Efficacy of botanicals and entomopathogenic fungi in controlling khapra beetle (Trogoderma granarium) infestations in stored wheat

[Eficácia de botânicos e fungos entomopatogênicos no controle de infestações do besouro Khapra (Trogoderma granarium) em trigo armazenado]

ABSTRACT

The khapra beetle (Trogoderma granarium) is a notorious pest causing significant damage to stored grains, leading to substantial economic losses globally. This study investigates the insecticidal efficacy of methanolic extracts derived from two indigenous plants, Cannabis sativa and Cymbopogon citratus, alongside two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae, against T. granarium. The study assesses mortality rates, weight loss in infected wheat seeds, and adult beetle emergence. Results revealed that higher concentrations and extended exposure periods of all extracts significantly increased mortality rates while simultaneously reducing seed weight loss and adult emergence. C. sativa at 25mg/mL achieved the highest mortality rate of 78.92% ± 5% after 72 hours, with C. citratus showing a 60.52% ± 2.43% mortality rate under similar conditions. The fungi extracts, particularly at concentrations of 10⁸ spores/mL, demonstrated substantial efficacy, with B. bassiana and M. anisopliae reducing adult emergence rates to 42.11% ± 1.92% and 41.17% ± 1.92%, respectively. These findings highlight the potential of these methanolic extracts as effective biopesticides for managing T. granarium infestations in stored grains, offering a natural alternative to chemical pesticides.

Keywords:
khapra beetle; biopesticides; C. sativa; C. citratus; B. bassiana; pest control

RESUMO

O besouro khapra (Trogoderma granarium) é uma praga notória que causa danos significativos aos grãos armazenados, levando a perdas econômicas substanciais em todo o mundo. Este estudo investiga a eficácia inseticida de extratos metanólicos derivados de duas plantas nativas, Cannabis sativa e Cymbopogon citratus, juntamente com dois fungos entomopatogênicos, Beauveria bassiana e Metarhizium anisopliae, contra o T. granarium. O estudo avalia as taxas de mortalidade, a perda de peso em sementes de trigo infectadas e a emergência de besouros adultos. Os resultados revelaram que concentrações mais altas e períodos de exposição prolongados de todos os extratos aumentaram significativamente as taxas de mortalidade e, ao mesmo tempo, reduziram a perda de peso das sementes e a emergência de adultos. O C. sativa a 25 mg/mL atingiu a maior taxa de mortalidade de 78,92% ± 5% após 72 horas, com o C. citratus apresentando uma taxa de mortalidade de 60,52% ± 2,43% em condições semelhantes. Os extratos de fungos, especialmente em concentrações de 108 esporos/mL, demonstraram eficácia substancial, com B. bassiana e M. anisopliae reduzindo as taxas de emergência de adultos para 42,11% ± 1,92% e 41,17% ± 1,92%, respectivamente. Esses resultados destacam o potencial desses extratos metanólicos como biopesticidas eficazes para o manejo de infestações de T. granarium em grãos armazenados, oferecendo uma alternativa natural aos pesticidas químicos.

Palavras-chave:
besouro khapra; biopesticidas; C. sativa; C. citratus; B. bassiana; controle de pragas

INTRODUCTION

Wheat Triticum aestivum L., a cereal crop belonging to the genus Triticum, family Poaceae, is of considerable importance as a worldwide staple food (Nyaupane et al., 2024). Approximately 732 million tons of wheat are cultivated globally, with major producers including the European Union, China, India, Russia, and the United States (Houssni et al., 2024). Almost two-thirds of the world's cultivated area is dedicated to wheat (Devadoss and Ridley, 2024). Pakistan ranks among the top ten wheat-producing countries, contributing 2.6% to the total 26% of gross domestic product (GDP) from agriculture (Ashraf et al., 2020).

