PROTECTANTS OF Spodoptera littoralis ...

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OF Spodoptera littoralis NUCLEOPOLYHEDROVIRUS, (BACULOVIRIDAE). By. A. El-Helaly, S. El-Salamouny, M. Khattab*, M. El-Sheikh and S. Elnagar.
4 th Conference on Recent Technologies in Agriculture, 2009………………………………………………………

(2) PRELIMINARY EVALUATION OF NATURAL ANTIOXIDANTS AS UV- PROTECTANTS OF Spodoptera littoralis NUCLEOPOLYHEDROVIRUS, (BACULOVIRIDAE) By A. El-Helaly, S. El-Salamouny, M. Khattab*, M. El-Sheikh and S. Elnagar Department of Economic Entomology and Pesticides, Faculty of Agriculture, Cairo University, and * Cutworms and Mole Cricket Research Department, Plant Protection Research Institute, Agriculture Research Center, Giza, Egypt. ABSTRACT Twenty natural plant derived material containing different groups of antioxidants were tested as UV- protectant additives of Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) suspension, to prolong the virus activity. Screening results were based on bioassays, to test Originally Activity Remaining (OAR) of the virus after exposure to UV source. The different sources of antioxidants were tested as groups in four bioassays. Preliminary results showed that, “Carob” Ceratonia siliqua and “Cacao”, Theubromo cacao additives provided 49.5 and 49 OAR%, respectively, while the virus alone treatment was only 2 % OAR five hours after exposure to simulated UV source. The green and red cabbage, Brassica oleracea treatment gave 53.05& 50.9 OAR %, respectively, compared to the virus alone treatment which provided only 6.88 OAR%. Coffee, Coffea arabia and lemon grass, Cymbopogoen germinea achieved 47.96 & 44.18 OAR % , respectively compared to virus alone treatment (16.62 OAR%) . Seeds and fruits of “grapevine” Vitaceae juss gave 72.5 and 51.66 OAR % , respectively, compared to (17.5 OAR %) for the virus alone treatment. Further screening was made to compare short listed additives that gave best results after ten hours exposure to UV irradiation. The obtained result demonstrated that green cabbage, cacao and coffee additives gave the higher rates of virus persistence than grapevine fruits, lemon grass and seeds of grapevine. The tested additives are currently undergoing further investigations including spectrophotometer studies, as well as pilot tests in the open field under Egyptian conditions. Key words: antioxidants, virus protection, nucleopolyhedrovirus, Spodoptera littoralis NPV, ultraviolet. oxidative enzyme support the hypothesis of inactivation (Ignoffo and Garcia, 1994). Antioxidants reported to provide the protection of entomopathogenic viruses from UV in sunlight, such as Dilodin and Inol (Zarin and Eglite, 1985), Vitamins (Murahabaskaran et al., 2000), natural antioxidants folic acid (Deotale et al., 2007), riboflavin, Pyridoxine, eucalyptus and mango leaf extract (Deotale et al., 2003). It was found that antioxidants prevent UV induced apoptosis by inhibiting mitochondrial cytochrome c release caspase activation in Spodoptera frugiperda (Sf 9) cells (Manjari et al., 2003). Other antioxidants derived from plants are green tea (Shapiro et al., 2008), Black tea (El Salamouny, et al., 2009a) and Cocoa & coffee (El Salamouny et al., 2009b). The present study, evaluated twenty locally available and inexpensive natural products for their suitability as UV protectants to SpliNPV.

1. INTRODUCTION The loss of infectivity of baculovirus by exposure to the UV portion of sunlight is well documented (Elnagar 1982; Ignoffo et al., 1989; Jones et al., 1993 and El Salamouny et al., 2009 a). Many efforts have been devoted to protect baculoviruses by various chemical compounds as UV-protection additives. Several dyes have been tested as UV protectants to baculoviruses (Shapiro, 1989; Ramakrichnan and Chaudhary, 1991; Reddy and Divakar, 2001), Copper ammonium nitrate and copper sulphate (Arivudainambi et al., 2000). The viral enhancers optical brighteners UV protectants (Baskaran, 2001; Kamaldeep & Battu, 2008). Natural products like Lignin derivatives as a side product of paper industries showed also a high rate of protection (Jacques, 1977, TamezGuerra et al., 2000; El Salamouny et al., 2002; Elnagar et al., 2003). The increase UV stability of baculovirus provided by an antioxidant or

