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Article

Behavioral Suppression and Rapid Lethality: Beauveria bassiana B4 Targets Adult Monochamus alternatus for Sustainable Management of Pine Wilt Disease

1
College of Forestry, Henan Agricultural University, Zhengzhou 450046, China
2
Henan Academy of Forestry, Zhengzhou 450002, China
3
Xixia Forestry Development Service Center, Nanyang 474500, China
4
School of Plant Protection, Anhui Agricultural University, Heifei 230036, China
5
Puyang Forestry Technology Workstation, Puyang 457099, China
*
Author to whom correspondence should be addressed.
Insects 2025, 16(10), 1045; https://doi.org/10.3390/insects16101045 (registering DOI)
Submission received: 23 August 2025 / Revised: 9 October 2025 / Accepted: 10 October 2025 / Published: 12 October 2025

Simple Summary

Pine wilt disease is a devastating forest disease that kills millions of pine trees worldwide. It is spread by the pine sawyer beetle. While previous efforts have targeted beetle larvae, they live deep inside the wood and are hard to reach. Instead, we focused on controlling the adult beetles that fly freely in the forest. Control of these beetles has relied on chemical insecticides, which can harm the environment. In this study, we tested a natural and environmentally friendly alternative: a fungus that kills insects. We found a specific type, called Beauveria bassiana strain B4, that is very effective at quickly killing the adult beetles. We also observed that infected beetles stop eating and moving long before they die, which means they likely stop spreading the disease. To use this in the forest, we created a special bag that slowly releases the fungus over time. When we hung these bags in the forest, they successfully attracted and killed beetles, reducing their numbers by over 66% in one month, and the effect lasted for more than a year. This method provides a sustainable and effective way to protect our forests without harmful chemicals.

Abstract

Pine wilt disease, transmitted primarily by Monochamus alternatus (Hope, 1842) adults, causes severe ecological and economic losses globally. Conventional chemical controls face challenges of resistance and non-target toxicity. This study identified Beauveria bassiana (Bals.-Criv.) Vuill. strain B4 as a high-virulence biocontrol agent against adult M. alternatus. Laboratory bioassays compared four strains (B1–B4), with B4 exhibiting rapid lethality (LT50 = 6.61 days at 1 × 108 spores/mL) and low median lethal concentration (LC50 = 9.63 × 105 spores/mL). Critically, B4 infection induced significant behavioral suppression, including reduced appetite and mobility prior to death. In forest trials, pheromone-enhanced nonwoven fabric bags impregnated with B4 spores reduced trap catches by 66.4% within one month, with effects persisting for over a year without reapplication. The slow-release carrier system enabled continuous spore dissemination and sustained population suppression. These results demonstrate that B4’s dual action—rapid lethality and behavioral disruption—provides an effective, eco-friendly strategy for sustainable pine wilt disease management.

1. Introduction

Monochamus alternatus is the main vector of nematode-carried pine wilt disease, which primarily affects Pinus massoniana Lamb., among other conifers [1]. By feeding on or ovipositing in healthy pine phloem, M. alternatus transmits the pathogenic pine wood nematode Bursaphelenchus xylophilus (Steiner and Buhrer, 1934) Nickle, 1970 to the host xylem, which results in the large-scale wilting or death of pine trees [2]. Pine wilt disease has devastated global pine forest ecosystems, and the direct economic losses caused by pine wood nematodes in China alone exceed billions of yuan per year (equivalent to approximately hundreds of millions of U.S. dollars based on the average exchange rate) [3]. At present, the prevention and control of M. alternatus is highly dependent on chemical insecticides (such as thiacloprid and beta-cypermethrin), but problems caused by long-term insecticide use, such as increased resistance, non-target mortality, and environmental pollution, have become increasingly prominent [4,5,6,7]. Therefore, the development of an environmentally friendly and sustainable biological control is urgently needed.
Entomopathogenic fungi are natural regulators of insect populations and have shown unique advantages in the comprehensive management of agricultural and forestry pests [8,9,10,11]. Beauveria bassiana is a broad-spectrum entomogenous fungus [12] that can cause insect mortality through surface infection, toxin secretion, and nutrient competition. It has been successfully applied to prevent and control locusts, aphids, Anastrepha ludens (Loew), and numerous Coleoptera [13,14,15,16]. Studies have shown that B. bassiana has significant pathogenicity to M. alternatus larvae, and the corrected mortality of larvae treated with a spore suspension (1 × 108 spores/mL) for 7 days can reach more than 80% [17]. Studies on the use of B. bassiana and Metarhizium anisopliae (Metschn.) Sorokīn have focused on the prevention and control of M. alternatus larvae [18,19,20,21,22]. M. alternatus larvae are mostly active in the deeper xylem of pine trees. Even if fungicides have a significant effect in the laboratory, it is difficult to cause substantial damage to the larvae in the xylem of trees. However, M. alternatus adults feed and mate on needles and other parts of the tree. An Unmanned Aerial Vehicle can cover a large range and provide a means to control M. alternatus adults.
To disrupt the transmission of the pathogenic nematode B. xylophilus, the causative agent of pine wilt disease, this study targeted its key vector insect, M. alternatus, using the entomopathogenic fungus B. bassiana. We first screened high-efficiency B. bassiana strains and determined their median lethal concentration (LC50) and median lethal time (LT50) against M. alternatus adults in the laboratory. Subsequently, a forest experiment was designed based on the occurrence period of M. alternatus adults to evaluate the control efficacy of the most virulent strain applied with nonwoven fabric. Our findings provide a foundation for the biological control of M. alternatus and contribute to optimizing the integrated management of pine wilt disease.

