Acta agriculturae Slovenica, 120/1, 1–11, Ljubljana 2024 doi:10.14720/aas.2024.120.1.13554 Original research article / izvirni znanstveni članek Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata [Hewitson, 1865]) in walnut (Juglans regia L.) under laboratory conditions Shaziya GULL 1 , Ajaz RASOOL 1 , Tariq AHMAD 1, 2 Received May 09, 2023; accepted January 02, 2024. Delo je prispelo 9. maja 2023, sprejeto 2. januarja 2024. 1 Entomology Research Laboratory, Postgraduate Department of Zoology, University of Kashmir, Srinagar, India 2 Corresponding author, e-mail: drtariqento@kashmiruniversity.ac.in Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata [Hewitson, 1865]) in walnut (Juglans regia L.) under laboratory conditions Abstract: Biological control nowadays is rapidly growing to reduce the incessant use of chemical insecticides for control of various insect pests. In the present study, entomopathogenic fungi are used to determine insecticidal activity against walnut blue butterfly under laboratory conditions. The experimental setup was completely randomized design (CRD) with two treat- ments along with control with different concentrations of ento- mopathogenic fungi. The bioassay was carried out by spraying second larval instar of Chaetoprocta odata [Hewitson 1865] with 1, 2, 3 & 4 % conidial concentration of Beauveria bassiana (Balsamo) Vuill. (1912) and Isaria fumosorosea Wize (1904). The results of this study showed that all the concentrations showed remarkable pathogenic activity but I. fumosorosea was highly pathogenic and recorded the highest mortality rate of 93.33 % after 144 hours compared to B. bassiana where 73.33 % mortality was reported. LC 50 values for B. bassiana (4.15) was higher than that of I. fumosorosea (3.34) which indicates that I. fumosorosea was more effective against C. odata population. Among different concentrations of I. fumosorosea, 4 % concen- tration was the most effective with lowest LC 50 values. Key words: Juglans regia; biological control; Chaeotoproc- ta odata;, virulence; Beauveria bassiana; Isaria fumosorosea Učinkovitost entomopatogenih gliv za zatiranje gosenic ore- hovega modrina (Chaetoprocta odata [Hewitson 1865]) na orehu (Juglans regia L.) v laboratorijskih razmerah Izvleček: Biotično varstvo rastlin danes pridobiva na pomenu, kar vpliva na manjšo rabo kemičnih insekticidov pri zatiranju različnih vrst škodljivih žuželk. V pričujoči raziskavi sta bili uporabljeni dve vrsti entomopatogenih gliv za določi- tev njunega insekticidnega delovanja na orehovega modrina v laboratorijskih razmerah. Poskus je bil zasnovan kot popolni naključni poskus z dvema obravnavanjema in kontrolo. Poskus z glivama je bil izveden s škropljenjem druge razvojne stopnje gosenic modrina z 1, 2, 3 & 4 % koncentracijo konidijev gliv Be- auveria bassiana (Bals.-Criv.) Vuill. (1912) in Isaria fumosoro- sea Wize (1904). Rezultati raziskave so pokazali, da so vse kon- centracije konidijev imele opazno patogeno aktivnost, a je bila gliva I. fumosorosea bolj patogena in je bila v obravnavanjih z njo dosežena največja smrtnost gosenic, 93,33 % po 144 urah, v primerjavi z glivo B. bassiana, kjer je bila smrtnost 73,33 %. LC 50 vrednosti so bile za glivo B. bassiana večje (4,15) kot pri glivi I. fumosorosea (3,34), kar kaže, da je bila gliva I. fumoso- rosea bolj učinkovita pri zatiranju gosenic orehovega modrina. Med različnimi koncentracijami konidijev glive I. fumosorosea je bila 4 % koncentracija najbolj učinkovita z najmanjšimi vre- dnostmi LC 50 . Ključne besede: Juglans regia, biotično varstvo rastlin, Chaeotoprocta odata, učinkovitost, Beauveria bassiana, Isaria fumosorosea Acta agriculturae Slovenica, 120/1 – 2024 2 S. GULL et al. 1 INTRODUCTION The caterpillars of walnut blue butterfly, Chaeto- procta odata [Hewitson 1865] Lepidoptera: Lycaenidae are severe leaf defoliators which feed on parenchyma tis- sue of leaves resulting in scleretonizing of new leaflets. The neonate larvae feed on the succulent leaves and are voracious feeders (Butani, 1979; Masoodi & Trali, 1987; Abbas, 2013). It is one of the pests feeding on walnut trees of Kashmir Valley with devastating potential to defoliate the whole host tree (Mir & Wani, 2005). Chemical in- secticides are being used since ages to protect plants due to which insects are developing resistance against them. One of the environmentally friendly techniques is bio- logical control especially, the use of entomopathogenic fungi, forming a key part of integrated pest management (IPM) program (Ren & Chen, 2012). They cause cuticu- lar infection resulting in toxin formation in pest after in- vasion. Additionally, they can infect any developmental stage of pests (Wang et al., 2010). Around 750 species of fungi are highly effective against various insect pests and offer great potential to manage insects with least hazard effects on environment and human health (Rabindra & Ramanujam, 2007; Laznik et al., 2012). This is the first attempt to use two strains of entomopathogenic fungi, Beauveria bassiana and Isaria fumosorosea for control- ling pests on walnut trees. 2 MATERIALS AND METHODS 2.1 BIOLOGICAL MATERIAL C. odata proceeding larvae were collected from the walnut orchards of Central Kashmir Viz., Srinagar (34̊ 04̍ 54.36̎ N, 74̊ 48̍ 33.00̎ E, 1587 m), Budgam (34̊ 01̍ 2.05̎ N, 74̊ 43̍ 6.71̎ E, 1610 m) and Ganderbal (34̊ 13̍ 39.11̎ N, 74̊ 46̍ 19.78̎ E, 1619 m) and rearing was done in glass jars placed in an incubator maintained at temperature of 25 (± 1) °C and relative humidity of 65 (± 5) %. Two fungal strains viz., Beauveria bassiana and Isaria fumosorosea in four different concentrations (1 %, 2 %, 3 % & 4 %) prepared from a stock solution of 1 × 10 9 conidiaml -1 by diluting with double distilled water obtained from Green Life Biotech Laboratory, Somanur, Coimbatore, India were tested against them to know the efficacy. 2.2 BIOASSAYS In the experiment, freshly cut leaves of walnut trees were placed in 11 cm diameter petri- dishes and leaf petiole was covered with water swabbed cotton. Second instar larvae of C. odata were inoculated by immersing them in different conidial suspensions for 30 seconds and in case of the control, larvae were dipped in the dis- tilled water. C. odata larvae were relocated to these leaf discs and all the petri-dishes were covered with the white muslin cloth tied with a rubber band for proper ventila- tion. The experiment design was a randomized complete block where three replicates were taken and each repli- cate had 10 larvae. Mortality was observed daily for six days at 23 ( ± 2) °C and 65 ( ± 5) % RH with a photoperi- od of 12:12h (Irigaray et al., 2003). Mycosis test was done on the dead larvae and was transferred to petri-dishes in which moist filter paper was placed for 10 days. Micro- scopic examination confirmed the cause of mortality by fungal entomopathogens (Cherry et al., 2005). 2.3 MYCOSIS TEST To know whether insect mortality had occurred due to fungal infection, mycosis test was done on the dead cadavers of insects. In this experiment, three petri plates were taken; two containing distilled water and one hav- ing 70 % ethanol. The process started by dipping cadavers of insects one by one in water followed by ethanol and then again in water to kill fungus present on the insect cuticle. Different isolates were dipped in separate Petri dishes. The procedure was repeated for all replicates of different isolates. Further, if fungi showed their growth again, the conidia of different isolates would penetrate the insect cuticle which subsequently enables us to know that death has occurred due to fungal infection (Grund & Hirch, 2010). The collected insect specimens were exam- ined under a Leica M205A stereo zoom trinocular mi- croscope and were photographed with a Leica DFC295 camera having automontage software version 4.10 (Leica Microsystems, Germany). 