2. MATERIAL AND METHODS 2. 1. Insect colony Laboratory colony of the cotton leaf worm, Spodoptera littoralis (Boisduval), was established

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Preliminary evaluation of natural antioxidants as………………………………………………………… surface of 50 ml semi-artificial diet poured in a special bioassay plate measures 5 x 10 x 1 cm (LICEFA, Germany) divided to 50 cells. Untreated control by using either distilled water or additive only was used as a negative control. Neonate larva was confined each to a well of a microbial plate covered by two layers of tissue paper and glass cover fixed with rubber band. Virus without additives (in distilled water) was used for comparison. The plates were incubated at 26±2ºC and 60±5% R.H. under the laboratory conditions. Bioassay experiment was examined at 5, 10 and 15 days (Fritsch and Huber, 1985). 2.6. Statistical analysis Concentration-mortality regressions were calculated to determine the effectiveness of plantderived material as UV protectants for SpliMNPV. Slope and LC 50s were calculated according to the method described by Finney (1971). Original Activity Remaining percentages were determined for each tested treatment using both parameters to insure the potential of the material to prolong the virus persistence as described by Muro and Paul (1985).

as the test insect species on a semi synthetic diet of Shorey and Hale (1965). 2.2. Virus inoculum A Local isolate of Spodoptera littoralis multiple embedded nucleopolyhedrovirus isolate (SpliMNPV) originally isolated in Egypt by Abul Nasr (1956) was used. 2.3. UV protectant additives in the virus inoculum Twenty different plant derived materials were used. One gram of each dry plant material was soaked in 50 ml distilled water for 24 hours to get a final stock of 2% then blended and the filtrate extract was added to virus inoculum according to the method described by Shapiro et al. (2008). The final mixture used in the UV-irradiation tests was adjusted to get a final additive/virus mixture of final 1% concentration additive and 2×10 6 PIB's/ ml SpliMNPV enough to cause 90% mortality (LC 90 ) as a control treatment. The hydrogen ion concentration at 1% of all natural products was measured using Beckman Instruments, Fullerton, and CA. 2.4. UV Irradiation test For both screening trials, sunlight UV was simulated (SUV) using a set of four UV lamps (Ultra-Vitalux, OSRAM, Germany) that were vertical with distance 160 cm from the samples and 60 cm between the center of each two lamps. A wetting agent (2.5% Teepol) was added to the viral suspension (with or without the additive). Standardization of virus concentrations was based on the number of polyhedral inclusion bodies (PIB's / ml) of aqueous suspension. Fifty micro liters of virus with or without additives resembling 200 fold LC90 (4x10 8 PIB's ml) were spread inside a Petri dish (10 cm in diameter) using a fine pipette (Jencons of Hemel Hempstead). After air drying, the dishes with the virus film on surface were exposed to the UV irradiation sources. For the first screening trial, the periods of exposure were 5, 30, 60, 120, 180, 240 and 300 minutes extended to 600 minutes for the second screening. The virus after irradiation was resuspended in 10 ml distilled water and kept refrigerated in a glass test tube until bioassayed. For treatments, the twenty tested material were divided to four groups, of five each and was evaluated in four bioassays. Untreated virus alone treatment was used for comparison. 2.5. Surface Treatment Bioassay Laboratory bioassay was performed using the diet surface contamination technique described by, Cisnero et al. (2002). Two ml of collected PIBs suspension after irradiation were spread on the P

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3. RESULTS Preliminary bioassay determined the virus activity rate by LC 90 value of 2.3x10 5 PIBs/ ml. Table (1) shows that activity of SpliMNPV without additive was lost soon after treatment. The calculated lethal inactivation time for 50% of the tested S. littoralis neonate larvae (LIT 50 ) value was 24.939 minutes . This activity increased to 1001.119 minutes by adding Cocoa protectant and 480.403 min by Green tea and 376.765 min with Carob additive. Vanilla and Sage showed low rates of protection with LIT 50 value of 132.472 and 109.028 minutes, respectively. The original activity remaining (OAR %) after 5 hours (Table2), recommended both of carob and cocoa for the second screening at a longer exposure to UV light. Screening of the second plant group at the concentration of 1% showed that SpliMNPV without additive has lost 50% of its activity after only 15 minutes. Addition of coffee and lemon grass to the virus provided the best virus protection and extended the persistence (LIT 50 ) up to 286.629 and 160.452 minutes for coffee and lemon grass, respectively. However, wheat, Doum palm and coriander provided low rate of protection. The calculated LIT 50 values were 146.003, 60.587 and 53.796 min, respectively. B