Wheat grain production faces threats from insect pests, diseases, and environmental variables, resulting in 10-25 percent annual losses worldwide (Iqbal et al., 2024). These challenges contribute to low productivity, as numerous insect pests and diseases cause significant damage to wheat crops in the field and during storage, leading to substantial losses in the quality and quantity of stored wheat grains (Berhe et al., 2024). The most devastating insect pests of stored wheat grains, including T. granarium, Sitotroga cerealella, Tribolium castaneum, Tribolium confusum, Sitophilus oryzae, and Ryzopertha dominica, cause 10-40% post-harvest losses globally (Mir et al., 2023; Sharma et al., 2023).

The khapra beetle, T. granarium (Coleoptera: Dermestidae), is one of the most important insect pests of stored grains, feeding on a wide range of stored products such as rice, barley, oats, and wheat (Ahmedani et al., 2007; Athanassiou et al., 2019). It can survive in hot and dry conditions, making it one of the most devastating storage pests (Morrison et al., 2020). Their feeding and infestation often cause 30-70 percent spoilage of the product, creating favorable conditions for microbial growth by producing heat and increasing humidity, which renders the grain unfit for human consumption and incurs damage expenses in the hundreds of millions of dollars (Ahmedani et al., 2007). Severe infestations in storage facilities can result in weight losses of up to 5-40%, or more than 60%, depending on the environmental conditions (Wilches et al., 2019).

In Pakistan, the management of T. granarium and other stored grain insect pests is heavily reliant on synthetic chemicals, such as permethrin, deltamethrin, spinosad, indoxacarb, and synthetic fumigants like aluminum phosphide, sulfonyl fluoride, and methyl bromide, for the management of T. granarium and other stored grain insect pests (Kavallieratos et al., 2023). While chemical pesticides remain the most efficient means of controlling significant storage pests, their use has proven hazardous for both individuals and the environment (Gad et al., 2023). Moreover, the widespread and often unnecessary application of chemical pesticides has led to increased resistance in target species (Collins and Schlipalius, 2018).

Recognition of the health and environmental hazards associated with chemical pesticides has driven a transition towards biopesticides and botanicals with insecticidal and repellent effects. Botanical insecticides offer various benefits, such as lower toxicity to mammals, environmental safety, efficacy, anisopliae and a reduced likelihood of generating resistance in arthropods (Diaz, 2016; Rizvi et al., 2024). Consequently, they have garnered significant attention for their potential in pest control, particularly in managing pests that infest stored products (Spochacz et al., 2018).

Entomopathogenic fungi have a significant impact on biological pest control worldwide (Bahadur, 2018). Metarhizium and Beauveria are commonly found in several taxonomic groups as entomopathogenic organisms (Deshpande, 1999).

Therefore, the current investigations aim to evaluate the insecticidal potential of two indigenous plant extracts C. sativa, C. citratus and two entomopathogenic fungi B. bassiana and M. anisopliae as potential biopesticides for the innovative management of T. granarium.

MATERIALS AND METHODS

The khapra beetle (T. granarium) was sourced from the Entomology Laboratory at the Uniersity of Arid Agriculture in Rawalpindi. Laboratory colonies were established and maintained on wheat grains in glass jars (15 x 6 cm) at 25 ± 2 °C and 70 ± 5% relative humidity (RH). Rubber bands secured the muslin cloth over the jar mouths to prevent the beetles from escaping.

The B. bassiana (Bb111) and M. anisopliae (Ma441) spores were obtained from the Department of Plant Pathology Arid Agriculture University, Rawalpindi, Pakistan. The spores were initially cultured by using Potato dextrose agar (PDA) at 25 °C, and the multiplication of the spores was done on an orbital shaker at 200 rpm for one week in a PDA LB broth by following the method described Iqbal et al., 2018). The conidial suspension was filtered with Whatman paper, and the spores were counted with a Neubauer hemocytometer (TIEFE 0.100 mm 1/400 9 mm), and serial concentrations of each isolate were prepared by dilution. To evaluate the pathogenicity, the following concentrations: 1×10⁸, 1×10⁷, 1×10⁶, 1×10⁵, and 1×10⁴ spores/ml suspension in 0.05% Tween 80 (v/v) were made.

