Journal of the Saudi Society of Agricultural Sciences xxx (xxxx) xxx

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Comparative efficacy of essential oil nanoemulsions and bioproducts as

alternative strategies against root-knot nematode, and its impact on the
growth and yield of Capsicum annuum L.
Eman A. Hammad, Atef M. El-Sagheer ⇑
a
Nematode Diseases Research, Plant Pathology Research Institute, Agricultural, Research Center (ARC), Giza, Egypt
b
Agricultural Zoology and Nematology Department, Faculty of Agriculture, Al-Azhar, University, Assiut, Egypt

a r t i c l e i n f o a b s t r a c t

Article history: In a study to compare the effectiveness of some essential oil nanoemulsion and bioproducts as an alter-
Received 2 November 2021 native to chemical nematicide in control of root-knot nematode, Meloidogyne incognita parasitizing on
Revised 25 May 2022 chili pepper, Capsicum annuum L. plants, in parallel with the plant growth and yield under two varying
Accepted 9 June 2022
conditions; in vitro, greenhouse and plastic house (intended for commercial production). In an in vitro
Available online xxxx
study, basil and mint essential oil nanoemulsions (at a rate of 5%) achieved the best effects on morality
reduction percentage (100%) at droplet sizes of 48.10 nm and 97.3 nm, respectively, compared with the
Keywords:
non-treated control. In the greenhouse, Basil and Mint essential oil nanoemulsions (at 5%) (82.59% and
Essential oil nanoemulsion
Root-knot nematode
81.52%) and at 3% (77.6% and 72.15%), respectively, ranked second on population reduction, with approx-
Bio-agnates imately the effect of Bio-arc (at 2.5 g/L) (74.81%), after Nemaphose (92.7%). A similar pattern was found in
Chili pepper a plastic house with a noted positive effect on the increase in yield, where Basil and Mint essential oil
Capsicum annuum L. nanoemulsions (at 5%) achieved 113.42 and 108.23, respectively.
Ó 2022 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an
open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

1. Introduction 2014). Therefore, the combat strategy is of great importance in

Among a wide range of plant pathogens, plant-parasitic nema-
todes (PPNs) are one of the most important obligate parasitic mul-
ticellular pathogens belonging to the animal kingdom and rank
second after fungi as plant pathogens(Quist et al., 2015). Globally,
over 4,100 species of plant-parasitic nematodes have been identi-
fied, including the root-knot nematodes (RKN) (Meloidogyne sp.)
occupy an advanced rank with crop losses of about 80$US billion
annually in various crops, especially vegetables (Sikora and
Fernandez, 2005; Nicol et al., 2011). RKN species, Meloidogyne
incognita (Kofoid and White, 1919) Chitwood (1949), reported that
one of the dominant species of the Meloidogyne genus parasitizes
on the pepper crop at most production locations (Aguiar et al.,
⇑ Corresponding author.
E-mail addresses: dreman.hammad@yahoo.com (E.A. Hammad), atefelsagheer@-
azhar.edu.eg (A.M. El-Sagheer).
Peer review under responsibility of King Saud University.
Production and hosting by Elsevier
the light of the global challenges of food production (Hassan
et al., 2013; Atolani and Fabiyi, 2020). Chemical control of RKN is
the most effective treatment (Ralmi et al., 2016). Typically, nemati-
cides are applied before transplanting to suppress RKN in pepper
soil (Oka et al., 2012). But, besides effectiveness, there are some
troubling aspects; high costs, unsustainable, other than the nega-
tive consequences for humans and their domesticated animals,
groundwater, and the environment (Renn et al., 2020). Therefore,
it has become imperative to shift towards alternative strategies
that adopt a comprehensive and sustainable vision that guarantees
the reduction of nematode damage while adhering to avoiding all
the previous negatives (Singh and Mathur, 2010). Lately, it has
become clear that holistic pest management is a promising alterna-
tive, by adopting multiple applications of biological control, not
only for PPNs but for all plant pests (Noling, 2014; Barrera,
2019). In some previous studies, essential oils such as basil and
mint in normal particle size have appeared to have some effective-
ness against some species of PPNs as an alternative control strategy
(Oka, 2001; Pérez et al., 2003; Mamusa et al., 2021). However, this
form may be expensive and consume large quantities, Therefore,
the hypothesis of using essential oils as nanoemulsion forms,
which enables them to take advantage of physical properties such
as the ability to spread in the water of irrigation, rapidly adhesion