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4 th Conference on Recent Technologies in Agriculture, 2009………………………………………………………

Table (1): Screening of additives at 1% concentration of for the persistence of Spodoptera littoralis nucleopolyhedrovirus (SpliMNPV) exposed to UV radiation. Treatment

LIT 50

OAR%

Group 1 Virus alone Virus + Sage garden Virus + Carob tree Virus + Green tea Virus + Cacao Virus + Vanilla beans

24.939 109.028 376.765 480.403 1001.1193 132.472

2 35 50 53 49 28

Group 2 Virus alone Virus + Doum palm Virus + Lemon grass Virus + Coriander Virus + Green coffee Virus + Wheat

15.634 60.587 160.452 53.796 286.629 146.003

17 23 44 14 48 38

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Group 3 Virus alone Virus + Green Cabbage Virus + Red Cabbage Virus + Parsley Virus + Rucola Virus + Coriander Group 4 Virus alone Virus + Grape fruit Virus + Grape seed Virus + Punica Virus + Garlic Virus + Onion

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28.8 199 329 48 72 41 20.235 477.353 366.305 283 80 79

OAR%= Original Activity Remaining. LIT 50% = lethal inactivation time for 50% of the tested population.

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15 86 61 59 53 49 19 68.33 43 36 31 28.8

Preliminary evaluation of natural antioxidants as…………………………………………………………

Fig. (1): Percentage of Original Activity Remaining (OAR%) values of SpliMNPV exposed to UV light for 10 hours in presence or absence of plant additives. 70 60

Virus alone Cacao

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Red Cabbage GreenCabbage

OAR%

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Coffee Grap fruit

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Carob tree Green tea

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Table (2): Second screening of the best UV protection additives to Spodoptera littoralis NPV (SpliNPV) obtained from the first screening at 1% irradiated for 10 hours.

Serial No. 1 2 3 4 5 6 7 8 9

Additive Virus alone Virus + Cacao Virus + Red Cabbage Virus + Green Cabbage Virus + Coffee Virus + Grape fruit Virus + Carob tree Virus + Green tea Virus + Lemon grass Virus + Grape seed

OAR%= Original Activity Remaining. LIT 50% = lethal inactivation time for 50% of the tested population.

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LIT 50 value B

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222.143 821.443 711.163 686.054 594.423 590.833 530.018 505.446 449.383 439.316

4 th Conference on Recent Technologies in Agriculture, 2009………………………………………………………

OAR% recommends coffee and lemon grass to run the second screening. Results of screening of the third plant group showed that red and green cabbage provided the highest rate of virus protection from the UV. Virus alone treatment lost 50% of its activity after 28.8 min. The addition of Red and Green Cabbage extended the activity time up to 329 and 199 min, respectively. Rucola, Parsley and Coriander provided a relatively lower rate of protection and LIT50 values were 72, 48, and 41 min, respectively. Grape fruit and Grape seed recorded the highest protection among the tested fourth group with LIT 50 values of 477.353 and 366.305 min., respectively compared to SpliMNPV alone treatment which lost half of its activity after 20 minutes. On the other hand, Punica, garlic and onion provided a lower rate of UV protection and LIT 50 values were 283, 79 and 80 min, respectively. The best two ultraviolet protectants from each group were subjected to run the second screening to test ultraviolet exposure time of 10 hours to select the best four candidates. As shown in Fig. (1), Cacao, Red and Green Cabbage and Green Coffee were the best four candidates among the tested 10 products compared to virus alone treatment . The extended persistence of virus measured by LIT 50s values were 821.443, 711.163, 686.054 and 594.423 minutes, respectively after irradiation period of 10 hours. B

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the highest antioxidant activity and phenolic contents compared to the other cruciferous vegetables. Beverages, especially coffee, contributed to a large share of the consumption of polyphenoles, as antioxidants, in the Japanese diet (Yoichi et al., 2009). The protection effect on the baculovirus is possibly due to the preventions of virus anti-oxidation ass well as the absorbance of UV light. El Salamouny et al. (2009a, b) attributed the protection effect of teas, coffee, cocoa and lignin to the high rate of UVB absorbance. Further measurements of the tested material by the spectrophotometery may be necessary to confirm the ratio of absorbance antioxidant effect responsible of UV protection. Nevertheless, these promising groups of virus protectants are worth considered in field test.

