Bioscience Biotechnology Research Communications

An Open Access International Journal

P-ISSN: 0974-6455 E-ISSN: 2321-4007

Bioscience Biotechnology Research Communications

An Open Access International Journal

Masoud Radmanesh

PhD Agriculture and Plant Breeding, Mazandaran, Iran

Corresponding author email:


Article Publishing History


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The contaminative effect of the two organic pesticides, Spinosad and NPVs, on the newborn worms of Spodoptera littoralis (Bosiduval) (Lepidoptera: Noctuidae) was examined within lab examination  circumstances to discover their competitive capacity. The capability of Spinosad to guard the Split NPV against Ultra Violet impacts under manufactured examination room circumstances was demonstrated, and various biological features of both organic pesticides and their combination were examined. In an attempt to discover whether or not there is a  coordinated impact  when the two of these organic insecticides are mixed with each other, six particular Spinosad composites (1, 2, 5, 10, 15 and 30 ppm) in isolation and in connection with a dangerous mixture of split NPV (1×103) were examined. As the Ultra Violet impact was recognized, the LC90 of NPVs was combined with LC10 of Spinosad in an attempt to inspect the capacity of Spinosad in extending the virus life. A study was taken place in the Department of Entomology (Virology Unit) Faculty of Agriculture, Cairo University, between July 2012 and May 2013.

The mortality rate multiplied, as it was 11.66, 19.33, 33.33, 55.00, 71.66, and 85.00 % in comparison with 11.66, 13.33, 15.00, 26.66, 36.66, and 63.33 % in isolated Spinosad and 20.11% in isolated NPVs analysis. A virtually unmitigated protection was observed after 30 minutes of exposure to manufactured ultraviolet light and showed 47 %  mortality rate 5 hours after the procedure, in contrast with the 2.8 % mortality rate which is exhibited when NPVs is employed in isolation. The larval stage was only impacted by Spinosad; pupal stage and adult lifespan were not impacted by the entire examination conditions. The sum of eggs laid by each female and their fertility rate were impacted in Spinosad and Spinosad NPVs coordinative functions in comparison with of the number that was reported by the control group. The discoveries of this study signify that Spinosad and NPVs, the organic pesticides, can be employed in affiliation with each other, which in turn lead to propitious results when it comes to annihilating the Mediterranean Brocade. The results of the research suggest that Spinosad and NPVs introduce a significant option to be employed in combined efforts in insect tackling in which Spodoptera littoralis is the primary insect.


Spodoptera littoralis, Spinosad, Nuclear Polyhydrosis Viruses, Coordination, Biological Traits.

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Radmanesh. M. The Combined and Isolated Effect of Spinosad and Nuclear Polyhedrosis Virus on the Mediterranean Brocade Spodoptera littoralis in laboratory Conditions. Biosc.Biotech.Res.Comm. 2021;14(2).

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Radmanesh. M. The Combined and Isolated Effect of Spinosad and Nuclear Polyhedrosis Virus on the Mediterranean Brocade Spodoptera littoralis in laboratory Conditions. Biosc.Biotech.Res.Comm. 2021;14(2). Available from: <a href=”“></a>


Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) is one of the most hostile insects in Africa, Asia, and Europe (Khayatnezhad and Gholamin, 2021a, Gholamin and Khayatnezhad, 2020d, Karasakal et al., 2020a, Si et al., 2020, Aletor, 2021). Its negative impact on the vegetables and decorative plants undermines their marketability. The extensive use of various insecticides to annihilate this insect has led to obstruction of many approved insecticides (Alayi et al., 2020, Esmaeilzadeh et al., 2020).

The makeup of entomopathogens could highly impact their effectiveness as organic  nanoparticle pesticides (Arjaghi et al., 2021). The makeup in specific can affect the immutability of the pathogen in when it comes to its storing ability and the effectiveness of its utilization on the crop. Furthermore, specific makeup assistants can improve the function of the pathogen and enhance its ecological resistance (Khayatnezhad and Nasehi, 2021, Gholamin and Khayatnezhad, 2020c, Sun et al., 2021).

In an attempt to enhance the function of the pathogen, one can mix them with limited amounts of cooperative components such as visual enhancers (Hewitt, 2021), mineral acids (Fataei, 2017, Ghomi Avili and Makaremi, 2020) or moderately deadly combinations of chemical pesticides (Gholamin and Khayatnezhad, 2020b, Huang et al., 2021). Nonetheless, the cooperation between a pathogen and other composites might as well bring about adverse results because of the cutback in nourishment or the alteration of pH in the intestines (Karasakal et al., 2020b, Li et al., 2021), or the autonomous function of each substance which may cause an increase in the mortality rate (Fataei et al., 2018).