https://doi.org/10.1016/j.jssas.2022.06.002
1658-077X/Ó 2022 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Please cite this article as: E.A. Hammad and A.M. El-Sagheer, Comparative efficacy of essential oil nanoemulsions and bioproducts as alternative strategies
against root-knot nematode, and its impact on the growth and yield of Capsicum annuum L., Journal of the Saudi Society of Agricultural Sciences, https://
doi.org/10.1016/j.jssas.2022.06.002
E.A. Hammad and A.M. El-Sagheer
and penetration of roots, and stay in the soil for a long time,
besides its ability to suppress PPNs (Walia et al., 2017; Mokrini
and Bouharroud, 2019; Yang et al., 2021). In contrast to Trifolium
incarnatum L. (crimson clover or carnation), which has not received
sufficient studies, despite its proven effectiveness against some
plant pests or as an organic fertilizer (Michael Jackson and
Harrison Jr, 2008; Ralmi et al., 2016).
Therefore, the current study aims to compare some alternative
strategies to suppress RKN, M. incognita infection in chili pepper
plants by two essential oils (nanoemulsions and normal particle
size) and two bio agnates as commercial bio products nematicides,
and to estimate its reflection on growth and crop production under
greenhouse and commercial plastic house conditions.
2. Materials and methods
2.1. Propagation of tested materials
2.1.1. Preparation of essential oils
Leaves of Mint (Mentha spcata L), Basil (Ocimum basilicum L) and
flowers of carnation (Trifolium incarnatum) were collected from
experimental farms, Plant Pathology Research Institute, Agricul-
tural Research (Giza, Egypt), during 2021 and authenticated by
the Ornamental, Medicinal, and Aromatic Plant Disease Depart-
ment, Agricultural Research Center, Egypt. After collecting and
identifying, the leaves were air-dried in a dark room (26 ± 5).
500 g of the leaves were powdered, wetted by 1 L of ethanol and
water solution (70:30) (v: v). Then, using percolation equipment,
a full extraction was performed for 48 h. According to Sadraei
et al. (2003) protocol, the extract was shaken and filtered, then
using a rotary evaporator, the extract was evaporated under
reduced pressure until a semiliquid extract was obtained.
2.2. Generate of essential oil nanoemulsions
The generation of essential oil as nanoemulsion has been done
in the laboratories of the Medicinal and Aromatic Plants Research
Department, Horticulture Research Institute, Agricultural Research
Center, Egypt. 5 ml of both basil and mint essential oils, non-ionic
surfactant, tween 80 (5 ml) were slowly added. Until a uniform
mixture was formed under gentle stirring, 85 ml of water was
added. added to bring the final mixture to 100 ml, to aid in the dis-
tribution and completeness of the combination. After that, a mag-
netic stirrer was used to integrate both Basil and Mint essential oils
for 30 min. An ultrasonicator was used to sonicate the mixture,
which was separated into two halves. For 15 and 60 min at 700 W.
 
