Biodegradation study by Pseudomonas sp. of flexible polyurethane foams derived from castor oil

The synthesis and biodegradation of polyurethane foams obtained from environmentally benign processes were studied.Flexible polyurethane foams based on castor oil modified with maleic anhydride (MACO) were synthesized. The synthesis involved a single-stage process by mixing castor oil/MACO (weight ratios 75:25 and 25:75) and 2-4 toluene diisocyanate (TDI) in stoichiometric amount of OH:NCO. The biodegradability studies with cultures of a Pseudomonas sp. strain (DBFIQ-P36) involved incubation periods of 2 months at 37 °C. Polymers were characterized before and after biodegradation by Fourier Transform Infrared Spectroscopy (FT-IR), INSTRON mechanical tester, and Scanning Electron Microscopy (SEM). The results showed that the addition of MACO produces a considerable increase in the rate of degradation and an important change in the chemical and morphological structures. This is due to the presence of ester groups that are vulnerable to chemical hydrolysis and enzymatic attack. The eco-toxicity after the biodegradation was evaluated. Toxic compounds such as primary amines were identified by Gas Chromatography–Mass Spectrometry (GC–MS) in combination with Nuclear Magnetic Resonance (NMR) as degradation products.     

Sequential interpenetrating polymer networks produced from vegetable oil based polyurethane and poly(methyl methacrylate)

Sequential interpenetrating polymer networks (IPNs) were prepared using polyurethane produced from a canola oil based polyol with primary terminal functional groups and poly(methyl methacrylate) (PMMA). The properties of the material were studied and compared to the IPNs made from commercial castor oil using dynamic mechanical analysis, differential scanning calorimetry, as well as tensile measurements. The morphology of the IPNs was investigated using scanning electron microscopy and transmission electron microscopy. The chemical diversity of the starting materials allowed the evaluation of the effects of dangling chains and graftings on the properties of the IPNs. The polymerization process of canola oil based IPNs was accelerated because of the utilization of polyol with primary functional groups, which efficiently lessened the effect of dangling chains and yielded a higher degree of phase mixing. The mechanical properties of canola oil based IPNs containing more than 75 wt % PMMA were comparable to the corresponding castor oil based IPNs; both were superior to those of the constituent polymers due to the finely divided rubber and plastic combination structures in these IPNs. However, when PMMA content was less than 65 wt %, canola oil based IPNs exhibited a typical mechanical behavior of rigid plastics, whereas castor oil based IPNs showed a typical mechanical behavior of soft rubber. It is proposed that these new IPN materials with high performance prepared from alternative renewable resources can prove to be valuable substitutes for existing materials in various applications.     

Improved tensile strength of glycerol-plasticized gluten bioplastic containing hydrophobic liquids

The aim of the present work has been to study the influence of hydrophobic liquids on the morphology and the properties of thermo-molded plastics based on glycerol-plasticized wheat gluten (WG). While the total amount of castor oil and glycerol was remained constant at 30 wt%, castor oil with various proportions with respect to glycerol was incorporated with WG by mixing at room temperature and the resultant mixtures were thermo-molded at 120 degrees C to prepare sheet samples. Moisture absorption, morphology, dynamic mechanical properties, and tensile properties (Young's modulus, tensile strength and elongation at break) of the plastics were evaluated. Experimental results showed that the physical properties of WG plastic were closely related to glycerol to castor oil ratio. Increasing in castor oil content reduces the moisture absorption markedly, which is accompanied with a significant improvement in tensile strength and Young's modulus. These observations were further confirmed in 24 wt% glycerol-plasticized WG plastics containing 6 wt% silicone oil or polydimethylsiloxane (PDMS) liquid rubber.     

