Application of statistical experimental design for optimisation of Bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium

In order to overproduce Bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on Bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L -1 starch, 30 g L -1 soya bean an d9gL -1 sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch) when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch). Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis Bioinsecticides at industrial scale which is of great importance in practical point of view.

Improvement of Photorhabdus temperata strain K122 Bioinsecticide production by batch and fed-batch fermentations optimization

Optimization of a fermentation process for Bioinsecticides production by Photorhabdus temperata strain K122 was investigated into fully controlled 3-L fermenter using an optimized medium (OM). Development of large-scale inocula showed that the composition of the growth medium greatly influenced the physiological state of P . temperata cells. The effect of pH, agitation and dissolved oxygen concentration (DO) on the growth, culturability and oral toxicity of P . temperata cells were also investigated. Indeed, maintaining the pH at 7 and controlling DO concentration at 50 % saturation throughout the fermentation process, improved biomass production, CFU counts and oral toxicity by 41.1, 35 and 32.1 %, respectively, as compared to cultures carried out in 500 mL shake flasks. At such conditions, 8 g/L glucose fed-batch fermentation, enhanced cell lysis and variants small colony (Vsm) polymorphism appearance. To overcome such limitations, glucose concentration should be maintained at 4 g/L. In this case, P . temperata cells were produced at high cell density and culturability reaching 4.5 and 1.2 × 10^9 cells/mL, respectively. In addition, the stability of the primary form was maintained for a long period in the stationary growth phase and Vsm polymorphism was completely avoided that can be crucial for scale-up the bioprocess of P. temperata Bioinsecticide.

Improvement of Photorhabdus temperata Bioinsecticides production in low‐cost media through adequate fermentation technology

To develop a cost effective process for Bioinsecticides production by Photorhabdus temperata, dissolved oxygen (DO) requirements were investigated in both the complex and the optimized media using diluted seawater as a source of micronutrients. By varying DO concentrations, tolerance to hydrogen peroxide was shown to be medium dependant. Indeed, P. temperata cells grown in the complex medium, exhibited higher tolerance than cells grown in the optimized medium (OM). Tolerance to H2O2 was shown to be related to intracellular reactive oxygen species (ROS) accumulation during soya bean meal or glucose assimilation, as shown by flow cytometry analysis. To avoid oxidative stress damages in P. temperata cells cultured in the OM, DO concentration should be constant 50% saturation throughout the fermentation. However, a DO-shift control strategy was demonstrated to be beneficial for P. temperata Bioinsecticide production in the complex medium. By using such a strategy biomass, culturability, and oral toxicity reached 16.5 × 108, 1.15 × 108 cells/mL and 64.2%, respectively, thus was 16.19, 26.37, and 12.2% more than in the cultures carried out at a constant 50% saturation. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012

Herbivore insects and predatory arthropods in freshwater swamp rice field in South Sumatra, Indonesia sprayed with Bioinsecticides of entomopathogenic fungi and abamectin

Abstract. Herlinda S, Prabawati G, Pujiastuti Y, Susilawati, Karenia T, Hasbi, Irsan C. 2020. Herbivore insects and predatory arthropods in freshwater swamp rice field in South Sumatra, Indonesia sprayed with Bioinsecticides of entomopathogenic fungi and abamectin. Biodiversitas 21: 3755-3768. Herbivore insect population and predatory arthropods in rice field may be effected by the application entomopathogenic fungi or synthetic insecticide. The objective of this research was to analyze individual quantity of herbivore insects and predatory arthropod inhabiting freshwater swamp rice fields treated with Bioinsecticides and abamectin (commercial insecticide). This research was conducted in the freshwater swamp rice field located in Village Pelabuhan Dalam, Sub District Pemulutan, District Ogan Ilir, and South Sumatra. The experiment was arranged in a Completely Randomized Block Design consisted of four plots of treatment. The research used the Bioinsecticides made from entomopathogenic fungi (Beauveria bassiana, Metarhizium anisopliae, and Cordyceps military), and abamectin. Sampling to collect arthropods inhabiting rice canopy was also conducted using an entomological net in the study locations. This research found 12 families of herbivore insects with 22 species dominated by Nilaparvata lugens and Leptocorisa acuta and 32 species of spider belonged to eight families dominated by Tetragnatha virescens and Oxyopes matiensis. The species diversity of spider was higher in the plots of the Bioinsecticide compared to that of the abamectin. Predatory insects found belonged to 14 species belonged to eight families dominated by species of Ophionea nigrofasciata, Verania discolor, and Paedorus fuscipe. The abundance of predatory insects in plots sprayed with the Bioinsecticides was higher compared to that of the abamectin plot. The abundance and species diversity of predatory arthropod sprayed with Bioinsecticide of B. bassiana, M. anisopliae, and C. militaris did not decrease, while the population of herbivore insect tended to decrease.