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5. REFERENCES Abul Nasr S. (1956). Polyhedrosis virus disease on cotton leaf worm Prodenia litura. Bulletin Entomological Society Egypt, Economic Series .40: 321-332. Arivudanainambi S., Selanarayanon V. and Vikash A.(2000). Enhancing the efficacy and persistency of Spodoptera litura (Fab.) nuclear polyhedrosis virus using UV irradiation protectants. Indian Journal of Experimental Biology. 38 (11):1175-1176. Baskaran R.K. M. (2001): In vivo screening of optical brighteners as UV protectants and their efficacy in enhancing the virulence of Maduri isolate of NPV of Amascata albistriga (Walker).Lepidoptera: Arctiidae. Journal of Biological Control.2 (1):169-174. Chiho M., Tomoko Y., Hitoshi T. and Terugoshi M. (2008). Changes in radical scavengering activity on sliced red and green cabbages during storage.Biofactors.21:297-299. Cisneros J. Peterz J. A. , Penagos, D. I., Ruiz, D., Goulson, P. Caballero, P. and Williams , T. (2002). Formulation of baculovirus with boric acid for control of Spodoptera frugiperda(Lepidoptera:Noctuidae). Biological Control.23:87-95. Deotale R. O., Dawane P. N. Biswane K. D. and Borker S. L. (2007). Effectiveness of UV protectants on the activity of NPV against Helicoverpa armigera (Hubner) on chickpea. Journal of Entomology Research. 31(1): 33-35. Deotale R. O., Dawane P. N., Underwide D. B., Deshmukh I. B. and Bisane K. D.( 2003). Performance of UV protectants for the effectivness of HaNPV against Helicoverpa

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4. DISCUSSION The first record of the use of plant extracts to increase the persistence of insect viruses was by Shapiro et al. (2007a, b). Both of green tea and black tea were reported to be UV protectants to the beet armyworm nucleopolyhedrovirus (Shapiro et al., 2008 and El Salamouny et al., 2009). This effect could be due to the antioxidants present in tea. Also, mango leaf extract provided a protection effect to baculovirus from the ultraviolet light (Deotale et al., 2007). The obtained results that coffee and cocoa acted as UV protectant to SpliMNPV agree with that recently found by El Salamouny et al. (2009 b) for the beet armyworm nucleopolyhedrovirus. It is known that vegetables such as cabbages are rich of antioxidant compounds and polyphenoles, which scavenge free radicals and reactive oxygen species (Chiho et al., 2008). Our result of the red cabbage that improved the persistence of SpliMNPV from the ultraviolet light could be explained by the finding by Lee et al. (2007) that red cabbage has