Identification and Association Analysis of Castor Bean Orthologous Candidate Gene-Based Markers for High Oil Content in Jatropha curcas

Jatropha (Jatropha curcas) and castor bean (Ricinus communis) possess several taxonomic similarities, and their seeds contain a high proportion of oil (up to 40%) which has been used in various industrial products, including diesel oil. Thirty-two candidate genes responsible for fatty acid biosynthesis were identified in the castor bean genome sequence. Testing of 48 primer pairs from candidate gene regions, including 12 SSRs from castor bean on 54 genotypes of J. curcas, 65% amplified successfully on Jatropha out of which 20% showed polymorphisms. Jatropha genotypes, categorized for oil content, were used in association analysis of candidate gene regions with high oil content. One marker–trait association for the oil trait was identified. Stearoyl desaturase amplicon (700 bp) consisting of intron and exon (P?=?0.00013) showed association with high oil content in Jatropha genotypes. Sequencing of the 1.3-kb amplicon, including the 700-bp fragment of stearoyl desaturase, which had shown association with the high oil content, revealed SNPs in the exonic region. The SNPs resulted in substitution of leucine with glutamine in the open reading frame of stearoyl desaturase of low oil content genotypes. The molecular marker is expected to be useful in marker-assisted breeding of high oil content genotypes in Jatropha.     

Identification and functional expression of a type 2 acyl-CoA:diacylglycerol acyltransferase (DGAT2) in developing castor bean seeds which has high homology to the major triglyceride biosynthetic enzyme of fungi and animals

Seed oil from castor bean (Ricinus communis) contains high amounts of hydroxy fatty acid rich triacylglycerols (TAGs) that can serve as raw material for production of bio-based products such as nylon, cosmetics, lubricants, foams, and surfactants. Diacylglycerol acyltransferase (DGAT) catalyses the terminal reaction in the acyl-CoA dependent Kennedy pathway of triglyceride biosynthesis. There is still some debate whether there are three or four enzymes in yeast that have DGAT activity and catalyse the synthesis of TAG but of these the DGAT2 homologue Dga1 contributes in a major way to TAG biosynthesis. Here we report on the cloning of a cDNA for DGAT2 from castor bean and prove its biological activity following expression in yeast and enzymatic assays using diricinolein as the acceptor and ricinoleoyl-CoA as the donor. Previous reports of DGAT in castor have focussed on DGAT1 which has little amino acid sequence homology to DGAT2. Expressional studies demonstrate that DGAT2 is 18-fold more highly expressed in seeds than in leaves and shows temporal specific expression during seed development. In contrast, DGAT1 shows little difference in expression in seeds versus leaves. We conclude that in castor bean DGAT2 is more likely to play a major role in seed TAG biosynthesis than DGAT1.     

Current and Potential Biofuel Production from Plant Oils

Environmental concerns and depletion of fossil fuels along with government policies have led to the search for alternative fuels from various renewable and sustainable feedstocks. This review provides a critical overview of the chemical composition of common commercial plant oils, i.e., palm oil, olive oil, rapeseed oil, castor oil, WCO, and CTO and their recent trends toward potential biofuel production. Plant oils with a high energy content are primarily composed of triglycerides (generally >?95%), accompanied by diglycerides, monoglycerides, and free fatty acids. The heat content of plant oils is close to 90% for diesel fuels. The oxygen content is the most important difference in chemical composition between fossil oils and plant oils. Triglycerides can even be used directly in diesel engines. However, their high viscosity, low volatility, and poor cold flow properties can lead to engine problems. These problems require that plant oils need to be upgraded if they are to be used as a fuel in conventional diesel engines. Biodiesel, biooil, and renewable diesel are the three major biofuels obtained from plant oils. The main constraint associated with the production of biodiesel is the cost and sustainability of the feedstock. The renewable diesel obtained from crude tall oil is more sustainable than biofuels obtained from other feedstocks. The fuel properties of renewable diesel are similar to those of fossil fuels with reduced greenhouse gas emissions. In this review, the chemical composition of common commercial plant oils, i.e., palm oil, olive oil, rapeseed oil, castor oil, and tall oil, are presented. Both their major and minor components are discussed. Their compositions and fuel properties are compared to both fossil fuels and biofuels.     

