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Effects of some entomopathogenic fungi on the aphid species, Aphis gossypii Glover (Hemiptera: Aphididae)



Effects of the entomopathogenic fungi Beauveria bassiana, Verticillium alfalfae, and Trichoderma viride; secondary metabolites of MS1 (B. bassiana) and MS2 (V. alfalfae); and Dimethoate active substances on the aphid species, Aphis gossypii Glover (Hemiptera: Aphididae), were tested.

Main body of the abstract

Fungus isolates were prepared as 107 conidia ml−1 of spore suspensions and applied on the 2nd instar nymphs of A. gossypii. After the applications, evaluations were made on the 1st, 3rd, 5th, and 7th days by counting the live individuals. Obtained data were 100 and 93% mortality rate at MS1 (B. bassiana) and MS2 (V. alfalfae), and secondary metabolites were recorded in the 3rd day count results. On the 5th day counts, the highest mortality rates after secondary metabolites were statistically at the same group with B. bassiana, T. viride, and dimethoate. On the 7th day, counting results of all experiment groups were analyzed statistically and were found effective.

Short conclusion

Obtained results showed that the fungal secondary metabolites might be useful when utilized as a biocontrol agent against the aphids.


Aphids are one of the most important insect groups that cause economic damages in the agricultural fields. Of them, the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is a cosmopolitan species, widely spread in tropical, subtropical, and temperate regions of the world (Leclant and Deguine 1994). Due to its wide host range and as vector of many plant viruses, it is an important pest. Nowadays, synthetic chemical insecticides are generally used to pest control (Wang et al. 2002). However, the widespread usage of chemical insecticides causes pest resistance, non-target organisms, and negative effects on the environment (Antwi and Reddy 2015). However, these negative effects on human health, environment, and non-target organisms could be reduced with IPM implementations (Lopes et al. 2009). Using the entomopathogenic microorganisms as biological control agents is an important application in the field (Lacey and Shapiro-Ilan 2008). It is estimated that 1000 entomopathogenic fungi (EPF) (Shang et al. 2015) and more than 100 mycoinsecticides are used as biological agents worldwide (Jaronski 2010). It has been recorded that fungi are effective in pre-adult and adult periods in insects that belong to orders of Lepidoptera, Hemiptera, and Diptera (Herlinda 2010). EPF used as controlling agents of aphids and some other pests are as follows: Beauveria bassiana (Kılıç and Yıldırım 2008), Metarhizium anisopliae (Inanlı et al. 2012), Lecanicillium lecanii (Ujjan and Shahzad 2012), Isaria fumosorosea (Jandricic et al. 2014), Paecilomyces (Shi and Feng 2004), and Nomuraea rileyi (Devi et al. 2003). EPF usually attack the pest through penetrating the insect cuticle and secrete toxins. Furthermore, they produce secondary metabolites effective on their hosts, such as pathogenic fungi (Sentürk and Abacı-Günyar 2019). The secondary metabolites produced by the fungi have low molecular weights and are beneficial bioactive compounds. These compounds are important in the field of agriculture, medicine, and several industrial sectors (Demain and Fang 2000).

In this study, the effects of B. bassiana, V. alfalfae, and T. viride isolates, and the secondary metabolites of MS1 (B. bassiana) and MS2 (V. alfalfae) on the 2nd instar nymphs of A. gossypii were investigated.

Materials and methods

Production of host plants and aphids

Cotton seed, Flash (Gossypium hirsutum L.) (ProGen®), variety was used by sowing the seeds in 1.5-l plastic pots that measured 170 mm × 140 mm. When the cotton plants had 5–6 leaves, the pots were transferred to a climate chamber (25 ± 1 °C, 60 ± 5% RH, and 16:8 h light-dark period conditions for aphid production). The aphid production was accomplished by infesting the cotton plants with the aphids.

