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European Journal of Applied Sciences – Vol. 11, No. 2

Publication Date: April 25, 2023

DOI:10.14738/aivp.112.14356.

Ghoneim, K., Tanani, M., Hassan, H. A., & Bakr, N. A. (2023). Pathogenicity of the Entomopathogenic Nematodes, Steinernema

carpocapsae and Heterorhabditis bacteriophora, against the Black Cutworm Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae).

European Journal of Applied Sciences, Vol - 11(2). 526-562.

Services for Science and Education – United Kingdom

Pathogenicity of the Entomopathogenic Nematodes, Steinernema

carpocapsae and Heterorhabditis bacteriophora, against the Black

Cutworm Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae)

Ghoneim, K.

Faculty of Science, Al-Azhar University, Cairo, Egypt

Tanani, M.

Faculty of Science, Al-Azhar University, Cairo, Egypt

Hassan, H. A.

Institute of Plant Protection Research,

Agricultural Research Center, Giza, Egypt

Bakr, N. A.

Faculty of Science, Al-Azhar University, Cairo, Egypt

ABSTRACT

The black cutworm Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is

worldwide in distribution. It is a polyphagous insect attacking nearly all vegetables

and many economic field crops in the world. The objective of the present study was

to investigate the pathogenicity of two entomopathogenic nematodes (EPNs),

Steinernema carpocapsae (Weiser) and Heterorhabditis bacteriophora (Poinar)

against 4th and 5th instar larvae of A. ipsilon. The newly moulted larvae were infected

with five concentrations of each EPN and the data were recorded at 24, 48, 72 & 96

hr post-infection. The most important results could be summarized as follows. After

infection of the 4th or 5th instar larvae with five concentrations of S. carpocapsae or

H. bacteriophora, a dose-dependent course of the total mortality rate of infected

larvae was recorded. After infection of larvae with S. carpocapsae or H.

bacteriophora, the mortality rate of infected larvae increased with the increasing

concentration of infective juveniles (IJs), at certain times of exposure. After

infection of the 4th or 5th instar larvae with S. carpocapsae, the mortality rate of

infected larvae did not increase with the increasing time interval of exposure. On

the other hand, infection of the 4th or 5th instar larvae with H. bacteriophora resulted

in an increasing mortality rate with the increasing time interval of exposure. After

infection of 4th instar larvae, LC50 values were 16 IJs/ml and 48 IJs/ml, for S.

carpocapsae and H. bacteriophora, respectively. After infection of 5th instar larvae,

LC50 values were 21 IJs/ml and 62 IJs/ml, for S. carpocapsae and H. bacteriophora,

respectively. Thus, S. carpocapsae was more pathogenic against A. ipsilon larvae

than H. bacteriophora. Also, the 4th instar larvae were more susceptible to the

nematode pathogenicity than the 5th instar larvae. A few nematode-infected 4th

instar larvae could pupate at the lower two concentrations of S. carpocapsae and H.

bacteriophora. A similar result was recorded after infection of 5th instar larvae with

S. carpocapsae. Moreover, the infected larvae with H. bacteriophora successfully

pupated at all concentrations except the highest one. In conclusion, S. carpocapsae

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Ghoneim, K., Tanani, M., Hassan, H. A., & Bakr, N. A. (2023). Pathogenicity of the Entomopathogenic Nematodes, Steinernema carpocapsae and

Heterorhabditis bacteriophora, against the Black Cutworm Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae). European Journal of Applied

Sciences, Vol - 11(2). 526-562.

URL: http://dx.doi.org/10.14738/aivp.112.14356.

may be an effective microbial control agent in the integrated pest management

program against the destructive insect pest, A. ipsilon.

Keywords: biocontrol, infectivity, mortality rate, pupae, symbiotic bacteria, virulence.

INTRODUCTION

The black cutworm Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae) is globally distributed

(Kononenko, 2003; Harrison, and Lynn, 2008; Binning et al., 2015) including tropical and

subtropical regions throughout North, Central and South America, Europe, Asia, Australasia,

Oceania, Africa, and the Middle East (Harrison, and Lynn, 2008; Binning et al., 2015; Mishra,

2020; Rodingpuia and Lalthanzara, 2021; CABI, 2021). It is a migrant and polyphagous species

known to feed on many economically important plants (Picimbon, 2020; Moustafa et al., 2021)

causing serious losses, especially in industrial plants and vegetables worldwide (Mahmoud et

al., 2016; Abdel-Hakim and El-Mandarawy, 2017; Muştu et al., 2021; Ren et al., 2023). In

addition to its obligate migratory behavior and strong flight performance, A. ipsilon has high

reproductive capacity (Mustu et al., 2021). This pest is a nocturnal insect and larvae remain

buried in the ground during the day, which hinders its viewing field is difficult (Bento et al.,

2007). Therefore, A. ipsilon is one of the most challenging economic pests when it comes to

control and management (Andersch and Schwarz, 2003; Zhang et al., 2022).

Although the IPM strategies are increasingly being developed in different parts of the world

(Veres et al., 2020), the majority of treatments for pest insects still rely exclusively on the

application of traditional insecticides (Jeschke et al., 2011; Meslin et al., 2021). The intensive

use of many broad-spectrum currently marketed insecticides usually causes serious

toxicological problems to the ecosystems (Ibarra et al., 2006; Tiryaki and Temur, 2010; Gill et

al., 2012; Chowański et al., 2014) and drastically affect the natural enemies, allowing an

exponential increase of pest populations (Demok et al., 2019) as well as adverse effects on

human health and domestic animals (Vattikonda and Sangam, 2017; Shahzad et al., 2020).