We collected the leaves and stems of hemp (C. sativa) and lemongrass (C. citratus) plants from Islamabad, Pakistan. We identified the collected plants by comparing the voucher specimen submitted in the herbarium of the National Agricultural Research Centre, Islamabad Pakistan. We washed the plants with fresh water and dried them in an oven at 50°C for 24 hours. We ground the dried plant materials into a powder using an electric grinder and then sieved them to remove unwanted granules. We soaked the resulting powder in 500 mL of 70% methanol and subjected the mixture to ultrasonic shaking for 30 minutes. We then stored the mixture in a dark place for 72 hours. We filtered the plant extract solution three times using Whatman filter paper (Sigma Aldrich) to remove any solid residues. We then evaporated the filtered solution using a rotary evaporator under reduced pressure at 55°C until we obtained the crude extract. The crude extracts were transferred to glass vials and stored at 4°C until use. Material extracts were prepared by re-dissolving the crude extracts in warm distilled water. Various concentrations of 25, 12.5, 6.25, 3.125, and 1.56 mg/mL were prepared.

Uninfected wheat seeds were obtained from the National Agricultural Research Centre, Islamabad, Pakistan, and One kilogram was kept in the deep freezer at -80°C for 48 hours to avoid hidden infestation. Then, 30 grams were submerged for 30 minutes in 50 mL of each of the five concentrations of plant and fungal extracts. Seeds were dried on filter papers in Petri dishes before use, while control treatments were submerged in distilled water. A total of 30 third instar larvae were introduced from the laboratory colony to the treated seeds under conditions of 25 ± 2°C and 70 ± 5% RH. The experiment, conducted in a factorial design with three replicates per concentration, measured larval mortality at 24, 48, and 72 hours post-exposure using the formula:

$Mortality\left(\%\right)=\frac{Numberofin\sec tsdiedintreatedjars}{Totalnumberofin\sec tsreleasedinjars}X100$

Mortality was corrected using the following equation:

$CorrectedMortalityRate=\frac{(MortalityRatetreatment-MortalityRatecontrol}{1-MortalityRatecontrol}X100$

We immersed 50 grams of preserved wheat seeds in the same concentrations as before for 30 minutes, then dried them on filter paper and placed them in plastic cups. We inserted ten pairs of T. granarium into the cups, covered them with white muslin cloth, and secured them with rubber bands. We replicated each treatment three times, each containing ten pairs of insects, and left them undisturbed for four weeks under controlled conditions of 25 ± 2°C and 70 ± 5% relative humidity (RH). We then calculated the weight loss rate using the following equation:

$Weightloss\left(\%\right)=\left[\left(Initialweight-Finalweight\right)\right]/Initialweight)*100$

$EmergenceRate\left(\%\right)=\frac{TotalNumberofadultsemerged}{Totalnumberoflarvaeinitially\mathrm{int}roduced}X100$

The data were analyzed with ANOVA variance test using the SPSS-26 software (SPSS Inc., Chicago, IL). Tukey’s honestly significant difference test achieved the mean separation. The statistical difference was considered a substantial variation at P < 0 .05.