2.4 STATISTICAL ANALYSES Experimental data was analysed by using Origin Pro software (Version 15). The data derived on means of per - centage mortality and corrected mortality of M. fotedari adults in different treatments were analysed by ANOVA at 0.05 % level of significance. Tukey’s Honest Square Difference test was used to separate means of treatment. Regression Analysis was done to estimate Lethal Con- centration 50 (LC50) values at different concentrations. Abbott’s formula (Abbott, 1925) was used for correction mortality data with that in control. CM (%) = T (%) - C (%) / 100 – C (%) Acta agriculturae Slovenica, 120/1 – 2024 3 Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata ... Where, CM (%) - Corrected mortality T - Mortality in treatment C - Mortality in control 3 RESULTS On treating larvae with B. bassiana at 1% concentra- tion, it was found that mortality of larvae occurred on the 4 th day after treatment. The corrected mortality after 120 hrs and 144 hrs was 3.33 % and 7.03 % respectively (Fig. 6 and Fig. 1). One way ANOV A depicted that mor- talities after 96 hrs, 120 hrs and 144 hrs were statistically similar (p > 0.05) with each other. The regression equa- tion for the suspension was calculated as Y=2.09X - 3.999 having a regression coefficient (R 2 ) value of 0.864 while LC 50 value of the B. bassiana at 1 % concentration was 25.78 whereas on treating larvae with I. fumosorosea at 1 % concentration, the average corrected mortality after 120 hrs and 144 hrs was 13.70 % and 17.77 % respec- tively (Fig. 6 & Fig. 1). The calculated regression equation was Y=5.141X - 9.109 having a regression coefficient (R 2 ) value of 0.880. On subjecting data to one way ANOVA, insignificant difference was between the mortality rate at 96 hrs and 144 hrs while significant difference was be- tween 144 hrs and 120 hrs at p ≤ 0.05. LC 50 value of I. fumosorosea at 1 % concentration was 11.49 % At 2 % concentration, the corrected mortality by Abbott’s formula after 120 hrs and 144 hrs was 41.11 % and 52.96 % respectively (Fig. 7 & Fig. 2) for B. bassiana with LC 50 value 5.65 (Fig. 5). On subjecting the data to one way ANOVA, the results showed that the larvicidal activity at 72 hrs, 96 hrs, 120 hrs and 144 hrs was sig- nificantly different with p ≤ 0.05 (Fig. 2). The regression equation was intended as Y=12.38X - 19.997 with an evaluated regression coefficient value (R 2 ) 0.950 (Fig. 5). On treating with 2 % I. fumosorosea the corrected mor- tality after 120 hrs and 144 hrs was 54.8 % and 71.47 % respectively (Fig. 7& Fig. 2). The evaluated LC 50 value at 2 % concentration was 4.69. The regression equation at 2 % concentration was Y=15.808X - 24.221 having a regression coefficient (R 2 ) value of 0.966. From the data analysis, it was revealed that mortalities at 72 hrs, 96 hrs, 120 hrs and 144 hrs were significantly different among themselves as shown by ANOV A at p ≤ 0.05. When larvae were inoculated with a spore concen- tration of 3 %, larvicidal activity of B. bassiana against 2 nd larval instar depicted that average mortality increased from 72 hrs to 144 hrs. The toxicity of B. bassiana was recorded showing the corrected mortality percentages at 120 hrs and 144 hrs were 58.51 and 64.07 respectively (Fig. 8) while LC 50 value at 3 % was 4.71. Linear regres- sion equation (Y=15.142X - 21.332) showed the value of regression coefficient (R 2 ) equal to 0.961 (Fig. 5 & Fig. 3). One way ANOV A results of the data showed that mortal- ity rates at 3 % concentration were statistically different at 72 hrs, 96 hrs and 120 hrs although, percent