Effect of sucrose on nisin production in batch and fed‐batch culture by Lactococcus lactis

The effects of sucrose on cell growth and nisin production by Lactococcus lactis were investigated in batch and pH feed‐back controlled fed‐batch cultures. In batch cultures, nisin titer reached its maximum, 2658 IU cm^?3, at the initial sucrose concentration of 30 g dm^?3. With sucrose concentrations higher than 30 g dm^?3, nisin production decreased while the biomass was not influenced significantly. By using the pH feed‐back controlled method, residual sucrose concentration could be controlled well in fed‐batch cultures and three conditions (sucrose maintained at 2, 16, 20 g dm^?3, respectively) were evaluated. Maintaining a low sucrose concentration at 2 g dm^?3 during feeding favored nisin biosynthesis, and the maximum nisin titer obtained was 4961 IU cm^?3 compared with 3370 IU cm^?3 (16 g sucrose dm^?3)and 3498 IU cm^?3 (20 g sucrose dm^?3), respectively. Copyright © 2005 Society of Chemical Industry     

In‐line mixing for production of citric acid by Candida lipolytica grown on n‐paraffins

This study reports on the effects of internal fermenter and external in‐line agitation and fed‐batch mode of operation on citric acid production from Candida lipolytica using n‐paraffin as the carbon source. An optimum range of fermenter agitation speeds in the range 800–1000 rpm corresponding to Reynolds numbers of 50433–62947 (based on initial batch conditions) seemed to give the best balance between substrate utilization for biomass growth and citric acid production. Proof of concept evidence is presented that indicates that an external in‐line agitator could be used in place of high speed internal agitation to increase citric acid production. However, more work is required to optimize the external agitator concept. Application of multiple fed‐batch feedings can be used to extend the batch fermentation and increase final citric acid concentrations and product yield. Experiments were conducted implementing a three‐cycle fed‐batch process which increased overall citric acid yields to 0.8–1.0 g citric acid g^?1 n‐paraffin, approximately 200% improvement from those found in the normal batch process. The three‐cycle fed‐batch mode of operation also increased the final citric acid concentration to 42 g dm^?3 from about 6 g dm^?3 for normal batch operation. Increased citric acid concentrations in three‐cycle fed‐batch mode was achieved at longer fermentation times. Copyright © 2004 Society of Chemical Industry     

Xylanase production by Bacillus polymyxa

Bacillus polymyxa produced high levels (12–13 U cm^?3) of extracellular xylanases when grown in a complex medium containing yeast extract and oat spelt xylan as nitrogen and carbon sources respectively. Substantially lower yields of enzyme were produced during growth on the monosaccharides glucose, arabinose and xylose. Meagre growth occurred when ammonium sulphate, instead of yeast extract, was used as nitrogen source. When assayed in culture broth supernatants, xylanase showed an optimum activity in 48°C and at pH values in the range 5.0–6.5. Under such conditions, the half-life of this xylanase preparation was 8 h. Mn²⁺ showed a strong inhibitory effect on the enzyme, but inhibition by EDTA (27% w/v) suggested dependence on a metallic ion. SDS-PAGE and zymogram overlay showed that up to five separate xylanases in the range of 20 to 116 kDa were produced.     

Continuous production of poly‐β‐hydroxybutyrate by high‐cell‐density cultivation of Wautersia eutropha

BACKGROUND: Poly‐β‐hydroxybutyrate (PHB) accumulation is triggered by limitation of a nutrient other than carbon. The production cost of PHB is very high. In order to reduce this cost, continuous cultivation for the accumulation of PHB was investigated. The culture was first allowed to grow under fed‐batch conditions to yield a significant increase in biomass and PHB accumulation. Thereafter this high‐cell‐density biomass containing PHB was allowed to grow and maintained under conditions of continuous cultivation so that the overall process could be simplified and economised. RESULTS: For continuous cultivation a medium containing 90 g L^?1 fructose and 2.5 g L^?1 nitrogen (as urea) was fed continuously at a dilution rate of 0.1 h^?1. A steady state biomass of 27.7 g L^?1 with a PHB concentration of 5.5 g L^?1 was established in the bioreactor. This resulted in a continuous PHB productivity of 0.55 g L^?1 h^?1. CONCLUSION: The experiments have resulted in the development of a novel production technology involving the integration of batch, fed‐batch and continuous processes. At the same time the production of PHB under continuous cultivation increases the overall industrial importance of the system. Copyright © 2008 Society of Chemical Industry     

