Background: SPINOSAD (a mixture of spinosyns A and D) is a unique natural pesticide produced by Saccharopolyspora spinosa. With regard to attempts to improve S. spinosa by classical mutagenesis, we propose that the bottleneck of screening out high-SPINOSADproduction strains is probably caused by the fermentation media. Objectives: The current study aimed to identify a new medium to extensively investigate the potential of S. spinosa strains to produce SPINOSAD. Materials and Methods: Statistical and regressive modeling methods were used to investigate the effects of the carbon source and to optimize the production media. Results: The SPINOSAD production of S. spinosa Co121 increased 77. 13%, from 310. 44 21. 84 g/mL in the initial fermentation medium (with glucose as the main carbon source) to 549. 89  38. 59  g/mL in a new optimized fermentation medium (98. 0 g of mannitol, 43. 0 g of cottonseed flour, 12. 9 g of corn steep liquor, 0. 5 g of KH2PO4, and 3. 0 g of CaCO3 in 1 L of H2O; pH was adjusted to 7. 0 before autoclaving). After screening 4, 000 strains, an overall 3. 33-fold increase was observed in SPINOSAD titers, starting from the parental strain Co121 in the original fermentation medium and ending with the mutant strain J78 (1035  34  g/mL) in the optimized medium. Conclusions: The optimized fermentation medium developed in this study can probably be used to improve SPINOSAD production in screening industrial strains of S. spinosa.

Background: Several compounds obtained from plants have potential insecticidal, growth deterrent or repellent characteristics. The control of hard ticks by non-chemical substances was targeted in this study.Methods: The effect of 36 materials on in-vitro ticks was studied, including 2 absolute controls (water only or absolute ethyl alcohol only), 6 conventionally used SPINOSAD preparations (aqueous solutions), 12 Nigella sativa (N. sativa) preparations (aqueous and alcoholic solutions), and 12 Thyme preparations (aqueous and alcoholic solutions). The engorged ticks were tested in-vitro for mortality and oviposition ability using the studied materials.Results: The final mortality after 48 hours of application in N. sativa aqueous preparations began from 10.0% concentration, 1.0% to 100% by concentration preparations ³10%. In addition, N. sativa alcoholic preparations began from 50.0% concentration, 2% to 100% by concentration ³5%. Meanwhile, Thyme aqueous and alcoholic preparations began from 70.0% concentration, 5% to 90% by concentration 10–20%. Additionally, SPINOSAD aqueous preparations and both of control preparations (Water and Alcohol) resulted in no mortality. All differences were statistically significant. The oviposition was stopped in N. sativa (aqueous ³10% and alcoholic ³5%) and in SPINOSAD (aqueous³25%). The aqoues dilution of the used matters killed B. annulatus larvae beginning from the concentration 5%.Conclusion: Nigella sativa alcohol 20% was the best of studied preparations being the lowest concentration (20%) that could achieve the highest lethal (100%) effect in shortest time (12 hours). Moreover, Thyme oil and SPINOSAD could not kill 100% of adult but did on larvae.