RESULT AND DISCUSSION

Some research has demonstrated that cannabis-based and lemongrass extracts possess insecticidal and repellent properties against phytophagous arthropods, although their mechanisms of action remain unclear (Benelli et al., 2018; McPartland, 1997). The current literature lacks extensive information on the biological activity of hemp and lemongrass against storage pests. This study addresses this gap by examining their effects on T. granarium, a significant pest of stored wheat grain. The results indicated that the mortality rate increased as the concentration and exposure period increased. The highest corrected mortality rate for hemp extract was 78.92% ± 5% at the highest concentration of 25 mg/ml after 72 hours of exposure, while the lowest rate was 2.43% ± 1.21% at the lowest concentration of 1.157 mg/ml after 24 hours of exposure (Table 1). Similarly, the methanolic extract of C. citratus showed protective effects on wheat seeds against T. granarium beetle infestation. Higher concentrations and longer exposure periods led to an increase in effectiveness. The highest corrected mortality rate reached 60.52% ± 2.43% at the highest concentration of 25 mg/ml after 72 hours of exposure, while the lowest rate was 6.90% ± 0.17% at the lowest concentration of 1.125 mg/ml after 24 hours of exposure (Table 2). Statistical analysis revealed significant differences between different concentrations and treatment periods. However, the C. sativa extract was more effective than the C. citratus extract in protecting wheat seeds from insect infestation. A t-test for two independent samples showed statistically significant differences between the LC50 values of the two extracts. The LC50 for C. sativa extract was 8.08 mg/ml, while the LC50 for C. citratus extract was 16.48 mg/ml (Table 3).

Table 1
Mortality rate of Khapra beetle T. granarium when treated with different concentrations of methanolic extract of C. sativa

Concentrations (mg/ml) /Days	% Mortality			F	df
	24	48	72
25	23.87±1.16a3	46.37±3.23a2	78.92±5.23a1	56.23	2
12.50	16.72±1.47b3	39.91±2.10ab2	69.46±2.85ab1	142.71	2
6.25	11.96±1.43bc3	32.22±1.93bc2	61.45±2.67bc1	143.83	2
3.13	6.00±1.28cd3	25.81±1.81cd2	50.78±2.31cd1	147.39	2
1.57	2.43±1.21d3	21.96±1.73d2	41.43±2.22d1	121.39	2
0	0±0.00e	0±0.00e	0±0.00e
F	50.137	63.55	88.35
df	5	5	5

• Mean values with different alphabets as superscript in a column are significant (P< 0.05).
• Mean values with different numbers as superscript in a row are significant (P < 0.05).

Table 2
Mortality rate of Khapra beetle T. granarium when treated with different concentrations of methanolic extract of C. citratus

Concentrations (mg/ml) /Days		Mortality%		F	df
	24	48	72
25	21.90±1.44a3	35.06±7.53a2	60.52±2.43a1	109.74	2
12.50	16.14±1.41b3	36.30±1.22a2	49.75±4.10b1	42.41	2
6.25	12.69±1.35bc2	31.53±2.04a1	37.25±1.31c1	64.54	2
3.13	9.24±1.29cd2	26.64±1.66a1	30.48±2.36cd1	44.76	2
1.57	6.90±0.17d2	20.68±1.70a1	23.54±1.61d1	42.97	2
0	0±0.00e	0±0.00b	0±0.00e
F	45.92	16.54	82.465
df	5	5	5

• Mean values with different alphabets as superscript in a column are significant (P< 0.05).
• Mean values with different numbers as superscript in a row are significant (P < 0.05).

Table 3
LD₅₀ of T. granarium when treated with plant and fungal extracts

Extracts		LD50
plant extracts	C. sativa	8.08*
	C. citratus	16.48
fungal extracts	B. bassiana	2.95 × 106*
	M. anisopliae	6.76 × 106

Other studies have reported similar results. Mantzoukas et al. (2020) Identified cannabidiol oil as the most promising substance in hemp, showing potential insecticidal effects against 4th instar larvae of Tribolium confusum and Oryzaephilus surinamensis, on wheat and rice seeds. Additionally, ethanolic hemp extract caused 42.5% mortality in Plodia interpunctella larvae after 72 hours of exposure (Prvulović et al., 2023). Moutassem et al. (2024) also demonstrated the toxicity of C. citratus plant extract against Sitophilus zeamais and T. castaneum, two significant post-harvest maize pests. Additionally, the effectiveness of C. citratus against Dermestes maculatus in fish farming was evaluated, with the highest mortality recorded for larvae at 2.5 and 5.0g (72.2% and 77.8%, respectively) and adults at 5.0 and 10.0g (22.2% and 16.7%) respectively (Ishaya et al., 2021). Mentha spicata oil showed the highest fumigant toxicity against T. granarium, causing 71.67% and 91.67% larval mortality. Ocimum basilicum and Cymbopogon shoenanthus oils showed remarkable contact toxicity against T. granarium, nearly half the efficacy of deltamethrin, a recommended insecticide for controlling stored insect pests ( Zoghroban et al., 2023), supporting our results.