Components of complex lipid biosynthetic pathways in developing castor (Ricinus communis) seeds identified by MudPIT analysis of enriched endoplasmic reticulum

Ricinoleic acid is a feedstock for nylon-11 (N11) synthesis which is currently obtained from castor (Ricinus communis) oil. Production of this fatty acid in a temperate oilseed crop is of great commercial interest, but the highest reported level in transgenic plant oils is 30%, below the 90% observed in castor and insufficient for commercial exploitation. To identify castor oil-biosynthetic enzymes and inform strategies to improve ricinoleic acid yields, we performed MudPIT analysis on endoplasmic reticulum (ER) purified from developing castor bean endosperm. Candidate enzymes for all steps of triacylglycerol synthesis were identified among 72 proteins in the data set related to complex-lipid metabolism. Previous reported proteomic data from oilseeds had not included any membrane-bound enzyme that might incorporate ricinoleic acid into oil. Analysis of enriched ER enabled determination of which protein isoforms for these enzymes were in developing castor seed. To complement this data, quantitative RT-PCR experiments with castor seed and leaf RNA were performed for orthologues of Arabidopsis oil-synthetic enzymes, determining which were highly expressed in the seed. These data provide important information for further manipulation of ricinoleic acid content in oilseeds and peptide data for future quantification strategies.     

Nano essential oils against the red palm weevil,Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae)

The red palm weevil, Rhynchophorus ferrugineus, is the biggest pest of various palm species. The aim of this study was to evaluate the impact of the essential oils of purslane, mustard and castor (bulk and nano) on R. ferrugineus and to evaluate the oviposition deterrent effect to prevent new infestations. Prepared concentrations of essential oils (bulk and nano) were tested for their impact on R. ferrugineus larvae, pupae and adults. The most effective oil was purslane oil (bulk and nano), followed by mustard and the least effective was castor. The percent mortality of larvae was 75.2, 45.3 and 17.9% (bulk phase) and 92.5, 84.4 and 65.5% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The percent mortality of adults was 67.2, 40.0 and 15.6% (bulk phase) and 83.5, 72.9 and 60.0% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The number of eggs laid decreased with increasing concentration of the tested oils. Purslane essential oil showed good impact on R. ferrugineus larvae and adults followed by mustard and castor oils as bulk and as nano.     

Preparation and characterization of polyurethane foams using a palm oil-based polyol

Polyurethane (PU) foams were prepared using a palm oil-based polyol (PO-p). At the first stage, palm oil was converted to monoglycerides as a new type of polyol by glycerolysis. A yield of the product reached 70% at reaction temperature of 90 degrees C by using an alkali catalyst and a solvent. At the second stage, PU foams were prepared from mixtures of the polyol and polyethylene glycol (PEG) or diethylene glycol (DEG) and an isocyanate compound. Characterization of the foams was carried out by thermal and mechanical analyses. The analyses showed that the chain motion of polyurethane becomes more flexible at the higher PO-p content in the whole polymer, which indicates that the monoglyceride molecules work as soft segments. The study here may lead to a development of a new type of polyurethane foams using palm oil as a raw material.     

1,8-cineole and castor oil in sodium lauryl ether sulphate disrupt reproduction and ovarian tissue of Rhipicephalus (Boophilus) microplus