Introduction: Trichogramma wasp is one of the most successful parasitic species in the world and the widespread use of these wasps has taken control of many pests in more than 30 countries. Approximately 32 million hectares of agricultural land and forests have been covered by the Trichogramma every year. This parasitoid is reared on Sitotroga cerealella (Lep.: Gelechiidae) and Ephestia kuehniella (Lep.: Pyralidae). The superiority of these bees is their ability to destroy the egg pest before its damage to the host plant. Nonselective insecticides can reduce the potential of biological control agents. In order to determine the most suitable insecticide for effective pest control of Trichogramma wasps, the present study evaluated the persistence toxicity of SPINOSAD, indoxacarb and imidacloprid after exposing of T. brassicae and T. evanescens, the egg parasitoids of tomato leaf miner, according to IOBC procedure. Materials and Methods: The experiments were carried out in the Laboratory of Plant Protection at Agricultural and Natural Resources Research and Education Center of Markazi Province, Arak, Iran. The egg parasitoids T. brassicae and T. evanescens (Hymenoptera: Trichogrammatidae) were provided from Biological Control Department of Plant Protection Research Institute (Tehran, Iran). The parasitoids were reared on the Angoumois grain moth, Sitotroga cerealella (Lepidoptera: Gelechiidae) at the laboratory under the controlled conditions of 25± 1° C, 65± 10% RH and photoperiod of 16: 8 h (L: D). Insecticides: The recommended doses (RD) of the insecticides of SPINOSAD (SC 22. 8% Spintor, DowAgroscience Co. ), Imidacloprid (SC35% Confidor, Bayer) and Indoxacarb (SC15% Avant, Aria Shimi) were tested. Persistent tests: The insecticides were applied at RD on tomato plants by a hand sprayer to the point of run off. Plants were maintained under a transparent polyethylene rain cover in the field. Leaves of the treated tomato plant were sampled and transferred to the laboratory at different time intervals of 3, 5, 16 and 31 days after insecticide application. These intervals were proposed by the IOBC/WPRS Working Group. The samples were placed in ventilated plastic Petri dishes for exposure to the adult parasitoids. The experiment was repeated six times on 30 adult individuals of each species. Assessment of mortality was made on the basis of 24 h post exposure time. Persistence rate of wasps against each insecticide was evaluated according to IOBC/WPRS Working Group. The categories under laboratory conditions include: A, short lived (<5days); B, slightly persistent (5-15days); C, moderately persistent (16-30days); D, Persistent (>30 days) Results and Discussion: Persistence tests for 30% mortality at each period of time revealed that imidacloprid with less than five days persistence classified as short lived insecticides (class A) for T. brassicae and T. evanescens wasps, indoxacarb with more than five days and less than 16 days effect was slightly persistent (class B) and SPINOSAD which persisted more than 16 days but less than 30 days was moderately persistent for the parasitoid T. brassicae and T. evanescens (class C) (Table 1, 2). This research clearly showed that imidacloprid which persisted less than five days at field conditions were compatible with both egg parasitoids, T. brassicae and T. evanescence as a part of integrated management program of pests. This study also revealed that both parasitoids responded at the same level of susceptibility after exposure to the insecticide. There is no evidence for effect of imidacloprid on Trichogramma parasitoids, however our finding was the same as the results of Ferná ndez et al. (2017) who reported imidacloprid was safe for predatory mite Amblyseius swirskii adults. Some researchers also emphasized on computability of this insecticide with a large number of natural enemies, including the predatory mites Amblyseius cucumeris and Phytoseiulus persimilis. The insecticide was compatible with all stages of phytoseiid mites, reported by Lefebvre et al. (2011) and Beers and Schmidt (2014) in Galendromus occidentalis. Garzon et al., (2015) also reported this insecticide was nontoxic to last instar larvae and adults of Chrysoperla carnea and Adalia bipunctata so they emphasized that it is a good candidate to be incorporated into IPM programs. On the contrary, Doker et al., (2015) reported a significant reduction in oviposition of Iphiseius degenerans after exposure to imidacloprid. Conclusion: This research focused on the impact of insecticide exposure on Trichogramma species in Markazi Province. Pesticides that control pests without severe side effects on substantial natural enemies are always a necessity for integrated control programs. The application of the short-lived insecticides can allow a rapid recolonization of the parasitoids. We found imidacloprid is non-harmful for both T. brassicae and T. evanescence wasps. By contrast, SPINOSAD and indoxacarb should be used with care as a part of IPM procedure.