Preparation of fungi cultures and spore suspensions

B. bassiana, V. alfalfae, and T. viride were defined morphologically and isolated from the soil of a wheat field at the National Plant Protection Institute (INPV-National Institute of Plant Protection of Constantine, Constantine, Algeria) following the method of Vinayaga Moorthi et al. (2015) and Abdelaziz et al. (2018). To isolate the fungi, 1 g of the soil was diluted in 9 ml of sterile distilled water; then, 100 μl from the 10−3, 10−4, and 10−5 dilutions of these suspensions was planted on potato dextrose agar (PDA: 200 g potatoes, 20 g glucose, and 20 g agar), supplemented with chloramphenicol (10 mg l−1). Petri dishes were incubated at 28 °C for 2 weeks.

Preparation of entomopathogenic fungal secondary metabolites

According to the method of Gurulingappa et al. (2011), secondary metabolite preparation was carried out at 4 steps. EPF were first placed in a 250-ml flask containing 100 ml of PDB and incubated at 28 °C for 21 days. The suspension was filtered in a Whatman no. 1 paper. Then, ethyl acetate was used for extraction purposes; later, the solvent was evaporated, using an evaporator. At last, extract was mixed by a sterile water to recuperate of secondary metabolites of MS1 (B. bassiana) and MS2 (V. alfalfae).

Application of entomopathogenic fungi and secondary metabolites

Fungus and secondary metabolite suspensions were used in the experiments by diluting with Tween 80 (0.05%) sterile distilled water containing 107 conidia ml−1. The experiments were carried out with 5 replicates. Blotting paper and untreated cotton leaves were placed on the floor of the Petri dishes. Cells with a size of 5 × 4 cm and 4 cm space were placed on the leaf surface, and 5 individuals in the 2nd instar nymphs of A. gossypii were transferred to this space. Fungus suspensions were then sprayed on the nymphs 3 times, using a hand sprayer from about 20 cm distance. The control group contained only Tween 80. After the applications, Petri dishes were incubated at 25 ± 1 °C, 60 ± 5% RH, 16:8 h light-dark period. Live individuals were counted and recorded on the 1st, 3rd, 5th, and 7th days of the experiment. Experiments were carried out with 5 replications.

Analysis of the data

One-way ANOVA was applied to the data obtained, and the data were evaluated using IBM SPSS® Statistics (version 20.0, August 2011, SPSS Inc., Chicago, IL, USA) package statistics program. The difference between the means was determined by using the Tukey (1949) multiple comparison test (P < 0.05), and the mortality rates (%) were calculated by using an Abbott formula (Abbott 1925).

$$ \mathrm{Abbott}'\mathrm{s}\ \mathrm{corrected}\ \mathrm{mortality}\%=\frac{\left(\mathrm{living}\ \mathrm{in}\ \mathrm{control}-\mathrm{living}\ \mathrm{in}\ \mathrm{treatment}\right)}{\mathrm{living}\ \mathrm{in}\ \mathrm{control}}\times 100 $$

Results and discussion

The highest mortality rate (53.33%) was recorded at the secondary metabolite of MS2 (V. alfalfae) on the 1st day counts of the experiment, while the mortality rate for V. alfalfae group was not determined. On the 3rd day counts, 100 and 93% mortality rates were recorded for MS1 (B. bassiana) and MS2 (V. alfalfae) secondary metabolites, respectively, showing that they were statistically at the same group. On the 5th day counts, the highest mortality rates after secondary metabolites were statistically at the same group with B. bassiana and T. viride and dimethoate. On the 7th day counts, all experiment groups were recorded statistically in the same group and were found effective (Table 1).

Table 1 Mortality rates resulting from applications of EPF and secondary metabolites at a concentration of 107 conidia ml−1 to 2nd instar nymphs of Aphis gossypii

The results in Table 1 show that the 2nd instar nymphs of A. gossypii, treated with EPF and secondary metabolites, were highly affected and mortality rates ranged 80–100% on the 7th day of the experiment. The results revealed that the secondary metabolites of MS1 (B. bassiana) and MS2 (V. alfalfae) had the most efficient pathogenicity (100%), followed by T. viride (93.33%), dimethoate (90.00%), B. bassiana (80.00%), and V. alfalfae (73.33%).