Essential and fixed oils have been researched as alternatives to chemical acaricides. The activity of volatile compounds from essential oils (1,8-cineole, citral and eugenol) at 1.0% (w/v) and fixed oil (castor oil) at 0.3% (w/v) dissolved in 2.0% (v/v) dimethyl sulfoxide (DMSO) + 0.2% (w/v) Tween 80^® was assessed against Rhipicephalus microplus using immersion tests. 1,8-cineole (29.0%) and castor oil (30.2%) had the highest reproductive inhibition rate. A second experiment was performed to verify the effect of the 1,8-cineole (10.0% w/v) and, or castor oil (0.3% w/v) on tick reproduction using different solubilizing agents. The highest reproductive inhibition was observed for the combination of 1,8-cineole/castor oil (94.1%) and 1,8-cineole in 2.0% (w/v) sodium lauryl ether sulphate (SLES) (92.8%). A third experiment showed morphological changes in R. microplus oocytes at different stages of development, as well as in pedicel cells. The most intense effects were observed when ticks were immersed in the formulation containing 1,8-cineole (10.0% w/v) and castor oil (0.3% w/v) dissolved in 2% (w/v) SLES. These findings highlight the potential of this formulation as an alternative for managing cattle ticks as their cytotoxic effects can reduce R. microplus reproductive success.     

Genetic variability and population structure in a collection of inbred lines derived from a core germplasm of castor

Castor (Ricinus communis L.) is an industrially important oilseed crop and is the only source of an unusual fatty acid, ricinoleic acid in plant species. The castor oil and its products have numerous industrial uses including biofuel; hence, the demand for castor oil is ever increasing globally. Current productivity levels in castor are inadequate to meet the requirement, which underscores the need for breeding high yielding cultivars with better adaptability by exploiting diverse genetic resources. This study reports development and characterization of a set of inbred lines derived from a core germplasm collection of castor. The panel of 144 inbreds exhibited an excellent phenotypic diversity for morpho-agronomic traits related to plant architecture and yield components. However, SSR allelic diversity appears to be only moderate. The average number of alleles per SSR locus in the genotype panel was 3.0 and mean gene diversity was 0.38. Nevertheless, a majority of the inbred pairs (77 %) had very less estimated kinship coefficients (<0.05) suggesting that they were not related by pedigree. A very low level of genetic relatedness among the genotypes and absence of population structure suggest that this genotype panel consists of ideal set of materials for association mapping studies aiming at molecular breeding of key traits in castor. To our knowledge, this is the first report on the development and characterization of a large inbred collection representing the bulk of genetic diversity in castor, which can be further exploited for genetic, physiological and molecular studies towards achieving higher productivity.     

Castor and camphor essential oils alter hemocyte populations and induce biochemical changes in larvae of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)

Two plant essential oils; camphor and castor were tested for insecticidal and antifeedant activity against the 4th instar larvae of Spodoptera littoralis, a serious pest on cotton in Egypt. Also the impact of LC₁₀ of both oils on some physiological parameters in larvae was studied by using leaf dipping technique. Analysis of both oils using GC–MS revealed several insecticidal and antifeedant compounds. Our results showed higher insecticidal activity and antifeedant index of camphor oil against S. littoralis. The LC₅₀ and the antifeedant indices were 163.1, 246.8?mg/ml and 12.69, 6.62% for camphor and castor bean oil, respectively. The total hemocyte count (THC) and differential hemocyte count (DHC) were reduced significantly after 48?h of treatment compared to controls. Both oils reduced all types of hemocytes except plasmatocytes which were reduced only by castor oil. Camphor oil decreased total proteins and carbohydrates while castor oil targeted only carbohydrate content. Both oils didn't affect the amount of total lipids. Lipase, α-amylase and glucose-6-phosphate dehydrogenase (G6PD) enzyme activities were increased significantly in larvae treated with camphor oil than other treatments. These results clearly indicate that castor and camphor oils can affect the nutritional status in S. littoralis larvae, thereby changing the internal metabolic processes in the larvae which make them as potential control agents in IPM programs against S. littoralis.     