mortalities at 120 hrs and 144 hrs were significantly similar among Figure 1: Mean percent mortality (± SD) at 1% concentration of (a) Beauveria bassiana (b) Isaria fumosorosea Acta agriculturae Slovenica, 120/1 – 2024 4 S. GULL et al. themselves at p ≤ 0.05. Similarly,when larvae were treated with 3 % concentration of I. fumosorosea, the correct- ed mortalities at 120 hrs and 144 hrs were recorded as 75.92 % and 82.21 % (Fig. 3 & Fig. 8). One way analysis of variance depicted that mortalities at 48 hrs, 72 hrs, 96 hrs and 120 hrs were statistically significant with each other although, mortality at 144 hrs was insignificant to mor- tality shown at 120 hrs. The obtained regression equation was Y=17.999X - 20.222 with regression coefficient (R 2 ) value 0.983 while the calculated LC 50 value was 3.90. At 4 % concentration, the larvicidal activity of B. bassiana was observed soon after 48 hours f treatment. Figure 2: Mean percent mortality (± SD) at 2 % concentration of (a) Beauveria bassiana (b) Isaria fumosorosea Figure 3: Mean percent mortality (± SD) at 3 % concentration of (a) Beauveria bassiana (b) Isaria fumosorosea Acta agriculturae Slovenica, 120/1 – 2024 5 Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata ... The mortality percentages at 120 hrs & 144 hrs corrected by Abbott’s formula were 65.55 and 71.10 correspond- ingly (Fig. 8 & Fig. 4) whereas the calculated regression equation was Y=16.095X - 16.889 having a regression coefficient (R 2 ) value of 0.960 (Fig. 5) in contrast to I. fumosorosea at 4 % concentration, the corrected mortal- ity percentages acquired through Abbott’s formula at 120 hrs and 144 hrs were 86.29 and 92.96 respectively (Fig. 9). One way ANOVA results exhibited that mortality at 24 hrs was significant to the mortalities at 48 hrs, 72 hrs, 96 hrs, 120 hrs and 144 hrs. The mortality at 48 hrs and 72 hrs was insignificant with each other but significant to Figure 4: Mean percent mortality (± SD) at 4 % concentration of (a) Beauveria bassiana (b) Isaria fumosorosea Figure 5: LC 50 of second larval instar of Chaetoprocta odata treated with different suspensions of entomopathogenic fungi Acta agriculturae Slovenica, 120/1 – 2024 6 S. GULL et al. others while the mortalities at 120 hrs and 144 hrs were statistically similar at p ≤ 0.05. The calculated regression equation was Y=18.38X - 11.556 with a regression coeffi- cient (R 2 ) value of 0.980. LC 50 value at 4 % concentration was 3.34 (Fig. 5). The calculated F values for 1, 2, 3 and 4% concen- Figure 6: Corrected mortality percent and toxic effects of entomopathogenic fungi (1 % suspension) against second larval instar of Chaetoprocta odata\ *Mean of 10 second instar larvae of Chaetoprocta odata/ replication/ treatment; means followed by identical letters in lower case each column are not significantly different by Duncan’s test at 5 %; Mean mortality % of individuals at the end of experiment corrected for mortality in control using Abbott formula Figure 7: Corrected mortality percent and toxic effects of entomopathogenic fungi (2 % suspension) against second larval instar of Chaetoprocta odata *Mean of 10 second instar larvae of Chaetoprocta odata/ replication/ treatment; means followed by identical letters in lower case each column are not significantly different by Duncan’s test at 5 %; Mean mortality % of individuals at the end of experiment corrected for mortality in control using Abbott formula Acta agriculturae Slovenica, 120/1 – 2024 7 Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata ... tration was high for 4% concentration which means significant mortality had occurred at this concentration predicting that the variance between different mortalities isn’t due to the random chance of all the variables used. The calculated regression equation for each concentra- tion