193 Arab Univ. J. Agric. Sci., Ain Shams Univ., Cairo, 16(1), 193-198, 2008

BIOEFFECTS OF THE ENTOMOPATHOGENIC FUNGI BEAUVERIA BASSIANA (BALS.) ON MUSHROOM OCELLARIS* (COMS.) (DIPTERA: ) [13] Dawalibi1, W.A.M.; S.M.T. Khoja1; M.M. Abou- Shaar2 and N.A. Kaake2 1. General Commission for Scientific Agricultural Research (Gcsar), Aleppo, Syria 2. Faculty of Agriculture, Aleppo University, Aleppo, Syria

Keywords: Beauveria bassiana, Bradysia sound method for the management of pests. ocellaris. Mushroom fly Hyphomycete fungi are the most promising candi- dates (Prior, 1990). ABSTRACT Entomopathogenic fungi have been intensively studied as potential microbial insecticides (Gilles- Laboratory experiments were carried out to de- pie 1988; Ferron et al 1991; Malsam et al 1998). termine the bioeffects of an isolate of ento- Beauveria spp. has been investigated because of mopthogenic fungi Beauveria bassiana (Bals.) and their ability to infect a wide range of . Biofly a commercial product of B. bassiana, on There are more than 400 species recorded as en- the 1st instar larvae of mushroom fly, Bradysia tomopathogenic fungi, from which only about 20 ocellaris (coms). The bioactivity of B. bassiana species have the potential to be used in microbial was tested, using five concentrations of B. bassi- control of insect pests (Zimmerman, 1986). ana on some biological criteria of the mushroom The present work was carried out with the en- fly, by calculating LC50 values after three and sev- deavor of evaluating the bioassay of Beauveria en days of treatment. Results indicated that the bassiana against the mushroom fly Bradysia ocel- mortality rates percentage increased with the in- laris and to evaluate the bioactivity effect of crease of the concentrations used and the period Beauveria bassiana on some biological criteria of after treatment. The highest percentage of mortali- mushroom fly B. ocellaris. ty occurred within the first seven days following treatment. Statistical analysis of the obtained lar- MATERIALS AND METHODS val-pupal and adults period and weight revealed significant differences between treated and non- Rearing mushroom fly treated insects.

INTRODUCTION Mushroom fly B. ocellaris was reared under C in ventilated plasticهlaboratory condition at 23±1 In recent years, an increasing consumer de- boxes (28×16×9 cm) filled with peat to a depth of mand for commercial mushroom products, as nat- about 5 cm. About 10 ml of water was mixed into ural healthy foods has raised concerns about prob- the peat so that the mixture will remain moist to lems in mushroom production. Mushroom the touch and 1g of wheat was distributed over the proved to be a serious pest facing mushroom surface of peat as food source for the larvae. Adult growers (Finley et al 1984; Ishitani, et al 1997; mushroom flies B. ocellaris were put into the box- Jess and Kilpatrick, 2000). The existing control es and an additional 1g of wheat grain was added measures against the fly still rely on chemical in- every 3-4 days together with a cotton plug soaked secticides, which are not always appropriate. The in a 20% glucose to boost the number of eggs laid use of pathogens may offer an environmentally by females (Gouge 1994).

*Bradysia ocellaris was identified at the Department of Entomology in the Natural History Museum, London (Received September 26, 2007) (Accepted November 10, 2007) 194 Dawalibi; Khoja; Abou-Shaar and Kaake