Efikasi Bioinsektisida Bacillus thuringiensis Barliner terhadap Gryllus bimaculatus De Geer (Orthoptera: Gryllidae) pada Tanaman Padi Utama dan Ratun

Hanif et al, 2017. Efficacy of Bacillus thuringiensis Barliner Bioinsecticide against  Gryllus bimaculatus De Geer (Orthoptera: Gryllidae) in Major Rice Crops and Ratooning Rice. JLSO 6(1):95-105.

Crickets ( Gryllus bimaculatus ) are an important pests on rice that can reduce productivity. The application of Bacillus thuringiensis Bioinsecticide is expected to be an alternative methode to control these pest. The objective of the study was to investigate optimal dose of B. thuringiensis in suppressing rice damage was caused by G. bimaculatus . The observation were conducted on rice cultivation in Pelabuhan Dalam Village, Pemulutan Subdistrict of Ogan Ilir Regency. The results showed that application of B. thuringiensis Bioinsecticide had significant effects in suppressing rice crop damage by G. bimaculatus , but did not had significant effect on growth of rice plants.

Efikasi Bioinsektisida Beauveria bassiana (Bals.) Vuill Terhadap Gryllus bimaculatus De Geer (Orthoptera: Gryllidae) Pada Padi Ratun

Soleha, et al.  Efficacy of Bioinsecticide from Beauveria bassiana (Bals.) Vuill on  Gryllus bimaculatus De Geer (Orthoptera: Gryllidae) on Ratooning Rice in Fresh Swamps. JLSO 5(2):189-197. The use of chemical insecticide to control pest of crop often causes the bad impact on nontarget organisms. B. bassiana is an entomopathogenic fungi has depeloved to control pest of crop as chemical insecticide alternative. This study aimed to measure mortality and damages rate by crickets to rice of the applications of Bioinsecticide containing B. bassiana. The Method of this study used Random Block Designs with 5 treatments and 4 replications. The treatments applications were 1 L.ha -1 , 2 L.ha -1 , 3 L.ha -1 , commercial B. bassiana as recommended, and 0 L.ha -1 as control. The applications Bioinsecticide containing B. bassiana was significant to nymph of crickets mortality. Mortality of nymph of the applications of B. bassiana was significant with nontreatment (0 L.ha-1). The highest mortality was found in the Bb 3 L .ha-1 about 52,50% and the lowest mortality on 0 L.ha -1 . The shortest LT50  was occurred in Bb 3 L.ha -1 about 7,00 days, while the longest LT50 occurred in Bb 0 L.ha -1 about 50,47 – 64,80 days. The highest damages was found in Bb 0 L.ha -1 about 11,55% – 34,14% and the lowest damages was found in Bb 3 L.ha -1 about 5,94 % – 19,82%. Application Bioinsecticide containing B. bassiana is significant to crickets mortality and will make an important contribution to the biological control for insect pests.

Herbivore insects and predatory arthropods in freshwater swamp rice field in South Sumatra, Indonesia sprayed with Bioinsecticides of entomopathogenic fungi and abamectin