According to the literature, applications of I. fumosorosea strain (Ifu13a) (Bugti et al. 2018), L. lecanii (Mohammed et al. 2018), and L. lecanii 41185 isolates were found effective by 100% on A. gossypii individuals at 108 conidia ml−1concentration (Vu et al. 2007). Similar studies reported the mortality rate of 100% for B. bassiana (IBCB 66) and M. anisopliae (IBCB 121) isolates applied to A. gossypii individuals (Loureiro and Moino 2006). Tesfaye and Seyoum (2010) recorded the mortality rates of 73.33–93.33% for isolates of Beauveria and Metarhizium. In other studies reported, Beauveria ARSEF 5493 isolate was found effective (Jandricic et al. 2014).

EPF are an important regulatory factor in biological control of insects. Earlier studies have been conducted with different species or isolates of EPF against different host species, which showed different pathogenicity. For instance, as a result of the application of B. bassiana (BB-72 and BB-252) and L. lecanii (V-4) isolates to Myzus persicae individuals, 95, 91, and 87% mortality rates were recorded (Nazir et al. 2018). In another study, B. bassiana had been reported to have an effect over 75%, as a result of application of BAU004, BAU018, and BAU019 isolates to the same aphid species (Al-alawi and Obeidat 2014). Another study was conducted on Aphis craccivora (Koch) individuals; 77.50 to 100% mortality rates were reported in application of B. bassiana, M. anisopliae, V. lecanii, Hirsutella thompsonii, and Cladosporium oxysporum isolates at the concentration of 108 conidia ml−1 (Saranya et al. 2010). According to Ekesi et al. (2000), mortality rates at the 4 different concentrations of B. bassiana CPD 11, and M. anisopliae CPD 4 and 5 isolates were recorded as 58–91, 64–93, and 66–100%, respectively. In a study with other aphids and as a result of the application of C. oxysporum isolate at 108 conidia ml−1 concentration against Aphis fabae individuals, the mortality rate was 67.90% (Bensaci et al. 2015). Mortality rate recorded was (86%) for the applications of V. lecanii IBCB 473 isolate on Cinara atlantica individuals at a concentration of 108 conidia ml−1 (Loureiro et al. 2004). Mortality rates were reported as 95.83, 63.98, and 51.83%, respectively, as a result of the application of B. bassiana, C. cladosporioides, and V. alfalfae isolates to Metopolophium dirhodum (Walker) individuals (Abdelaziz et al. 2018).


The obtained results showed that the fungal secondary metabolites might be useful when utilized as a biocontrol agent against the aphids. Among them, B. bassiana and V. alfalfae were the most promising ones. However, the present work indicated the potentiality of V. alfalfae, as a new resource of secondary metabolite, which may suggest that these metabolites could be used in the selection of candidates of aphid biological control.

Availability of data and materials

All data generated or analyzed during this study are included in this manuscript.



Potato dextrose agar


Days after application


Analysis of variance


Secondary metabolite—Beauveria bassiana


Secondary metabolite—Verticillium alfalfae


  1. Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol 18:265–267

    CAS  Article  Google Scholar 

  2. Abdelaziz O, Senoussi MM, Oufroukh A, Birgücü AK, Karaca İ, Kouadr F, Naima B, Bensegueni A (2018) Pathogenicity of three entomopathogenic fungi, to the aphid species, Metopolophium dirhodum (Walker) (Hemiptera: Aphididae), and their Alkaline protease activities. Egypt J Biol Pest Control 28(24):1–5

    Google Scholar 

  3. Al-alawi MS, Obeidat M (2014) Selection of Beauveria bassiana (Balsamo) Vuillemin isolates for management of Myzus persicae (Sultzar) (Hom.: Aphidae) based on virulence and growth related characteristics. Am J Agric Biol Sci 9(1):94–100

    Article  Google Scholar 

  4. Antwi FB, Reddy GVP (2015) Toxicological effects of pyrethroids on non-target aquatic insects. Environ Toxicol Pharmacol 40:915–923