Two ELISA dishes were used for each treat- Brewer, 1999). Each concentration test included 3 ment; every well of the bottom ELISA dish con- replicates each of 24 larvae (72 larvae/ concentra- tained one wheat grain covered with spawn and tion, 432 larvae /treatment). one larvae of Bradysia ocellaris . The dishes were treated with different concentrations of B. bassi- Biological studies: 1st instar larvae were treated ana (102, 103, 104, 105, 106) conidia/ml, control in the same manner with the determined LC50 to treatment was treated with 0.1%Tween alone. In study the effect of the entomopathogenic fungus the well of the upper dish, a piece of cotton mois- B. bassiana and Biofly on certain biological as- tened with similar concentrations of B. bassiana pects of the mushroom fly. Five replicates of 1st and with sterilized water + 0.1%Tween 80 in the instar larvae (40 larvae/replicate) were used for control treatment. Dishes were then put inside each compound. Larval and pupal mortalities, moist boxes containing moistened filter paper. The their durations, weight and moth emergence were C, 70 ± 5 % RH recorded. The experiments were carried out under هdishes were incubated at 23± 1 and 24h (dark). Mortality was recorded daily for laboratory conditions of 23±1Co and 80±5 RH. about 14 days. Dead larvae were removed and maintained in a moist chamber to allow for fructi- Statistical analysis fication of the relevant fungus ANOVA was performed using SAS software Bioassay experiments program.

In this study, the entomopathogenic fungus B. RESULTS AND DISCUSSION bassiana, (S1), isolated from adults of Eurygaster integriceps pest and (Biofly), a commercial prod- Bioeffect of an isolates of the entomopathogenic uct of B.bassiana, were assayed for their bioinsec- fungus B. bassiana and Biofly on mushroom fly ticidal activities against 1st instar larvae of the B. B. ocellaris larvae ocellaris. The first isolate was grown and main- tained on Potato Dextrose Antibiotic Agar Results in Table (1) show the mortality per- (PDAA) and incubated in the dark for 20 days at centage of 1st instar larvae of mushroom fly after 25±1°C. for the actual bioassay, conidial suspen- treatment with different concentrations of B. bas- sions of each of the isolates were prepared by siana. The tested B. bassiana had a great efficacy scraping conidia from the surface of 20 days old against mushroom fly larvae. Also results indicate cultures in 0.1% Tween 80. The suspension was that the mortality rates increased with the increase diluted in sterilized water + 0.1% Tween 80 to get of the concentration used and the period after suspensions ranging from 1×102 to 1×106 conidia / treatment. ml. A total of 10 ml suspension was prepared for The corrected mortality percentages after 3 each dilution. days for S1 isolate treatment ranged from 8.9% using the lowest concentration (102 conidia/ml), to Larvae treatment 38.82% using the highest concentration (106 co- nidia/ml). As treatment, the corrected percentages The experiments were carried out under labor- of mortality ranged from 4.3 to 29.34 % for the atory conditions of 23±1oC and 80±5 RH. Five lowest and highest concentrations, respectively for concentrations of B. bassiana 102, 103, 104, 105 Biofly. These percentages increased to 50.6, and and 106 conidia/ml were used. In all experiments 19.06% after 7 days of treatment for S1 and bio percentage mortality was calculated after 3, 5, 7, fly, respectively. In contrast, the mortality per- 10, and 14 days. Percentage of mortality of each centages recorded for the highest concentration treatment was corrected use using Abbott,s formu- (106 conidia/ml) were 82.06, 64.61% after 7 days, la (Abbott, 1925). The percentages of mortality for the treatment with S1 and bio fly) respectively. were statistically computed according to (Finney, Also the highest cumulative mortality (85.06, 1952) and computed mortality percentages were 69.6%) was recorded after 14 days of treatment plotted versus log concentrations on logarithmic with 106 conidia /ml of (S1, biofly) respectively probit paper to obtain the corresponding regres- Table (1). The concentration mortality lines are sion mortality lines. The concentrations required graphically illustrated in Figs. (1 and 2) and to give 50% mortality (LC50) were estimated showed a positive relationship between larval from the established regression lines (Jyoti and mortality and the concentrations used. It is worth

Arab Univ. J. Agric. Sci., 16(1), 2008

Bioeffects of Beauveria bassiana on mushroom fly 195

st Table 1. Corrected mortality percentages for the 1 instar of mushroom fly Bradysia ocellaris larvae treated with an isolate (S1) of the entomopathogenic fungus Beauveria bassi- ana and Biofly

