Research Article

Efficacy of Beauveria bassiana and Metarhizium anisopliae for the Management of Tomato Fruit Borer, Helicoverpa armigera (Lepidoptera: Noctuidae)

Abdul Basit1*, Usman Khalique2, Muhammad Salim1 and Ahmad Ur Rahman Saljoqi1

1Department of Plant Protection Faculty of Crop Protection Sciences, The University of Agriculture Peshawar, Pakistan; 2Plant Protection Division, Nuclear Institute for Food and Agriculture, Peshawar, Khyber Pakhtunkhwa, Pakistan.

Received | Oct 04 2024; Accepted | Jul 10, 2025; Published | November 05, 2025

*Correspondence | Abdul Basit, Department of Plant Protection Faculty of Crop Protection Sciences, The University of Agriculture Peshawar, Pakistan. Email: [email protected]

Citation | Basit, A., U. Khalique, M. Salim and A.U.R. Saljoqi. 2025. Efficacy of Beauveria bassiana and Metarhizium anisopliae for the management of tomato fruit borer, Helicoverpa armigera (Lepidoptera: Noctuidae). Sarhad Jurnal of Agriculture, 41(4): 1698-1709.

DOI | https://dx.doi.org/10.17582/journal.sja/2025/41.4.1698.1709

Keywords | Beauveria bassiana, Damage assessment, Helicoverpa armigera, Metarhizium anisopliae, Ovipositional preference, Tomato yield.

Copyright: 2025 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Tomato is one of the most valuable short-term vegetable crops in the world. Tomatoes are a nutrient-dense food that contains vitamins A, B, and C, minerals, amino acids, sugars, and dietary fiber (Khokhar and HRI, 2013). In 2022, the global tomato planting area was 05 million hectares, with a total output of 186.82 million tons (FAOSTAT, 2022). China is the main producer of tomatoes accounting for 34.72% of the total global production followed by India, Turkey, United States, and Egypt. Pakistan has been ranked as the 26th largest tomato producer in the world (Anonymus, 2022; Tiwari et al., 2022).

In Pakistan, tomatoes are grown in all provinces in spring and autumn, but unfortunately the net productivity is very low compared to the rest of the world. The tomato is a tropical plant that is well adapted to almost all climate zones in the world; however, environmental stress significantly reduces yield potential due to environmental stress. Abiotic stresses including high temperature, chilling, excessive light, toxic ions, water logging, wounding, osmotic shock, drought, salinity, exposure to ozone and radiation (UV-B) and biotic stresses including arthropod pests, fungal, bacterial and viral diseases are responsible for low yield in tomato crops (Kubienova et al., 2013).

Among arthropod pests, the most important pests infesting tomato crop are the fruit borer (Helicoverpa armigera), leafminer moth (Tuta absoluta Meyrick), whiteflies (Bemisia tabaci Gennadius), thrips (Frankiliniella occidentalis Pergade), aphids (Myzus persicae) and mites (Tetranychus spp) (Wakil et al., 2018). The fruit borer, H. armigera (Hübner) is a destructive pest of tomato plants severely hinders crop yield and productivity, damages developing fruit and causes 20 to 60% fruit loss (Talekar et al., 2006; Saljoqi et al., 2022).

H. armigera is a globally important pest affecting many crops, causing severe economic losses to pigeon pea, soybean, sorghum, maize, tomato, cotton, flaxseed, pea, chickpea and many vegetables (Ali et al., 2020). As it is polyphagous in nature and due to lack of environmental friendly different managing strategies, many farmers around the world especially in Pakistan rely on synthetic pesticides to cope with this destructive pest (Dagne et al., 2018). Beta-cyhalothrin and dimethoate are commonly used insecticides for the control of H. armigera in many countries (Fite et al., 2024). However, excessive use of pesticides may cause severe environmental pollution, can pose very harmful effects on beneficial organisms and human beings, increased production costs and ecological imbalances (Macharia, 2015; Khalique et al., 2018). In addition, H. armigera become resistant to nearly all commonly used conventional insecticides (Sun et al., 2019).