Number of dead juveniles in treatment  Number of jdead juveniles in control
Mortality inhibition % ¼
100  mortality percentage in the negative controlÞ
100 Number ofdead juveniles in control
2.3. Measurement of nanoemulsion droplet size:
At room temperature, a dynamic light scattering analysis (Zeta-
NanoZS, Malvern Instrs, UK) was used to determine the size of
nanoemulsion droplets. At 25 °C, 30 L of nanoemulsion was diluted
with 3 ml of water before testing. The mean of the Z-average of
three different batches of nanoemulsions (Basil and Mint) was
used to calculate particle size (Fig. 1a and b). This stage was carried
out in the Nanotechnology Laboratory, Regional Center for Food
and Feed, ARC, Giza, Egypt.
2
Journal of the Saudi Society of Agricultural Sciences xxx (xxxx) xxx
2.4. Droplet size by Transmission electron microscopy (TEM):
For both Basil and Mint, 20 ll of diluted samples were depos-
ited on a film-coated 200-mesh copper specimen grid for 10 min,
and the excess fluid was filtered out using filter paper using
TEM. The grid was then stained with a drop of 3% phosphotungstic
acid and allowed to dry for three minutes. The coated grid was
examined under a TEM after drying (Tecnai G20, Super twin, dou-
ble tilt, FEI, The Netherlands). At a voltage of 200 kV, the samples
were evaluated (as shown in Fig. 2a and b).
2.5. Tested bioproducts
– Bio-arc: a commercial formulation of phosphorus soluble bacte-
ria, Bacillus megaterium (25
106 cfu/g) at recommended rate of
4L./fed. (2.5 g/L of water).
– Bio-Nematon (1.15% WP) a commercial formulation of filamen-
tous fungi, Paecilomyces lilacinus containing 108 units/cm3 at
recommended rate of 2L./fed.
2.6. Chemical nematicide
A commercial product Nemaphos(organophosphate) with
active ingredient Fenamiphos at recommended rate of 6 ml/100L
water/ acre.
2.7. In vitro experiments
The inhibitory effect of the normal particle size of Mint and
Basil essential oil nanoemulsions was determined using five rates
(0.25%, 0.50%, 1%, 3%, and 5% v/v) against the second stage juveniles
(J2) of M. incognita. Hand-picked egg-masses from galled pepper
roots were incubated in sterile distilled water at room temperature
(26 + 2 °C) for 48 h, second stage juveniles were extracted, counted,
and concentrated in suspension until they reached a content of
1 ml of distilled water, approximately 100 J2 according to the pro-
tocol described by Demeure and Freckman (1981). 1 ml of juve-
niles suspension (100 J2) poured in screw-capped test tubes
which contained 5 ml of different rates of tested materials timely
prepared and incubated in room conditions at 26 ± 2 °C for 4 days
and the numbers of active and inactive juveniles were counted at
24,48 and 72 h, using nematode counting slide (Hussey, 1973).
Each treatment was replicated three times. Distilled water served
as a control. The corrected mortality percentages were calculated
according to the following formula (Schneider, and Orelli, 1947).

100
2.8. Greenhouse experiments
The nematicidal characteristics of Mint and Basil essential oil
nanoemulsions at 3% and 5% v/v rates and bioproducts (Bionema-
ton and BioArc), essential oil of Carnation at a rate of 11% (v/v)
against the root-knot nematode, M. incognita, and the influence
on plant growth parameters of chili pepper plants under green-
house conditions during the spring season 2019. One seedling of
green pepper cv. Serrano chile (20 days old) was planted in pots
diam. 20
40 cm, containing a mixture of clay and sandy soil
E.A. Hammad and A.M. El-Sagheer Journal of the Saudi Society of Agricultural Sciences xxx (xxxx) xxx

Fig. 1. Mean droplet size (nm) of essential oil nanoemulsions. (a) Basil, (b) Mint.

Fig. 2. Transmission electron micrographs of tested essential oils nanoemulsion, where shows the Droplet size of Basil (at 48.10 nm) and Mint (at 97.3 nm).

(1:1 w/w) (soil organic matter 1.61%). The pepper seedlings were
infected with 1000 newly hatched juveniles of M. incognita for
each plant after one week. Each application was repeated five
times, with the application of each Mint and Basil essential oil
nanoemulsion concentrations and essential oil of Carnation being
applied three times after inoculation (first, third, and fifth weeks),
essential oil of Carnation, as well as the same applications of Bione-
maton and BioArc, being applied three times, with nematicides
Nemaphose (6 ml/100L.watre). All treatments were applied as a
soil drench. Five inoculated pots were left without adding any
materials as a negative control, in addition to another five repli-
cated healthy seedlings without inoculation with nematodes as a
positive control. All pots were arranged in the greenhouse at
27 ± 5 °C in a randomized block design. After fifty days of inocula-
tion, the plants were harvested. Juveniles of M. incognita were
extracted from soil by sieving and the modified Baeman technique
(Seinhorst, 1962). Roots were stained by acid fuchsin (Bybd et al.,
1983) and examined under a stereomicroscope for counting devel-