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Preliminary evaluation of natural antioxidants as………………………………………………………… armigera on chickpea. Journal of Soils and Crop. 13(1): 154-157. Elnagar S. (1982). The inactivation of nuclear polyhedrosis virus by ultraviolet radiation. Bulletin Entomology Society Egypt Economic Series. 13:171-174. Elnagar S., El-Sheikh, M. A. K., El-Salamouny, S., Amin, A., and Khattab, M. (2003). Screening of four lignin products as UV protectants to baculovirus. Bulletin Entomological Society Egypt, Economic Series . 29:165-178. El Salamouny S., Herz A., and Huber J. ( 2002 ). Suitability of three lignin products as UV protectants to baculovirus. Bulletin Entomological Society Egypt, Economic Series 28, 103-111. El Salamouny S., Shapiro M., Ling K. S. and Shepard B. M. (2009a). Black tea and lignin as Ultraviolet protectants for the beet armyworm nucleopolyhedrovirus. Journal of Entomological Science. 44(1): 50-58. El Salamouny S., Ranwala D., Shapiro M., Shepard M. and Farrar R. (2009b). Tea, coffee, and cocoa as ultraviolet radiation protectants for the beet armyworm nucleopolyhedrovirus. Journal of Economic Entomology. 102(5): (In Press). Finney D. J. (1971). Probit analysis. 3 rd ed., Cambridge Univ., Cambridge U.K. Fritsch E. and Huber J. (1985). Inactivation of codling moth granulosis viruses by ultraviolet radiation and temperature. Nachrichtenblatt des deutshen pfanzenchutzdientes, 37(6),84-88. Ignoffo C. M, Garcia, C. (1994): Antioxidant and oxidative enzyme effects on the inactivation of inclusion bodies of the Heliothis baculovirus by simulated sunlight-UV. Environmental Entomology. 23 (4): 1025-1029. Ignoffo, C. M., Rice, W. C. and Mcintoch, A. H. (1989): Inactivation 0f non occluded and occluded baculovirus and baculovirus-DNA exposed to simulated sunlight .Environmental Entomology.18(1)177-183. Jacques R. P. (1977). Stability of entomopathogeneic viruses. Miscell publication of the Entomological Society of America.10 (3): 99-116. Jones K. A., Moawad G. M., Makinley D. J. and Grzywacz D. (1993): The effect of natural sunlight on Spodoptera littoralis nuclearpolyhedrosis virus. Biologicl Science and Technology. 3:189-197. Kamaldeep S. and Battu G. S.(2008). Residual persistence of the HearNPV against

Helicoverpa armigera (Hubner) on tomato as affected by two selected optical brighteners. Indian Journal of Entomology. 10(1):26-30. Lee W. Y., Emmy H. K. I., Abbe M. U. J. and Amin I. (2007). Antioxidant capacity and phenolic contents of selected commercially available cruciferous vegetables. Journal of Nutrients.13 (1): 71-80. Manjari M., Tanejo T. K., Sudhir S., Krishnoveni M., Rashidonisa B., Mohammed A., Sah N. K. and Hassanin S. E. (2003). Antioxidant prevent UV induced apoptosis by inhibiting mitochondrial cytochromoe release and caspase activation in Spodoptera frugiperda (Sf9) cells . Journal of Cell Biology International . 27(6): 483-490. Muro E. M. and Paul J. I. (1985). Laboratory evaluation of new ultraviolet absorbers for protection of Douglas-fir tussock moth (Lepidoptera: Lymantriidae) baculovirus. Journal of Economic Entomology. 78. 951957. Murohabaskaran R. K., Venugopal M. S. and Mahadevan N. R. (2000). Potential of vitamins as ultraviolet screens for Spodoptera litura (Lepidoptera; Noctuidae) nuclear polyhedrovirus. Indian Journal of Agriculture Science.10 (4): 264-266. Ramakrichnan N. and Chaudhary S.(1991).Effectiveness of some ultraviolet light protectants on the activity of nuclear polyhedrosis virus of Spodoptera litura (Fabricius). Indian Journal of Entomology. 53(4): 548-551. Reddy A. R. and Divakar B. J (2001). Effect of Rubin blue on the efficiency of nuclear polyhedrosis for management of Helicoverpa armigera (Hub)in ground nut crop. National perspectives in biotechnology Proceedings of a national symposium wrangler, India.26-27 Febrauary-1999-2001; 41-43. Shapiro M.(1989). Congo red as an ultraviolet protectant for the gypsy moth (Lipidoptera: Lymantriidae) nuclear polyhedrosis virus. Journal of economic Entomology.82(2):548550. Shapiro M., El Salamouny S., and Shepard B. M. (2008). Green tea extracts as ultraviolet protectants for the beet armyworm, Spodoptera exigua nucleopolyhedrovirus. Journal of Biocontrol Science and Technology. 18(6): 591-603. Shapiro M., B. M. Shepard and R. Lopez. (2007a). Effect of spices upon the activity of the gypsy moth (Lepidoptera: Lymantriidae)