Lovastatin production by Aspergillus terreus in a two‐staged feeding operation

BACKGROUND: Lovastatin is known to inhibit its own synthesis in the fungus Aspergillus terreus. Therefore, the use of a fermentation strategy that continuously removes some of the lovastatin produced from the bioreactor can enhance its productivity. This paper reports on the effects of dilution rate and the composition of the feed medium on lovastatin production by A. terreus. RESULTS: The feeding strategy consisted of an initial batch/fed‐batch phase and a semi‐continuous culture phase in which the pelleted biomass was retained inside a slurry bubble column. A nitrogen‐free medium was fed at various fixed dilution rates in the semi‐continuous phase. In experiments that were designed to assess the effects of the composition of the medium, the dilution rate was held at 0.42 d^?1, but different feed media were used in separate runs. The best two‐staged production strategy was shown to consist of a 96 h batch/fed‐batch phase that used a nutritionally complete medium. This was followed by a semi‐continuous operation using a medium that was free of both nitrogen and carbon sources. CONCLUSION: Semi‐continuous operation enhanced productivity of lovastatin by 315% compared with a conventional batch operation. The optimal dilution rate in semi‐continuous operation was about 0.42 d^?1. Copyright © 2008 Society of Chemical Industry     

Xylitol production by Candida tropicalis from corn cob hemicellulose hydrolysate in a two‐stage fed‐batch fermentation process

BACKGROUND: Xylitol, a sugar alcohol widely used in food and pharmaceutical industries, can be produced through biological reduction of xylose present in hemicellulose hydrolysates by Candida tropicalis. However, the aeration rate and by‐products originating from hemicellulose hydrolysis strongly inhibit the production of xylitol in a fermentation process. A two‐stage fed‐batch fermentation system was developed to reduce these inhibitory effects and to improve xylitol production from corn cob hemicellulose hydrolysates by C. tropicalis. RESULTS: Results of batch fermentations indicated that high xylitol production could be obtained from C. tropicalis at an initial xylose concentration of 80 g L^?1 in corn cob hydrolysate medium at an aeration rate of 0.4 vvm at the micro‐aeration stage. In the two‐stage fed‐batch fermentation process, 96.5 g L^?1 xylitol was obtained after 120 h, giving a yield of 0.83 g g^?1 and a productivity of 1.01 g L^?1 h^?1, which were 12.16% and 65.57% higher than those in a batch fermentation. CONCLUSION: High xylitol production can be achieved in a two‐stage fed‐batch fermentation process, in which the negative effects of aeration rate and inhibitory compounds on xylitol formation can be considerably reduced. Copyright © 2011 Society of Chemical Industry     

Pilot‐plant production of xylo‐oligosaccharides from corncob by steaming,enzymatic hydrolysis and nanofiltration

This paper reports a pilot‐plant production process for xylo‐oligosaccharides (mainly xylobiose and xylotriose) from corncob meal by steaming treatment followed by enzymatic hydrolysis and nanofiltration. The effects of corncob meal pretreatment, steaming temperature and time were investigated in order to obtain maximum extraction of xylan and to minimize the autohydrolysis of xylan into xylose. The enzymatic reaction was carried out using Aspergillus niger AN‐1.15 endo‐xylanase at 55 °C and the optimum enzymatic hydrolysis time was 5 h. The conventional downstream processing for purification of xylo‐oligosaccharides was incorporated with nanofiltration technology, giving benefits of energy saving and removal of monosaccharides. The final product from 40 kg of corncob meal was 10 dm³ of xylo‐oligosaccharide syrup (800 g dm^?3 total sugar), containing 74.5% xylobiose and xylotriose. Copyright © 2004 Society of Chemical Industry     

Performance evaluation of batch and unsteady state fed‐batch reactor operations for the production of a marine microbial surfactant