3
E.A. Hammad and A.M. El-Sagheer Journal of the Saudi Society of Agricultural Sciences xxx (xxxx) xxx

opmental stages, females, galls, and egg-masses. Root galling or 3. Results

egg masses were rated on a scale of 0–5, with 0 indicating no galls
or egg masses, 1 indicating 1–2 galls or egg masses, 2 indicating 3– 3.1. Evaluation of basil and mint essential oil nanoemulsion on the
10 galls or egg masses, 3 indicating 11–30 galls or egg masses, 4 mortality of second- stage juveniles of M. incognita in vitro
indicating 31–100 galls or egg masses, and 5 indicating more than
100 galls or egg masses per root system. (Taylor and Sasser, 1978). Data in Table 1 and Fig. 3, represented the effect of Mint and
Basil essential oil nanoemulsion solutions at five rates (0.25%,
0.50%, 1%, 3%, and 5%) on the mortality of M. incognita juveniles.
2.9. Productive plastic houses experiment All the tested treatments were found to cause significant inhibition
in juvenile mortality percentage to various extents compared to
The nematicidal characteristics of Mint and Basil essential oil the non-inoculated control.
nanoemulsions at 3% and 5% v/v rates and bioproducts (Bionema- Generally, the effect of the studied essential oil nanoemulsion
ton and BioArc), essential oil of Carnation at a rate of 11% (v/v). varies depending on the concentrations and the period of exposure.
The influence on plant growth parameters of chili pepper plants Where all treatments were characterized by a stable line in terms
under greenhouse conditions was evaluated under greenhouse of the effect of the relationship between concentration and expo-
infested soil by the root-knot nematode, M. incognita, during the sure time as a steady relationship, as this provided a stable vision
spring season of 2019. of the effective value, especially in higher concentrations.
The findings revealed that the higher rates (5%) of Basil and
Mint essential oil nanoemulsions achieved the best results in the
2.10. Experimental design reduction percentage of morality (100 and 100%) at the end of
the 72-hour test. But considering the medium effect of the lowest
In the plastic house (18
40 m3), located in Qelin in Kafr El- rates (0.25%), which was the opposite of what was expected as a
Sheikh governorate. It is described as a newly reclaimed land and biomaterial, it achieved a reduction percentage of morality up to
a dry semi-desert region (sandy soil and 30 ± 5 °C) with heavily 67.94 and 69.96 respectively.
infested chili pepper plants by M. incognita.
Before the test began, the number of M. incognita in soil and
roots was estimated as the initial population independently for
each treatment according to methods described by Christie & Table 1
Evaluation of basil and mint essential oil nanoemulsion solutions on the mortality
Perry (1951), and Southey (1970). Before planting, all treatments
percentage of second stage juveniles of M. incognita at different exposure periods
were set up as a split-plot. The chemical control (Nemaphose) in vitro.
was applied at the recommended rate, and two rows of plants were
Treatments Conc.% Mortality%
lifted without being treated as a negative control. After seedling
planting, each treatment was applied three times as a soil drench m/L 24 h. 48 h. 72 h.
at two-week intervals. From seedling to the end of the plant’s eco- Basil 0.25 34.55f 55.18de 67.94i
nomic production period (95–100 days), the treatment was 0.5 45.73de 65.85cd 75.16g
1 50.20d 62.45cd 86.31e
repeated three times with both essential oil nanoemulsions and
3 75.00b 82.11b 96.60c
bio-agents. 5 100a 100a 100.0a
The number of juveniles in soil, developmental stages, galls, and
Mint 0.25 35.07f 48.63e 69.96h
egg-masses per root was counted. Eggs often randomly selected 0.5 40.46ef 60.20cd 76.66f
egg masses of each root system were also counted. The rate of 1 65.56c 60.50cd 87.30d
nematode build-up and percentages of reduction in nematode final 3 74.89b 82.00b 98.33b
population (PF) were calculated by the following formula: 5 100a 100a 100.0a
Control – 2.33i 2.60j 3.13j

Pf ¼ no: Egg  masses per root
no: Eggs per Egg  masses
þ ½ Developmental stage=root þ ½Juveniles in soil
þ ½ Adult females=root:
Each value presented the mean of three replicates.
Mean values followed by the same letter(s) did not significantly different at the 5%
level by DMRT.

 
Pf in control  Pf in treatment
Reduction % ¼
100
Pf in control

Pf
Nematode build  up ¼
Pi
Fruits progressively produced on five chili pepper plants were
picked and weighted to calculate the final yield per plant for yield
determination.

2.11. Data analysis

The data were subjected to analysis of variance (ANOVA)

(Gomez and Gomez, 1984) and Duncan’s multiple range tests in Fig. 3. Effect of progress exposure time in five tested rates of basil and mint
the form of probability (P  0.05) (Duncan, 1955) using Costas essential oil nanoemulsion solutions on juveniles of M. incognita at different
software. exposure periods in vitro.