Background: Metarhiziumanisopliae strain IRAN 437C is one of the most virulent fungal isolates against house fly, Muscadomestica. The objective of this study was to determine the interaction of this isolate with sublethal doses of spino­sad against housefly.Methods: In adult bioassay, conidia of entomopathogenic fungus were applied as inoculated bait at 105 and 107 spore per gram and SPINOSAD at 0.5, 1 and 1.5 µg (A.I.) per gram bait. In larval bioassay, conidia were applied as combina­tion of spore with larval bedding at 106 and 108 spore per gram and SPINOSAD at sublethals of 0.002, 0.004 and 0.006 µg (AI) per gram medium.Results: Adult mortality was 48% and 72% for fungus alone but ranged from 66–87% and 89–95% in combination treat­ments of 105 and 107 spore/g with sublethal doses of SPINOSAD respectively. The interaction between 105 spore/g with sublethals exhibited synergistic effect, but in combination of 107 spore in spite of higher mortality, the interac­tion was additive. There was significant difference in LT50 among various treatments. LT50 values in all combination treat­ments were smaller than LT50 values in alone ones. Larval mortality was 36% and 69% for fungus alone but ranged from 58%–78% and 81%–100% in combination treatments of 106 and 108 spore/g medium with sublethals of spino­sad respectively. The interaction was synergistic in all combination treatments of larvae.Conclusion: The interaction between M. anispliae and SPINOSAD indicated a synergetic effect that increased the house fly mortality as well as reduced the lethal time.

Toxicity of SPINOSAD was primarily evaluated with three bioassay methods: feeding, spraying (Residual exposure) and combination with larval medium against one susceptible and one field population. Feeding method was selected as efficient method for survey of susceptibility or resistance of seven field populations. In feeding bioassay method, SPINOSAD LD50 of susceptible strain at 24h and 72h were 3.78 and 1.54 mg (AI) per gram bait and for LD95 were 5.59 and 3.35 mg (AI) /g, respectively. LD50 of field populations at 24h ranged  from 3.974- 4.303 and for LD95 from 7.33- 8.30 mg (AI)/gr. LD50 at 72h ranged from 1.54-1.72 mg/gr and LC95 were 3.31- 3.93 mg /g, respectively. Determination of lethal dose ratios with lower and upper limits indicated no significant difference between SPINOSAD LD50 of susceptible and field population at 24h and 72h. In Residual method, SPINOSAD LD50 of susceptible and field population (AHDS) at 24h were 0.015 and 0.016 and for LD95 were 0.03, 0.033 g (AI) per m2, respectively. At 72h, LD50 were 0.0065, 0.007 and LD95 were 0.014, 0.015 g (AI)/ m2 for the above populations. In combination of SPINOSAD with larval medium, LD50 and LD95 of susceptible population were 9.79 and 29.5 mg (AI) per kg medium. For field population (AHDS), LD50 and LD95 were 9.95 and 56.6 mg (AI)/kg. There was no significant difference at LD50 of susceptible and field population with these two methods. Totally, the result of this study indicated that LD50 values decrease approximately 2-3-fold between 24 and 72 h and all field populations were susceptible to SPINOSAD.

Orius albidipennis Reuter is an important predator of economic pests in greenhouse. In this studu, investigations were performed on toxicity values of abamectin, SPINOSAD and chlorpyrifos in laboratory (Petri dish) and greenhouse (clip cage) conditions. LC50 and LC90 values were determined to both sexes, male and female, by using SAS software, and comparisons were made according to ratio of LC50 values and 95% confidence limits. For males, toxicity of the insecticides was more than females in laboratory and greenhouse conditions. Toxicity rates were significantly different between the insecticides. Chlorpyrifos had the most toxicity in laboratory (LC50=0.06 ppm for males and LC50= 0.05 ppm for females) and greenhouse conditions (LC50=83 ppm for males and LC50= 99.3 ppm for females) and SPINOSAD had the least one. Laboratory and greenhouse trials showed that toxicities of these insecticides were lower in greenhouse bioassay than in laboratory bioassay but this difference was notable for SPINOSAD. It is concluded that multiple testing methods should be used in evaluating the effects of pesticides on beneficial arthropods.

