Bio-processing of agro-byproducts to animal feed.

Agricultural and food-industry residues constitute a major proportion (almost 30%) of worldwide agricultural production. These wastes mainly comprise lignocellulosic materials, fruit and vegetable wastes, sugar-industry wastes as well as animal and fisheries refuse and byproducts. Agro-residues are rich in many bioactive and nutraceutical compounds, such as polyphenolics, carotenoids and dietary fiber among others. Agro residues are a major valuable biomass and present potential solutions to problems of animal nutrition and the worldwide supply of protein and calories, if appropriate technologies can be used for their valorization by nutrient enrichment. Technologies available for protein enrichment of these wastes include solid substrate fermentation, ensiling, and high solid or slurry processes. Technologies to be developed for the reprocessing of these wastes need to take account of the peculiarities of individual wastes and the environment in which they are generated, reprocessed, and used. In particular, such technologies need to deliver products that are safe, not just for animal feed use, but also from the perspective of human feeding. This review focuses on the major current applications of solid-state fermentation in relation to the feed sector.

Lignin analysis using microwave digestion.

Lignin quantification in apple pomace residues was carried out using a microwave oven to replace traditional refluxing during the mild acidolysis step to augment the selectivity of this step towards cleavage of lignin-carbohydrate bonds and to reduce the time needed to quantify lignin. The pressure, temperature and time were optimized by response surface methodology and the results were compared to the Klason lignin methodology. Temperature and pressure had a significant positive effect (p < 0.05) on the determination of lignin. However, the time was also significant (p < 0.05) on lignin quantification. The optimal conditions of digestion were: 30 bar, 170 °C for 15 min. The digestion using microwave (lignin content = 33 % w/w) was more accurate (p < 0.05) than the the traditional refluxing (lignin content = 27 % w/w).

Extraction and analysis of polyphenols: recent trends.

In recent years, there has been an increasing interest in diets rich in fruits and vegetables and this is mostly due to their presumed role in the prevention of various degenerative diseases, such as cancer and cardiovascular diseases. This is mainly due to the presence of bioactive compounds, such as polyphenols, carotenoids, among others. Polyphenols are one of the main classes of secondary metabolites derived from plants offering several health benefits resulting in their use as functional foods. Prior to the use of these polyphenols in specific applications, such as food, pharmaceutical, and the cosmetic industries, they need to be extracted from the natural matrices, then analyzed and characterized. The development of an efficient procedure for the extraction, proper analysis, and characterization of phenolic compounds from different sources is a challenging task due to the structural diversity of phenolic compounds, a complex matrix, and their interaction with other cellular components. In this light, this review discusses different methods of extraction, analysis, and the structural characterization of polyphenolic compounds.

Bioproduction of hydrolytic enzymes using apple pomace waste by A. niger: applications in biocontrol formulations and hydrolysis of chitin/chitosan.

Studies were carried out for ß-glucosidase production using apple pomace (AP) in solid-state fermentation using 2(4) factorial design and response surface methodology. The influence of four independent variables including initial moisture level and inducers [veratryl alcohol (VA), lactose (LAC) and copper sulfate (CS)] was studied. The experimental design showed that initial moisture level had significant negative effect on the response. Higher ß-glucosidase activity of 64.18 IU/gram fermented substrate (gfs) was achieved in solid-state tray fermentation with optimum conditions having initial moisture level 55% (v/w), pH 4.5, 2 mM/kg VA, 2% (w/w) LAC and 1.5 mM/kg CS concentration, respectively,. The non-specific chitinase 70.28 ± 6.34 IU/gfs and chitosanase activities 60.18 ± 6.82 to 64.20 ± 7.12 IU/gfs were observed. The study demonstrated that AP can be potentially used for the ß-glucosidase production by Aspergillus niger. Moreover, ß-glucosidase can be used for the hydrolysis of chitin/chitosan to depolymerized products and in the formulation of biocontrol agents for enhanced entomotoxicity levels.

Optimization of spray drying process for Bacillus thuringiensis fermented wastewater and wastewater sludge.