Pesticide selection pressure can be reduced by substituting synthetic pesticides with biopesticides in IPM strategies pointing this pest (Majeed et al., 2018). The use of EPFs (entomopathogenic fungi) as insect biocontrol agents has received worldwide attention (Majeed et al., 2018). Numerous studies have evaluated EPFs-biomolecules in integrated pest management strategies as an alternative to synthetic pesticides (Batool et al., 2022).

Biopesticides from Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metchnikoff) Sorokin (Ascomycota: Hypocreales) have been used to reduce important agricultural insect pests worldwide (Ahmad et al., 2019; Batool et al., 2022). M. anisopliae and B. bassiana were characterized for nematodes (Reinbacher et al., 2021), tomato fruit worm (Kary et al., 2022), Agrotis ipsilon (El-Hawary, 2019), and several other pests (Ahmed et al., 2023). In view of the current challenges in managing H. armigera in tomato crops, the use of entomopathogenic fungi, namely M. anisopliae and B. bassiana presents a best control. The objectives of the current research were focused on: i) to evaluate the efficacy of M. anisopliae and B. bassiana against H. armigera larvae under controlled conditions. ii) to determine the effect of M. anisopliae and B. bassiana on ovipositional preference of H. armigera on tomato plants.

Materials And Methods

The current research was conducted in bio-control laboratory, Plant Protection Division at Nuclear Institute for Food and Agriculture (NIFA), Peshawar during 2021-22. Laboratory conditions were maintained at 25±2°C temperature and 65±5% R.H. with 08:16 hrs. (L:D) photoperiod.

Insect culture

An initial stock of fruit worm larvae were collected from the chickpea fields (34.0155° N and 71.7129° E) during the months of December-February, 2021-22 from the field area of Nuclear Institute for Food and Agriculture and reared on natural diet in controlled laboratory conditions (Temp. 25±2°C; 65±5% R.H.; 08:16 h. (L:D) photoperiod) for the culture maintenance before conducting the experiment. After adult emergence, these were paired to get eggs and neonate larvae. Counted number of 3rd instar larvae or newly emerged adults (un-sexed) were used/ released in the experiments.

Plant material

Tomato plants (Lariqa cultivar) were selected for the experiment. For this, tomato nursery was obtained from Tarnab farm nursery area, Peshawar and kept under controlled conditions. Nursery was covered with transparent plastic bags to prevent any interruption. Plants with 5-7 leave stage were used for the experiment.

Fungus culture

M. anisopliae (PACER®) and B. bassiana (RACER®) manufactured by AgriLife SOM Phytopharma (India) Limited (www.agrilife.in) were obtained from University of Agriculture, Faisalabad and evaluated against fruit worm larvae and adults. Different concentrations (1×108, 5×107 and 1×107 spores/ml) of M. anisopliae and B. bassiana were prepared in distilled water and were used against 3rd instar larvae of H. armigera and ovipositional preference of their adults.

Experimental layout

Evaluation of EPF’s against tomato fruit worm larva

Laboratory experiment was carried out during April-June, 2022 to determine the efficacy of M. anisopliae, and B. bassiana against fruit worm larvae. Different concentrations of both entomopathogenic fungi were formed in distilled water and water alone was considered as a control treatment (Table 1).

 

Table 1: List of treatments and their concentrations used in the experiment.

Treatment	Active ingredient	Concentration (No. of Spores)
T1	Metarhizium anisopliae	1 × 108
T2	Metarhizium anisopliae	5 × 107
T3	Metarhizium anisopliae	1 × 107
T4	Beauveria bassiana	1 × 108
T5	Beauveria bassiana	5 × 107
T6	Beauveria bassiana	1 × 107
T7	Water

 

Each treatment were applied on five 3rd instar larvae of tomato fruit worm by topically inoculation with fresh conidial suspensions. Each larva was topically inoculated with 10 μl of conidial suspensions by using a micropipette (Eppendorf pipettes, 1-20 μl). In control treatment, larvae were treated with distilled water only. Soon after treatment application, larvae were fed with fresh natural diet which was changed daily. Completely randomized design (CRD) was followed with three replications of each treatment.