Cannabis sativa and C. citratus are renowned for their bioactive compounds, making them valuable for both medicinal and commercial purposes (Oladeji et al., 2019; Pellati et al., 2018). These plants make flavones, luteolin derivatives, and terpenoids. These chemicals are essential for controlling the movement of auxin, the interactions between plants and pathogens, UV protection, and coloration (Mathesius, 2018; Oladeji et al., 2019). Notably, these plants synthesize a variety of terpenoids such as limonene, linalool, and pinene, which are effective as insecticides (García et al., 2005; Moore et al., 2014; Owolabi et al., 2020). A study by Andre et al. (2016) found that the main phytocannabinoids in industrial hemp are Δ9-tetrahydrocannabinol (THC) and cannabidiol. On the other hand, Moutassem et al. (2024), Found that C. citratus essential oil is full of geraniol, limonene, and camphene (Table 4).

Table 4
Chemical composition of C. sativa and C. citratus reported literature

No	Name	Bioactive Compounds	References
1	C. sativa	δ9-tetrahydrocannabinol, δ-8-tetrahydrocannabinol (δ8-THC), cannabidiol (CBD), and cannabinol (CBN).	(Barrales-Cureño et al., 2020; Kornpointner et al., 2021; Xu et al., 2022)
2	C. citratus	Myrcene, Neral, Geraniol, Linalool, (E)-Anethole	(Sefatlhi et al., 2024; Tazi et al., 2024a; Tazi et al., 2024b; Tazi et 2024c

The methanolic extracts of B. bassiana and M. anisopliae significantly increased mortality rates, with the highest concentration (104 mg/ml) resulting in a mortality rate of 59.09 ± 2.47% and 60.47 ± 4.66%, and the lowest concentration (10⁴ spores/ml) 32.18 ± 3.14% and 24.48 ± 2.98%, respectively, suggesting that these extracts could be beneficial for controlling T. granarium populations (Table 5 and 6). These findings are consistent with previous studies showing that these fungal strains can induce high mortality rates in the larval and adult stages of stored grain insects. For instance, Lawrence et al. (Lawrence et al., 1995) observed a significant increase in the mortality rate of Japanese beetles after three days of treatment with both B. bassiana and M. anisopliae. T. harzianum was given to adults of A. obtectus at a concentration of 2.1 × 10⁷ spores/ml, leading to 57.3% mortality after 7 days and 68.0% mortality after 21 days when given to adults of S. oryzae (Gad et al., 2020). After 14 days, 86.67% of T. granarium larvae treated with 1.0 × 10⁹ conidia/ml of local and imported isolates of M. anisopliae died, while 66.67% of larvae treated with 1.0 × 10⁹ conidia/ml of local and imported isolates of B. bassiana died (Mehdi and Al-Fadili, 2021). Another study compared M. anisopliae to T. granarium larvae and discovered that at a concentration of 1.0 × 10⁸ conidia/ml, the fungus killed 98.3% of the larvae after 14 days (Iqbal et al., 2021). The insect mortality observed may be due to secondary metabolites produced by the fungal strains, such as peptaibols, which are known to be made by T. harzianum and exhibit significant insect toxicity (Charnley and Collins, 2007; Rahim and Iqbal, 2019). Similarly, antifeeding properties of natural compounds isolated from fungal strains like T. citrinoviride have been reported against Corcyra cephalonica and Schizaphis graminum (Evidente et al., 2008; Vijayakumar and Alagar, 2017; Vijayakumar et al., 2016). The cause of insect death may also be attributed to the ability of entomopathogenic fungi to penetrate the insect cuticle and utilize the insect body as a nutrient source for growth and propagation (Iqbal et al., 2021).