2. Materials and Methods

2.1. Tested Insects and Fungal Strains

Adult M. alternatus were collected from Pinus massoniana forest in Xixia County (111°01′–111°46′ E 33°05′–33°48′ N and 181–2212.5 m above sea level), Nanyang City, Henan Province, China. The insects were collected from a BF-8 pine sawyer trap (Hangzhou Feiluomeng Biotechnology Co., Ltd., Hangzhou, China) every 2 days, A total of approximately 1600 individuals were captured. These insects were transported to the laboratory and acclimated for 5 days with fresh pine branches. After this period, healthy and active individuals were selected based on morphological identification, resulting in a final cohort of 1440 insects for the experiment. Four strains of B. bassiana, identified as B1, B2, B3, and B4, were provided by Baiyun Biological Company of Jiyuan City, Henan Province. These strains were selected for this study because they displayed varying phenotypic characteristics (e.g., colony morphology, growth rate) in preliminary observations, indicating their potential for differential virulence, which was then empirically tested against M. alternatus.

2.2. Preparation of Culture Medium and Fungal Suspensions

The B. bassiana strains were cultured in PDA medium containing streptomycin sulfate and kanamycin at 25 °C for 7 d. The surface of the colony was then washed with sterile water to obtain a conidial suspension. Conidial suspensions with concentrations of 1.0 × 108, 1.0 × 107, 1.0 × 106, 1.0 × 105, and 1.0 × 104 spores/mL were prepared with 0.05% Tween-80 solution for the inoculation test.

2.3. Nonwoven Bags and Attractants

We manufactured nonwoven bags for B. bassiana strain B4 and an F8 attractant (Hangzhou Feiluomeng Biotechnology Co., Ltd.).

2.4. Preparation and Inoculation of Nonwoven Bags

Nonwoven fabric bags (25 cm × 30 cm × 5 cm) were filled with sponge blocks of the same size, sterilized, and cooled. A conidial suspension with a concentration ≥ 1 × 108 spores/mL was prepared, and the sterilized nonwoven bags and sponges were immersed in the solution for 24 h. The bags were then dried in the shade.

2.5. Determination of the pathogenicity of Beauveria bassiana to Monochamus alternatus

Healthy adult M. alternatus of similar size were selected as test insects. Each B. bassiana strain was assessed at five concentrations with three replicates of 20 insects each (n = 60 per concentration). Conidial suspensions were poured into a clean Petri dishes, and adult M. alternatus were allowed to crawl freely in different concentrations of conidial suspension for 20 s. After the surface of the insect’s body was fully contaminated with spore suspension, the insects were placed in an incubator (25 °C ± 1 °C, 65 ± 5% relative humidity). The control group was treated with sterile water containing 0.05% Tween-80 solution. The mortality of M. alternatus was recorded every 24 h. When stiff insects appeared, they were removed to observe subsequent mycelial growth.

2.6. Forest Test of the B4 Strain Combined with Nonwoven Fabric

The forest experimental site was located in a pine wood nematode epidemic area in Xixia County, Nanyang City, Henan Province (111°28′26″ E 33°18′25″ N and 409 m above sea level). Two 20 ha test plots were selected that contained P. massoniana mixed forest. The experiment began at the end of May 2023. The treatment (T1) area was distributed according to a 20 m × 20 m grid, and two nonwoven bags (1.5 m from the ground) were hung at each point. One bottle of F8 attractant was hung 5 cm above the mouth of each bag. A schematic diagram of this field deployment setup is shown in Figure 1. Four traps were hung in the T1 area and the control area at intervals of 50 m. The number of insects in the traps was recorded every 10 d, and the lures were replaced every 2 mo. In 2024, traps were hung at the same locations and the same methods were used to collect data.