First mortality

After the application of tested EPFs (B. bassiana and M. anisopliae), the mortality data was recorded for ten days. The first mortality of tomato fruit worm larva was recorded in replications of each treatment. Larva was considered dead when it was disturbed with camel hairbrush and no sign of movement was observed.

Total corrected mortality

Mortality data of tomato fruit worm larva was recorded in replications of each treatment and corrected mortality data was designed by using Abbott formula (Abbott, 1925) as follows:

Where: n = Insect population, T = Treated, Co = Control

Fungal growth

For confirmation, dead larvae were placed in sterile sealed Petri dishes with moist filter paper to promote fungal growth. As a result of the experimental treatment, signs of mycelial growth and conidia formation on the cadavers were observed.

Pupal recovery and pupal weight

Surviving larvae were kept in petri dishes to form pupae. Sex differentiation was determined at the pupal stage based on the presence or absence of small black dots on top of the abdomen of males and females, and pupal weights were also recorded. The percentage of pupae recovery was calculated using the following formula:

Adult emergence

After pupation, pupae were placed in separate clean Petri plates using forceps and were kept in plastic containers for eclosion. Percent adult emergence (eclosion) was calculated as:

Lethal concentration and lethal time to calculate 50% mortality of fruit worm larva

All larval mortality counts were analysed by Probit using SPSS v16 software. The efficacy of tested entomopathogenic fungi was compared on the basis of median lethal concentration (LC50) and lethal time (LT50).

Regression analysis between mean mortality and tested M. anisopliae and B. bassiana

Data obtained on mortality percentage of tomato fruit worm larvae by the two fungi (M. anisopliae and B. bassiana) were used for plotting a linear relationship between the mortality percentage and the days after treatments, using different concentrations of tested fungi. Regression equations were calculated by conducting regression analysis.

Evaluation of entomopathogenic fungi against ovipositional preference

Free choice bioassay was conducted to assess the ovipositional preference of fruit worm moths on tomato plants with 5-7 leave stage. Plants were sprayed with given concentrations of M. anisopliae and B. bassiana as shown in Table 1 using suitable hand pump sprayer. Plant treated with water alone was considered as controlled treatment. Counted number of fruit worm adults were released in each replication to assess the ovipositional preference. In this bioassay, treated and untreated plants were kept in same cage. Completely randomized design was followed with three replications of each treatment. After three days, plants with eggs were removed from the cage and each plant was kept in separate cage for the data recording of following parameters:

Egg hatch rate (%)

Data regarding egg laying numbers of tomato fruit worm adults on treated and untreated plants were recorded on daily basis till the death of the adults. Egg hatch rate (%) was calculated by using the below mentioned formula:

Tomato yield

On maturity of tomato plants, data regarding number of total fruits and damaged fruits per plant were recorded for percent damage assessment of each treated and untreated tomato plants by using the following formula:

Data regarding percent yield increase over control treatment was calculated by following formula:

Where, Tt = Treated and T0 = Control.

Statistical analysis

Recorded data were analyzed by using statistical software “Statistix 8.1” through ANOVA and mean were compared by using Least Significance Difference (LSD) test at 5% level of significance. Probit analysis was performed to calculate LC50 and LT50 of tested entomopathogenic fungi by using statistical software “SPSS v16”.

Results

First mortality

M. anisopliae and B. bassiana were evaluated against the first mortality of tomato fruit worm, Helicoverpa armigera in laboratory. Experimental results showed that the treatments caused significant mortality of H. armigera larvae under controlled conditions (P < 0.05; F = 17.7). Comparison of each treatment with mean values (days) and standard errors showed that the earliest mean mortality of H. armigera i.e., 3.33 days was recorded in B. bassiana @ 1×108 spores/ml followed by B. bassiana @ 5×107 spores/ml (5.00 days), M. anisopliae @ 1×108 spores/ml (7.33 days), M. anisopliae @ 5×107 spores/ml (9.33 days), B. bassiana @ 1×107 spores/ml (10.67 days) and M. anisopliae @ 1×107 spores/ml (12.67 days) while late mortality was recorded in control treatment i.e., 15.67 days (Table 2).

 

Table 2: Mean table for first mortality (days) of H. armigera in different entomopathogenic treatments.