    CAS  PubMed  Article  Google Scholar 

  5. Bensaci OA, Daoud H, Lombarkia N, Rouabah K (2015) Formulation of the endophytic fungus Cladosporium oxysporum Berk. & M.A. Curtis, isolated from Euphorbia bupleuroides subsp. luteola, as a new biocontrol tool against the black bean aphid (Aphis fabae Scop.). J Plant Prot Res 55(1):80–87

    Article  Google Scholar 

  6. Bugti GA, Na C, Bin W, Feng LH (2018) Control of plant sap-sucking insects using entomopathogenic fungi Isaria fumosorosea strain (Ifu13a). Plant Prot Sci 54(4):258–264

    CAS  Article  Google Scholar 

  7. Demain AL, Fang A (2000) The natural functions of secondary metabolites. In: Fiechter A (ed) History of Modern Biotechnology I, vol 69. Springer, Heidelberg, pp 1–39

    Chapter  Google Scholar 

  8. Devi PS, Prasad YG, Chowdary DA, Rao LM, Balakrishnan K (2003) Identification of virulent isolates of the entomopathogenic fungus Nomuraea rileyi (F) Samson for the management of Helicoverpa armigera and Spodoptera litura (identification of virulent isolates of N. rileyi). Mycopat 156(4):365–373

    CAS  Article  Google Scholar 

  9. Ekesi S, Akpa AD, Onu I, Ogunlana MO (2000) Entomopathogenicity of Beauveria bassiana and Metarhizium anisopliae to the cowpea aphid, Aphis craccivora Koch (Homoptera: Aphididae). Arch Phytopathol Plant Protect 33(2):171–180

    Article  Google Scholar 

  10. Gurulingappa P, McGee PA, Sword GA (2011) Endophytic Lecanicillium lecanii and Beauveria bassiana reduce the survival and fecundity of Aphis gossypii following contact with conidia and secondary metabolites. Crop Prot 30(3):349–353

    Article  Google Scholar 

  11. Herlinda S (2010) Spore density and viability of entomopathogenic fungal isolates from Indonesia, and their virulence against Aphis gossypii Glover (Homoptera: Aphididae). Trop Life Sci Res 21(1):11–19

    PubMed  PubMed Central  Google Scholar 

  12. Inanlı C, Yoldaş Z, Birgücü AK (2012) Effects of entomopathogenic fungi, Beauveria bassiana (Bals.) and Metarhizium anisopliae (Metsch.) on larvae and egg stages of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae). Ege Univ J Agri Fac 49(3):239–242

    Google Scholar 

  13. Jandricic SE, Filotas M, Sanderso JP, Wraight SP (2014) Pathogenicity of conidia-based preparations of entomopathogenic fungi against the greenhouse pest aphids Myzus persicae, Aphis gossypii, and Aulacorthum solani (Hemiptera: Aphididae). J Invertebr Pathol 118:34–46

    CAS  PubMed  Article  Google Scholar 

  14. Jaronski ST (2010) Ecological factors in the inundative use of fungal entomopathogens. Biol Control 55(1):159–185

    Google Scholar 

  15. Kılıç E, Yıldırım E (2008) The use of entomopathogen fungi in control of whiteflies (Homoptera: Aleyrodidae). Atatürk Univ J Agri Fac 39(2):249–254

    Google Scholar 

  16. Lacey LA, Shapiro-Ilan DI (2008) Microbial control of insect pests in temperate orchard systems: potential for incorporation into IPM. Annu Rev Entomol 53:121–144

    CAS  PubMed  Article  Google Scholar 

  17. Leclant F, Deguine JP (1994) Cotton aphids. In: Mathews GA, Tunstall JP (eds) Insect Pests of Cotton. CAB International, Wallingford, pp 285–323

    Google Scholar 

  18. Lopes C, Spataro T, Lapchin L, Arditi R (2009) Optimal release strategies for the biological control of aphids in melon greenhouses. Biol Control 48(1):12–21

    Article  Google Scholar 

  19. Loureiro ES, Moino JA (2006) Pathogenicity of hyphomycete fungi to aphids Aphis gossypii Glover and Myzus persicae (Sulzer) (Hemiptera: Aphididae). Neotrop Entomol 35(5):660–665