2.7. Data Analysis

This study included an indoor virulence determination and an outdoor forest prevention and control experiment. In the laboratory, the cumulative mortality and corrected mortality of M. alternatus treated with different concentrations of B. bassiana spore solution were calculated as follows: η = (μt − μc)/(1 − μc) × 100%, where η is corrected mortality, and the unit is percentage (%); μt is the mortality of the treatment group; and μc is the mortality of the control group. Probit regression analysis was performed using IBM SPSS Statistics (version 26.0, IMB Corp., Armonk, New York, NY, USA) software to establish the virulence equation and calculate LC50, LT50, and the 95% confidence intervals. The cumulative mortality data, expressed as percentages, were subjected to an arcsine square root transformation to stabilize variances and meet the assumptions of normality and homogeneity of variances for parametric tests. One-way analysis of variance (ANOVA) and Duncan’s multiple range test were then performed on the transformed data. Graphs were made with GraphPad Prism10 (GraphPad Software Inc., Boston, MA, USA) based on the untransformed data for clarity of presentation.

3. Results

3.1. Screening the Virulence of Beauveria bassiana Strains Following Infection of Monochamus alternatus Adults

At a concentration of 1 × 108 spores/mL, the virulence of the four strains of B. bassiana against M. alternatus differed significantly (Table 1). The B4 strain was the most pathogenic, and the cumulative corrected mortality rate of M. alternatus was 87.18% after 20 d of treatment. The mortality rate induced by the B3 strain was 84.98%. The effects of the B1 and B2 strains were weaker, and the mortality rates were 60.07% and 17.21%, respectively. The virulence of strain B2 was only 19.74% that of B4. The differences among the strains were highly significant, which indicates that genetic background had a decisive influence on the expression of virulence. The above results demonstrated that at the same concentration, B. bassiana strains B3 and B4 were the most lethal.

3.2. Symptoms of Beauveria bassiana Infection of Monochamus alternatus Adults

In the early stage of infection (1−5 d), M. alternatus adults showed symptoms such as decreased appetite and slowness, and mortality soon occurred. In the middle stage of infection (5−10 d), white hyphae began to appear on the surface, particularly around joints and appendages. At the middle-to-late stage of infection (10−15 d), extensive proliferation of mycelia covered large areas of the cuticle. At the late stage of infection (15−25 d), complete coverage of the cadaver by a dense layer of fungal mycelia was observed, producing a powdery layer of white conidia (Figure 2). The progression from exposure to infection, to disease, to death, and to sporulation took a minimum of 5 d and a maximum of 30 d. In most cases, this cycle lasted 10–25 d.

3.3. Determining the Toxicity of Strains B3 and B4 to Monochamus alternatus Adults

The virulence of B. bassiana strains B3 and B4 against adult M. alternatus was evaluated. The LC50 values (Table 2) indicate that strain B4 was significantly more virulent than strain B3. Similarly, the LT50 and LT90 values at different spore concentrations are summarized in Table 3. In general, the LT50 values for both strains gradually decreased with increasing spore concentration, and at all tested concentrations, strain B4 exhibited a lower LT50 than strain B3, indicating a faster speed of kill. The cumulative corrected mortality over time is shown in Figure 3. For statistical analysis, the mortality percentage data were subjected to an arcsine square root transformation to meet the assumptions of normality prior to ANOVA and DMRT; the figure presents the untransformed means (±SE) for graphical clarity. These results demonstrate the high efficiency of strain B4, even at low doses, supporting its selection for the targeted biological control of M. alternatus adults.

3.4. Forest Test of Strain B4 Combined with a Nonwoven Bag

The T1 group, in which strain B4 was combined with a nonwoven fabric bag and lure, significantly reduced the number of M. alternatus trapped at multiple time points (all p < 0.05, Table 4). In the short term (in the first month of 2023), the number of M. alternatus trapped in the T1 group (9.5 ± 1.9) was 66.4 % lower than that in the control (CK) group (28.3 ± 4.1) (p < 0.001). There was no overlap in the confidence interval (T1: 6.5 ± 12.5; CK: 21.7 ± 34.8), which demonstrates that the treatment effect was stable. In the long term (treatment for 1 year), the number of M. alternatus trapped in the T1 group (14.5 ± 2.4) was 30.9 % of that in the CK group (CK: 47.0 ± 2.8); p < 0.001). Although B. Bassiana spore suspensions were not reapplied in the second year, the number of M. alternatus adults trapped in the T1 group (5.75 ± 2.75) was still significantly lower than that in the CK group (37.25 ± 10.08), which indicates that strain B4 had established a stable local population.