Sr. No.	Treatment	First mortality (days) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	7.33 ± 0.40 de
T2	M. anisopliae (5 × 107 spores/ml)	9.33 ± 0.67 cd
T3	M. anisopliae (1 × 107 spores/ml)	12.67 ± 0.88 ab
T4	B. bassiana (1 x 108 spores/ml)	3.33 ± 0.99 f
T5	B. bassiana (5 × 107 spores/ml)	5.00 ± 1.73 ef
T6	B. bassiana (1 × 107 spores/ml)	10.67 ± 0.77 bc
T7	Control (water)	15.67 ± 1.20 a
	LSD value	3.10

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Corrected mortality

M. anisopliae and B. bassiana were evaluated against the maximum corrected mortality of tomato fruit worm, H. armigera in laboratory. Experimental results showed that the treatments caused significant mortality of H. armigera larvae under controlled conditions (P < 0.05; F = 7.80). Comparison of each treatment with mean values (% mortality) and standard errors showed that the minimum corrected mortality of H. armigera i.e., 26.80% was recorded in M. anisopliae @ 1×107 spores/ml followed by B. bassiana @ 1×107 spores/ml (40.07%), M. anisopliae @ 5×107 spores/ml (46.80%), B. bassiana @ 5×107 spores/ml (66.93%) and M. anisopliae @ 1×108 spores/ml (73.53%) while maximum mean corrected mortality was recorded in B. bassiana @ 1×108 spores/ml i.e., 86.86% (Table 3).

 

Table 3: Mean table for corrected mortality (%) of H. armigera in different entomopathogenic treatments.

Sr. No.	Treatment	Corrected mortality (%) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	73.53 ± 8.67 a
T2	M. anisopliae (5 × 107 spores/ml)	46.80 ± 6.60 bc
T3	M. anisopliae (1 × 107 spores/ml)	26.80 ± 6.80 c
T4	B. bassiana (1 x 108 spores/ml)	86.86 ± 8.63 a
T5	B. bassiana (5 × 107 spores/ml)	66.93 ± 9.02 ab
T6	B. bassiana (1 × 107 spores/ml)	40.07 ± 8.60 c
	LSD value	24.99

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Pupal recovery

M. anisopliae and B. bassiana were evaluated against the pupae recovery (%) of tomato fruit worm, H. armigera from alive larvae in laboratory. Experimental results showed that the treatments have significant pupae recovery of H. armigera under controlled conditions (P < 0.05; F = 5.37). Comparison of each treatment with mean values and standard errors showed that the minimum pupae recovery (%) of H. armigera i.e., 6.67% was recorded in B. bassiana @ 1×108 spores/ml followed by M. anisopliae @ 1×108 spores/ml (20.00%), B. bassiana @ 5×107 spores/ml (26.67%), M. anisopliae @ 5×107 spores/ml (40.00%), B. bassiana @ 1×107 spores/ml (53.33%) and M. anisopliae @ 1×107 spores/ml (66.67%) while maximum pupae recovery was recorded in control treatment (water) i.e., 93.33% (Table 4).

 

Table 4: Mean table for pupae recovery (%) of H. armigera in different entomopathogenic treatments.

Sr. No.	Treatment	Pupae recovery (%) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	20.00 ± 11.55 cd
T2	M. anisopliae (5 × 107 spores/ml)	40.00 ± 11.55 bcd
T3	M. anisopliae (1 × 107 spores/ml)	66.67 ± 13.33 ab
T4	B. bassiana (1 x 108 spores/ml)	6.67 ± 6.67 d
T5	B. bassiana (5 × 107 spores/ml)	26.67 ± 17.63 cd
T6	B. bassiana (1 × 107 spores/ml)	53.33 ± 17.63 bc
T7	Control (water)	93.33 ± 6.67 a
	LSD value	38.97

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Pupal weight

M. anisopliae and B. bassiana were evaluated against the pupae weight (gm) of tomato fruit worm, H. armigera in laboratory. Experimental results showed that the treatments have significant pupae weight of H. armigera under controlled conditions (P < 0.05; F = 18.5). Comparison of each treatment with mean values and standard errors showed that the minimum pupae weight (gm) of H. armigera i.e., 0.19 gm was recorded in B. bassiana @ 1×108 spores/ml followed by B. bassiana @ 5×107 spores/ml (0.24 gm), B. bassiana @ 1×107 spores/ml (0.25 gm), M. anisopliae @ 1×108 spores/ml (0.27 gm), M. anisopliae @ 5×107 spores/ml (0.27 gm) and M. anisopliae @ 1×107 spores/ml (0.30 gm) while maximum pupae weight was recorded in control treatment (water) i.e., 0.32 gm (Table 5).