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‫‪nucleopolyhedrovirus. Journal Entomological‬‬ ‫‪Science. 42: 84-91.‬‬ ‫‪Shapiro M., B. M. Shepard and R. Lopez.‬‬ ‫‪(2007b). Effects of medicinal herbs on the‬‬ ‫‪biological activity of the gypsy moth‬‬ ‫‪nucleopolyhedrovirus. Journal Entomological‬‬ ‫‪Science. 42: 426-429.‬‬ ‫‪Shorey H. and Hale R. L. (1965). Mass rearing of‬‬ ‫‪the larvae of nine noctuid species on a simple‬‬ ‫‪artificial medium. Economic Entomology,‬‬ ‫‪58:522-524.‬‬ ‫‪Tamez-Guerra P., McGuire M., Behle R. W.,‬‬ ‫‪Hamm J. J., Sumner H. S. and Shasha B.‬‬ ‫‪S.(2000). Sunlight persistence and rain fastness‬‬ ‫‪of spray-dried formulations of baculovirus‬‬ ‫‪isolate from Anagrapha falcifera (Lepidoptera:‬‬

‫‪Noctuidae). Journal of Economic Entomology.‬‬ ‫‪93(2): 210-218.‬‬ ‫‪Yoichi F., Takashi Y. J., Suhiko Y., Kohel‬‬ ‫‪Y.,Hiroki A., Yoko M.,Makto W., Masato T.,‬‬ ‫‪Malko H.,Chie T., and Kazuo K. (2009).‬‬ ‫‪Coffee and green tea as large source of‬‬ ‫‪antioxidant polyphenoles in the Japanese‬‬ ‫‪population. Journal of Agriculture Food and‬‬ ‫‪Chemistry. 57(4): 1253-1259.‬‬ ‫‪Zarin Sh. I. and Eglite G. (1985). Antioxidant‬‬ ‫‪promising compounds for the protection of‬‬ ‫‪entomopathogeneic viruses from UV rays:‬‬ ‫‪Trudy- latviskoni-Sel. Skokhozyaistvennoi.‬‬ ‫‪Akademii. 222: 12-15.‬‬