4. Discussion

The management of mobile insect vectors requires strategies that transcend mere lethal toxicity. Our study demonstrates that the entomopathogenic fungus B. bassiana B4 suppresses critical behaviors in adult M. alternatus prior to death and that this sublethal effect can be harnessed through an innovative slow-release system to achieve sustained population suppression. This work therefore contributes to the growing field of behavioral manipulation in integrated pest management. Strain B4 showed significant advantages in lethal concentration (LC50 = 9.63 × 105 spores/mL) and lethal time (LT50 = 6.61 d, the highest concentration treatment), and it was significantly more virulent than strain B3 (LC50 = 4.93 × 107 spores/mL, LT50 = 11.36 d). This difference in pathogenicity is likely attributable to variations in key physiological characteristics among the strains, such as spore production, germination rate, and cuticle-degrading enzyme activity. Previously, it was shown that the pathogenicity of entomogenous fungi was affected by the combined effects of spore production, the spore germination rate, and body wall penetration ability [23]. In this study, strain B4 caused rapid mortality of M. alternatus adults at a low concentration (1 × 107 spores/mL) (LT50 = 0.947 d), which demonstrates that its spore germination efficiency and toxin synthesis ability were strong and may have been related to its smaller spore size (2–3 μm) [24]. Small spores are more likely to adhere to the surface of an insect and penetrate the body wall. In addition, the B4 strain could sporulate on the surface of M. alternatus adults after infection and form a secondary transmission effect, which increases its potential for continuous prevention and control in a forest environment.
Compared with M. alternatus larvae, the prevention and control of M. alternatus adults is more challenging. Adults have a wide range of activities and can migrate. Traditional chemical control is difficult to apply accurately and may lead to insecticide resistance. In this study, an indoor toxicity test confirmed that B. bassiana had a direct lethal effect on adults, and the cumulative corrected mortality rate reached 90% under a high concentration treatment (1 × 108 spores/mL). Larvae experienced greater than 80% mortality. Previous studies have consistently demonstrated [25,26] that feeding by Dendrolimus pini (Linnaeus, 1758) or M. alternatus infected with B. bassiana decreased sharply (daily food intake decreased by 16.2–16.7%), and their life span was reduced to less than half that of the control group. Although the number of eggs laid by adults was not significantly affected [26], the inhibition of feeding behavior and a shorter lifespan could significantly reduce the transmission of B. xylophilus, thus indirectly reducing the spread of pine wilt disease.
A key consideration for the environmental application of entomopathogenic fungi like B. bassiana is their potential impact on non-target organisms. A recent study demonstrated that a field-realistic concentration of B. bassiana caused increased mortality, altered behavior, reduced reproduction, and induced colony failure in the predatory social wasp Polistes dominula (Christ, 1791), a valuable natural enemy of agricultural pests [27]. This inherent challenge underscores the importance of developing targeted application strategies that maximize pest control efficacy while minimizing ecological non-target exposure. In previous studies on the release of biocontrol agents in the forest, nonwoven fabric was bound directly to the trunk of trees, and the biocontrol was not combined with an attractant [28,29]. In this experiment, a nonwoven bag and attractant were combined, and the attractant was used to draw M. alternatus adults to the nonwoven bag to become infected by B. bassiana, which significantly improved control over M. alternatus in the forest. In the treatment with nonwoven bags containing a solution of strain B4, a significantly lower number of M. alternatus were trapped; this produced a short-term (within one month) reduction of 66.4%. The prevention and control effect lasted into the next year, which indicates that the B4 strain established a stable population in the forest. Therefore, the targeted nature of our "lure-and-infect" system presents a methodological framework that lends itself to future empirical evaluation of non-target effects under field conditions. Therefore, while the potential for non-target effects exists, our application method is designed to proactively mitigate such risks by limiting environmental dispersal. Future research directly quantifying the impact of this specific strain and application technique on key non-target species in pine forests will be crucial to fully validate its environmental safety and optimize its use in integrated pest management.
It is important to note that the effects of environmental factors on the virulence of B. bassiana require further exploration. The laboratory portion of this study was conducted under constant conditions (25 °C, 65% relative humidity), but fluctuations in field temperature and humidity may affect spore germination and infection processes [30,31]. For example, it was found that the LT50 of M. anisopliae under natural temperature and humidity was extended by 9–15 d, which was significantly different from the laboratory results [21]. It is important to optimize the application of microbial agents for different geographical and climatic conditions, and this may be done by enhancing the environmental tolerance of spores via microcapsule embedding technology [32] or by combining microbial agents with trapping devices to improve targeting.
This study demonstrates the promise of the B4 strain for the biological control of M. alternatus; however, several avenues for further research remain to support its practical application. Firstly, while our field trials showed efficacy, the influence of key environmental factors such as ultraviolet radiation and rainfall on fungal persistence was not directly quantified, and potential synergies with other management techniques await exploration [33,34,35]. Secondly, the molecular mechanisms underlying the pathogenicity of strain B4 and the optimization of its large-scale production are crucial areas for future investigation to enhance efficacy and economic viability. Finally, an important consideration for any vector-control strategy is the potential for pathogen transmission during the infection period. Although the short LT50 of B4 (6.61 days) likely reduces the window for transmission, future work should directly quantify nematode transmission rates by beetles at different stages of mycosis to fully validate the strategy’s capacity to disrupt the disease cycle.
In summary, we identified B. bassiana strain B4 as having high virulence against M. alternatus. The use of an attractant and a nonwoven bag in combination with B. bassiana strain B4 in the forest was effective, and this method provides a foundation for the biological control of pine wilt disease.