Adult emergence

M. anisopliae and B. bassiana were evaluated against the adult emergence of tomato fruit worm, H. armigera in laboratory.

 

Table 5: Mean table for pupae weight (gm) of H. armigera in different entomopathogenic treatments.

Sr. No.	Treatment	Pupae weight (mg) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	0.27 ± 0.01 b
T2	M. anisopliae (5 × 107 spores/ml)	0.27 ± 0.02 b
T3	M. anisopliae (1 × 107 spores/ml)	0.30 ± 0.00 a
T4	B. bassiana (1 x 108 spores/ml)	0.19 ± 0.00 d
T5	B. bassiana (5 × 107 spores/ml)	0.24 ± 0.01 c
T6	B. bassiana (1 × 107 spores/ml)	0.25 ± 0.00 bc
T7	Control (water)	0.32 ± 0.01 a
	LSD value	0.03

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05

 

Experimental results showed the significant adult emergence of H. armigera under controlled conditions (P < 0.05; F = 5.92). Comparison of each treatment with mean values and standard errors showed that the minimum or no adult emergence (%) of H. armigera i.e., 0.00 % was recorded in B. bassiana @ 1×108 spores/ml followed by B. bassiana @ 5×107 spores/ml (11.11%), B. bassiana @ 1×107 spores/ml (38.89%), M. anisopliae @ 1×108 spores/ml (50.00%), M. anisopliae @ 5×107 spores/ml (83.33%) and M. anisopliae @ 1×107 spores/ml (86.67%) while maximum mean adult emergence was recorded in control treatment (water) i.e., 88.89% (Table 6).

 

Table 6: Mean table for adult emergence of H. armigera in different entomopathogenic treatments.

Sr. No.	Treatment	Adult emergence ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	50.00 ± 28.87 abc
T2	M. anisopliae (5 × 107 spores/ml)	83.33 ± 8.33 ab
T3	M. anisopliae (1 × 107 spores/ml)	86.67 ± 6.67 a
T4	B. bassiana (1 x 108 spores/ml)	0.00 ± 0.00 d
T5	B. bassiana (5 × 107 spores/ml)	11.11 ± 11.11 cd
T6	B. bassiana (1 × 107 spores/ml)	38.89 ± 20.03 bcd
T7	Control (water)	88.89 ± 11.11 a
	LSD value	45.79

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Regression analysis between mean mortality and tested entomopathogenic fungi

Linear regression analysis was conducted between mean mortality of tomato fruit worm and different concentrations of M. anisopliae. The data indicates that the calculated regression equations and R2 values of the tested entomo-pathogenic i.e., M. anisopliae as follows: for T1, y = 4.5057x, R2 = 0.5938; T2, y = 3.3498x, R2 = 0.8677; T3, y = 1.6749x, R2 = 0.5867 at intercept = 0 (Figure 1). Linear regression analysis was also conducted between mean mortality of tomato fruit worm and different concentrations of B. bassiana. The data indicates that the calculated regression equations and R2 values of the tested entomo-pathogenic i.e., B. bassiana as follows: for T1, y = 4.9787x, R2 = 0.7621; T2, y = 3.422x, R2 = 0.7226; T3, y = 1.931x, R2 = 0.613 at intercept = 0 (Figure 2). Probit analysis showed that the lethal concentration (LT50) recorded for M. anisopliae and B. bassiana was 3.966 x 107 and 1.721 x 107 spores/ ml, respectively, while lethal time (LT50) recorded for M. anisopliae and B. bassiana was 10.496 and 10.230 days, respectively (Table 7).

 

Table 7: Lethal concentration (LC50) and lethal time (LT50).