Spinosad is a combination of spinosyns A and D generated while the soil actinomycete Saccharopolyspora spinosa is being dissolved. Spinosad is fundamentally an intestine toxin which exhibits various external activities and is specifically poisonous when it comes to Lepidoptera and Diptera. Nonetheless, contamination examinations suggest that Spinosad possesses literally no contamination risks to birds and mammals and possesses approximately low contamination risks to specific bug assailants in the nature (Omrani and Fataei, 2018), while a number of insect hunters and freeloaders seem to be vulnerable to Spinosad contamination ( Jia et al., 2020, Gholamin and Khayatnezhad, 2020a, Huma et al., 2021).

It is rather significant to examine the contamination of Spinosad and NPVs as organic pesticides and the effects of their combination on the new born larvae of Spodoptera littoralis, alongside the impact of such procedures on specific biological features in an attempt to understand whether Spinosad has an impact on guarding the NPVs or not. The purpose of this study is to improve the effectiveness of NPVs by mixing them with moderately deadly composites of Spinosad in an attempt to achieve a higher level of command over Spinosad littoralis.

Empirical Data

Insect Colony :A colony of the Mediterranean Brocade, Spodoptera littoralis (Boisduval), was set up as the trial insect group on a somewhat unnatural daily food intake of Shorey and Hale (1965) under the pursuing circumstances in the laboratory: Thermal reading of 25 ± 2°C and 65 ± 5%R.H. and the physiological response to the light and dark periods of 16:8 (L: D).

Virus Inoculation: A separate group of Spodoptera littoralis combined with another separate group of Nuclear Polyhedrosis Virus (Split MNPV) which was previously secluded in Egypt was employed within the study.

Chemical tested: The insecticide upon which the examination was carried was Tracer (24% Spinosad, liquid formulation containing solid pesticide active components that need to be mixed with water before use; Dow Agro Sciences, Alexandria, Egypt). 

The Biological Assessment: The effect of sunlight UV rays (SUV) was replicated by a set of four UV lamps (Ultra-Vitalux, OSRAM, Germany), which were vertically placed 160 cm away from the susceptible virus samples, and there was a 60-cm space in the middle of the two lamps. The organic impact is estimated nearly 6-7 times more than the actual sun-light when the space between unnatural sunlight, lamp, and moisture less solid is determined exactly 50 cm (Huber and Ludcke, 1996).

A surfactant (Teepol 2.5%) was intermixed with the virus composite, and 50 μl  of it was diffused within a Petri dish (10 cm in diameter) through a fine pipette . When the moisture from the surfaces was removed using air, the dishes containing the virus films were subjected to the lightening source of the experiment which entailed the subjection of virus combination to UV rays (2000 fold LC 90 PIB’s). The polyhedra solids in the Petri-dish were ejected into in 10 ml distilled water with 2.5% Teepol for the second time in order to achieve the desired state for the application in biological assessments.

The virus infected Split NPV was diffused in distilled purified water and the virus composite was altered to be composed of 108 OBs/ml (=LC 90-95 %). Virus endurance was estimated by % OAR (percentage of Original Activity Remaining) according to 100% destruction rate at ‘0’ day after the examination was taken place. Analytical assessments were once more taken place in three simulated experiment groups with 10 larvae used for each examination (Shapiro et al., 2008).

Biological Features : The juvenile worms that withstood all conditions within the experiment were gathered and some biological features of them were documented in order for them to be compared with the larvae (control) that were not employed within the experiment. These biological features involved larval stage, pupal stage, adult life-span, the number of the eggs each female larva laid and their fertility rate.

Numerical Assessment: Concentration-dependent mortality lapse were estimated in order to discover the impact of plant-based substances, which might act as Split MNP UV shielding supplements. Slopes and LC50s were evaluated based on the design introduced by Finney (Finney, 1971). Initial motion remaining percentages were designated for each demonstrated experiment by the application of both factors to guarantee the substances’ capacity to extend the virus endurance as detailed by Muro and Paul (Martignoni and Iwai, 1985, Kabir et al., 2021).


The United Impact: In a study on organic insecticides that was taken place recently researchers located in the countries with less developed industrial base demonstrated that composition was the most significant problem in the production of organic pesticides ( Radmanesh, 2021). Because of the fact that spinosyns are developed through fermentation of an actinomycete, Spinosad has been categorized as an organic insecticide (Huang et al., 2021), despite the fact that it evidently possesses features that affords it to be used as a pesticide which can be distinguished from most of entomopathogen-based organic insecticides (Gholamin and Khayatnezhad, 2021, Rodríguez, 2021).