of B. bassiana demonstrated positive correlation Figure 8: Corrected mortality percent and toxic effects of entomopathogenic fungi (3 % suspension) against second larval instar of Chaetoprocta odata *Mean of 10 second instar larvae of Chaetoprocta odata / replication/ treatment; means followed by identical letters in lower case each column are not significantly different by Duncan’s test at 5 %; Mean mortality % of individuals at the end of experiment corrected for mortality in control using Abbott formula Figure 9: Corrected mortality percent and toxic effects of entomopathogenic fungi (4 % suspension) against second larval instar of Chaetoproct aodata *Mean of 10 second instar larvae of Chaetoprocta odata /replication/ treatment; means followed by identical letters in lower case each column are not significantly different by Duncan’s test at 5 %; Mean mortality % of individuals at the end of experiment corrected for mortality in control using Abbott formula Acta agriculturae Slovenica, 120/1 – 2024 8 S. GULL et al. between concentration and mortality i.e., with an in- crease in the independent variable, the dependent vari- able also increases. The derived R 2 values range from 0 to 1 which signifies 0 to 100 % and the derived values in the experiment were close to 1 which depicted that increasing concentration of B. bassiana caused more larvicidal activity. The results of LC 50 values calculated through Probit analysis are given in Fig. 5. Among the two entomopathogenic fungi viz.,I. fumosorosea and B. bassiana, I. fumosorosea was recorded to have maximum mean percent mortalities at different concentrations (Fig. 5). The lowest LC 50 value (3.34) was calculated for I. fu- mosorosea at 4 % concentration which indicated its high toxicity to kill the larvae as lower LC 50 values show acute pathogenicity. All the concentrations caused significantly higher mortality rates compared to control (distilled wa- ter). However, during the present study, no concentration of both entomopathogens caused 100 % mortality to lar- vae although; there was an upsurge in the mortality rates when concentration increased. 4 DISCUSSION C. odata is one of the potential pests defoliating wal- nut trees in Kashmir valley (Abass, 2013). The present work is the first attempt to know the pathogenicity caused by two entomopathogenic fungi viz., I. fumosorosea and Figure 10: Entomopathogenic fungi Isaria fumosorosea and Beauveria bassiana infesting larvae of Chaetoprocta odata (A and B) Disruption of insect body tissue due to mycelial growth after infestation (C) Infestation by Beauveria bassiana (D) Infestation by Isaria fumosorosea Acta agriculturae Slovenica, 120/1 – 2024 9 Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata ... B. bassiana against second larval instar of C. odata (Fig. 10). Various entomopathogens have been used to control Lepidopteran pests especially I. fumosorosea, B. bassiana and Metarhizium robertsii (Metchnikoff) Sorokin (1883) (Hussain et al., 2009). Present results were in line with the observations of Gopalakrishnan & Narayan (1988) who evaluated the mortality rate of different larval in- stars of Helicoverpa armigera (Hübner, 1808), at differ- ent concentrations ranging from 1×10 10 to 1×10 7 with a mortality rate ranging from 60-100  %. During the experimental study, no emergence of adult took place from the treated larvae which was in agreement with the study carried by Hafez et al.