4
E.A. Hammad and A.M. El-Sagheer Journal of the Saudi Society of Agricultural Sciences xxx (xxxx) xxx

3.2. Efficacy comparison of essential oils (nanoemulsion and normal
drop size), bioproducts and chemical nematicide, with estimation of
their reflection on growth parameters of chili pepper infested by M.
incognita under greenhouse and plastic house conditions.
3.2.1. Greenhouse assay
This study was conducted in the normal growing season (2019)
to evaluate the effect of the tested materials (Basil and Mint at 5%
and 3% Carnation, Bio-nematon, bio-arc, and Nemaphose) on the
population of M. incognita infesting chili pepper plants.
Generally, the current work showed that all tested treatments
caused a significant reduction in the root-knot nematode criteria;
numbers of root galls, build-up, and nematode reduction at varying
levels compared to control and chemical nematicide. Data pre-
sented in Tables 2 and 3 showed that all treatments resulted in a
considerable reduction in the total number of root-knot juvenile
nematodes in the soil and on the roots of chili pepper plants, with
nematode build-up rates ranging from 0.24% for Nemaphose to
1.12% for Bio-nematon. However, some notable disparities
between applications have emerged, Nemaphose provided the
greatest reduction in the nematode population (92.7%). Followed
by Basil and Mint essential oil nanoemulsions at concentration
5% (82.59 and 81.52), Basil (77.6%) essential oil nanoemulsions at
contraction 3%, carnation (75.12%), Mint essential oil nanoemul-
sions at rate 0.20, and Bio-arc (72.15 and 74.81%) in the second
and third rank, respectively. On the other hand, Bio-nematon,
had the lowest nematode population levels, with a reduction of
67.18%. A similar pattern of treatments showed in the root gall
index of M. incognita infested chili pepper roots (Table 2).
On the other side, data shown in Table 2 revealed that all of the
used treatments recorded significant increases in plant growth
(P < 0.05) compared with infected plants. As expected, the chemi-
cal nematicide (Nemaphose) ranked the highest growth parame-
ters. As expected, the chemical nematicide (Nemaphose) ranked
highest in effect on growth parameters compared with infested
and non-treated control.
However, the results of the essential oil nanoemulsions test
were promising (Table 3). Based on increasing percentages in
growth parameters; shoot and root (length and weight), statisti-
cally significant differences were noted between Basil and Mint
essential oil nanoemulsion rates in all plant growth parameters.
The highest increase in shoot dry weight was achieved through
the use of 5% of basil and mint (172.28% and 228.22%), respectively.
Nevertheless, the lower rate of Basil and Mint essential oil
nanoemulsions (3%) achieved a better reflection on plant growth
parameters (68.81% and 119.8%) compared to Bio-nematon and
bio-arc (63.86% and 76.73%), respectively.
3.2.2. Plastic house (production) assay
Table 4 compares the effectiveness of essential oil nanoemul-
sions (5%), carnation essential oil, bio-agates, and Nemaphose on
RKN population density and yield of chilli pepper plants grown
in plastic houses. Nemaphose had the greatest nematode popula-
tion reduction (89.73%), followed by Basil and Mint essential oil
nanoemulsions at 5% (80.7 and 78.56%), with the rate of nematode
build-up being the slowest (0.87 & 1.01%). On the other side, BioArc
and Bionematon had the highest rates of least value loss (70.90 and
65.21%), as well as the highest rate of nematode build-up (1.33%
and 1.56%). In terms of the root-gall index, all sprays significantly
reduced the quantity of M. incognita galls on chili pepper plants
as compared to untreated plants (control). Table 3 shows that
the tested essential oil nanoemulsions, bio-agents, and essential
oils have substantial differences in chilli pepper yield (Kg/plant).
Nemaphose essential oil nanoemulsions (131.18%) had the highest

Table 2
Efficacy comparative of essential oils nanoemulsions solutions, bioproducts and chemical nematicide on M. incognita infesting chilli pepper roots under greenhouse conditions.

Treatments R.G.I J2 in soil In root Build-up Reduction%

DS Females
3.0c 615.3 Basil (3%) 68.4 77.3 0.76 77.6
2.0d 492.3 Basil (5%) 49.3 50.2 0.59 82.59
3.2bc 782.4 Mint (3%) 76.8 87.3 0.95 72.15
2.3d 513.2 Mint (5%) 52.4 62.4 0.63 81.52
3.7abc 965.1 Bio-nematon 82.3 67.9 1.12 67.18
4.5ab 736.2 Bio-Arc 62.4 57.3 0.86 74.81
4.9a 746.2 Carnation 82.3 67.9 0.85 75.12
2.3e 193.7b Nemaphose 21.5b 32.9b 0.24 92.7
5.0a 3127.3a Control 192.50a 78.4 a 3.39 –

Each value presented the mean of three replicates.