Table 1. The Impact of Spinosad and NPVs in Isolation and Their Combination on the New Born Larvae of Spodoptera littoralis under Examination Room Circumstances.

Spinosad concentrations Mortality percent
Spinosad NPVs 1×103 Spinosad+ NPVs 1×103
R1 R2 R3 Mean R1 R2 R3 Mean R1  



R3 Mean
30 ppm

15 ppm

10 ppm

5 ppm

2 ppm

1 ppm





























































The cooperation of Spinosad with entomopathogens has not been studied before, and the attempt of doing scientic research on SlMNPV–Spinosad intermixtures was thought to be  worthwhile since Spinosad exhibits no antimycotic, antimicrobial or antiviral capacity (Khayatnezhad and Gholamin, 2021b, Wan, 2021).

Table 2. The Impact of LC10 Spinosad Additive on the Endurance of Spodoptera littoralis NPV Split NPV Subjected to Unnatural UV Light Rays and Biological Assessment on S.  littoralis New Born Larvae.

Irradiation period (min) Mortality (%) due to Spli NPV alone Mortality (%) due to virus + Spinosad at LC10
Control (D. W.) 0 0
300 min 2.8 47
240 min 4 50
180 min 11.6 56.5
120 min 14.28 66
60 min 38 96
30 min 70 99
Zero time 89.4 90

The LC50 rate which was estimated for newborn larvae of S. littoralis subjected to Spinosad under the diet of Shorey and Hale through surface intoxication method was literally entirely dissimilar to the 3 ppm rate (95% C.L.: 1.10–6.60) of spinosyn A documented for S. littoralis larvae in the undetermined stages which were submerged in water..The rate was documented while employing composites ranging from 1-100 ppm and it was reported to be 27.23 ppm.

Table 3. The Impact of NPVs, Spinosad in Isolation and NPVs Spinosad Intermixture on various biological

Aspects Pesticides Larval Duration/Day Pupal Stage/Day Adult longevity/Day No. of Laid Egg/Female Hatchability percentage


NPVs Spinosad Mixture






















The destruction rate of larvae subjected to SlMNPV 1× 103 combined with the smallest amassing of Spinosad (1ppm) proved to be less than the anticipated rate in isolation (i.e., a level of hostility was noticed between these items). A small level of cooperation was noticed in the larvae subjected to SLMNPV+ 30 ppm Spinosad, while the biological reason for such interplay is not known at the very moment. The sequence of insect destruction during different periods of time was considered significant because of the notable dissimilarities in the mortality rate of SlMNPV and Spinosad (Gholamin and Khayatnezhad, 2020b).

The normally approved quantile function or the logarithm which are used in biological assessments do not provide us with the appropriate assessment mediums when it comes to the analysis of the  concentration–dependent destruction rate in virus–pesticide combinations; first, since the binomial distribution of reactions does not normally abide from logistic or Gaussian distributions because of the dissimilarities in the activity patterns and/or interplay among the virus and pesticide, and second, because of the fact that the sum of reactions of entities involved in an examined group are associated with the elements of time (Khayatnezhad and Gholamin, 2020a,).

Nevertheless, the combination of Spinosad exhibited a significant impact on the mortality rate which in turn entailed a notable reaction in less than 48 hours which was then trailed by virus-caused destruction of the larvae that withstood the Spinosad examination a few days later. The control rate of S. littoralis larvae was estimated due to the decline in larval rehabilitation within the plants that were used in the trial (probably because of the Spinosad-caused destruction) and the destruction rate that was noticed in the examination room caused by the virus. In comparison to the virus in isolation, the level of insect control was increased to a great degree by adding the 30 ppm Spinosad to the virus combination (Khayatnezhad and Gholamin, 2020b).

The Preservative Impact: Spinosad was examined as a preservative supplement in order to discover whether the intermixture of these two organic insecticides have improved their effects as preservative substances from the ultraviolet light, which might be taken into account as one of the most mitigating forces in employment of the virus. The results exhibit (Table 2) that moderately deadly quantities of Spinosad LC10 provided total preservation effect 30 min after being employed in the trial and in 5 hours exhibites 47% destruction rate in contrast with 2.8 in the examination of the virus in isolation. No sufficient data is at hand on the effect of Spinosad and NPVs on Spodoptera littoralis within this study; it was been discovered that examining different kinds of pesticides that destruct or mitigate the biological procedures of the examined insect provide valuable insight on the various options on the combined insect control methods.