‫ﺘﻘﻴﻴﻡ ﻤﺒﺩﺌﻲ ﻟﻤﻀﺎﺩﺍﺕ ﺍﻷﻜﺴﺩﺓ ﺍﻟﻁﺒﻴﻌﻴﺔ ﻜﻤﻭﺍﺩ ﺤﺎﻤﻴﺔ ﻤﻥ ﺍﻷﺸﻌﺔ ﻓﻭﻕ ﺍﻟﺒﻨﻔﺴﺠﻴﺔ‬ ‫ﻟﻔﻴﺭﻭﺱ ﺩﻭﺩﺓ ﻭﺭﻕ ﺍﻟﻘﻁﻥ ﺍﻟﺒﻭﻟﻴﻬﺩﺭﻭﺯﻱ ﺍﻟﻨﻭﻭﻯ‬ ‫ﺃﻟﻜﺴﻨﺩﺭﺍ ﺍﻟﻬﻼﻟﻰ ‪ -‬ﺴﻌﻴﺩ ﺍﻟﺴﻼﻤﻭﻨﻰ‪ -‬ﻤﺎﺠﺩﺓ ﺨﻁﺎﺏ* ‪ -‬ﻤﺤﻤﺩ ﺍﻟﺸﻴﺦ – ﺼﻼﺡ ﺍﻟﺩﻴﻥ ﺍﻟﻨﺠﺎﺭ‬ ‫ﻗﺴﻡ ﺍﻟﺤﺸﺭﺍﺕ ﺍﻻﻗﺘﺼﺎﺩﻴﺔ ﻭﺍﻟﻤﺒﻴﺩﺍﺕ ‪ -‬ﻜﻠﻴﺔ ﺍﻟﺯﺭﺍﻋﺔ ‪ -‬ﺠﺎﻤﻌﺔ ﺍﻟﻘﺎﻫﺭﺓ‬ ‫* ﻗﺴﻡ ﺍﻟﺩﻭﺩﺓ ﺍﻟﻘﺎﺭﻀﺔ ﻭﺍﻟﺤﻔﺎﺭ‪ ،‬ﻤﻌﻬﺩ ﺒﺤﻭﺙ ﻭﻗﺎﻴﺔ ﺍﻟﻨﺒﺎﺕ‪ -‬ﻤﺭﻜﺯ ﺍﻟﺒﺤﻭﺙ ﺍﻟﺯﺭﺍﻋﻴﺔ‪ -‬ﺍﻟﺩﻗﻰ‬ ‫ﺍﻟﺠﻴﺯﺓ ‪ -‬ﻤﺼﺭ‪.‬‬ ‫ﻤﻠﺨﺹ‬ ‫ﺘﻡ ﺘﻘﻴﻴﻡ ﻋﺸﺭﻭﻥ ﻨﺒﺎﺕ ﻁﺒﻴﻌﻰ ﻤﺤﺘﻭﻴﻪ ﻋﻠﻰ ﻤﺠﺎﻤﻴﻊ ﻤﺨﺘﻠﻔﺔ ﻤﻥ ﻤﻀﺎﺩﺍﺕ ﺍﻻﻜﺴﺩﺓ ﻜﻤﻭﺍﺩ ﻤﻀﺎﻓﺔ ﻟﻔﻴﺭﻭﺱ ﺩﻭﺩﺓ ﻭﺭﻕ‬ ‫ﺍﻟﻘﻁﻥ ﺍﻟﺒﻭﻟﻴﻬﺩﺭﻭﺯﺱ ﺍﻟﻨﻭﻭىﻼﻁﺎﻟﺔ ﺍﻟﻤﺩﺓ ﺍﻟﺘﻰ ﻴﻅل ﻓﻴﻬﺎ ﺍﻟﻔﻴﺭﻭﺱ ﻤﺤﺘﻔﻅﺎ ﺒﻜﻔﺎﺀﺘﻪ‪ .‬ﻜﺎﻨﺕ ﻨﺘﺎﺌﺞ ﺍﻟﻤﺴﺢ ﻤﺒﻨﻴﺔ ﻋﻠﻰ ﺍﻻﺨﺘﺒﺎﺭﺍﺕ‬ ‫ﺍﻟﺤﻴﻭﻴﺔ ﺍﻟﻤﻌﻤﻠﻴﺔ ﻻﺨﺘﺒﺎﺭﺍﻟﻜﻔﺎﺀﺓ ﺍﻷﺼﻠﻴﺔ ﺍﻟﻤﺘﺒﻘﻴﺔ ﻟﻠﻔﻴﺭﻭﺱ ﺒﻌﺩ ﺍﻟﺘﻌﺭﻴﺽ ﻟﻤﺼﺩﺭ ﺼﻨﺎﻋﻰ ﻟﻸﺸﻌﺔ ﻓﻭﻕ ﺍﻟﺒﻨﻔﺴﺠﻴﺔ‪ .‬ﺘﻡ ﺍﺨﺘﺒﺎﺭ‬ ‫ﺍﻟﻤﺼﺎﺩﺭ ﺍﻟﻤﺨﺘﻠﻔﺔ ﻟﻤﻀﺎﺩﺍﺕ ﺍﻷﻜﺴﺩﺓ ﻓﻰ ﻤﺠﺎﻤﻴﻊ ﻓﻰ ﺃﺭﺒﻊ ﺘﻘﺩﻴﺭﺍﺕ ﺤﻴﻭﻴﺔ‪ .‬ﺃﻅﻬﺭﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺍﻟﻤﺒﺩﺌﻴﺔ ﺃﻥ ﺍﻟﻤﻌﺎﻤﻼﺕ ﺍﻟﺘﻰ‬ ‫ﺍﺴﺘﺨﺩﻡ ﻓﻴﻬﺎ ﺍﻟﺨﺭﻭﺏ ﻭ ﺍﻟﻜﺎﻜﺎﻭ ﺃﻋﻁﺕ ‪ % OAR ٤٩ ، ٤٩,٥‬ﻋﻠﻰ ﺍﻟﺘﺭﺘﻴﺏ‪ ،‬ﺒﻌﺩ ﺨﻤﺱ ﺴﺎﻋﺎﺕ ﻤﻥ ﺍﻟﺘﻌﺭﻴﺽ ﻟﻠﻤﺼﺩﺭ‬ ‫‪، % OAR‬ﻋﻠﻰ ﺍﻟﺘﺭﺘﻴﺏ‬ ‫ﺍﻟﺼﻨﺎﻋﻰ ﻻﺸﻌﺔ ﺍﻟﺸﻤﺱ ﻓﻭﻕ ﺍﻟﺒﻨﻔﺴﺠﻴﺔ‪ .