In Egypt, the control strategy of this insect pest depends mainly on the application of

conventional insecticides, particularly organophosphates, carbamates and pyrethroids

(Vattikonda and Sangam, 2017; Abd-El-Aziz et al., 2019; Ismail, 2021), to which this pest

quickly develops resistance and cross resistance (Yu et al., 2012; Fahmy, 2014; Mahmoud et al.,

2016; Ahmed et al., 2022). Also, the chemical insecticides are often not effective and remain

inadequate for the control of A. ipsilon because of its larval hiding behavior during the daylight

hours causing hidden damage in the fields (Capinera, 2001; Takeda, 2008). Moreover, the latter

instar larvae remain hidden in cracks and crevices during most of the life cycle, so chemical

control is often ineffective (Kumar et al., 2022). However, soil-applied insecticides can cause

serious ecological problems (Kumari et al., 2002; Mosleh et al., 2003) and, obviously, have no

effect on migrating adults (Zhang et al., 2022).

Therefore, various researchers and research institutions in the world have searching for new

control tools as alternatives to synthetic insecticides (Laznik and Trdan, 2012; Glare et al.,

2016). These alternative agents should be eco-environmentally safe (Liao et al., 2017; Kunbhar

et al., 2018), effective at low concentrations (Walkowiak et al., 2015) and biodegradable into

harmless compounds (Tiryaki and Temur, 2010; Li et al., 2017). One of the eco-friendly

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European Journal of Applied Sciences (EJAS) Vol. 11, Issue 2, April-2023

management strategies is the biological control of insect pests by natural enemies (parasitoids,

predators and pathogens). It is highly promising (Amutha et al., 2021; Devi et al., 2021) because

these agents are safe for humans and the environment. They have little or no effect on other

non-targeted organisms (Jagodič et al., 2019).

Among the biological control agents, entomopathogenic nematodes (EPNs) have broad

potential to kill the cutworms in soil itself (Kumar et al., 2022). They are natural enemies of soil

insect pests whose effects as a biocontrol agent against many harmful pests have been recorded

by many studies in the world (Peçen and Kepenekci, 2022). The parasitism of these beneficial

nematodes leads to the suppression of the immune system of the insect host (Lewis and Clarke,

2012; Shapiro-Ilan and Brown, 2013; Lacey et al., 2015; Kaliaskar et al., 2022). Moreover, EPNs

have an association with certain symbiotic bacteria which are carried in the intestine of

infective juveniles (IJs) of EPNs (Arthurs et al., 2004; Lewis and Clarke, 2012; Chaston et al.,

2013). The EPNs can be used individually as a biological control agent or in combination with

other biocontrol agents, such as entomopathogenic bacteria and fungi in order to improve their

efficacy for controlling the insect pests (Laznik et al., 2012). For reviews, see Vashisth et al.,

2013; Sujatha and Jeyasankar, 2018; Jagodič et al., 2019; Askary and Abd-Elgawad, 2021;

Kumar et al., 2022; Shaurub, 2023).

Out of 23 EPNs families, Steinernematidae and Heterorhabditidae are the two most famous

families living naturally in the soil (Kumar et al., 2015) and used as biocontrol agents against

diverse insect pests (Lacey and Georgis, 2012; Abd-Elgawad, 2020; Koppenhöfer et al., 2020;

Yüksel et al., 2022). Their association with symbiotic bacteria is found to be the primary cause

of insect mortality (Leonar et al., 2022). There are the genera Xenorhabdus and Photorhabdus

associating with the families Steinernematidae and Heterorhabditidae, respectively, which

produce natural products with insecticidal potential (Vicente-Díez et al., 2021) for the

suppression of the immune system of the insect host (Lewis and Clarke, 2012; Shapiro-Ilan and

Brown, 2013; Lacey et al., 2015; Kaliaskar et al., 2022). To date, more than 100 Steinernema and

16 Heterorhabditis have been described in the world (Shapiro-Ilan et al., 2017; Koppenhöfer et

al., 2020; Bhat et al., 2020).

In general, EPNs have been reported to exhibit pathogenicity against many Lepidopteran insect

pests, such as the tobacco cutworm Spodoptera litura (Rajkumar et al., 2003; Yadav et al., 2017;

Thakur et al., 2022; Javed et al., 2022), the diamondback moth Plutella xylostella (Somvanshi et

al., 2006; Kumar et al., 2022), the pink bollworm Pectinophora gossypiella (Shairra et al., 2016),

the cotton bollworm Helicoverpa armigera (Vashisth et al., 2019; Yan et al., 2021; Thakur et al.,

2022; Srivastava et al., 2022), the greater was moth Galleria mellonella (Khashaba et al., 2020;

Ali et al., 2022), the cabbage butterfly Pieris brassicae (Pervez and Rao, 2020; Tomar et al.,

2022), the tomato leaf miner Tuta absoluta (Dlamini et al., 2020), the eggplant fruit borer

Leucinodes orbonalis (Hussaini et al., 2002), the fall armyworm Spodoptera frugiperda (Caccia

et al., 2014; Acharya et al., 2020; Lalramnghaki et al., 2021; Yan et al., 2021; Fallet et al., 2022;

Mohamed and Shairra, 2023), and the Egyptian cotton leafworm Spodoptera littoralis (Yağcı et

al., 2022; Abd El Azim, 2022; Yağcı et al., 2022; Abd El Azim, 2022).

Also, EPNs have been reported to exhibit pathogenicity against different insects of various

orders, such as the Mediterranean fruit fly Ceratitis capitata (Diptera)(Malan et al., 2006;

Shaurub et al., 2021), the silverleaf whitefly Bemesia tabaci (Hemiptera)(Qiu et al., 2008), the