BACKGROUND: Biosurfactants are microbially derived surface‐active and amphipathic molecules produced by various microorganisms. These versatile biomolecules can find potential applications in food, cosmetics, petroleum recovery and biopharmaceutical industries. However, their commercial use is impeded by low yields and productivities in fermentation processes. Thus, an attempt was made to enhance product yield and process productivity by designing a fed‐batch mode reactor strategy. RESULTS: Biosurfactant (BS) production by a marine bacterium was performed in batch and fed‐batch modes of reactor operation in a 3.7 L fermenter. BS concentration of 4.61 ± 0.07 g L^?1 was achieved in batch mode after 22 h with minimum power input of 33.87 × 10³ W, resulting in maximum mixing efficiency. The volumetric oxygen flow rate (K_La) of the marine culture was about 0.08 s^?1. BS production was growth‐associated, as evident from fitting growth kinetics data into the Luedeking‐Piret model. An unsteady state fed batch (USFB) strategy was employed to enhance BS production. Glucose feeding was done at different flow rates ranging from 3.7 mL min^?1 (USFB‐I) to 10 mL min^?1 (USFB‐II). USFB‐I strategy resulted in a maximum biosurfactant yield of 6.2 g l^?1 with an increment of 35% of batch data. The kinetic parameters of USFB‐I were better than those from batch and USFB‐II. CONCLUSION: Comparative performance evaluation of batch and semi‐continuous reactor operations was accomplished. USFB‐I operation improved biosurfactant production by about 35% over batch mode. USFB‐I strategy was more kinetically favorable than batch and USFB‐II. © 2012 Society of Chemical Industry     

A hybrid neural model (HNM) for the on‐line monitoring of lipase production by Candida rugosa

A mechanistic model was proposed by Gordillo for the representation of lipase production by Candida rugosa, with the bioreactor in batch and fed‐batch operation. However, the model was not able to represent the lipolytic activity. The objective of the present study is to propose an efficient hybrid neural‐phenomenological model (HNM) for this process. The experimental data used corresponded to fed‐batch operation with constant substrate feed rate at 2.8 × 10^?7; 5.6 × 10^?7 and 9.7 × 10^?7 kg s^?1. Artificial neural networks (ANNs) were trained to represent the aqueous and intracellular lipase activity and were further associated with a reduced version of the mechanistic model of the proposed HNM. When compared to the experimental data, the HNM exhibited higher accuracy. The HNM can be employed in process monitoring using only on‐line measurements of CO₂ and substrate feed rate to infer enzyme activities and also substrate and biomass concentrations. Copyright © 2007 Society of Chemical Industry     

Continuous perfusion culture of encapsulated hybridoma cells

BACKGROUND: The production of Monoclonal antibodies (mAbs) is often performed in batch or fed‐batch operations where low cell densities and low volumetric productivities are achieved. The main bottleneck of both processes is the short operating time with productive cells at maximum cell concentration. RESULTS: The process studied in this work is based on a fluidized‐bed bioreactor culture of encapsulated KB26.5 cells in a liquid core of calcium alginate microcapsules as a culture strategy to produce IgG₃. First, DMEM medium was modified in order to protect the microcapsules from degradation, and later, the optimal operating conditions were set. Under these conditions encapsulated KB26.5 cells reached cell densities of 1.05 × 10⁸cells mL^?1 or 9.8 × 10⁶ cells mL^?1 (referred to the inner capsule volume or total bioreactor volume, respectively), and a mAb volumetric productivity of 2.75 µg mL^?1 h^?1. CONCLUSIONS: The productivity of encapsulated KB26.5 cells in perfusion culture was enhanced significantly in comparison with batch and fed‐batch processes. Continuous operation of the perfusion culture for periods longer than 35 days, represented a volumetric productivity about five‐fold higher than conventional operations. However, the fluidized‐bed also showed limitations such as low cell viability at high cell densities due to the mass transfer limitations of large molecules inside the microcapsules. Copyright © 2011 Society of Chemical Industry     

Clonal selection of high producing,stably transfected HEK293 cell lines utilizing modified,high‐throughput FACS screening