Author Contributions

S.L. designed the research. Y.Z. (Yaqi Zhang) conducted the indoor experiments and data analysis and wrote the manuscript. Y.Z. (Yaqi Zhang) and J.W., conducted the indoor experiments. X.Z., D.G. and L.C. conducted the forest experiments. Y.Z. (Yi Zheng), L.A. and H.B. conducted the data analysis. All authors have read and agreed to the published version of the manuscript.

Funding

This study was sponsored by the Staring Foundation for the Henan Academy of Agricultural Science (grant number 2025ZC136) and the Henan Provincial Forestry Technology Work Station (grant numbers 221612414).

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors upon request.

Acknowledgments

We are gratefully acknowledge the staff from the local Forestry Pest Control Station for their invaluable assistance in setting up the field experiments.

Conflicts of Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to have influenced the work reported in this paper.

Abbreviations

The following abbreviations are used in this manuscript:
ANOVAAnalysis of variance
PDAPotato Dextrose Agar
RHRelative Humidity
SEStandard Error
DMRTDuncan’s Multiple Range Test

References

  1. Kiyohara, T.; Tokushige, Y. Inoculation Experiments of a Nematode, Bursaphelenchus Sp., onto Pine Trees. J. Jpn. For. Soc. 1971, 53, 210–218. [Google Scholar] [CrossRef]
  2. Futai, K. Pine Wood Nematode, Bursaphelenchus Xylophilus. Annu. Rev. Phytopathol. 2013, 51, 61–83. [Google Scholar] [CrossRef] [PubMed]
  3. Yan, J.; Sun, H.; Wang, Y.; Chen, Y.; Ma, H.; Yu, Z. The occurrence of major forestry pests in China in 2024 and the trend forecast for 2025. For. Pest Dis. 2025, 44, 52–56. [Google Scholar] [CrossRef]
  4. Deng, J.; Zhuang, W.; Liu, Y.; Song, L.; Zhang, L. Pathogenicity of white muscardine fungus Beauveria bassiana against Japanese pine sawyer beetle Monochamus alternatus and its compatibility with ectoparasitic beetle Dastarcus helophoroides. J. Plant Prot. 2021, 48, 602–609. [Google Scholar] [CrossRef]
  5. Luo, L.; Cai, Z.; Lin, T. Research progress on natural enemies against Monochamus alternatus Hope and its bio-control. China Plant Prot. 2015, 35, 21–25. [Google Scholar]
  6. Dang, Y.; Wang, X.; Yang, Z. Advances in biological control of forest insect pests by using natural enemies in China. J. Environ. Entomol. 2018, 40, 242–255. [Google Scholar]
  7. Lewis, S.M.; Jusoh, W.F.A.; Walker, A.C.; Fallon, C.E.; Joyce, R.; Yiu, V. Illuminating Firefly Diversity: Trends, Threats and Conservation Strategies. Insects 2024, 15, 71. [Google Scholar] [CrossRef]
  8. Wei, Y. Study on the Function of Dopamine Signaling Pathway in the “Tree Top Disease” of Lymantria Dispar. Master’s Thesis, Northwest A & F University, Xianyang, China, 2023. [Google Scholar]
  9. Rosenheim, J.A.; Kaya, H.K.; Ehler, L.E.; Marois, J.J.; Jaffee, B.A. Intraguild Predation Among Biological-Control Agents: Theory and Evidence. Biol. Control 1995, 5, 303–335. [Google Scholar] [CrossRef]
  10. Panwar, N.; Szczepaniec, A. Endophytic Entomopathogenic Fungi as Biological Control Agents of Insect Pests. Pest Manag. Sci. 2024, 80, 6033–6040. [Google Scholar] [CrossRef]
  11. Kim, J.-C.; Lee, M.R.; Yu, J.S.; Park, S.E.; Ha, P.; Kim, J.S. Management of Overwintering Pine Sawyer Beetle, Monochamus alternatus with Colonized Beauveria bassiana ERL836. PLoS ONE 2022, 17, e0274086. [Google Scholar] [CrossRef]
  12. Wei, Q. The Inhibiting Effect of Beauveria bassiana Induced Solanum lycopersicum Defense on Bemisia tabaci. Ph.D. Thesis, Southwest University, Chongqing, China, 2021. [Google Scholar]
  13. Angel-Ruiz, N.A.; Zavala-Izquierdo, I.; Pérez-Staples, D.; Díaz-Fleisher, F.; Andrade-Torres, A.; Guillén-Navarro, G.K.; Colunga-Salas, P. Bioprospecting of Four Beauveria bassiana Strains and Their Potential as Biological Control Agents for Anastrepha Ludens Loew 1873 (Diptera: Tephritidae). PLoS ONE 2025, 20, e0324441. [Google Scholar] [CrossRef]
  14. Zhang, Y.; Liu, B.; Zhang, J.; Lu, Y. Toxic efficiency of biocontrol fungi Beauveria bassiana and Metarhizium anisopliae jointly with four insecticides on controlling cotton aphid Aphis gossypii. J. Plant Prot. 2024, 51, 1457–1465. [Google Scholar] [CrossRef]