The Impact on the Biological Features of the Insect : The larval stage, pupal stage, and adult life-span were not impacted in the tested group as opposed to the control group (Table 3). No dissimilarities were observed within all the tested groups (Table 3) in larval stage, pupal stage, and adult life-span. The number of eggs laid by each female was impacted in all the examined groups in comparison with the control group, specifically while spinosad and SpLiNPV were employed as a mixture. The fertility rate was declined in all the tested groups, and specifically within the SplitNPV Spinosad combination (45.2%) as opposed to the control group (60.2%).

Wang et al. (2009) discovered that Spinosad in moderately deadly combinations notably prolonged the growth age of Helicoverpa armigera and reduced the development rate, fertility, and life-span of the adults. Pesticides that were incorporated together present a significant area to explore in S. littoralis management for the public health purposes. Diverse effects of such mixtures go hand in hand with that of conventional pesticides, which entails that endurance is definitely anticipated to develop when the pesticide is employed during a long period of time and one cannot exactly determine to what extent combinations such as this one are going to remain effective since the worms have developed resistance to other combinations before, and continue to develop in spite of them.

However, pesticide mixtures with varying roles could contribute  to a great deal in S. littoralis management methods, specifically in areas where S. littoralis already exhibits high levels of endurance to normally used pesticides. The accessibility of new groups of pesticides has been limited with the past 10 years and depending upon the emergence of new pesticides is not a viable alternative for the management of enduring insects within the upcoming days or even years. However, the choice of incorporating different insecticides with various functions is within reach at this very moment.


The contamination rate of the two organic pesticides, Spinosad and NPVs, on the new born larvae of Spodoptera littoralis (Bosiduval) (Lepidoptera: Noctuidae) was examined within the examination room circumstances with an attempt to examine their antagonistic features. The capacity of Spinosad to guard the Split NPV against Ultra Violet light rays within cooperative examination room circumstances was discovered, and various biological features of the two organic pesticides and their combination was examined. The results actively suggest that organic pesticides Spinosad and NPVs can be employed as a mixture which in turn exhibit potential impact on the destruction of Mediterranean Brocade. The findings of this study indicate that Spinosad and NPVs provide a significant option for application in combined insect control methods when S. littoralis is the insect of interest.


Alayi, R., Sobhani, E. & Najafi, A. 2020. Analysis Of Environmental Impacts On The Characteristics Of Gas Released From Biomass. Anthropogenic Pollution Journal, 4, 1-14.

Aletor, S. 2021. Environmentally Induced Alternative Livelihood Strategies Among The Artisanal Fishers Of The Kainji Lake Basin, Nigeria. Water And Environmental Sustainability, 1, 1-7.

Arjaghi, S. K., Alasl, M. K., Sajjadi, N., Fataei, E. & Rajaei, G. E. 2021. Retraction Note To: Green Synthesis Of Iron Oxide Nanoparticles By Rs Lichen Extract And Its Application In Removing Heavy Metals Of Lead And Cadmium. Biological Trace Element Research, 1-1.

Esmaeilzadeh, H., Fataei, E. & Saadati, H. 2020. Nh3 Removal From Sour Water By Clinoptilolite Zeolite: A Case Study Of Tabriz Refinery. Chemical Methodologies, 4, 754-773.

Fataei, E. 2017. Soil Carbon, Nitrogen And Phosphorus Pools Under Exotic Tree Plantations In The Degraded Grasslands Of Iran. Agricultural & Biological Research, 33, 113-127.

Fataei, E., Varamesh, S. & Seiied Safavian, S. T. 2018. Effects Of Afforestation On Carbon Stocks In Fandoghloo Forest Area. Pakistan Journal Of Agricultural Sciences, 55.

Finney, D. 1971. Probit Analysis, Cambridge University Press. Cambridge, Uk.

Gholamin, R. & Khayatnezhad, M. 2020a. Assessment Of The Correlation Between Chlorophyll Content And Drought Resistance In Corn Cultivars (Zea mays). Helix, 10, 93-97.

Gholamin, R. & Khayatnezhad, M. 2020b. The Effect Of Dry Season Stretch On Chlorophyll Content And Rwc Of Wheat Genotypes (Triticum durum L.). Bioscience Biotechnology Research Communications, 13, 1833-1829.

Gholamin, R. & Khayatnezhad, M. 2020d. The Study Of Path Analysis For Durum Wheat (Triticum durum Desf.) Yield Components. Bioscience Biotechnology Research Communications, 13, 2139-2144.

Gholamin, R. & Khayatnezhad, M. 2021. Impacts Of Peg-6000-Induced Drought Stress On Chlorophyll Content, Relative Water Content (Rwc), And Rna Content Of Peanut (Arachis hypogaea L.) Roots And Leaves. Bioscience Research, 18, 393-402.