BACKGROUND: Human embryonic kidney‐293 (HEK‐293) cells are commonly used as a transient expression host but their application in stable therapeutic protein production is limited. This is presumably due to the absence of a suitable amplifiable expression system and hence limited protein output compared with other mammalian cells such as Chinese hamster ovary cells. This paper describes a rapid clonal selection method for isolating HEK293 cell lines with high specific productivity, for a non‐amplifiable expression system, to achieve high‐level, scalable expression of recombinant antibodies. RESULTS: Flow cytometry utilizing cold capture of secreted protein on the cell surface was applied to isolate high expressing clones from a stable antibiotic resistant pool. The top three isolated clones showed a five‐ to seven‐fold improvement in volumetric outputs compared with the initial resistant pool (～20 mg L^?1) under batch conditions. In fed‐batch conditions using commercially available hydrolysate supplements, the final titre was further increased to 500–600 mg L^?1 in shaker flasks. One clone was scaled up to 25 L bag production using a similar hydrolysate feeding regime. The antibody titre reached 655 mg L^?1, and 12 g of antibody was recovered after purification, demonstrating scalability of the process. The process of clonal selection through to fed‐batch production of gram quantities was completed within 4 months. CONCLUSION: HEK‐293 cells can be used as a stable host for the production of biopharmaceuticals, producing gram quantities of recombinant proteins for preclinical development. Copyright © 2011 Society of Chemical Industry     

Simultaneous organic carbon and nitrogen removal in an SBR controlled at low dissolved oxygen concentration

Simultaneous organic carbon and nitrogen removal was studied in a sequencing batch reactor (SBR) fed with synthetic municipal wastewater and controlled at a low dissolved oxygen (DO) level (0.8 mg dm^?3). Experimental results over a long time (120 days) showed that the reactor achieved high treatment capacities (organic and nitrogen loading rates reached as high as 2.4 kg COD m^?3 d^?1 and 0.24 kg NH₃‐N m³ d^?1) and efficiencies (COD, NH₃‐N and total nitrogen removal efficiencies were 95%, 99% and 75%). No filamentous bacteria were found in the sludge even though the reactor had been seeded with filamentous bulking sludge. Instead, granular sludge, which possessed high activity and good settleability, was formed. Furthermore, the sludge production rate under low DO was less than that under high DO. Significant benefits, such as low investment and less operating cost, will be obtained from the new process. © 2001 Society of Chemical Industry     

Biodiesel production from Stichococcus strains at laboratory scale

BACKGROUND: This paper reports the results of an experimental campaign of autotrophic cultures of Stichococcus strains aiming at selecting the most promising strain for biofuel production. The strain selected—S. bacillaris 158/11—was cultivated in 1 L lab‐scale bubble column photobioreactors under fed‐batch and semi‐continuous conditions. A Bold basal medium supplemented with NaNO₃ as nitrogen source was adopted. Tests were carried out at 23 °C, 140 µE m^?2 s^?1, and air flow rate ranging between 0.4 and 4 vvm. Cultures were characterized in terms of pH, concentration of total nitrogen, total organic carbon, total inorganic carbon, biomass, lipid fraction and methyl‐ester distribution of transesterified lipids. RESULTS: S. bacillaris 158/11 proved to be the best strain to produce biodiesel. Methyl‐ester distribution was characterized by a large fraction of methyl palmitate, methyl linolenate, methyl linoleate, and methyl oleate along with phytol. The process photosynthetic efficiency—fraction of available light stored as chemical energy ‐ was about 1.5%. Specific biomass productivity was ～60 mg_DM L^?1 day^?1 under the semi‐continuous conditions tested. Total lipid productivity was 14 mg L^?1 day^?1 at a dilution rate of 0.050 L day^?1. CONCLUSION: S. bacillaris 158/11 is a potential strain for massive microalgae cultures for biofuel production. Higher biomass/total‐lipid productivity could be obtained in sunlight. Copyright © 2011 Society of Chemical Industry     

Exploiting olive oil mill effluents as a renewable resource for production of biodegradable polymers through a combined anaerobic–aerobic process

BACKGROUND: The performance of a three‐stage process for polyhydroxyalkanoate (PHA) bioproduction from olive oil mill effluents (OME) has been investigated. In the first anaerobic stage OME were fermented in a packed bed biofilm reactor into volatile fatty acids (VFAs). This VFA‐rich effluent was fed to the second stage, operated in an aerobic sequencing batch reactor (SBR), to enrich mixed cultures able to store PHAs. Finally, the storage response of the selected consortia was exploited in the third aerobic stage, operated in batch conditions. RESULTS: The anaerobic stage increased the VFA percentage in the OME from 18% to ～32% of the overall chemical oxygen demand (COD). A biomass with high storage response was successfully enriched in the SBR fed with the fermented OME at an organic load rate of 8.5 gCOD L^?1 d^?1, with maximum storage rate and yield (146 mgCOD gCOD^?1 h^?1 and 0.36 COD COD^?1, respectively) very similar to those obtained with a synthetic VFA mixture. By means of denaturing gradient gel electrophoresis (DGGE) analysis, different bacterial strains were identified during the two SBR runs: Lampropedia hyalina and Candidatus Meganema perideroedes, with the synthetic feed or the fermented OMEs, respectively. In the third stage, operated at increasing loads, the maximum concentration of the PHA produced increased linearly with the substrate fed. Moreover, about half of the stored PHAs were produced from substrates other than VFAs, mostly alcohols. CONCLUSION: The results obtained indicate that the process is effective for simultaneous treatment of OME and their valorization as a renewable resource for PHA production. Copyright © 2009 Society of Chemical Industry     