Mean of corrected mortality % Treatment Concentration Days after treatment conidia/ml Isolate(S1) 3 5 7 10 14 10+2 8.9 32.86 50.6 56.7 56.7 10+3 16.4 40.27 53.70 64.12 64.12 10+4 31.26 58.21 68.66 71.65 71.65 10+5 37.27 61.11 73.14 76.04 76.04 10+6 38.82 67.13 82.06 82.06 85.06 LC50 0.95×102 0.9×102 R 0.52 0.55 Bio fly* 10+2 4.3 14.6 19.06 24.89 24.8 10+3 7.30 17.58 26.37 29.3 29.34 10+4 14.6 26.37 30.8 33.79 33.79 10+5 13.18 35.16 45.5 48.4 48.4 10+6 29.34 47.03 64.61 69. 6 69.6 LC50 5.7×105 4.55×105 R 0.83 0.79

*Biofly: Commercial protect of B.bassiana

7

6

5

Probit Mortality %

4

3 1e+2 1e+3 1e+4 1e+5 1e+6

Log Concentration (conidia/ml)

Fig.1. Toxicity lines of isolate S1 and Biofly on mushroom fly Bradysia ocellaris (Coms.) after 7 days following treatment Arab Univ. J. Agric. Sci., 16(1), 2008

196 Dawalibi; Khoja; Abou-Shaar and Kaake

Strain S1 Strain S1 Bio Fly 7 Bio Fly

6

5

Probit Mortality %

4

3 1e+2 1e+3 1e+4 1e+5 1e+6 Log Concentration (conidia/ml.)

Fig. 2. Toxicity lines of isolate S1 and Bio fly on mushroom fly Bradysia ocellaris (Coms.) after 10 days following treatment

mentioning that the highest percentage of mortali- Biofly which killed 50% of the treated larvae (af- ty occurred within the first 7days following treat- ter 7 days of treatment) had great effects on lar- ment, while Larval mortality was relatively lower val-pupal periods and adult longevity. Statistical after 10 days (Table 1). analysis of the obtained larval-pupal periods and pupal weight revealed significant differences be- Effect of LC50 concentrations of an isolate of tween treated and non-treated insects. The larval, the entomopathogenic fungi B. bassiana and pupal periods and adult longevity were 11.2, 5, Biofly on larval, pupal and adult stages mortal- 6.1, days respectively (non-treated).Application of ity of mushroom fly Bradysia ocellaris the entomopathogenic fungi B.bssiana and Biofly shortened larval and pupal development and adult Data in Table (2) show that the LC50 concen- longevity, which were 8.2, 4.2, 4.2- 8.4, 4.4, 4.4 trations of the entomopathogenic B. bassiana and days after treatment respectively.

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Bioeffects of Beauveria bassiana on mushroom fly 197

st Table 2. Effect of treating 1 instar larvae of the mushroom fly Bradysia ocellaris with LC50 concen- tration of the Beauveria bassiana (Bals.) and Biofly on larval and pupal periods, pupal weight and adult longevity

Mean larval Mean larval Mean pupal Mean pupal Mean No of Treatment duration weight duration weight adults long pupal* period days (mg) period (days) (mg) (days)

S1 8.2b 0.35b 4.2b 1.08b 4.2b 22.4b Biofly 8.4b 0.29b 4.4ab 0.84b 4.4b 18.8c control 11.2a 1.54a 5a 3.55a 6.1a 38.6a CV 6.8 21.1 12.7 12.9 13.4 9.4 F 35.2 105.7 2.6 204.7 12.6 88.7 SE ± 0.4 0.02 0.33 0.06 0.43 6.2 LSD.50 0.87 0.21 0.8 0.32 0.91 3.4

 No. of larvae = 40 larvae.

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Arab Univ. J. Agric. Sci., 16(1), 2008