vitro and greenhouse experiment. Pseudomonas isolates
also caused greater increase in the growth of seedlings of
pea.
Pseudomonas fluorescens can achieve M. incognita
juveniles mortality to 100% at 72 hrs in vitro tests. The
bacterium was the most effective against M. incognita
reproduction and enhanced the plant growth in vivo tests
(Ashoub and Amara, 2010). P. fluorescens and B. subtilis
caused the maximum inhibition of root knots in Vigna
mungo plants inoculated with M. incognita and
simultaneously treated with 20 ml of bio-control agent
suspension. The treatments also increased the growth
parameters viz. shoot length, root length, shoot fresh and
dry weights, root fresh and dry weights (Akhtar et al.,
2012). T. harzianum significantly decreased nematode
populations in treated plots as well as significantly
increased the growth and yield parameters of Cowpea
(Izuogu et al., 2014).In pot experiment, T. harzianumplusT.
Harzianum were the most effective in reduction of
nematode reproduction over control as well as improving
plant growth in cowpea infected with M. incognita (Nama
et al., 2015).T.harzianum, T. viride and P. fluorescens, as
seed treatments, revealed the minimum number of
galls/plant, egg masses/plant, eggs/egg mass and total
nematode population and the maximum growth
parameters of Cowpea in pot experiment (Nama and
Sharma, 2017).Some of Bacillus caused J2 mortality
greater than 80% in vitro after 24 h. Seed treatment of
soybean with Bacillus caused a reduction in root-knot
galling and egg mass counts and also caused a significant
increase in shoot weight , when compared to the control
(Chinheya et al.,2017).This work is aimed to study the
nematicidal activity of certain bacterial and fungal species
for controlling Meloidogyne incognita in Cowpea in pot
experiment.
MATERIALS AND METHODS
Plant species
Seeds of Cowpea cv. baladi (Agriculture Research Center)
were sown in 15 cm diameter (five seeds per pot) pots
filled with autoclaved soil. Prior to sowing the seeds were
disinfected by 1% NaOCl for five minutes. After
germination, the seedlings were thinned to two per pot.
Extraction and Identification of M. incognita
The second stage juveniles (J2) of root-knot nematode
used throughout the study were extracted according to
Young,1954 from a pure culture maintained in eggplant
roots in the greenhouse of the National Research Centre
and identified as Meloidogyne incognita according to
Taylor and Sasser (1979).
Bio-control agents
Three of bacterial bio-control agents viz., Bacillus subtilis
(Bs), Bacillus megaterium (Bm) and Pseudomonas
fluorescens (Pf) as well as three species of fungal genus,
Trichoderma viz. T. harziamum (Th) ,Trichoderma vierns
(Tvr) and T. viride (Tv) isolated from Egyptian soil were
applied. These treatments were compared with the
nematicide, Furadan (Carbofuran 10%G ).
Preparation of bio-control agents inoculums
For the preparation of bacterial bio-control agent's
inoculums, pure culture of Bs, Bm and Pf were isolated
from the fields. The culture tubes were incubated for 48 h
at (30±2) °C for the multiplication of bacteria. For mass
production, one-liter conical flasks containing 500 ml
nutrient glucose (2%) medium (Broth) [3 g beef extract; 5
g peptone, 20 g glucose, in 1000 ml distilled water and pH
at 7.2) were autoclaved. After the flasks were cooled, each
flask was separately inoculated with 1.0 ml of the tested
bacteria cultured broth. The flasks were kept at (30±2) °C
for 48 h and were shaken two times a day. Inoculum
cultures of bacteria were mixed and adjusted to 107-109
colony forming unit (CFU)/ml using dilution method
adjusted to 107-109 colony forming unit (CFU)/ml using
dilution method.