Lethal concentration (LC50)
Entomo-pathogenic fungi (EPF)	n†	LC50	Fiducial Limits 95 %
Metarhizium anisopliae	45	3.97 x 107	1.17 x 107 - 1.54 x 108
Beauveria bassiana	45	1.72 x 107	1.57 x 106 - 3.56 x 107
Lethal time (LT50)
Entomo-pathogenic fungi (EPF)	n†	LT50 (days)	Fiducial Limits 95 %
Metarhizium anisopliae	45	10.49	8.28 - 16.08
Beauveria bassiana	45	10.23	7.11 - 21.69

 

† = Number of larvae used in bioassay.

 

Ovipositional preference

M. anisopliae and B. bassiana were evaluated against the ovipositional preference of tomato fruit worm, H. armigera on treated tomato plants in the laboratory. Experimental results showed that significant egg laying was recorded on treated tomato plants (P < 0.05; F =5.93). Comparison of each treatment with mean values and standard errors showed that the minimum mean egg laying i.e., 21.00 eggs was recorded in Beauveria bassiana @ 1×108 spores/ml followed by M. anisopliae @ 1×108 spores/ml (22.00 eggs.), B. bassiana @ 5×107 spores/ml (35.00 eggs), B. bassiana @ 1×107 spores/ml (49.00 eggs.), M. anisopliae @ 5×107 spores/ml (59.67 eggs) and

 

 

M. anisopliae @ 1×107 spores/ml (77.00 eggs) while maximum mean egg laying was recorded in control treatment i.e., 103.33 eggs (Table 8).

Egg hatch rate (%)

M. anisopliae and B. bassiana were evaluated against egg hatching percentage of tomato fruit worm, H. armigera in laboratory. Experimental results showed the significant egg hatch rate of H. armigera under controlled conditions (P < 0.05; F = 7.44). Comparison of each treatment with mean values and standard errors. The result showed that the minimum mean egg hatch rate i.e., 49.58% of H. armigera was recorded in B. bassiana @ 1×108 spores/ml followed by B. bassiana @ 5×107 spores/ml (56.62%.), M. anisopliae @ 1×108 spores/ml (70.37%.), B. bassiana @ 1×107 spores/ml (76.85%), M. anisopliae @ 5×107 spores/ml (79.64%.) and M. anisopliae @ 1×107 spores/ml (81.05%.) while maximum mean egg hatch rate was recorded in control treatment i.e., 86.30% (Table 9).

 

Table 8: Mean table for ovipositional preference of H. armigera on tomato plants treated with different entomopathogenic treatments.

Sr. No.	Treatment	Oviposition (Eggs) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	22.00 ± 7.21 d
T2	M. anisopliae (5 × 107 spores/ml)	59.67 ± 5.36 bc
T3	M. anisopliae (1 × 107 spores/ml)	77.00 ± 8.74 ab
T4	B. bassiana (1 x 108 spores/ml)	21.00 ± 6.56 d
T5	B. bassiana (5 × 107 spores/ml)	35.00 ± 15.09 cd
T6	B. bassiana (1 × 107 spores/ml)	49.00 ± 15.50 bcd
T7	Control (water)	103.33 ± 20.22 a
	LSD value	37.63

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Table 9: Mean table for egg hatching rate of H. armigera on tomato plants treated with different concentrations of entomopathogenic fungi.

Sr. No.	Treatment	Egg hatching (%) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	70.37 ± 2.45 bc
T2	M. anisopliae (5 × 107 spores/ml)	79.64 ± 3.88 ab
T3	M. anisopliae (1 × 107 spores/ml)	81.05 ± 7.25 ab
T4	B. bassiana (1 x 108 spores/ml)	49.58 ± 4.68 d
T5	B. bassiana (5 × 107 spores/ml)	56.62 ± 8.34 cd
T6	B. bassiana (1 × 107 spores/ml)	76.85 ± 2.29 ab
T7	Control (water)	86.30 ± 1.89 a
	LSD value	15.12

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Damage assessment (%)