(1994) who observed various life stage parameters in potato tuber moth (Ph- thorimaea operculella [Zeller, 1837] when treated with B. bassiana and found no emerge of adults at a concentra- tion from 1×10 4 to 1×10 10 sporesml -1 .Entomopathogens are larvicidal as they contain extra-cellular secondary metabolites that have biocidal properties (Vey et al., 1985;Omura, 2011). As reported by other workers, the metabolites of entomopathogens use glycogen and lipid reserves of insects resulting in disruption of insect tissue due to mycelial growth that further leads to loss of ap- petite in them (Thomas et al., 1997) (Fig. 5). Similar ob- servations were recorded during the experimental study when larvae were treated with entomopathogenic fungi and they refrained from the food as no notches were ob- served on the fresh walnut leaves. Our results were fur- ther affirmed with the findings of Shelton et al. (1998) who treated 2 nd larval instar of diamondback moth, Plu- tella xylostella [Linnaeus, 1758] with entomopathogenic fungi, Beauveria spp. And found similar results. In ad- dition, Hatting (2012) determined the pathogenicity of three entomopathogens when treated against ball worm, Helicoverpa armigera and found that Nomuraea rileyi (Farl.) Samson showed the highest toxicity followed by I. fumosorosea and B. bassiana respectively. During the present investigation, it was observed that mortality due to entomopathogens was time and dose dependent. Initially, mortality was low after treat- ment which then gradually augmented from day 3 to day 6. Similarly, the finding of Wright et al. (2005) found that pathogenicity of the infected insect is dose and time dependent. Further, the study conducted by Bashir et al. (2018) reported that B. bassiana leads to 79 % mortality of Corcyra cephalonica [Stainton, 1866] larvae in in-vit- ro conditions. Likewise, our observations concurred with the finding of Nguyen et al. (2007) who found that the treatment of Helicoverpa armigera with M. anisopliae, B. bassiana and P . fumosoroseus, resulted in 68 to 100 % mortality,in laboratory investigations. Besides, it was ob- served that increased concentration showed higher mor- tality rates with amplified conidial growth which was in line with the findings of Safaviet al. (2007) who found that the virulence of entomopathogenic fungi is depend- ent on concentration, conidial growth and mostly on the nutritional content of pest especially carbon: nitrogen content. Thus, the availability of host food is one of the prime factors for the development of fungal pathogens (Tefera & Pringle, 2004). On the other hand, pathogenic- ity of I. farinosa against larval stages of Harmonia axy- ridis [Pallas, 1773] was studied by Steenberg & Harding (2009) and found that larval stages were most vulner- able and resulted in high mortality rates. In the current study, it was recorded that the highest mortality occurred due to I. fumosorosea which was in corroboration with findings of Zimmermam (2008) who found that I. fu- mosorosea has a broad host range and is highly infective to several insect orders especially Lepidoptera. Sabbour (2015) revealed that I. fumosorosea played a significant role in controlling the pests of the corn crop. Likewise, Schemmer et al. (2016) found I. fumosorosea as most pathogenetic fungi with a median lethal concentration of 0.09×104 conidiaml-1 against Camerariao hridella (De- schka & Dimic, 1986) (Lepidoptera: Gracillariidae). Thus from the above inferences and the results of the current study, it can be concluded that I. fumosorosea contains pathogenic characters that can be used as a bio-control agent against larvae of C. odata and may provide a practi- cable alternative to commercial insecticides used to con- trol larvae in the early season. 