  15. Liu, H.; Liu, L.; Liu, X.; Wang, D. The Control Efficacy of Two Fungal Strains against Locusts of Grassland in Yulin, Shaanxi. Chin. J. Biol. Control 2021, 37, 380–384. [Google Scholar] [CrossRef]
  16. Wang, Y. Basic Research on the Control of Xylotrechus rusticus L. by Beauveria Spp. and Dastarcus helophoroides. Ph.D. Thesis, Northeast Forestry University, Harbin, China, 2021. [Google Scholar]
  17. Chen, Y.; Cai, S.; Lin, Y.; Zeng, L.; Zhan, F. Isolation and Idetification of a Beauveria bassiana Strain from the Infected Larvae of Monochamus alternatus in the Pine Forest. J. Fujian For. Sci. Technol. 2024, 51, 9–13,32. [Google Scholar] [CrossRef]
  18. He, X.; Huang, J.; Cai, F.; Yang, X.; Chen, D.; Kang, W. Bioassay of pathogenicity of different Beauveria bassiana isolates to longihorn beetle. In Proceedings of the Research and Application of Entomogenous Fungi in China (Volume V); Fujian Academy of Forestry Sciences: Fuzhou, China, 2003; p. 6. [Google Scholar]
  19. He, X.; Chen, S.; Huang, J. Preliminary screening of virulent strains of Metarhizium anisopliae against Monochamus alternatus. Acta Entomol. Sin. 2005, 975–981. [Google Scholar] [CrossRef]
  20. Liu, H.; Piao, C.; Wang, L.; Shen, X.; Zheng, R.; Shu, Q. Biocontrol of Monochamus alternatus by Beauveria bassiana and Scleroderma guani. Sci. Silvae Sin. 2007, 43, 64–68. [Google Scholar]
  21. Zhang, Y.; Wang, X.; Yang, Z.; Wei, K.; Cao, L. Research Progress on Natural Enemies and Their Application of the Vector Insects of Bursaphelenchus Xylophilus. For. Pest Dis. 2022, 41, 21–29. [Google Scholar] [CrossRef]
  22. Zhang, Z. Study on Screening and Cultural Condition Optimization for Pathogenic Fungi Against Longhorn Beetle. Master’s Thesis, Northwest A & F University, Xianyang, China, 2023. [Google Scholar]
  23. Cai, S.; Liu, J.; He, X.; Li, Z.; Wu, L. Bioassay of different strains of Beauveria bassiana and Metarhizium anisopliae to Anoplophora chinensis adults. For. Pest Dis. 2008, 27, 1–3. [Google Scholar]
  24. Guo, H.; Liu, Z.; Sun, J. Effects of spore suspension concentration and host body size on the pathogenicity of Beauveria bassiana against Monochamus alternatus (Coleoptera: Cerambycidae) larvae. Acta Entomol. Sin. 2020, 63, 835–842. [Google Scholar] [CrossRef]
  25. Kovač, M.; Lacković, N.; Pernek, M. Effect of Beauveria bassiana Fungal Infection on Survival and Feeding Behavior of Pine-Tree Lappet Moth (Dendrolimus Pini L.). Forests 2020, 11, 974. [Google Scholar] [CrossRef]
  26. Liu, H.; Shi, J.; Shu, Q.; Meng, Z.; Dong, G.; Fang, J. Influence of Beauveria bassiana on the life habits of Monochamus alternatus larvae. J. Anhui Agric. Univ. 2010, 37, 196–199. [Google Scholar] [CrossRef]
  27. Cappa, F.; De Fazi, L.; Baracchi, D.; Cervo, R. Adverse Effects of the Fungal Biopesticide Beauveria bassiana on a Predatory Social Wasp. Sci. Total Environ. 2024, 908, 168202. [Google Scholar] [CrossRef] [PubMed]
  28. Li, W. Control Trail of Monochamus alternatus (Hope) in the Forest. Biol. Disaster Sci. 2013, 36, 202–205. [Google Scholar]
  29. Wang, B.; Fan, M.; Li, Z. Control Forest Beetles with Beauveria bassiana Nonwoven Fabric Sheet in Combination with Beetle Attractants. Chin. J. Biol. Control 2003, 91–92. [Google Scholar] [CrossRef]
  30. Zhang, L.; Liu, J.; Wu, H. The Screening Virulent Strain of Beauveria bassiana to Monochamus alternatus. J. Nanjing For. Univ. (Nat. Sci. Ed.) 2000, 33–37. [Google Scholar]
  31. Bai, Y. Effecfs of Humidity Temperature and Initial Infection Ratio on Fungal Disease of B. bassiana Against Stephanitis nashi and Locusta Migratoria Manilensis and Tts Model Construction. Master’s Thesis, Anhui Agricultural University, Hefei, China, 2017. [Google Scholar]
  32. Lu, L. Scleroderma sichuanensis Carrying B. bassiana Microcapsules to Control M. alternatus. Master’s Thesis, Sichuan Agricultural University, Chengdu, China, 2023. [Google Scholar]
  33. Wu, S.; Gao, Y.; Smagghe, G.; Xu, X.; Lei, Z. Interactions between the Entomopathogenic Fungus Beauveria bassiana and the Predatory Mite Neoseiulus Barkeri and Biological Control of Their Shared Prey/Host Frankliniella Occidentalis. Biol. Control 2016, 98, 43–51. [Google Scholar] [CrossRef]