The common Pistachio psylla, Agonoscena pistaciae, is one of the most important pests that feeding on the sap both in nymphal and adult stage. In this study, the effects of spirodiclofen, SPINOSAD, azadirachtin and Thymol soap were studied on the psyllid nymphs. At the same time, the impact of these pesticides on the activity of esterase and glutathione S-transferase (GST) enzymes were studied. In this research, three doses of each pesticide were selected (Recommended dose, 2-fold recommended dose and 1/2 recommended dose). The results showed that thymol soap had better performance in controlling pistachio psylla with concentration of 2000 mg/l compared to other pesticides. The populations of nymphs were reduced to 95.40, 94.12, and 82.30, 55.51 and 29.48 percent at 1, 3, 7, 14 and 21 days after application. The biochemical results of this study showed that spirodiclofen, SPINOSAD, azadirachtin and Thymol soap, had the greatest impact on the inhibition of esterase enzyme in concentrations 120, 120, 200 and 2000 mg/l, respectively in different days after spraying. Most of the inhibition in glutathione S-transferase was observed in spirodiclofen, SPINOSAD, azadirachtin and Thymol spraying soap in concentrations of 180, 240, 200 and 1000 mg/l, respectively. Because of the effectiveness of Thymol spraying soap, its competitiveness with other pesticides, and reduction in production costs (cost-effectiveness) and compatibility with environment, it seems that this insecticide could be a good candidate for psylla control.

BACKGROUND: Blowfly (Lucilia sericata,Diptera: Calliphoridae) is an external parasites of domestic animals and the causative agent of myiasis, which is involved in the transmission of a number of bacterial pathogens. Application of chemical insecticides is one of the most common methods for controlling this pest, which have recently been reported resistant populations of L. sericata to some insecticides. Therefore, identifying and recommending new and low-risk insecticides for humans and environment is of prime importance. OBJECTIVES: This research aimed to investigate the toxicity of insecticides deltamethrin, SPINOSAD, pyridalyl, and pirimiphos-methyl on the third instar larvae of L. sericata. METHODS: In the present study, the bioassay tests were performed by immersion of larvae in concentrations of insecticides that caused 10 to 90% mortality of larvae, as a factorial experiment in a randomized complete block design. Additionally, the median lethal concentration (LC50) was determined from the probit analysis of bioassay data for the studied insecticides. RESULTS: The results of analysis of variance and the comparison of the mean mortality (%) of third instar larvae were significant at the level of 1% probability. According to bioassay tests, the estimated LC50 values for deltamethrin, SPINOSAD, pyridalyl, and pirimiphos-methyl were 186. 49, 26. 88, 85. 08, and 134. 97 mg L-1, respectively. Overall, the results revealed that SPINOSAD and pyridalyl had the highest lethal effect on blowfly larvae while deltamethrin had less advers effects on this pest. CONCLUSIONS: SPINOSAD, as a relatively new insecticide, with very low toxicity towards mammals and other non-target organisms, showed the highest lethal effect on blowfly larvae and can be an appropriate alternative to older insecticides in controlling this pest.

To examine the differences between spinetoram and SPINOSAD in their insecticidal activity and stability on the stored wheat, bioassay tests were carried out for six consecutive months at 20 and 30 ± 1 ° C by adults of Rhyzopertha dominica (F. ) (Coleoptera: Bostrichidae). Samples were taken after zero, 2, 3, 5, 7, 15, 30, 60, 90 and 180 days of insecticides application. Results of median lethal concentrations (LC50s) determined after three days of treatment indicated that SPINOSAD was significantly more toxic than spinetoram against the adults. Results also revealed that mortality decline of R. dominica was correlated with temperature and the gradual degradation of the two insecticides over time during storage period. The half-life of SPINOSAD and spinetoram on stored wheat were 99. 02 and 69. 32 days at 20 ± 1 ° C, respectively. These were shortened at 30 ± 1 ° C to 49. 51 and 46. 21 days, respectively. However SPINOSAD displayed more bioactivity and stability than spinetoram. It can be concluded that SPINOSAD and spinetoram do not remain persistent and even a slow degradation of these insecticides is likely to occur during wheat storage. Residues of SPINOSAD and spinetoram persist on the wheat grains at levels well below the established tolerance levels.