For preparation the tested species of Trichoderma
inoculums, pure cultures of Th, Tvr and Tv isolated from
the field were maintained on PDA in Petri plates at (27±5)
°C. The propgules of each species were separately grown
on malt extract agar medium [30 g malt extract, 5 g
peptone, 15 g agar in 1000 ml distilled water] incubated at
28°C. Flasks with sorghum seeds were plugged with
cotton and sterilized by autoclaving for 30 min at 1 kg/cm²
pressure. The conical flasks containing sterilized sorghum
seeds were inoculated with 1 cm-diameter PDA discs
punched from the periphery of actively growing 5 d-old
cultures of species of Trichoderma and then, placed in an
incubator at (30±2) °C and the fungi were allowed to grow
with periodic shaking of the flasks. Species of Trichoderma
inoculums were adjusted to 3, 6X 108 propgules/ml.
Greenhouse experiment
The experiment was conducted to assess the effects of Bs,
Bm, Pf, Th, Tvr and Tv on M. incognita at greenhouse of
Department of Plant Pathology, National Research Centre,
Egypt. The plastic pots (20- cm diameter), containing 2 kg
of a sterilized mixture of sandy and loamy soil (1: 1, sandy:
loamy), were arranged according to a completely
randomized design on a bench in the glasshouse

3. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
Int. J. Entomol. Nematol. 072
maintained at 35 °C. After seed germination, each pot was
thinned to two plants. At the same time of nematode
inoculation, Cowpea plants were treated with each bio-
control agent inoculum as a mixture of bacterial cells and
cultural filtrates at the tested rate of 30 ml/pot (109 colony
forming unit (CFU)/ml). Then, the pots were inoculated (in
four holes made around the plant) with 1000 newly
hatched J2 of M. incognita. Six replicated pots were used
per treatment as well as for the control (Nematode
only).These treatments were compared with nematicide,
Furadan (Carbofuran 10% G), at the rate of 0.02g/pot
(equivalent to 10kg /Fed.).
Effects on M. incognita parameters
Effects of bio-control agents on nematode parameters
were recorded as numbers of J2 per 200g soil, they were
extracted using a sieving and decanting technique (Barker,
1985). For roots of cowpea, they were carefully uprooted
and washed thoroughly with running tap water and then
hatched numbers of J2 in roots (5g), they were extracted
by incubation method (Young, 1954). All J2 numbers of
nematodes were counted under a light microscope.
Numbers of galls and egg-masses per 5 g roots of cowpea
plants were counted (six plant roots per treatment), after 3
months of nematode inoculation.
Effects on growth parameters of Cowpea
Effects of bio-control agents on vegetative growth
parameters of cowpea, viz. shoot length, shoot fresh and
dry weights, dry root weight as well as leaf numbers were
recorded.
Statistical analysis
Data were subjected to analysis of variance using
Computer Statistical Package (CO-STATE) User Manual
Version 3.03, Barkley Co., USA, and means compared
with the Least Significant Difference (LSD) test at P = 0.05
(Snedecor and Cochran, 1980). The significance of the
factors treatment, concentration, exposure time and their
interactions were also analyzed. Nematode data were
normalized before analysis by log transformation.
RESULTS AND DISCUSSION
Application of the bacterial bio-control agents viz., Bs, Bp
and Pf , as single soil treatment together M. incognita
inoculation , reduced the nematode parameters viz.,
numbers of J2 in soil and roots, galls and egg-masses in
the ranges of 56.1-80.4% , 55.3-78.4% , 65.2-70.8% and
67.2-68.7% compared to untreated check, respectively.