Response surface methodology was used to optimize spray drying process for producing biopesticide powders of Bacillus thuringiensis by using fermented broth of starch industry wastewater and wastewater sludge. Analysis of variance was carried out using number of viable spores in the powder as dependent variable. The determination coefficients of models were 92 and 94% for fermented broth of starch industry wastewater and wastewater sludge, respectively. Under the optimal conditions of the operational parameters of spray drying, the numbers of viable spores were 2.2 × 10(8) and 1.3 × 10(8) CFU/mg in the dry powders for starch industry wastewater and wastewater sludge respectively, with a loss of viable spores of 18 and 13% when compared with their respective fermented broths. The entomotoxicity (measured by the bioassay method) of the powders obtained under optimal conditions showed a loss of 28 and 18% when compared with the fermented broth of starch industry wastewater and wastewater sludge, respectively. The optimized results of spray drying were used for field application calculations. The volume of fermented broth required to produce powder formulated product when compared with the volume required for liquid formulation product in order to treat 1 ha of balsam fir was less and offered several advantages.

Rheological profile of diets produced using agro-industrial wastes for rearing codling moth larvae for baculovirus biopesticides.

A rheological study of diets using the agro-industrial wastes (brewery wastewater and pomace waste) was carried out in order to obtain a diet most adapted to supply nutrients for growth of codling moth (CM) larvae. Nutritive capacity (g/L) of brewery wastewater (BWW) (25.5 ± 5.5 carbohydrates; 16.9 ± 2.1 proteins; 6 ± 1.6 lipids) and pomace waste (POM) (22.0 ± 0.03 carbohydrates; 11.3 ± 1.3 proteins; 2 ± 0.2 lipids) were essential and important as replacement or in association with other ingredients [soya flour (SF), wheat germ (WG), yeast extract (YE)] of the standard diet for the breeding of codling moth larvae. These diet additives also contributed to the preservation of texture and nutritive content of larvae diet. The eggs and CM larvae were grown on alternate diets under industrial conditions (16:8 h photoperiod; 25 ± 1 °C and 50 ± 0.5 % of humidity). The higher assimilation of nutrients of the diets in BWW and control diet was observed by calculating the rate of hatching of eggs (0.48 to 0.71); larvae growth (0.23 to 0.4) and fertility (1.33 to 3 for control diet). The excellent growth and fertility rates of codling moth larvae were attributed to variations in viscosity (varying from 50 to 266 mPa.s⁻¹), particle size (varying 24.3 µm in 88.05 µm with regard to 110 µm the control diet) and total solids (145.88 g/L POM + YE; 162.08 g/L BWW + YE; 162.2 g/L POM + WG; 173 g/L control; 174.3 g/L BWW + WG) diets. Lower viscosity favored improved diet due to ease of assimilation of nutrients. Thus, rheology is an important parameter during preparation of diets for growth of codling moth larvae as it will dictate the nutrient assimilation which is an important parameter of larvae growth.

Impact of different pH control agents on biopesticidal activity of Bacillus thuringiensis during the fermentation of starch industry wastewater.

Different pH control agents (NaOH/H(2)SO(4)--SodSulp, NaOH/CH(3)COOH--SodAcet, NH(4)OH/CH(3)COOH--AmmoAcet and NH(4)OH/H(2)SO(4)--AmmoSulp) were used to investigate their effects on growth, enzyme production (alkaline protease and amylase), and entomotoxicity of Bacillus thuringiensis var. kurstaki HD-1 (Btk) against eastern spruce budworm larvae (Choristoneura fumiferana) using starch industry wastewater (SIW) as a raw material in a 15-l fermentor. AmmoSulp and SodSulp were found to be the best pH control agents for alkaline protease and amylase production, respectively; whereas, the fermented broth obtained by using SodAcet as pH control agents recorded the highest delta-endotoxin production of 1043.0 mg/l and entomotoxicity value 18.4 x 10(9) SBU/l. Entomotoxicity of re-suspended centrifuged pellet in one-tenth of original volume in case of SodAcet as pH control agents was 26.7 x 10(9) SBU/l and was the highest value compared to three other pH control agents.

Starch industry wastewater for production of biopesticides--ramifications of solids concentrations.