  34. Fattorini, S. Upward and Poleward (but Not Phenological) Shifts in a Forest Tenebrionid Beetle in Response to Global Change in a Mediterranean Area. Insects 2024, 15, 242. [Google Scholar] [CrossRef] [PubMed]
  35. Fattorini, S.; Vitozzi, A.; Di Biase, L.; Bergamaschi, D. Macroecology of Dung Beetles in Italy. Insects 2024, 15, 39. [Google Scholar] [CrossRef] [PubMed]
Figure 1. Schematic diagram of the field deployment setup.
Figure 1. Schematic diagram of the field deployment setup.
Insects 16 01045 g001
Figure 2. Symptoms of Beauveria bassiana B4 infection in adult Monochamus alternatus. (a) Early infection stage; (b) middle infection stage; (c) middle-to-late infection stage; (d) late infection stage.
Figure 2. Symptoms of Beauveria bassiana B4 infection in adult Monochamus alternatus. (a) Early infection stage; (b) middle infection stage; (c) middle-to-late infection stage; (d) late infection stage.
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Figure 3. The corrected cumulative mortality of adult Monochamus alternatus treated with different concentrations of Beauveria bassiana strains B3 and B4.
Figure 3. The corrected cumulative mortality of adult Monochamus alternatus treated with different concentrations of Beauveria bassiana strains B3 and B4.
Insects 16 01045 g003
Table 1. The cumulative corrected mortality (%) of adult Monochamus alternatus caused by different Beauveria bassiana strains 1.
Table 1. The cumulative corrected mortality (%) of adult Monochamus alternatus caused by different Beauveria bassiana strains 1.
Days After
Inoculation
Fungal Strains
B1B2B3B4
1 d0.00 ± 0.00 b6.67 ± 11.55 b0.00 ± 0.00 b26.67 ± 5.77 a
3 d32.85 ± 15.00 a1.95 ± 8.36 b1.95 ± 8.36 b33.59 ± 18.70 a
5 d34.12 ± 12.23 ab1.18 ± 25.45 b9.54 ± 18.32 ab38.56 ± 12.61 a
10 d40.90 ± 12.19 ab−1.63 ± 29.30 b69.19 ± 28.83 a73.11 ± 28.17 a
15 d50.00 ± 7.14 ab7.74 ± 26.75 b86.31 ± 14.32 a80.95 ± 32.99 a
20 d60.07 ± 23.22 a17.21 ± 10.56 b84.98 ± 15.40 a87.18 ± 22.20 a
25 d66.00 ± 19.15 b19.56 ± 9.95 c89.10 ± 12.80 ab94.44 ± 9.62 a
30 d71.13 ± 22.25 b19.56 ± 9.95 c100 ± 0.00 a100 ± 0.00 a
1 The data show the means ± SE. In each column, the means with the same letter had no significant difference at the 0.05 level according to Duncan’s multiple range test. The concentration was 1.0 × 108 spores/mL.
Table 2. LC50 of Beauveria bassiana strains B3 and B4 against adult Monochamus alternatus.
Table 2. LC50 of Beauveria bassiana strains B3 and B4 against adult Monochamus alternatus.
StrainLC50 (spores/mL)95% FL 1 (spores/mL)Regression Equation2)
LowerUpper
B34.93 × 1077.01 × 106100.323y = 0.728x − 5.59711.574
B49.63 × 1053.18 ×1052.89 × 106y = 0.326x − 1.9490.847
1 FL, fiducial limits. Parameters were derived from probit regression analysis.
Table 3. Time to lethality for strains B3 and B4 tested against adult Monochamus alternatus.
Table 3. Time to lethality for strains B3 and B4 tested against adult Monochamus alternatus.
StrainSpore Concentration
(spores/mL)
LT50 (d)LT90 (d)Regression EquationCorrelation Coefficient
B31 × 10811.3619.93y = 0.150x − 1.6990.804
1 × 10724.9239.15y = 0.900x − 2.2430.896
1 × 10635.2360.86y = 0.050x − 1.7620.724
1 × 10537.7657.91y = 0.064x − 2.4010.799
1 × 10440.9562.41y = 0.060x − 2.4440.750
B41 × 1086.6119.67y = 0.980x − 0.6490.965
1 × 1077.4728.64y = 0.061x − 0.4520.879
1 × 10612.4041.32y = 0.044x − 0.5490.833
1 × 10526.2852.98y = 0.048x − 1.2620.786
1 × 10431.3360.64y = 0.044x − 1.3700.785
Table 4. Comparison of trap catches between the Beauveria bassiana-treated group and control group at different time points.
Table 4. Comparison of trap catches between the Beauveria bassiana-treated group and control group at different time points.
Study YearTime PointTreatment GroupMean ± SD95% FL 1 (mg/mL)p-Value
LowerUpper
202330 dT19.5 ± 1.96.512.5p < 0.001
-CK28.3 ± 4.121.734.8-
60 dT15.0 ± 2.41.18.9p < 0.001
-CK18.8 ± 3.213.723.8-
One yearT114.5 ± 2.410.718.3p < 0.001
-CK47.0 ± 2.842.551.5-
202430 dT13.3 ± 2.8−1.17.60.002
-CK22.5 ± 7.011.333.7-
60 dT12.5 ± 0.61.63.40.002
-CK14.8 ± 4.67.522.0-
One yearT15.75 ± 2.751.3710.13p < 0.001
-CK37.3 ± 10.121.2153.29-
1 FL, fiducial limits. Parameters were derived from probit regression analysis.
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MDPI and ACS Style