‬ﺃﻋﻁﺎ ﺍﻟﻜﺭﻨﺏ ﺍﻻﺤﻤﺭ ﻭ ﺍﻷﺨﻀﺭ ‪٥٠,٩ ، ٥٣,٠٥‬‬ ‫‪، % OAR‬ﻋﻠﻰ ﺍﻟﺘﺭﺘﻴﺏ‪ .‬ﻭﺃﻋﻁﺕ ﺒﺫﻭﺭ ﻭ ﺜﻤﺎﺭ‬ ‫‪.‬ﺃﻋﻁﻰ ﻜل ﻤﻥ ﻤﺴﺤﻭﻕ ﺍﻟﺒﻥ ﻭ ﺤﺸﻴﺸﺔ ﺍﻟﻠﻴﻤﻭﻥ ‪٤٤,١٨ ،٤٧,٩٦‬‬ ‫‪ % OAR‬ﻋﻠﻰ ﺍﻟﺘﺭﺘﻴﺏ‪.،‬ﻭ ﻗﺩ ﺘﻡ ﻋﻤل ﻏﺭﺒﻠﺔ ﻤﺭﺓ ﺜﺎﻨﻴﺔ ﻟﻌﻤل ﺘﺤﺩﻴﺩ ﺍﻜﺜﺭ ﻟﻠﻤﻭﺍﺩ ﺍﻟﺘﻰ ﺍﻋﻁﺕ‬ ‫ﺍﻟﻌﻨﺏ ‪٥١,٦٦ ،٧٢,٥‬‬ ‫ﺃﻓﻀل ﺍﻟﻨﺘﺎﺌﺞ ﺒﻌﺩ ﻋﺸﺭ ﺴﺎﻋﺎﺕ ﺘﻌﺭﻴﺽ ﻟﻼﺸﻌﺔ ﻓﻭﻕ ﺍﻟﺒﻨﻔﺴﺠﻴﺔ ﻭ ﺃﻅﻬﺭﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﺍﻟﻜﺭﻨﺏ ﺍﻷﺨﻀﺭ‪ ،‬ﺍﻟﻜﺎﻜﺎﻭ ﻭ ﺍﻟﻘﻬﻭﺓ‬ ‫ﺃﻋﻁﻭﺍ ﺍﻟﻤﻌﺩﻻﺕ ﺍﻻﻋﻠﻰ ﻻﻁﺎﻟﺔ ﻤﺩﺓ ﻜﻔﺎﺀﺓ ﺒﻘﺎﺀ ﺍﻟﻔﻴﺭﻭﺱ ﺒﻴﻨﻤﺎ ﺜﻤﺎﺭ ﺍﻟﻌﻨﺏ‪ ،‬ﺤﺸﻴﺸﺔ ﺍﻟﻠﻴﻤﻭﻥ‪ ،‬ﺒﺫﻭﺭ ﺍﻟﻌﻨﺏ ﻜﺎﻨﺕ ﺍﻻﺩﻨﻰ ‪.‬‬ ‫ﺘﺨﻀﻊ ﺍﻟﻤﻭﺍﺩ ﺍﻟﻤﺨﺘﺒﺭﺓ ﺍﻻﻥ ﻟﻤﺯﻴﺩ ﻤﻥ ﺍﻻﺨﺘﺒﺎﺭﺍﺕ ﻤﺜل ﺍﻟﺴﺒﻜﺘﺭﻭﻓﻭﺘﻭﻤﻴﺘﺭ ﻭ ﺍﺠﺭﺍﺀ ﺘﺠﺭﺒﺔ ﺤﻘﻠﻴﺔ ﻤﺼﻐﺭﺓ ﺘﺤﺕ ﺍﻟﻅﺭﻭﻑ‬ ‫ﺍﻟﺤﻘﻠﻴﺔ ﺍﻟﻤﺼﺭﻴﺔ‪.‬ﻭ ﺘﻭﺼﻰ ﺍﻟﺩﺭﺍﺴﺔ ﺒﺎﺴﺘﺨﺩﺍﻡ ﺍﻻﻀﺎﻓﺎﺕ ﺍﻟﻁﺒﻴﻌﻴﺔ ﻜﻤﻭﺍﺩ ﺤﺎﻤﻴﺔ ﻟﻤﺠﻤﻭﻋﺔ ﻓﻴﺭﻭﺴﺎﺕ ‪ Baculovirus‬ﻀﺩ‬ ‫ﺃﺸﻌﺔ ﺍﻟـ ‪.UV‬‬

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