The same bio-control agents, when applied in
combination, as soil treatments together to M. incognita
inoculation, reduced the above M. incognita parameters in
the ranges of 51.5-85.9%, 57.7-84.1%, 67.4-80.9% and
67.2-82.1%, respectively. Carbofuran 10%G as soil
treatment together to M. incognita inoculation, reduced
the nematode parameters by 87.8, 57.1, 79.9 and 97.1%
for J2 in soil, J2 in roots, galls and egg-masses,
respectively. Bm alone had nematicidal activity against J2
in soil, while Bs was the best against J2 in roots, galls and
egg-masses. In combined soil treatments, Bs + Bm highly
reduced the tested parameters of M. incognita, followed by
Pf+Bm, Pf+Bs+Bm and Pf+Bs, respectively. The
maximum reduction in M. incognita parameters was
obtained with Bs + Bm treatment, while the minimum
reduction was obtained with treatments of Pf + Bs for J2 in
soil and Bm for J2 in roots, galls and egg-masses. The
combined application of Bs, Bp and Pf significantly
increased the reduction of nematode parameters, than
those applied alone (Table, 1).
Treatments of Bs, Bm and Pf alone also increased the
growth parameters viz., shoot length, fresh and dry shoot
weights, and leaf numbers in the ranges of 27.2-42.1%,
5.6-33.9%, 28.6-78.6% and 16.7-25.0% compared to
untreated check, respectively, while combined treatments
increased the same growth parameters viz., 4.9-54.1% ,
0.5-95.4% , 25.0-121.4% and 33.3-66.7%, compared to
16.3 , 34.9 , 39.3 and 25.0 % with the Carbofuran 10%
treatment, respectively. Application Bs alone highly
increased the growth parameter, while combined
treatment of Pf + Bs highly increased the tested growth
parameters of Cowpea plants, than others (Table, 2).
These findings are in agreement with the results recorded
by Almaghrabi et al. (2013); Mukheriee and Sinha (2013)
and Munshid et al. (2013). They reported that P.
fluorescens suppressed M. incognita by enhancing
defense mechanism leading to induced systemic
resistance in Cowpea, where the soil treatment proved to
be more effective than foliar spray on root galling and eggs
in roots . B. subtilis and P. fluorescens reduced the
nematode parameters and improved the growth
parameters of tomato and onion plants. Pseudomonas and
Bacillus may produce the beneficial effects such as
mineralization process and specifically nitrogen uptake
that enhances the plant growth (Akhtar et al., 2012). It is
clear that, the plant growth promoting bacteria viz., Bs, Bm
and Pf play a protective role in controlling M. incognita in
the roots of Cowpea, comparing with the presence of
nematode in the soil in absence of beneficial bacteria.
These results revealed that beneficial bacteria suppressed
nematode reproduction in the rhizosphere of Cowpea
when bacteria were inoculated together with nematode.
Plants select those bacteria contributing most to their
fitness by releasing organic compounds through exudates
creating a very selective environment where diversity is
low. Bacteria are the most abundant microorganisms in the
rhizosphere, especially considering their competitiveness
in root colonization. It is may be due to plant growth

4. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
El-Nagdi and Abd-El-Khair 073
Table 1. Effect of Pseudomonas fluorescens, Bacillus subtilis and Bacillus megaterium as single or combined treatments on
densities of the root-knot nematode, Meloidogyne incognita (J2) in Cowpea plant.
Bacterial bio-control
agents
log10 numbers and reduction of M. Incognita parameters
J2 in soil J2 in roots Galls Egg-masses
log 10 Red.
%
log 10 Red.
%
log 10 Red.
%
log 10 Red.
%
Pseudomonas fluorescens (Pf) 2.77b 56.1 2.33b 61.6 1.47b 65.2 1.31b 68.7
Bacillus subtilis(Bs) 2.45c 78.7 2.08c 78.4 1.40b 70.8 1.31b 68.7
Bacillus megaterium (Bm) 2.41cd 80.4 2.39b 55.3 1.48b 66.2 1.34b 67.2
Pf + Bs 2.81b 51.5 2.37b 57.7 1.46b 67.4 1.34b 67.2
Pf + Bm 2.29de 85.6 1.92c 84.1 1.24c 80.9 1.09c 80.6
Bs +Bm 2.28de 85.9 1.92c 84.1 1.23c 80.9 1.07c 82.1
Pf +Bs +Bm 2.35cde 83.0 2.00c 82.0 1.23c 80.9 1.10c 80.6
Carbofuran 10%G 2.21e 87.8 2.37b 57.1 1.24c 79.9 1.12c 79.1
Inoculated plant 3.13a - 2.74a - 1.94a - 1.83a -
Each value presented the mean of five replicates. Means in each column followed by the same letter(s) did not significantly differ at P ≤ 0.05
according to Duncan`s multiple range test.