Table 5
Mortality rate of Khapra beetle T. granarium when treated with different concentrations of methanolic extract of B. bassiana

Concentrations (spores /ml)	% Mortality			F	df
	5 days	6 days	7 days
108	12.91±1.13a1	35.83±2.74a2	59.09±2.47a3	107.34	2
107	8.27±2.245ab1	31.02±1.64ab2	54.00±3.37ab3	81.92	2
106	7.13±1.91abc1	23.87±1.50bc2	46.30±3.20bc3	71.10	2
105	4.79±2.235bc1	16.72±1.35cd2	41.12±2.07cd3	92.10	2
104	2.45±1.07bc1	9.57±1.19d1	32.18±3.14d2	58.32	2
0	.0000±0.00d	0±0.00e	0±0.00e
F	7.78	69.23	64.63
df	5	5	5

• Mean values with different alphabets as superscript in a column are significant (P< 0.05).
• Mean values with different numbers as superscript in a row are significant (P < 0.05).

Table 6
Mortality rate of Khapra beetle T. granarium when treated with different concentrations of methanolic extract of M. anisoplie.

Concentrations spores /ml) /Days	Mortality			F	df
	5 days	6 days	7 days
108	12.87±6.45a3	37.71±4.33a2	60.47±4.66a1	20.72	2
107	10.60±5.31a2	32.86±4.30ab1	50.26±5.51ab1	15.38	2
106	9.42±4.67a2	26.72±4.35ab12	41.18±3.34bc1	14.63	2
105	8.29±4.20a2	21.87±2.17ab1	32.13±1.85c1	16.66	2
104	5.99±3.11a2	15.73±4.37bc12	24.48±2.98d1	6.81	2
0	0±0.00a	0±0.00c	0±0.00e
F	1.02	13.64	35.91
df	5	5	5

• Mean values with different alphabets as superscript in a column are significant (P< 0.05).
• Mean values with different numbers as superscript in a row are significant (P < 0.05).

Our study evaluated the effect of C. sativa and C. citratus extracts on the weight loss rate of wheat seeds infected with T. granarium. Fig. 1 displays the results. The rate of weight loss for seeds treated with C. citratus decreased from 54.67% at the control treatment to 9.33% at 25 mg/mL. Similarly, for C. sativa, the weight loss reduced from 56% at the control treatment mg/mL to 15.33% at 25 mg/mL. The statistical analysis results showed statistically significant differences between the different concentrations (Fig. 1). Various reports on the activity of hemp against phytophagous pests support these findings. For instance, Park et al. (2019) investigated the defensive role of cannabidiol in a feeding preference assay with Manduca sexta. The larvae preferred hemp tissue with low cannabidiol content over high cannabidiol content, actively avoiding the high cannabidiol diet. These results indicate that cannabidiol has a defensive role against pest insects, suggesting its potential use as both an insecticide and a repellent.

The distinct bioactive compounds present in each plant species may be responsible for the observed insecticidal activity and varying efficiencies of the tested essential oils (Moutassem et al., 2024). Our study highlighted the emergence rate of adult insects at various concentrations. For C. sativa, the emergence rate of adult T. granarium decreased from 82.051% ± 6.93 in the control to 42.11% ± 1.92 at 25 mg/mL. For C. citratus, the emergence rate decreased from 88.23% ± 10 in the control to 41.17% ± 1.92 at 25 mg/mL. Both extracts significantly inhibited the emergence of adult beetles, with higher concentrations proving more effective (Table 7). Previous studies have demonstrated that natural compounds can induce symptoms associated with neurotoxic activity, including hyperactivity, seizures, and tremors, often followed by paralysis and death (Kostyukovsky et al., 2002). Additionally, Abdelgaleil et al. (2016) reported that EOs and their significant components strongly inhibit acetylcholinesterase (AChE) in related insect pest species, such as S. oryzae.