Mean values followed by the same letter(s) did not significantly different at the 5% level by DMRT.

Table 3
Efficacy comparative of essential oils nanoemulsions solutions, bioproducts and chemical nematicide on growth parameters of chili pepper infested with M. incognita infesting
under greenhouse conditions.

Treatment Plant growth parameters

Length (cm) Plant fresh wt.(g) Shoot Dry wt. Increase%
Shoot Increase% Root Increase% Shoot Increase% Root Increase% (g)
Basil 3% 65.10e 24.42 12.95e 94.86 35.48c 99.33 8.64de 118.2 3.41d 68.81
Basil 5% 72.10c 46.54 13.72d 106.95 34.95cd 96.35 10.10b 122.8 5.5b 172.28
Mint 3% 62.2g 26.42 14.6c 120.54 33.21de 86.57 11.20a 119.63 4.44bc 119.8
Mint 5% 77.75a 58.03 16.1a 143.2 36.70b 106.18 9.6c 128.98 6.63ab 228.22
Bionematon 69.91d 42.1 12.40e 87.3 34.2d 92.13 8.91d 113.2 3.31d 63.86
Bio-Arc 73.6b 49.6 13.5d 103.93 36.31b 103.99 9.36cd 125.82 3.57c 76.73
Carnation 63.12f 28.29 15.5b 134.14 38.85a 118.26 10.38b 143.47 4.92bc 143.56
Nemaphose 69.5d 41.26 13.4d 102.42 32.67e 83.54 10.98b 115.88 7.37a 264.85
Control 49.2h – 6.62f – 17.8f 2.42e – 2.02e –

Each value presented the mean of three replicates.

Mean values followed by the same letter(s) did not significantly different at the 5% level by DMRT.