M. anisopliae and B. bassiana were evaluated to check the percent damage assessment of tomato fruit worm, H. armigera in laboratory. Experimental results showed that the H. armigera larvae caused significant damage under controlled conditions (P < 0.05; F = 4.23). Comparison of each treatment with mean values and standard errors showed that the minimum mean damage assessment (%) of H. armigera i.e.,17.58% was recorded in B. bassiana @ 1×108 spores/ml followed by M. anisopliae @ 1×108 spores/ml (23.58%), B. bassiana @ 5×107 spores/ml (27.58%), B. bassiana @ 1×107 spores/ml (35.06%), M. anisopliae @ 5×107 spores/ml (36.57%.) and M. anisopliae @ 1×107 spores/ml (51.21%) while maximum mean damage assessment (%) was recorded in control treatment (water) i.e., 60.01% (Table 10).

 

Table 10: Mean table for damage assessment (%) of H. armigera in different entomopathogenic treatments.

Sr. No.	Treatment	Damage assessment (%) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	23.58 ± 2.00 cd
T2	M. anisopliae (5 × 107 spores/ml)	36.57 ± 1.93 b
T3	M. anisopliae (1 × 107 spores/ml)	51.21 ± 2.63 a
T4	B. bassiana (1 x 108 spores/ml)	17.58 ± 2.10 d
T5	B. bassiana (5 × 107 spores/ml)	27.58 ± 1.30 bc
T6	B. bassiana (1 × 107 spores/ml)	35.06 ± 6.20 b
T7	Control (water)	60.01 ± 3.55 a
	LSD value	9.72

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05.

 

Increase yield

M. anisopliae and B. bassiana were evaluated to check the increase yield (%) of tomato over control treatment. Experimental results showed the significant increase yield of tomato fruits over control under controlled conditions (P < 0.05; F = 22.3). Comparison of each treatment with mean values and standard errors showed that the minimum yield increase (%) of tomato fruits i.e., 26.04% was recorded in B. bassiana @ 1×107 spores/ml followed by M. anisopliae @ 1×107 spores/ml (27.18%), M. anisopliae @ 5×107 spores/ml (32.32%), B. bassiana @ 5×107 spores/ml (42.01%) and M. anisopliae @ 1×108 spores/ml (45.73%) while maximum yield increase (%) of tomato fruits was recorded in B. bassiana @ 1×108 spores/ml i.e., 54.40% over control treatment (Table 11).

 

Table 11: Mean table for increase yield (%) of tomato in different entomopathogenic treatments.

Sr. No.	Treatment	Increase yield (%) ± Std. error
T1	M. anisopliae (1 × 108 spores/ml)	45.73 ± 4.73 ab
T2	M. anisopliae (5 × 107 spores/ml)	32.32 ± 3.95 bc
T3	M. anisopliae (1 × 107 spores/ml)	27.18 ± 3.37 c
T4	B. bassiana (1 x 108 spores/ml)	54.40 ± 2.42 a
T5	B. bassiana (5 × 107 spores/ml)	42.01 ± 1.76 abc
T6	B. bassiana (1 × 107 spores/ml)	26.04 ± 6.27 c
	LSD value	16.91

 

Means within columns followed by the different lowercase letters are significantly different at P < 0.05

 

Discussion

The present research was conducted to evaluate the effect of B. bassiana and M. anisopliae against tomato fruit worm, H. armigera and results revealed that both fungi were virulent to H. armigera. Among the tested entomopathogenic fungi, B. bassiana was more virulent than the M. anisopliae under controlled conditions. In current study, the fastest mortality of H. armigera larvae was observed in petri dishes treated with B. bassiana at higher concentration i.e., 1×108 spores/ml. These results are align with the findings of Swathi et al. (2017), who reported that B. bassiana requires minimum time to kill H. armigera larvae compared to M. anisopliae under controlled conditions.

The results of the current experiment revealed that the highest mean corrected mortality of H. armigera larvae was recorded in M. anisopliae and B. bassiana at higher concentration. These findings are consistent with those of Douro et al. (2012), who reported maximum larval mortality of H. armigera with B. bassiana and M. anisopliae at higher concentrations. Furthermore, our results are similar to some extent with the observations of Ana et al. (2018).