Zhang, Y.; Zhang, X.; An, L.; Gong, D.; Wang, J.; Bi, H.; Zheng, Y.; Cao, L.; Lu, S. Behavioral Suppression and Rapid Lethality: Beauveria bassiana B4 Targets Adult Monochamus alternatus for Sustainable Management of Pine Wilt Disease. Insects 2025, 16, 1045. https://doi.org/10.3390/insects16101045

AMA Style

Zhang Y, Zhang X, An L, Gong D, Wang J, Bi H, Zheng Y, Cao L, Lu S. Behavioral Suppression and Rapid Lethality: Beauveria bassiana B4 Targets Adult Monochamus alternatus for Sustainable Management of Pine Wilt Disease. Insects. 2025; 16(10):1045. https://doi.org/10.3390/insects16101045

Chicago/Turabian Style

Zhang, Yaqi, Xuejie Zhang, Liudi An, Dongfeng Gong, Jinsheng Wang, Huitao Bi, Yi Zheng, Lei Cao, and Shaohui Lu. 2025. "Behavioral Suppression and Rapid Lethality: Beauveria bassiana B4 Targets Adult Monochamus alternatus for Sustainable Management of Pine Wilt Disease" Insects 16, no. 10: 1045. https://doi.org/10.3390/insects16101045

APA Style

Zhang, Y., Zhang, X., An, L., Gong, D., Wang, J., Bi, H., Zheng, Y., Cao, L., & Lu, S. (2025). Behavioral Suppression and Rapid Lethality: Beauveria bassiana B4 Targets Adult Monochamus alternatus for Sustainable Management of Pine Wilt Disease. Insects, 16(10), 1045. https://doi.org/10.3390/insects16101045

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