5
E.A. Hammad and A.M. El-Sagheer
Table 4
Efficacy comparative of essential oils nanoemulsions solutions, bioproducts and chemical nematicide on M. incognita infesting chili pepper roots under plastic house (production)
conditions.
Treatment Initial popul. Final popul. Reprod.
(Pi) (Pf) (Pf/Pi)
Basil (5%) 462.34a 399.95f 0.87
Mint (5%) 440.89d 444.29e 1.01
Bio-nematon 462.34a 720.94b 1.56
Bio- Arc 450.32c 603.04c 1.33
Carnation 438.89e 517.03d 1.18
Nemaphose 450.32c 212.82g 0.47
Control 453.45b 2072.27a 4.57
Each value presented the mean of three replicates.
Mean values followed by the same letter(s) did not significantly different at the 5% level by DMRT.
percentage growth, followed by Basil and Mint essential oil
nanoemulsions at 5% (113.42 and 108.23%). Carnation and Bio-
arc essential oils exhibited considerable yield improvements
(87.10%), whereas Bio-nematon essential oil produced the lowest
yield increases (47.18%).
4. Discussion
In Vitro and in Vivo investigations, the results showed that all
treatments, including Basil and Mint essential oil nanoemulsions,
essential oils Carnation, and bioproducts Bionematon and Bio-
Arc, have a positive effect on RKN. The use of 5% Basil and Mint
essential oil nanoemulsions, as well as T. incarnatum and Carnation
essential oil, resulted in a greater reduction in the number of
nematode juveniles in the soil.
The major difference between emulsion and nanoemulsion is
the size of the nanoemulsion particles. When the size of the oil par-
ticles is reduced, the emulsion’s stability improves dramatically
(Zedan et al., 1994; Anton and Vandamme, 2011). Generally, essen-
tial oils (which have low water solubility) cannot easily interact
with cell membranes because of their greater size than nanoemul-
sion particles. However, nanoemulsion particles can deliver essen-
tial oils to the surface of nematode cell membranes. This could be
related to the ability of smaller particles to kill or hinder the nema-
tode at any stage of its life cycle (Pérez et al., 2003; Barbosa et al.,
2010).
However, the efficiency mechanisms of those essential oils as
nanoemulsions or essential oils as natural nematicides are varied
(Park et al., 2005; Laquale et al., 2015). According to Zambonelli
et al. (1996), the responsibility for its antifungal activity could
be caused by chitin penetration of the cell wall, which disrupts
the lipoprotein cytoplasmic membrane, allowing cytoplasm to
escape. Or, as Mendes et al., (2018) indicated, maybe antibacterial
activity of the nanoemulsion was boosted while cytotoxicity was
reduced. The nanoemulsion with Mint essential oil extract in the
presence of chitosan displayed nematicidal action against root-
knot nematodes with low cytotoxicity in a human cell line in this
investigation (Kumar et al., 2019). Because the ideal sample had
Mint essential oil nanoemulsion extract that contained chitosan,
it is possible to speculate that the presence of chitosan can regu-
late the size, nucleation, and nematicidal activity of the
nanoemulsion. (Feizi and Kazemi, 2019). Also, the concentration
of sabinene, myrcene, and trans-caryophyllene in basil essential
oil nanoemulsions, which is a group of terpenoid chemicals, sabi-
nene plays a role in nematicidal action (Santana et al., 2014;
Bahmani et al., 2020; Sarkar, 2020). Referring to the chemical
composition of basil essential oil nanoemulsion, it is noted that
the main compounds include sabinene, myrcene trans-
caryophyllene, which could be responsible for its properties in
6
Journal of the Saudi Society of Agricultural Sciences xxx (xxxx) xxx
Red. R.G.I Yields Icreas. In yield%
% (kg./plant)
80.7 1.0d 329.3b 113.42
78.56 2.0c 321.3c 108.23
65.21 3.0b 227.1f 47.18
70.9 3.0b 264.3e 71.29
75.05 2.0c 288.7d 87.1
89.73 1.0d 356.7a 131.18
– 5.0a 154.3g –
suppressing nematodes (Abdulrazaq, 2018; Oka et al., 2000). On
the other hand, the carnation in normal particle size reduced
nematode population because it included some of the volatile
compounds with some of the permeability such as (z)-3-
hexenyl acetates, (z)-3-hexane-1–01, and (E)-and (z)-ocinenes
(Lichtenstein et al., 1962).
On the other hand, in the view of comparing essential oil
nanoemulsion with bio-agent products (Bio-nemato and Bio-
Arc) and considering that they are easier in manufacture, our
study showed a convergence of their reduction effect on RKN as
a considerable reduction in root gall index was detected on the
roots of plants infected with root-knot nematodes treated with
bio-agents. But in a variety of modes of action mechanisms,
where nematostatic effects of Bio-nemato (Purpureocillium lilaci-
nus) against RKN could be attributed to the direct penetration
of fungal hypha into eggs, juveniles, and females (Abd-Elgawad,
2021). Because of this, penetration of the eggs by hypha is facil-
itated by mechanical and/or enzymatic activity, which causes
embryos death in a relatively short period of time (Leong et al.,
2021). Moreover, it could produce leucion toxin and other
nematicidal components, which have been interpreted in a vari-
ety of ways (Walia and Devakumar, 2012). While Bio-Arc as a
bio-product of nematophagous bacteria (B. megaterium) reported
in some of the literature reviews with many different ways of
affection on RKN through all stages of life (Niu et al., 2012;
Eissa and Abd-Elgawad, 2015), some of the most important;
maybe B. megaterium extracts crude extracellular protein from
bacteria culture as a method of action against RKN (Zhou et al.,
2021); production some of the nematicidal materials, like a paras-
poral crystal toxin, have been demonstrated to infect larvae and
eggs of plant-parasitic nematodes (Wei et al., 2003); also, as a
molecular mechanism of nematophagous bacteria, it was sug-
gested that hydrolytic enzymes participate in several steps of
nematode infection (Huang et al., 2004).
5. Conclusions
From this study, it was clear that essential oil nanoemulsions
inhibited nematode increase, and the success of treatments with
basil and mint essential oil nanoemulsions compared with bio
and chemical compounds resulted in increased chili pepper growth
and yields, indicating a promising usage as an eco-friendly strategy
for nematode control.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
E.A. Hammad and A.M. El-Sagheer
Acknowledgement
The authors would like to thank Dr. Mahmoud M. Hassanin,
Research Institute, Agricultural Research Center, Egypt, for his con-
tribution to the preparation and processing of essential oil
nanoemulsions.
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