Total solids (TS) concentrations ranging from 15 to 66 g L(-1) of starch industry wastewater (SIW) were tested as raw material for the production of Bacillus thuringiensis var. kurstaki HD-1 (Btk) biopesticide in shake flasks and a 15 L bench-scale fermenter. Shake flask studies revealed a higher delta-endotoxin concentration of Btk at 30 g L(-1) TS concentration and 2.5% (v v(-1)) volume of pre-culture. The fermenter experiments conducted using SIW at 30 g L(-1) TS concentration under controlled conditions of temperature, pH and dissolved oxygen showed higher spore count, enzyme production (protease and amylase) and delta-endotoxin concentration as compared with those of SIW at 15 g L(-1) TS concentration. The entomotoxicity, at the end of fermentation, with SIW at 30 g L(-1) solids concentration (17.8 x 10(9) SBU L(-1), measured against spruce budworm) was considerably higher as compared with entomotoxicity at 15 g L(-1) solids concentration (15.3 x 10(9) SBU L(-1)) and semi-synthetic medium (11.7 x 10(9) SBU L(-1)). The pellet, comprising spores and delta-endotoxin complex obtained after centrifugation and followed by resuspension (in supernatant) in one-tenth of the original volume, of SIW at 30 g L(-1) solids concentration media registered the highest potential for application (to protect forests against spruce budworm) than other media in term of entomotoxicity.

Bacillus thuringiensis fermentation of wastewater and wastewater sludge--presence and characterization of chitinases.

This study investigated the presence of chitinases in Bacillus thuringiensis ssp kurstaki HD-1 (Bt) fermented broths of wastewater sludge (non-hydrolyzed and hydrolyzed); starch industry wastewater and soyameal. Chitinase activity was absent in soyameal and present in others. Chitinase demonstrated peaks at pH 4.0 and temperatures 40 and 50 degrees C with higher activity between pH 4-5 and 10-11. The chitinase band on SDS-PAGE was found to be between 36 and 45 kDa for non-hydrolyzed (NH) and hydrolyzed sludge (TH) and starch industry wastewater. The chitinase profile during fermentation showed peaks at 15 and 30 h for non-hydrolyzed and hydrolyzed sludge and 15 and 24 h for starch industry wastewater. Chitinase retained 96-99 % activity after two weeks incubation at room temperature and pH 4. Bioassays with supplementation of Bt chitinases showed 1.2 fold increase in entomotoxicity of wastewater sludge and a small increase in starch industry wastewater. This study sheds light on production of Bt chitinases in alternative media which will have a long term effect on entomotoxicity of these formulations.

Chemical and biological conversion of crude glycerol derived from waste cooking oil to biodiesel.

In this study, crude, purified, and pure glycerol were used to cultivate Trichosporon oleaginosus for lipid production which was then used as feedstock of biodiesel production. The purified glycerol was obtained from crude glycerol by removing soap with addition of H3PO4 which converted soap to free fatty acids and then separated from the solution. The results showed that purified glycerol provided similar performance as pure glycerol in lipid accumulation; however, crude glycerol as carbon source had negatively impacted the lipid production of T. oleaginosus. Purified glycerol was later used to determine the optimal glycerol concentration for lipid production. The highest lipid yield 0.19g/g glycerol was obtained at 50g/L purified glycerol in which the biomass concentration and lipid content were 10.75g/L and 47% w/w, respectively. An energy gain of 4150.51MJ could be obtained with 1tonne of the crude glycerol employed for biodiesel production through the process proposed in this study. The biodiesel production cost estimated was 6.32US$/gal. Fatty acid profiles revealed that C16:0 and C18:1 were the major compounds of the biodiesel from the lipid produced by T. oleaginosus cultivated with crude and purified glycerol. The study found that purified glycerol was promising carbon source for biodiesel production.

Starch industry wastewater as a substrate for antagonist, Trichoderma viride production.