Table 2. Effect of Pseudomonas fluorescens, Bacillus subtilis and Bacillus megateriumas single or mixed treatments on vegetative
growth characteristics of Cowpea.
Bacterial bio-control
agents
Vegetative growth parameters
Shoot Leaf
No.
Root dry
Weight (g)Length (cm) Fresh weight (g) Dry weight (g)
Cm. Inc. % g Inc. % g Inc. % Count Inc. % g Inc. %
Pseudomonas fluorescens (Pf) 111.3bcd 27.2 20.6b 5.6 3.8 35.7 15.0 25.0 1.7 0.0
Bacillus subtilis(Bs) 124.3ab 42.1 26.1b 33.9 5.0 78.6 15.0 25.0 1.9 11.8
Bacillus megaterium (Bm) 121.0abc 38.3 21.9b 12.3 3.6 28.6 14.0 16.7 1.7 0.0
Pf + Bs 134.8a 54.1 38.1a 95.4 6.2 121.4 20.0 66.7 1.8 5.9
Pf + Bm 91.8de 4.9 25.2b 29.2 4.1 46.4 16.8 41.7 1.8 5.9
Bs +Bm 92.3de 5.5 25.3b 29.7 3.6 28.6 16.3 33.3 1.7 0.0
Pf +Bs +Bm 106.3bcde 21.5 19.6b 0.5 3.5 25.0 12.3 0.0 1.7 0.0
Carbofuran 10% 101.8cde 16.3 26.3b 34.9 3.9 39.3 15.0 25.0 2.3 35.3
Inoculated plant 87.5e - 19.5b - 2.8 - 11.5 - 1.7 -
Each value presented the mean of five replicates. Means in each column followed by the same letter(s) did not significantly differ at P ≤ 0.05 according
to Duncan`s multiple range test.
promoting rhizobacteria of Bacillus species and
Pseudomonas species had antagonistic activity against
phytopathogens and produced the plant growth promoting
substances in soil (Sivasakthi et al., 2014). Bakker et al.
(2007) and Siddiqui et al. (2009) showed that
fluorescent Pseudomonas plays a role in disease
suppression by producing siderophore-mediated
competition for iron, antibiosis, production of lytic enzymes
and induced systemic resistance. The volatile nematicidal
products of genus Bacillus also were effective against
juveniles and eggs (Huang et al., 2009).
Trichoderma species viz., Th, Tv and Tvr, as single soil
treatment together to M. incognita inoculation, reduced
the parameters of M. incognita by the ranges of 45.2-88.35
(J2 in soil) , 63.4-69.7% (J2 in roots) , 70.8-82.0% (Galls)
and 71.6-83.6% (Egg-masses). Trichoderma, in combined
treatments together to M. incognita inoculation, reduced
the same above parameters of M. incognita in the ranges
of 36.4-87.9%, 61.6-80.2%, 75.3-91.0% and 74.6- 91.1%,
respectively (Table,3). Carbofuran 10% G treatment
reduced the nematode parameters by 87.8, 57.1, 79.9 and
97.1% for J2 in soil, J2 in roots, galls and egg-masses,
respectively. Th alone was highly effective in reducing of
all tested nematode parameters, followed by Tvr and Tv
treatments. The combined treatment of Tv + Tvr highly
reduced the numbers of J2 in soil and roots , followed by
treatments of Th+Tv+Tvr, Th+Tvr and Th+Tv, respectively.