5 CONCLUSION Chemical insecticides are being used since ages to protect plants from pest attacks which resulted in insect resistance, environmental pollution and various health is- sues. Nowadays emphasis on biological control methods are increasing to manage insect pest population mostly entomopathogenic fungi. Current study demonstrated that, most important percent mortality was found in I. fumosorosea followed by B. Bassiana at all concentrations although both could be utilized as alternate tools to con- trol population of C. odata. Among different concentra- tions of I. fumosorosea, 4 % concentration was the most effective with the lowest LC 50 values. Therefore, further field study is necessary to evaluate the effectiveness of entomopathogenic fungal formulations in the manage- ment of C. odata in walnut orchards that can prove the most effective strategy in integrated pest management programs. Acta agriculturae Slovenica, 120/1 – 2024 10 S. GULL et al. 5.1 CONFLICT OF INTEREST Authors declare that there is no conflict of interest. 5.2 ACKNOWLEDGEMENT Authors are highly thankful to Head Department of Zoology, University of Kashmir for providing working facilities. 6 REFERENCES Abass, A. (2013). Studies on Life Cycle and Management of Some Important Insect Pests Infesting Walnut (Juglans regia Lin- naeus). Dissertation, Sher-e-Kashmir University of Agri- cultural Sciences & Technology of Kashmir. Bashir, N. H., Nazir, T., Nasir, M., Majeed, M. Z., Hanan, A., Sagheer, M., Hasan, H. (2018). In vitro entomopathogenic efficacy of Beauveria bassiana (Ascomycota: Hypocre- ales) against Corcyra cephalonica (Lepidoptera: Pyralidae) and Tribolium castaneum (Coleoptera: Tenebrionidae). Journal of Entomology, 15, 56-61. https://doi.org/10.3923/ je.2018.56.61 Butani, D. K. (1979). Insects and fruits. Periodical Export Book, Delhi. Cherry, A. J., Abalob, P., Hella, K. (2005). A laboratory assess- ment of the potential of different strains of the entomopath- ogenic fungi Beauveria bassiana (Balsamo) Vuilleum and Metarhizium anisopliae (Metschnikoff) to control Calloso- bruchus maculatus (F.) (Coleoptera: Bruchidae) in stored cowpea. Journal of Stored Products Research, 41, 295-309. https://doi.org/10.1016/j.jspr.2004.04.002 Gopalakrishnan, M., & Narayanan, K. (1988). Occurrence of the entomopathogenic fungi Nomuraea rileyi (Farlow) Samson on Acontia graellsii F. (Noctuidae: Lepidoptera) and Beauveria bassiana (Balsamo) Vuill. on Myllocerus sub- faciatus G. (Curculionidae: Coleoptera). Journal of Biologi- cal Chemistry, 2, 58-59. Grund, J., & Hirsch, L. (2011). The potential of entomopatho- genic fungal isolates as an environmentally friendly man- agement option against Acanthoscelide sobtectus. Hafez, M., Zaki, F. N., Moursy, A., Sabbour, M. (1994). Bio- logical effects of the entomopathogenic fungus, Beauveria bassiana on the potato tuber moth Phthorimaea operculella (Seller). Journal of Islamic Academy of Science, 7, 11-214. Hatting, J. L. (2012). Comparison of three entomopathogenic fungi against the bollworm, Helicoverpa armigera (Hüb- ner) (Lepidoptera: Noctuidae), employing topical vs per os inoculation techniques. African Entomology, 20, 91-100. https://doi.org/10.4001/003.020.0112 Hussain, A., Tian, M.Y., He, Y.R., Ahmed, S. (2009). En- tomopathogenic fungi disturbed the larval growth and feeding performance of Ocinara varians Walker (Lepidop- tera: Bombycidae) larvae. Insect Science, 16, 511-517. htt- ps://doi.org/10.1111/j.1744-7917.2009.01272.x Irigaray, F. J. S. C., Marco-Manceborn, V., Perez-Moreno, I. (2003). The entomopathgenic fungus Beauveria bassi- ana and its compatibility with triflumuron: Effects on the two spotted spider mite Tetranychus urticae. Biologi- cal Control, 26, 168-173. https://doi.org/10.1016/S1049- 9644(02)00123-8 Laznik, Z., Vidrih, M., Trdan, S. (2012). The effect of differ - ent entomopathogens on white grubs (Coleoptera: Scara- baeidae) in an organic hay-producing grassland. Archives of Biological Sciences, 64, 1235-1246. https://doi.org/10.2298/ ABS1204235L Masoodi, M. A., &Trali, A. R. (1987). Record of Chaetoprocta odata Hewitson (Lycaenidae: Lepidoptera) on walnut trees in Kashmir. Indian Journal of Plant Protection, 15, 213. Mir, G. M., & Wani, M. A. (2005). Severity