Starch industry wastewater was investigated to assess and improve its potential as a raw material for the conidia production of biocontrol fungi, Trichoderma viride. The wastewater was tested with and without supplements of glucose, soluble starch, meat peptone and probable conidiation inducer chemicals in shake flask culture. Addition of complex carbon source (soluble starch, 1% and 2% w/v) produced maximum conidia ( approximately 3.02 and 4.2 x 10(10)CFU/mL, respectively). On the other hand, glucose addition as a simpler carbon source was either ineffective or, reduced conidia production (from 1.6 x 10(8) in control to 3.0 x 10(7)CFU/mL in 5% w/v glucose supplement). Supplement of nitrogen source showed a small increase of conidia concentration. Propionic, maleic and humic acids, EDTA, pyridine, glycerol and CaCO(3) were examined as probable conidiation inducers and showed effect only on initial rate of conidiation with no increase in final conidia concentration. Intra and extracellular ATP correlation with spore production showed dependence on growth media used and conidia concentration at the end of fermentation. Addition of carbon and nitrogen sources showed an increase in protease activity (from 0.4985 to 2.43 IU/mL) and entomotoxicity (from 10448 to 12335 spruce budworm unit (SBU)/microL). Entomotoxicity was improved by 11% in fermenter over shake flask when starch industry wastewater was supplemented with meat peptone.

Bacillus thuringiensis fermentation of hydrolyzed sludge--rheology and formulation studies.

Rheology of Bacillus thuringiensis fermentation of hydrolyzed sludge was investigated in bench scale fermenter. Stable liquid formulations were developed and optimized for two-year based studies comprising various physical/chemical (viscosity, particle size, corrosion and suspendibility) and biological (microbial contamination, viable spores and entomotoxicity) parameters at different pHs and temperatures. The hydrolyzed sludge depicted non-Newtonian and pseudoplastic behaviour during fermentation with 90% to 96% confidence of fits into Casson, Power and IPC paste models. Higher values of consistency and flow index during exponential growth and stationary phase, respectively, affected downstream processing. The power law was also followed by stable formulations. Sorbitol, sodium monophosphate and sodium metabisulfite (2.2:1:1) as suspending agents produced suspendibility ranging from 69% to 94%. The stable formulation (FH-4) comprising sorbitol, sodium monophosphate and sodium metabisulfite deteriorated at pHs 6, 6.5 and temperatures, 40 and 50 degrees C, with no signs of corrosion and microbial contamination. The viscosity of FH-4 formulations decreased with shear rate which could improve handling and consequent spraying.

Efficient centrifugal recovery of Bacillus thuringiensis biopesticides from fermented wastewater and wastewater sludge.

Studies were conducted on harvesting of Bacillus thuringiensis (Bt)-based biopesticides from fermented broths of starch industry wastewater (SIW), wastewater sludge (raw and hydrolyzed-NH and TH, respectively) and semi-synthetic soyameal to enhance entomotoxicity (Tx) by centrifugation. Pertinent factors influencing Tx, solids concentration, pH, temperature and centrifugal force were investigated. The centrifugate solids concentration beyond 100 g/l did not enhance Tx, instead caused pellet formation. Centrifugation efficiency (Tx recovery) was higher at pH 4, and temperature 20 degrees C for starch wastewater (98%), wastewater sludge (98% and 97.8% for non-hydrolyzed and hydrolyzed, respectively) and soya broth (83%). For maximum Tx recovery (SIW-95%; NH-90%; TH-98% and soya-78%), the centrifugal force and time required was 48,000 g and 30 min, respectively. Losses in recovery efficiency were lower for SIW and wastewater sludge in comparison to soya on adopting commercially recommended centrifugal force of 9000 g. The settling velocity computations for different fermented broths enabled calculation of Sigma factor for continuous commercial centrifuge of a given capacity and hence simulation of power requirements. It was established that power requirements for a given Tx recovery efficiency were highest for conventional medium (soya) in comparison to other waste-based fermented broths.

Bioconversion of industrial wastewater and wastewater sludge into Bacillus thuringiensis based biopesticides in pilot fermentor.

