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.

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.

Background and objective: SACCHAROPOLYSPORA erythraea is a member of the aerobic and gram positive actinomycetes and the industrial microorganism for erythromycin production .It has shown that biomass composition of any microorganism and production of secondary metabolites strongly depends on composition of culture medium.Materials and methods: Spore suspensions of SACCHAROPOLYSPORA erythraea PTCC1685 were inoculated into flasks containing seeding medium and incubated at 30°C for 40-44 h with 180 rpm in aerobic condition. The appropriate amount of seeding cultures that showed suitable morphology, pH and growth were added into flasks containing fermentation medium. Flasks were inoculated at 30°C for for 11 days with 180 rpm. Biomass in stationary phase had separated by sentrifugation in 4000 rpm for 20 min. For lipid extraction, biomass is homogenized with chloroform/methanol (2/1) to a final volume 20 times the volume of the biomass .After dispersion, the whole mixture is agitated during 12h in an orbital shaker at room temperature. Then the mixture was sentrifuged at 4000rpm for 20 min. Upper phase removed by siphoning. The lower chloroform phase containing lipids evaporated under vacuum in rotary evaporator. Methyl esters of fatty acids of lipids prepared for GC-Mass analysis.Results: Eight fatty acids were detected which contained 15 to 18 carbon atoms. 14-methyl pentadecanoic acid (iC16:0) was in the highest percentage (36.25)% and pentadecanoic acid (aC17:0) were in the lowest percentage (1.64.)% Other fatty acids were C17:0(18.9)%, C18:2(14.99)%, C16:0(8.85)%, C18:1 (8.67)%, C18:0(5.47 )% and C15:0 (5.2.)% Fatty acids with less than 15 carbon atoms had not detected in this study. Conclusions: Fatty acid composition and erythromycin production in S.erythreae strongly depends on culture composition.

To find a suitable surfactant for enhancing the solubility of fatty acid, some nonionic surfactants including Tween 20, 40, 60, 80, 85; poly-ethylene glycol 200, 300, 400, 600; di-ethylene glycol di-methyl ether (diglyme), Triton X-WO, tetrahydrofuran (THF) and dioxan were added to the fermentation medium (with or without palmitic acid) and their effects were studied on the morphology of SACCHAROPOLYSPORA erythraea and erythromycin production. Tween 20 and Triton X-WO lysed the hyphae, but in the media containing other surfactants, the hyphae remained in a vegetative form. PEG 300, Tween 40 and THF had neither negative nor positive effect on the erythromycin production. But, addition of Triton X-100 and dioxan to the fermentation medium significantly decreased the concentration of erythromycin, 9.9 and 1.3 times less than that of control, respectively. In the media containing PEG 200, PEG 400, PEG 600, Tween 60, Tween 80, Tween 85 and diglyme, erythromycin concentration was 1.3, 1.5, 2.0, 1.4, 1.4, 1.3 and 1.3 times higher than that of control, respectively. Production of erythromycin in the palmitic acid containing medium was 1.62 times lower than that of control. However, the effect of palmitic acid on the erythromycin production was different when surfactants were added to media. Erythromycin production in the media containing palmitic acid plus PEG 400 and palmitic acid plus PEG 600 was more than that of palmitic acid containing medium (without any additional surfactant). However, erythromycin concentration in the media containing Tween 40, 60, 80 and 85, THF and diglyme was less than that of medium containing palmitic acid (without any surfactant).

Background & Objectives: Increase in the yield while reducing costs is one of the most important aspects of researches in biotechnology. Considering the amount of bioproducts consumed annually, antibiotics stand at second place, and therefore there has been considerable research to increase their production. This study was aimed to optimize the media used in fermentation process in terms of nitrogen source.Materials & Methods: This study was conducted in Tehran, based on a continuously sampling from the metabolites produced by SACCHAROPOLYSPORA erythraea. After sporulation, seeding and fermentation, the role of whey powder on several indicators such as pH, biomass production percentage morphological changes in the strain was analyzed. The concentration of the produced erythromycin were examined by spectrophotometry.Results: The results of this study showed the effectiveness of different concentrations of whey powder on fermentation indices. Also, according to the results, the use of soybean meal with whey powder had positive impacts on the amount of erythromycin produced, duration of fermentation and the production costs. The optimized concentration of nitrogen nitrogen for production of this antibody was a combination of 54g/l whey powder with 12g/l soybean meal.Conclusion: Since whey powder is an inexpensive source of nitrogen, it can be used to reduce costs, and increase the yield. Therefore, whey powder can be used as an alternative source of nitrogen in industrial production.

Hydroalcoholic extract of Pycnocycla SPINOSA Decne. exBoiss. var. SPINOSA has in vitro spasmolytic action, and at oral dose of 250µg/kg inhibits castor oil induced diarrhoea in mice. In this investigation, effects of P. SPINOSA var. SPINOSA extract in comparison with nifedipine on blood pressure and heart rate in animal model was studied. Injection of three bolus doses of P. SPINOSA var. SPINOSA extract (100µg/kg, 500µg/kg and 1mg/kg) into the jugular vein, temporary reduced blood pressure and heart rate. However, soon after completion of extract administration blood pressure and heart rate returned to normal. Nifedipine on the other hand caused a sustained reduction in blood pressure and decreased heart rate compared with the control group. From this study it was concluded that P. SPINOSA var. SPINOSA extract at antidiarrhoeal dose has no significant effect on blood pressure and heart rate.