Lipolytic enzyme production in batch and fed‐batch cultures of Ophiostoma piceae and Fusarium oxysporum

Lipase and esterase production by Ophiostoma piceae and Fusarium oxysporum were enhanced and extended by developing a fed‐batch process in stirred tank reactors. Fed‐batch strategy improved lipolytic enzyme production from Ophiostoma piceae in both 2 and 20 dm³ stirred tank reactors. However, fed‐batch fermentation of Fusarium oxysporum in the 2 dm³ reactor was more effective than both batch and fed‐batch fermentations in the 20 dm³ reactor. When a medium composed of only carbon and nitrogen source was intermittently fed to the cultures, the maximum specific lipase activity was improved by more than 80% and 35% in Ophiostoma piceae and Fusarium oxysporum cultures respectively. The maximum specific esterase activity was improved by 20% and 15% in Ophiostoma piceae and Fusarium oxysporum cultures respectively. The duration of production for both fungi extended from 144 to 216 h compared with a batch culture under the same condition. © 2000 Society of Chemical Industry     

Effect of Saccharomyces cerevisiae fermentation conditions on expanded bed adsorption of heterologous cutinase

The effect of culture media composition, and fermentation conditions and strategy on the growth and cutinase production of recombinant Saccharomyces cerevisiae and subsequent cutinase purification by expanded bed adsorption (EBA) was studied. The reduction in the amount of yeast extract used as nitrogen source from 20 g dm^?3 to 10 g dm^?3 in batch cultures led to a 29% decrease in the heterologous cutinase production, while the 5% cutinase dynamic adsorption capacity (q5%) on the cation Streamline SP XL was increased 6.7‐fold. By dilution of the whole fermentation broth, performed with the lowest yeast extract content, which reduces conductivity, the q5% was additionally increased by 1.9‐fold. After implementation of a fed‐batch strategy the cutinase concentration, cutinase yield on carbon source, cutinase yield on nitrogen source and productivity were increased by 10.8‐, 2.9‐, 5.3‐ and 6.4‐fold, respectively, in relation to the previously‐mentioned batch fermentation. However, the increased cutinase production was compromised by heterologous protein loss during the EBA recovery operation and the cutinase dynamic adsorption capacity and purification productivity decreased by 90% and 75%, respectively. Thus, target protein production by S cerevisiae fermentation and a downstream process with EBA cannot be considered as separate entities, where the understanding of the factors that affect the interactions among them are crucial towards optimization of the overall production process of heterologous proteins. © 2002 Society of Chemical Industry     

Start‐up and enrichment of a granular anammox SBR to treat high nitrogen load wastewaters

BACKGROUND: Landfill leachate is characterized by low biodegradable organic matter that presents difficulties for the complete biological nitrogen removal usually performed by conventional biological nitrification/denitrification processes. To achieve this, the anaerobic ammonium oxidation (anammox) process is a promising biological treatment. This paper presents an anammox start‐up and enrichment methodology for treating high nitrogen load wastewaters using sequencing batch reactor (SBR) technology. RESULTS: The methodology is based on the gradual increase of the nitrite‐to‐ammonium molar ratio in the influent (from 0.76 to 1.32 mole NO₂^?‐N mole^?1NH₄⁺‐N) and on the exponential increase of the nitrogen loading rate (NLR, from 0.01 to 1.60 kg N m^?3 d^?1). 60 days after start‐up, anammox organisms were identified by polymerase chain reaction (PCR) technique as Candidatus Brocadia anammoxidans. After one year of operation, NLR had reached a value of 1.60 kg N m^?3 d^?1 with a nitrogen (ammonium plus nitrite) removal efficiency of 99.7%. The anammox biomass activity was verified by nitrogen mass balances with 1.32 ± 0.05 mole of nitrite removed per mole of ammonium removed and 0.23 ± 0.05 mole of nitrate produced per mole of ammonium removed. Also, enrichment of anammox bacteria was quantified by fluorescence in situ hybridization (FISH) analysis as 85.0 ± 1.8%. CONCLUSIONS: This paper provides a methodology for the enrichment of the anammox biomass in a SBR to treat high nitrogen loaded wastewaters. Copyright © 2007 Society of Chemical Industry     