Figure 1
Rate of weight loss of wheat seeds infected with T. granarium and treated with plant extracts C. sativa and C. Citratus.

Table 7
Rate of emergence of adult T. granarium when raised on wheat seeds treated with plant extracts C. sativa and C. citratus

Concentration (mg/mL)	Emergence Rate (%)±S.E
	C. sativa	C. citratus
25	42.11±1.92 a	41.17±1.92 c
12.5	44±5.091 b	52.94±3.33bc
6.25	47.058±6.66bc	58.82 ±3.84abc
3.12	51.28±3.33bc	64.70±5.091abc
1.56	58.97±1.92bc	73.52±5.091ab
Control	82.051±6.93 c	88.23±10 a

• Mean values with different alphabets as superscript in a column are significant (P< 0.05).

The results also showed that methanolic extracts of B. bassiana and M. anisopliae significantly reduced weight loss at higher concentrations. At the highest concentration (10⁸ spores/ml), weight loss was reduced to 35% for B. bassiana and 33% for M. anisopliae, compared to 40% and 38%, respectively, in the control group (figure 2). Hassuba et al. (2024) and other studies before this one showed that M. anisopliae, T. citrinoviride, and T. harzianum significantly reduced the production of adult F1 and F2 spores of T. granarium at the highest concentration that was tested (2.0 × 10⁹ spores/ml). The present study supports these findings, with emergence rates reduced to 42.11% and 41.17% for B. bassiana and M. anisopliae, respectively, at 10⁸ spores/ml (Table 8). Furthermore, the highest concentration of tested fungi effectively protected wheat grains from T. granarium damage, with B. bassiana and M. anisopliae reducing weight loss to 12% and 10%, respectively, after 7 days. Similarly, studies by Gad et al., (2020); Abdelgaleil et al., (2021); Hassuba et al., (2024), have demonstrated the effectiveness of these fungi in preventing damage to stored commodities from other insect species.

Figure 2
Rate of weight loss of wheat seeds infected with T. granarium and treated with fungi extracts B. bassiana and M. anisoplie.

Table 8
Rate of emergence of adult T. granarium when raised on wheat seeds treated with fungi extracts B. bassiana and M. anisoplie

Concentration (spores/mL)	Emergence Rate (%)±S.E
	B. bassiana	M. anisoplie
108	42.11±1.92 a	41.17±1.92 c
107	44±5.091 b	52.94±3.33bc
106	47.058±6.66bc	58.82 ±3.84abc
105	51.28±3.33bc	64.70±5.091abc
104	58.97±1.92bc	73.52±5.091ab
Control	82.051±6.93 c	88.23±10 a

• Mean values with different alphabets as superscript in a column are significant (P< 0.05).

CONCLUSION

Finally, our research shows that C. sativa and C. citratus extracts, along with B. bassiana and M. anisopliae fungi, could be used as effective biopesticides against T. granarium. These biopesticides had high kill rates against larvae, stopped infested wheat seeds from losing weight, and stopped adult beetles from emerging. C. sativa was slightly more effective than C. citratus at controlling beetle populations, while B. bassiana and M. anisopliae were about the same. These findings highlight the promise of biopesticides in integrated pest management strategies, offering sustainable alternatives to synthetic chemicals with potential benefits for both agricultural productivity and environmental health. Future research should focus on optimizing application methods and exploring synergistic effects with other control measures to maximize their practical effectiveness in farm settings.

ACKNOWLEDGEMENTS

The authors sincerely thank the Researchers Supporting Project number (RSP2024R112), King Saud University, Riyadh, Saudi Arabia.

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• FUNDING
  This project was funded by Researchers Supporting Project number (RSP2024R112), King Saud University, Riyadh, Saudi Arabia.

Publication Dates

• Publication in this collection
  21 Feb 2025
• Date of issue
  Mar-Apr 2025

History

• Received
  13 Aug 2024
• Accepted
  16 Sept 2024