In current study, the lowest mean pupal recovery, reduced pupal weight and absence of adult emergence were recorded in B. bassiana at high concentration of 1×108 spores/ml. Jarrahi and Safavi (2016) also reported the similar findings that M. anisopliae and B. bassiana exhibit virulent and adverse-effects on the biological parameters of H. armigera and causing substantial larval mortality.

Results of the current study indicated that the lowest LC50 and LT50 values were recorded for B. bassiana at higher concentration as compared to M. anisopliae. These findings are consistent with those of Swathi et al. (2017) who demonstrated that B. bassiana had lowest LC50 and LT50 values against H. armigera larvae under laboratory conditions. Several studies have reported the insecticidal activity of B. bassiana against Rhynchophorus ferrugineus (El Kichaoui et al., 2017), Plutella xylostella (Sabbour and Sahab, 2005; Xia et al., 2013), Spodoptera exigua (Wraight et al., 2010), H. armigera (Douro et al., 2012) and Bemisia tabaci (Mascarin et al., 2013).

The mycelial outgrowth and conidial formation observed on the dead cadavers of treated larval instar of H. armigera confirmed that their mortality was occurred due to the application of entomopathogenic fungi. The results are align with the findings of Gabarty et al. (2014) and Lacey et al. (2015) who reported that fungal conidia is accountable for the infection by penetrating the host cuticle. Additionally, the yield data of the current experiment showed that the tomato plants treated with B. bassiana at higher concentration achieved the highest tomato yield increase over control compared to M. anisopliae. These findings are agreed with those of Patil et al. (2018), who demonstrated that tomato plants treated with B. bassiana produced higher tomato yields as compared to M. anisopliae.

Entomopathogenic fungi significantly affect the net reproductive rate of H. armigera moths. In our study, the highest egg laying (fecundity) and egg hatching percentage (fertility) were recorded in control treatment, where only water was sprayed. These results are similar to the findings of de Souza et al. (2020), who reported that the net reproductive rate (egg laying and hatching) of H. armigera was maximum in control treatment as compared to entomopathogenic fungal isolates such as M. anisopliae and B. bassiana.

Conclusions and Recommendations

It is concluded from the above experiments that both entomopathogenic fungi i.e., B. bassiana and M. anisopliae are highly effective in terms of pathogenicity against tomato fruit worm larvae, H. armigera. Beauveria bassiana at high concentration exhibited superior performance resulting in the earliest mortality, maximum corrected mortality, minimum pupal recovery and reduced adult emergence as compared to M. anisopliae. The lowest LC50 and LT50 values were also recoded for B. bassiana than M. anisopliae. Moreover, minimum fecundity and egg hatching percentage was recorded at higher concentrations of B. bassiana and M. anisopliae. Furthermore, maximum tomato yield (%) was also recorded in tomato plants treated with B. bassiana at higher concentration as compared to M. anisopliae. It is recommended that B. bassiana and M. anisopliae at a concentration of 1×108 spores/ml be utilized for the effective management of tomato fruit worm, H. armigera in vegetable crops.

Acknowledgments

We would like to thank the Nuclear Institute for Food and Agriculture (NIFA) for providing an outstanding laboratory facilities and the support of their dedicated team. Their resources and assistance have been instrumental in the success of this research.

Novelty Statement

The novelty of this research lies in the evaluation of two entomopathogenic fungal base products, RACER® (Beauveria bassiana) and PACER® (Metarhizium anisopliae Metarhizium anisopliae) against Helicoverpa armigera, a major pest of tomato crops. This study not only explores their efficacy in reducing pest populations but also provides insights into environmentally friendly and sustainable alternatives to chemical insecticides. The findings contribute to integrated pest management (IPM) strategies, potentially offering new avenues for pest control that minimize environmental impact and pesticide resistance.

Authors Contribution

Abdul Basit: Reviewed the literature, conducted the experiments and wrote the initial manuscript draft.

Usman Khalique: Conceived the basic idea, conducted the experiments, statistical analysis and improved the manuscript draft.

Muhammad Salim: Collaborated in drafting the article.

Ahmad Ur Rahman Saljoqi: Supervised the study, helped in improving the basic idea and suggested improvements in the manuscript draft.

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this article.

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