    Article  Google Scholar 

  20. Loureiro ES, Oliveira NC, Wilcken CF, Batista AB (2004) Pathogenicity of Verticillium lecanii to pine aphid. Rev Arvore 28(5):765–770

    Article  Google Scholar 

  21. Mohammed AA, Kadhim JH, Kamaluddin ZNA (2018) Selection of highly virulent entomopathogenic fungal isolates to control the greenhouse aphid species in Iraq. Egypt J Biol Pest Control 28(71):1–7

    Google Scholar 

  22. Nazir T, Basit A, Hanan A, Majeed MZ, Qiu D (2018) In vitro pathogenicity of some entomopathogenic fungal strains against green peach aphid Myzus persicae (Homoptera: Aphididae). Agronomy 9(7):1–12

    Google Scholar 

  23. Saranya S, Ushakumari R, Jacob S, Philip BM (2010) Efficacy of different entomopathogenic fungi against cowpea aphid, Aphis craccivora (Koch). J Biopest 3(1):138–142

    Google Scholar 

  24. Sentürk S, Abacı-Günyar Ö (2019) Fungal biocontrol agents and their secondary metabolites. J Fungus 10(1):70–83

    Google Scholar 

  25. Shang Y, Feng P, Wang C (2015) Fungi that infect insects: altering host behavior and beyond. PLoS Pathog 11(8):e1005037

    PubMed  PubMed Central  Article  Google Scholar 

  26. Shi WB, Feng MG (2004) Lethal effect of Beauveria bassiana, Metarhizium anisopliae, and Paecilomyces fumosoroseus on the eggs of Tetranychus cinnabarinus (Acari: Tetranychidae) with a description of a mite egg bioassay system. Biol Control 30(2):165–173

    Article  Google Scholar 

  27. Tesfaye D, Seyoum E (2010) Studies on the pathogenicity of native entomopathogenic funal isolates on the cotton/melon aphid, Aphis gossypii (Homoptera: Aphididae) Glover under different temperature regimes. Afr Entomol 18(2):302–312

    Article  Google Scholar 

  28. Tukey JW (1949) Comparing ındividual means in the analyses of variance. Biometrics 5(2):99–114

    CAS  PubMed  Article  Google Scholar 

  29. Ujjan AA, Shahzad S (2012) Use of entomopathogenic fungi for the control of mustard aphid (Lipaphis erysimi) on canola (Brassica napus L.). Pak J Bot 44(6):2081–2086

    Google Scholar 

  30. Vinayaga Moorthi P, Balasubramanian C, Selvarani S, Radha A (2015) Efficacy of sub lethal concentration of entomopathogenic fungi on the feeding and reproduction of Spodoptera litura. Springer Plus 4:681

    CAS  PubMed  Article  Google Scholar 

  31. Vu VH, Hong SI, Kim K (2007) Selection of entomopathogenic fungi for aphid control. J Biosci Bioeng 104(6):498–505

    CAS  PubMed  Article  Google Scholar 

  32. Wang KY, Liu TX, Yu CH, Jiang XY, Yi MQ (2002) Resistance of Aphis gossypii (Homoptera: Aphididae) to Fenvalerate and Imidacloprid and activities of detoxification enzymes on cotton and cucumber. J Econ Entomol 95(2):407–413

    CAS  PubMed  Article  Google Scholar 

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ABE, OA, AKB, MMS, AO, and İK collaborated in the creation of the manuscript. ABE carried out the experiments, recorded the data, interpreted the results, and wrote the manuscript. AKB supervised the manuscript. All authors read and approved the final manuscript during the present study.

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Correspondence to Alime Bayındır Erol.

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Bayındır Erol, A., Abdelaziz, O., Birgücü, A.K. et al. Effects of some entomopathogenic fungi on the aphid species, Aphis gossypii Glover (Hemiptera: Aphididae). Egypt J Biol Pest Control 30, 108 (2020).

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  • Aphis gossypii
  • Entomopathogenic fungi
  • Biological control
  • Secondary metabolite