Abstract. Herlinda S, Prabawati G, Pujiastuti Y, Susilawati, Karenia T, Hasbi, Irsan C. 2020. Herbivore insects and predatory arthropods in freshwater swamp rice field in South Sumatra, Indonesia sprayed with Bioinsecticides of entomopathogenic fungi and abamectin. Biodiversitas 21: 3755-3768. Herbivore insect population and predatory arthropods in rice field may be effected by the application entomopathogenic fungi or synthetic insecticide. The objective of this research was to analyze individual quantity of herbivore insects and predatory arthropod inhabiting freshwater swamp rice fields treated with Bioinsecticides and abamectin (commercial insecticide). This research was conducted in the freshwater swamp rice field located in Village Pelabuhan Dalam, Sub District Pemulutan, District Ogan Ilir, and South Sumatra. The experiment was arranged in a Completely Randomized Block Design consisted of four plots of treatment. The research used the Bioinsecticides made from entomopathogenic fungi (Beauveria bassiana, Metarhizium anisopliae, and Cordyceps military), and abamectin. Sampling to collect arthropods inhabiting rice canopy was also conducted using an entomological net in the study locations. This research found 12 families of herbivore insects with 22 species dominated by Nilaparvata lugens and Leptocorisa acuta and 32 species of spider belonged to eight families dominated by Tetragnatha virescens and Oxyopes matiensis. The species diversity of spider was higher in the plots of the Bioinsecticide compared to that of the abamectin. Predatory insects found belonged to 14 species belonged to eight families dominated by species of Ophionea nigrofasciata, Verania discolor, and Paedorus fuscipe. The abundance of predatory insects in plots sprayed with the Bioinsecticides was higher compared to that of the abamectin plot. The abundance and species diversity of predatory arthropod sprayed with Bioinsecticide of B. bassiana, M. anisopliae, and C. militaris did not decrease, while the population of herbivore insect tended to decrease.

Efikasi Bioinsektisida Bacillus thuringiensis Barliner terhadap Gryllus bimaculatus De Geer (Orthoptera: Gryllidae) pada Tanaman Padi Utama dan Ratun

Hanif et al, 2017. Efficacy of Bacillus thuringiensis Barliner Bioinsecticide against  Gryllus bimaculatus De Geer (Orthoptera: Gryllidae) in Major Rice Crops and Ratooning Rice. JLSO 6(1):95-105.

Crickets ( Gryllus bimaculatus ) are an important pests on rice that can reduce productivity. The application of Bacillus thuringiensis Bioinsecticide is expected to be an alternative methode to control these pest. The objective of the study was to investigate optimal dose of B. thuringiensis in suppressing rice damage was caused by G. bimaculatus . The observation were conducted on rice cultivation in Pelabuhan Dalam Village, Pemulutan Subdistrict of Ogan Ilir Regency. The results showed that application of B. thuringiensis Bioinsecticide had significant effects in suppressing rice crop damage by G. bimaculatus , but did not had significant effect on growth of rice plants.

The impact of Bioinsecticide overdoses of Beauveria bassiana on species diversity and abundance of not targeted arthropods in South Sumatra (Indonesia) freshwater swamp paddy

Abstract. Hanif KI, Herlinda S, Irsan C, Pujiastuti Y, Prabawati G, Hasbi, Karenina T. 2020. The impact of Bioinsecticide overdoses of Beauveria bassiana on species diversity and abundance of not targeted arthropods in South Sumatra (Indonesia) freshwater swamp paddy. Biodiversitas 21: 2124-2136. Bioinsecticides have been shown to reduce insect pest populations, but it is necessary to monitor the effect of Bioinsecticide overdoses on non-target arthropods because they are abundant in rice fields. This study aimed to analyze the impact of Bioinsecticide overdoses of B. bassiana on species diversity and abundance of non-target arthropods. Bioinsecticides derived from Beauveria bassiana were sprayed on the paddy every two weeks until the ripening stage, at a dose of 1, 2, 3, 4 L.ha-1, control (without insecticide), and abamectin as prescribed. The abundance of spiders and predatory arthropods did not decrease when Bioinsecticides were applied but the abundance of spiders, especially Lycosidae and Salticidae dropped significantly when abamectin was applied. The abundance of the parasitoid decreased significantly when abamectin was applied, but the abundance of parasitoids applied by Bioinsecticide was not significantly different from those of control. The abundance of insect pests started to decrease when Bioinsecticide was applied to 2, 3 or 4 L.ha-1. The diversity species of non-target arthropods (spiders, predatory arthropods, and parasitoids) tended to decrease when it was applied Bioinsecticides 3 and 4 L.ha-1 or abamectin, but their abundance remained high and no species dominance was found. For this reason, the correct dosage for suppressing insect pests abundance by not decreasing the abundance and diversity of non-target species is 2 L.ha-1 and applied when herbivore abundance is high.