On the other hand, the combined treatment of Th+Tv+Tvr
significantly reduced the galls and egg-masses numbers,
followed by Tv+Tvr, Th+Tvr and Th+Tvr, respectively. Our
results showed that three Trichoderma species
significantly reduced the nematode parameters than the
un -infected treated plants (Nematode only).
Trichoderma species viz., Th, Tv and Tvr, as single soil
treatment together to M. incognita inoculation, reduced
the parameters of M. incognita by the ranges of 45.2-88.35
(J2 in soil) , 63.4-69.7% (J2 in roots) , 70.8-82.0% (Galls)
and 71.6-83.6% (Egg-masses). Trichoderma, in combined
treatments prior to M. incognita inoculation, reduced the
same above parameters of M. incognita in the ranges of
36.4-87.9%, 61.6-80.2%, 75.3-91.0% and 74.6- 91.1%,
respectively (Table,3). Carbofuran 10% G treatment

5. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
Int. J. Entomol. Nematol. 074
Table 3. Effect of Trichoderma harzianum, Trichoderma viride and Trichoderma vierns as single or combined treatments on
densities of the root-knot nematode, Meloidogyne incognita in Cowpea plant.
Trichoderma spp.
log 10 of numbers and reduction of M. incognita parameters
J2 in soil J2 in roots Galls Egg-masses
log 10 Red. % log 10 Red. % log 10 Red. % log 10 Red. %
Trichoderma harzianum(Th) 2.18d 88.3 2.27b 69.7 1.17cd 82.0 1.01de 83.6
Trichoderma viride (Tv) 2.87b 45.2 2.31b 63.4 1.40b 70.8 1.26b 71.6
Trichoderma vierns (Tvr) 2.45c 79.0 2.24b 68.9 1.26bc 79.9 1.08cd 82.1
Th + Tv 2.92b 36.4 2.33b 61.6 1.34b 75.3 1.21bc 74.6
Th +Tvr 2.42c 80.2 2.29b 64.9 1.24bc 79.9 1.08cd 82.1
Tv +Tvr 2.21d 87.9 2.02c 80.2 1.07d 86.5 0.89e 88.1
Th+ Tv +Tvr 2.26d 86.4 2.10c 76.9 0.88e 91.0 0.73f 91.1
Carbofuran 10% 2.21d 87.8 2.37b 57.1 1.24bc 79.9 1.12bcd 79.1
Inoculated plant 3.13a - 2.74a - 1.94a - 1.83a -
Each value presented the mean of five replicates. Means in each column followed by the same letter(s) did not significantly differ at P ≤ 0.05 according
to Duncan`s multiple range test.
Table 4. Effect of Trichoderma harzianum, Trichoderma viride and Trichoderma vierns as single or combined treatments on
vegetative growth characteristics of Cowpea.
Trichoderma spp.
Vegetative growth parameters
Shoot Leaf
No.
Root dry
Weight (g)Length (cm) Fresh weight (g) Dry weight (g)
Cm. Inc. % g Inc. % g Inc. % Count Inc. % g Inc. %
Trichoderma harzianum(Th) 108.3a 25.2 29.7a 52.3 4.3a 53.6 14.8ab 28.7 1.9ab 11.8
Trichoderma viride (Tv) 108.8a 25.8 29.6a 51.8 4.2a 50.0 14.0abc 21.7 2.5a 47.1
Trichoderma vierns (Tvr) 102.3a 18.3 26.8a 37.4 4.2a 50.0 14.5ab 26.1 1.7b 0.0
Th + Tv 90.0b 4.1 25.0ab 28.2 3.9ab 39.3 13.8abc 20.0 1.7b 0.0
Th +Tvr 91.0b 5.2 24.3ab 24.6 4.0a 42.9 12.0bc 4.4 2.1ab 23.5
Tv +Tvr 91.0b 5.2 20.0b 2.6 3.7ab 32.1 13.0abc 13.0 1.8ab 5.9
Th+ Tv +Tvr 88.0b 1.7 19.6b 0.5 3.4ab 21.4 15.2a 32.2 1.8ab 5.9
Carbofuran 10%G 101.8a 17.7 26.3ab 34.9 3.9ab 39.3 15.0a 30.4 2.3ab 35.3
Inoculated plant 86.5b - 19.5b - 2.8b - 11.5c - 1.7b -
Each value presented the mean of five replicates. Means in each column followed by the same letter(s) did not significantly differ at P ≤ 0.05 according
to Duncan`s multiple range test.