Starch industry wastewater (SWW), slaughterhouse wastewater (SHWW) and secondary sludges from three different wastewater treatment plants (Jonquière--JQS, Communauté Urbaine de Québec--CUQS and Black lake-BLS) were used as raw materials for the production of Bacillus thuringiensis (Bt) based biopesticides in a pilot scale fermentor (100 L working volume). The slaughterhouse wastewater exhibited the lowest Bt growth and entomotoxcity (Tx) potential (measured against spruce budworm) due to low availability of carbon, nitrogen and other nutrients. Performance variation (growth, sporulation, proteolytic activity and Tx potential) within the three types of sludges was directly related to the availability of nitrogen and carbohydrates, which could change with sludge origin and methods employed for its generation. The Tx potential of Bt obtained in different secondary sludges (JQS: 12 x 10(9) SBU/L; CUQS: 13 x 10(9) SBU/L and BLS: 16 x 10(9) SBU/L) and SWW (18 x 10(9) SBU/L) was higher than the soybean based synthetic medium (10 x 10(9) SBU/L). The maximum protease activity was obtained in CUQ secondary sludge (4.1 IU/mL) due to its high complex protein concentration. Nevertheless, high carbohydrate concentration in SWW repressed enzyme production. The secondary sludges and SWW were found to be suitable raw materials for high potency Bt biopesticide production.

Screening of different adjuvants for wastewater/wastewater sludge-based Bacillus thuringiensis formulations.

Screening of different adjuvants, namely, suspending agents, phagostimulants, stickers, antimicrobial agents, and UV screens to develop aqueous biopesticidal suspensions of Bacillus thuringiensis (Bt) variety kurstaki HD-1 fermented broths, specifically, nonhydrolyzed sludge, hydrolyzed sludge, starch industry wastewater, and soya (commercial medium), were investigated. The selected suspending agents [20% (wt:vol)] included sorbitol, sodium monophosphate, and sodium metabisulfite with corresponding suspendibility of 74-92, 69-85, and 71-82%, respectively. Molasses [0.2% (wt:vol)] increased adherence by 84-90% for all fermented broths. The optimal phagostimulants [0.5% (wt:vol)], namely, soya and molasses, caused entomotoxicity increase of 3-13 and 7-13%, respectively. Sorbic and propionic acids showed high antimicrobial action [0.5% (wt:vol)], irrespective of fermentation medium. Sodium lignosulfonate, molasses, and Congo red, when used as UV screens [0.2% (wt:vol)], showed percent corresponding entomotoxicity losses of 3-5, 0.5-5 and 2-16, respectively. The Bt formulations, when exposed to UV radiation, showed higher half-lives (with and without UV screens) than the fermented broths or semisynthetic soya medium and commercial Bt formulation. UV screen-amended nonhydrolyzed, hydrolyzed, and starch industry wastewater formulations showed 1.3-1.5-fold higher half-lives than commercial Bt formulation. Thus, the recommended formulation comprises sorbitol, sodium monophosphate, sodium metabisulfite (suspending agents); molasses, soya flour (phagostimulants); molasses and skimmed milk powder (rainfasteners); sorbic and propionic acids (antimicrobial agents) and sodium lignosulfate; and molasses and Congo red (UV screens). These waste-based Bt formulations offer better UV resistance in comparison with commercial formulation.

Membrane processes for removal of pharmaceutically active compounds (PhACs) from water and wastewaters.

Pharmaceutically active compounds (PhACs), which find their way easily into the water sources, are emerging as a major concern for drinking water quality and aquatic species. Therefore, their removal from water sources is a priority from environmental point of view. During the past decade, different methods including membrane separation, adsorption systems and chemical transformation have been evaluated for removal of these compounds. This paper reviews different aspects of PhAC removal by using membrane separation processes, as they have been conventionally known to show high potential in the production of superior quality drinking and industrial water. In brief, osmosis membranes can efficiently remove almost all PhACs though its operational cost is relatively high and nanofiltration (NF) membranes are highly influenced by electrostatic and hydrophobic interaction. Moreover, the efficiency of membrane bioreactors (MBRs) is difficult to predict due to the complex interaction of compounds with microorganisms. To improve the performance and robustness of membrane technology, it is suggested to combine membranes with other systems, such as activated carbon and enzymatic degradation.

Adsorption study of environmentally relevant concentrations of chlortetracycline on pinewood biochar.