Ghomi Avili, F. & Makaremi, M. 2020. Predicting Model Of Arsenic Transport And Transformation In Soil Columns And Ground Water Contamination (Case Study: Gorgan Plain, Iran). Anthropogenic Pollution Journal, 4, 57-64.

Hewitt, E. 2021. Ecological Plunging And Wireless Filming For Science Education: A New Zealand Pilot Experimeent. Water And Environmental Sustainability, 1, 24-29.

Huang, D., Wang, J. & Khayatnezhad, M. 2021. Estimation Of Actual Evapotranspiration Using Soil Moisture Balance And Remote Sensing. Iranian Journal Of Science And Technology, Transactions Of Civil Engineering, 1-8.

Huma, Z., Lin, G. & Hyder, S. L. 2021. Promoting Resilience And Health Of Urban Citizen Through Urban Green Space. Water And Environmental Sustainability, 1, 37-43.

Jia, Y., Khayatnezhad, M. & Mehri, S. 2020. Population Differentiation And Gene Flow In Rrodium Cicutarium: A Potential Medicinal Plant. Genetika, 52, 1127-1144.

Kabir, K., Arefin, S. M. A. & Hosain, M. T. 2021. Analysis Of Momentary Variations In The Quality Of Water On Specific Criteria In Cole Mere. Water And Environmental Sustainability, 1, 8-12.

Karasakal, A., Khayatnezhad, M. & Gholamin, R. 2020a. The Durum Wheat Gene Sequence Response Assessment Of Triticum Durum For Dehydration Situations Utilizing Different Indicators Of Water Deficiency. Bioscience Biotechnology Research Communications, 13, 2050-2057.

Karasakal, A., Khayatnezhad, M. & Gholamin, R. 2020b. The Effect Of Saline, Drought, And Presowing Salt Stress On Nitrate Reductase Activity In Varieties Of Eleusine coracana (Gaertn). Bioscience Biotechnology Research Communications, 13, 2087-2091.

Khayatnezhad, M. & Gholamin, R. 2020a. A Modern Equation For Determining The Dry-Spell Resistance Of Crops To Identify Suitable Seeds For The Breeding Program Using Modified Stress Tolerance Index (Msti). Bioscience Biotechnology Research Communications, 13, 2114-2117.

Khayatnezhad, M. & Gholamin, R. 2020b. Study Of Durum Wheat Genotypes’ Response To Drought Stress Conditions. Helix, 10, 98-103.

Khayatnezhad, M. & Gholamin, R. 2021a. The Effect Of Drought Stress On The Superoxide Dismutase And Chlorophyll Content In Durum Wheat Genotypes. Advancements In Life Sciences, 8, 119-123.

Khayatnezhad, M. & Gholamin, R. 2021b. Impacts Of Drought Stress On Corn Cultivars (Zea mays L.) At The Germination Stage. Bioscience Research, 18, 409-414.

Khayatnezhad, M. & Nasehi, F. 2021. Industrial Pesticides And A Methods Assessment For The Reduction Of Associated Risks: A Review. Advancements In Life Sciences, 8, 202-210.

Martignoni, M. E. & Iwai, P. J. 1985. Laboratory Evaluation Of New Ultraviolet Absorbers For Protection Of Douglas-Fir Tussock Moth (Lepidoptera: Lymantriidae) Baculovirus. Journal Of Economic Entomology, 78, 982-987.

Omrani, M. & Fataei, E. 2018. Synthesizing Colloidal Zinc Oxide Nanoparticles For Effective Disinfection; Impact On The Inhibitory Growth Of Pseudomonas aeruginosa On The Surface Of An Infectious Unit. Polish Journal Of Environmental Studies, 27.

Radmanesh, M. 2021. Evaluation Of The Efficient Management Of Greenhouses For Healthy Items In The Province Of Alborz. Water And Environmental Sustainability, 1, 20-23.

Rodríguez, R. 2021. The Study Of Enzyme-Water Mutualism Theory. Water And Environmental Sustainability, 1, 44-49.

Si, X., Gao, L., Song, Y., Khayatnezhad, M. & Minaeifar, A. A. 2020. Understanding Population Differentiation Using Geographical, Morphological And Genetic Characterization In Erodium cicunium. Indian J. Genet, 80, 459-467.

Sun, Q., Lin, D., Khayatnezhad, M. & Taghavi, M. 2021. Investigation Of Phosphoric Acid Fuel Cell, Linear Fresnel Solar Reflector And Organic Rankine Cycle Polygeneration Energy System In Different Climatic Conditions. Process Safety And Environmental Protection, 147, 993-1008.