reduced the nematode parameters by 87.8, 57.1, 79.9 and
97.1% for J2 in soil, J2 in roots, galls and egg-masses,
respectively. Th alone was highly effective in reducing of
all tested nematode parameters, followed by Tvr and Tv
treatments. The combined treatment of Tv + Tvr highly
reduced the numbers of J2 in soil and roots , followed by
treatments of Th+Tv+Tvr, Th+Tvr and Th+Tv, respectively.
On the other hand, the combined treatment of Th+Tv+Tvr
significantly reduced the galls and egg-masses numbers,
followed by Tv+Tvr, Th+Tvr and Th+Tvr, respectively. Our
results showed that three Trichoderma species
significantly reduced the nematode parameters than the
un -infected treated plants (Nematode only).
Three species of Trichoderma viz., Th, Tv and Tvr, as
single treatment together to M. incognita inoculation,
increased the growth parameters of cowpea plants viz.,
shoot length (18.3-25.8%), fresh shoot weight (37.4-
52.3%), fresh shoot weight (50.0-53.6%) and leaf numbers
(21.7-28.7%). The combination of Trichoderma treatments
cleared that the shoot parameters increased to the ranges
of 1.7-5.2%, 0.5 – 28.2%, 21.4-42.9% and 4.4-32.2%,
compared to 17.7, 34.9, 39.3 and 30.4% with nematicide
treatment, respectively (Table, 4). Our results showed that
three species of Trichoderma significantly increased
growth parameters than inoculated plant.
These finding are in agreement with the results recorded
by Hasan et al. (2016). They reported that application of
Trichoderma spp. ,as seed dressing and seed coating;
compared to Carbofuran 10G reduced the number of galls
and egg masses of M. incognita in the roots of Indian
Spinach. Nama et al.(2015) indicated that T. harzianum +
T. harzianum as soil and seed treatments were found to
be the most effective in reducing M. incognita reproduction
and improving plant growth over control in Cowpea .T.
harzianum and P. Fluorescens, as soil treatment, reduced
the against root-knot nematode, M. incognita in green
gram (Haseeb et al., 2005).Trichoderma stimulated
growth of floricultural and horticultural plants .Trichoderma
increase phosphate solubility and the availability of micro
nutrients in the soil and it could promote growth of the
plants (Altmare et al., 1999).Trichoderma strains were able
to control M. javanica as separated eggs and the second
stage juveniles in sterile in vitro assays indicating that
improved proteolytic activity of the antagonist may be
important for the biological control of the nematodes. As
for plant growth promoting microorganisms, the possible
explanation of this phenomenon includes control of minor
pathogens which leads to stronger growth and nutrient
uptake (Ousley et al.,1993). It is noticeable to show that B.
subtilis, B. megaterium and P. fluorescens, as well as, T.
harziamum, T.vierns and T. viride may be useful sources
of potential biological control agents for M. incognita.

6. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
El-Nagdi and Abd-El-Khair 075
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7. Application of certain bacterial and fungal species for controlling Meloidogyne incognita parameters in cowpea
Int. J. Entomol. Nematol. 076
Accepted 11 September 2017
Citation: El-Nagdi WMA and Abd-El-Khair H (2017).
Potentials of Compost Tea of Certain Botanicals for
Minimizing Root- Knot and Reniform Nematodes Infection
and Altering Chemical Constituents in Eggplant.
International Journal of Entomology and Nematology 3(2):
070-076.
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