of infestation and damage to walnut plantation by important insect pests in Kashmir. Indian Journal of Plant Protection, 33, 188-193. Nguyen, N. T. H., Borgemeister, C.,Poehling, H., Zimmer- mann, G. (2007). Laboratory investigations on the poten- tial of entomopathogenic fungi for biocontrol of Helicov- erpa armigera (Lepidoptera: Noctuidae) larvae and pupae. Biocontrol Science and Technology, 17, 853-864. https://doi. org/10.1080/09583150701546375 Omura, S. (2011). Microbial metabolites: 45 years of wander- ing, wondering and discovering. Tetrahedron, 67, 6420- 6459. https://doi.org/10.1016/j.tet.2011.03.117 Rabindra, R. J., & Ramanujam, B. (2007). Microbial control of sucking pests using entomopathogenic fungi. Journal of Biological Chemistry, 21, 21-28. Ren, S. X., & Chen, X. X. (2012). Biological Control. Beijing: China Agric Press 227–262. Sabbour, M. (2015). Efficacy of Isaria fumosorosea and Metarhi- zium flavoviride against corn pests under laboratory and field conditions in Egyptian Journal of Global Agricultural and Ecology, 5, 42-47. Safavi, S. A., Shah, F. A., Pakdel, A. K., Rasoulian, G. R., Ban- dani, A. R., Butt, T . M. (2007). ‘Effect of nutrition on growth and virulence of the entomopathogenic fungus Beauveria bassiana’. FEMS Microbiology Letters, 270, 116-123. https:// doi.org/10.1111/j.1574-6968.2007.00666.x Schemmer, R., Chládeková, P ., Medo, J., Barta, M. (2016). Nat- ural prevalence of entomopathogenic fungi in hibernating pupae of Cameraria ohridella (Lepidoptera: Gracillariidae) and virulence of selected isolates. Plant Protection Science, 52, 199-208. https://doi.org/10.17221/110/2015-PPS Shelton, A. M., Vanderberg, J. D., Ramos, M., Wilsey, W. T. (1998). Efficacy and persistence of Beauveria bassiana and other fungi for control of diamondback moth (Lepidoptera: Plutellidae) on cabbage seedlings. Journal of Entomological Science, 33, 142-151. https://doi.org/10.18474/0749-8004- 33.2.142 Steenberg, T., & Harding, S. (2009). “Entomopathogenic fungi recorded from the harlequin ladybird, Harmonia axyridis. Journal of Invertebrate Pathology, 102, 88-89. https://doi. org/10.1016/j.jip.2009.07.002 Tefera, T., & Pringle, K. (2004). Evaluation of Beauveria bassiana and Metarhizium anisopliae, for controlling Chilopartellus (Lepidoptera: Cerambidae) in maize. Bio- Acta agriculturae Slovenica, 120/1 – 2024 11 Efficacy of entomopathogenic fungi for control of walnut blue butterfly (Chaetoprocta odata ... control Science and Technology, 14, 849–953. https://doi. org/10.1080/0958315041000172707 Thomas, M. B., Blanford, S., Lomer, C. J. (1997). Reduction of feeding by the variegated grasshopper, Zonocerus variega- tus, following infection by the fungal pathogen, Metarhi- zium flavoviride. Biocontrol Science Technology, 7, 327-334. https://doi.org/10.1080/09583159730730 Vey, A., Quiot, J. M., Vago, C., Fargues, J. (1985).Effetimmu- node´presseur de toxines fongiques: inhibition de la re´ac- tiondõencapsulement multicellulaire par les destruxines. Comptes Rendus de l’Académie des Sciences, 300, 647-651. Wang, L. D., Y ou, M. S., Huang, J., Zhou, R. (2010). Diversity of entomopathogenic fungi and their application in biologi- cal control. Acta Agriculturae Universitatis Jiangxiensis,32, 0920–0927. Wright, D. J., Peters, A., Schroer, S., Fife, J. P. (2005). Applica- tion technology. In “Nematodes as biocontrol agents.” (P. S. Grewal, R. U. Ehlers, and D. I. Shapiro-Ilan (Eds.) 91-106. https://doi.org/10.1079/9780851990170.0091 Zimmermann, G. (2008). The entomopathogenic fungi Isari- afarinosa (formerly Paecilomyces farinosus) and the Isaria fumosorosea species complex (formerly Paecilomyces fu- mosoroseus): biology, ecology and use in biological control. Biocontrol Science and Technology, 18, 865-901. https://doi. org/10.1080/09583150802471812