The presence of pharmaceutically active compounds (PhACs) in water and wastewater has raised concerns because of potential environmental impacts and thus their removal is of high importance. The adsorption behavior of chlortetracycline (CTC) from aqueous solution on raw and activated pinewood biochar was studied at 298 K. The effect of initial pH of the solution was studied by performing the experiment at three different pHs (1, 5 and 9). At each pH, CTC showed varied electrostatic charge (+1, 0 and -1, respectively) which affected its adsorption. The results indicated that CTC followed Langmuir isotherm and the related parameters were calculated. Also, it was observed that the maximum adsorption occurred at pH1. The adsorption capacity of CTC for raw and activated biochar was at least 2.1 and 208.3mg/g adsorbent, respectively. The characteristics of biochars were studied using zeta potential analyzer, laser size analyzer and scanning electron microscopy (SEM). The results showed that raw and activated biochars are promising candidates for removal of CTC from water due to the acidic character of pinewood that can result in better interaction with ionizable compounds at lower pHs.

Sludge based Bacillus thuringiensis biopesticides: viscosity impacts.

Viscosity studies were performed on raw, pre-treated (sterilised and thermal alkaline hydrolysed or both types of treatment) and Bacillus thuringiensis (Bt) fermented sludges at different solids concentration (10-40 g/L) for production of biopesticides. Correlations were established among rheological parameter (viscosity), solids (total and dissolved) concentration and entomotoxicity (Tx) of Bt fermented sludges. Exponential and power laws were preferentially followed by hydrolysed fermented compared to raw fermented sludge. Soluble chemical oxygen demand variation corroborated with increase in dissolved solids concentration on pre-treatments, contributing to changes in viscosity. Moreover, Tx was higher for hydrolysed fermented sludge in comparison to raw fermented sludge owing to increased availability of nutrients and lower viscosity that improved oxygen transfer. The shake flask results were reproducible in fermenter. This study will have major impact on selecting fermentation, harvesting and formulation techniques of Bt fermented sludges for biopesticide production.

Wastewater sludge as a potential raw material for antagonistic fungus (Trichoderma sp.): role of pre-treatment and solids concentration.

Feasibility of production of antagonistic Trichoderma sp. conidial spores using wastewater sludge as a raw material employing different suspended solids concentration (10-50 g/l) was investigated in shake flasks. Maximum conidial spore count obtained for raw sludge was 1.98 x 10(4) CFU/ml, which was enhanced by sludge pre-treatments (alkaline and thermal alkaline). Conidial spore count ranging from 1.3 x 10(6) to 2.8 x 10(7) CFU/ml was observed for alkaline and thermal alkaline treated sludges. Optimal suspended solids concentration was 30 g/l (10(7) CFU/ml) whereas, lower (<20 g/l) and higher (>30 g/l) solids concentration were less efficient. Thermal alkaline pre-treated sludge showed diauxic growth due to multiplicity of sludge biodegradability. A simple, modified CFU filtration technique was also developed for fungal spore assessment in sludge. Bioassay of fermented sludge against spruce budworm larvae showed entomotoxicity (15036 SBU/microl), on par with Bacillus thuringiensis biopesticides. This study successfully demonstrated potential of wastewater sludge as a raw material for production of value added product, aiding in sludge management and proliferation of eco-friendly and economical biocontrol agents.

Starch industry wastewater-based stable Bacillus thuringiensis liquid formulations.

Liquid formulations were developed from Bacillus thuringiensis (Bt)-fermented broths of starch industry wastewater (SIW) and of soya medium. Stability studies were carried out for 1 yr. Storage stability was tested by studying various physical and chemical (e.g., viscosity, particle size, corrosion, and suspendibility) and biological (e.g., microbial contamination, viable spores, and entomotoxicity) parameters at different pH levels and temperatures. Three suspending agents, sorbitol, sodium monophosphate, and sodium metabisulfite, were added to fermented broth in different concentrations. Sorbitol and sodium monophosphate in the ratio 3:1 was the best suspending agent combination for both formulations. Starch industry wastewater fermentation yielded cell and viable spore counts 10- and 4-fold greater than those from soya medium, respectively, and a 1.7-fold increase in entomotoxicity. However, both formulations started deteriorating at pH 6 and 6.5 and 40 and 50 degrees C. There were no signs of corrosion and microbial contamination in both types of formulations.