Experimentation of a new mode of batch culture for lactic acid bacteria: cell reuse with an initial period of cell reactivation at acidic pH

Cell reuse was compared with conventional batch culture for lactic acid fermentation, the objective was to simplify the batch process and to alleviate the need for added nitrogen. At high levels of nitrogen supplementation to the culture medium (20 g dm^?3 yeast extract and 5 g dm^?3 each of tryptic and pancreatic casein peptones), similar mean production rates were obtained with partial cell reuse and the conventional batch process, without any additional gain when cells were initially reactivated at acidic pH. On the other hand, cell reuse with an initial period without pH control appeared particularly effective for low levels of nitrogen supplementation (5 g dm^?3 yeast extract): a 57% increase in the mean production rate with regard to the conventional batch process was obtained. © 2001 Society of Chemical Industry     

Extraction of anthocyanins from Aronia melanocarpa skin waste as a sustainable source of natural colorants

Aronia melanocarpa (Michx.) Elliott (black chokeberry) skin wastes from the production of Aronia fruit juice were extracted using a batch extraction method and a novel integrated extraction‐adsorption process. Optimum conditions for batch extraction were as follows: 60 °C, 3 h, acid (0.1% v/v hydrochloric acid), biomass‐to‐solvent ratio of 1:16, and biomass‐to‐solid phase extraction resin ratio of 1:1. The integrated extraction‐adsorption process gave improved anthocyanin yields of higher quality when the process was performed for 3 h without cooling of the circulating liquid, and with a flow rate of 1.3 ml s^?1. Overall, the new method showed better anthocyanin yield and purity compared with the batch method, increasing the extraction yield by ca. 20% (5.25→6.34 mg g^?1 dry weight of pomace) and increasing anthocyanin content by ca. 40% (19.9%→28.4% w/w dry weight of extract). This method also simplified the process as three steps were eliminated saving time and energy. Furthermore, the integrated extraction‐adsorption method is industrially scalable to produce large quantities of anthocyanins. In the batch method, anthocyanins present in A. melanocarpa skins were identified as cyanidin‐3‐O‐galactoside (38.8%), cyanidin‐3‐O‐arabinoside (6.4%), cyanidin‐3‐O‐glucoside (3.6%), cyanidin‐3‐O‐xyloside (0.5%), and the cyanidin aglycon (50.7%); in the continuous method, anthocyanin content was cyanidin‐3‐O‐galactoside (45.7%), cyanidin‐3‐O‐arabinoside (16%), cyanidin‐3‐O‐glucoside (3.6%), cyanidin‐3‐O‐xyloside (2.7%), and the cyanidin aglycon (32%). The integrated extraction‐adsorption method was shown to be substantially less susceptible to acid‐catalysed anthocyanin decomposition processes. All anthocyanins were derived from only one anthocyanidin parent structure (cyanidin), and only monosaccharide glycosides were identified, which is unusual when compared with other berries that typically have more anthocyanidins and/or greater glycosylation diversity.     

Production of Bacillus thuringiensis‐based biopesticides in batch and fed batch cultures using wastewater sludge as a raw material

Bacillus thuringiensis subsp kurstaki was grown in batch and fed batch cultures using wastewater sludge as a raw material. A simple fed batch strategy based on dissolved oxygen measurement during the fermentation cycle was developed in this work. It was established that while shifting the process strategy from batch to fed batch, the maximum spore concentration was increased from 5.62 × 10⁸ to 8.6 × 10⁸ colony forming units per cm³ and resulted in an increase of entomocidal activity from 13 × 10⁹ to 18 × 10⁹ spruce budworm potency units per dm³. Higher entomotoxicity was recorded at low spore concentration using wastewater sludge as a raw material whereas low entomotoxicity was reported at high spore concentration in synthetic medium. Copyright © 2005 Society of Chemical Industry