Influence of fatty acid methyl esters from hydroxylated vegetable oils on diesel fuel lubricity

Current and future regulations on the sulfur content of diesel fuel have led to a decrease in lubricity of these fuels. This decreased lubricity poses a significant problem as it may lead to wear and damage of diesel engines, primarily fuel injection systems. Vegetable oil based diesel fuel substitutes (biodiesel) have been shown to be clean and effective and may increase overall lubricity when added to diesel fuel at nominally low levels. Previous studies on castor oil suggest that its uniquely high level of the hydroxy fatty acid ricinoleic acid may impart increased lubricity to the oil and its derivatives as compared to other vegetable oils. Likewise, the developing oilseed Lesquerella may also increase diesel lubricity through its unique hydroxy fatty acid composition. This study examines the effect of castor and Lesquerella oil esters on the lubricity of diesel fuel using the High-Frequency Reciprocating Rig (HFRR) test and compares these results to those for the commercial vegetable oil derivatives soybean and rapeseed methyl esters.     

γ-decalactone production by Yarrowia lipolytica and Lindnera saturnus in crude glycerol

Flavor compounds are commonly obtained from chemical synthesis or extracted from plants. These sources have disadvantages, such as racemic mixture generation, more steps to yield the final product, low yield, and high cost, making the microbial fermentation an alternative and potential way to obtain flavor compounds. The most important lactone for flavor application is γ-decalactone, which has an aroma of peach and can be obtained by ricinoleic acid biotransformation through yeast peroxisomal β-oxidation. The aim of this work was to use crude glycerol, a residual biodiesel industry, for the production of bioaroma from two different yeasts. Yarrowia lipolytica CCMA 0357 and Lindnera saturnus CCMA 0243 were grown at different concentrations (10, 20, and 30% w/v) of substrates (castor oil and crude glycerol) for γ-decalactone production. L. saturnus CCMA 0243 produced higher concentration of y-decalactone (5.8?g/L) in crude glycerol, whereas Y. lipolytica CCMA 0357 showed a maximum production in castor oil (3.5?g/L). Crude glycerol showed better results for γ-decalactone production when compared to castor oil. L. saturnus CCMA 0243 has been shown to have a high potential for γ-decalactone production from crude glycerol.     

Castor phospholipid:diacylglycerol acyltransferase facilitates efficient metabolism of hydroxy fatty acids in transgenic Arabidopsis

Producing unusual fatty acids (FAs) in crop plants has been a long-standing goal of green chemistry. However, expression of the enzymes that catalyze the primary synthesis of these unusual FAs in transgenic plants typically results in low levels of the desired FA. For example, seed-specific expression of castor (Ricinus communis) fatty acid hydroxylase (RcFAH) in Arabidopsis (Arabidopsis thaliana) resulted in only 17% hydroxy fatty acids (HFAs) in the seed oil. In order to increase HFA levels, we investigated castor phospholipid:diacylglycerol acyltransferase (PDAT). We cloned cDNAs encoding three putative PDAT enzymes from a castor seed cDNA library and coexpressed them with RcFAH12. One isoform, RcPDAT1A, increased HFA levels to 27%. Analysis of HFA-triacylglycerol molecular species and regiochemistry, along with analysis of the HFA content of phosphatidylcholine, indicates that RcPDAT1A functions as a PDAT in vivo. Expression of RcFAH12 alone leads to a significant decrease in FA content of seeds. Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12 restored FA levels to nearly wild-type levels, and this was accompanied by a major increase in the mass of HFAs accumulating in the seeds. We show the usefulness of RcPDAT1A for engineering plants with high levels of HFAs and alleviating bottlenecks due to the production of unusual FAs in transgenic oilseeds.     

Design and development of low cost polyurethane biopolymer based on castor oil and glycerol for biomedical applications

In the current study, we present the synthesis of novel low cost bio‐polyurethane compositions with variable mechanical properties based on castor oil and glycerol for biomedical applications. A detailed investigation of the physicochemical properties of the polymer was carried out by using mechanical testing, ATR‐FTIR, and X‐ray photoelectron spectroscopy (XPS). Polymers were also tested in short term in‐vitro cell culture with human mesenchymal stem cells to evaluate their biocompatibility for potential applications as biomaterial. FTIR analysis confirmed the synthesis of castor oil and glycerol based PU polymers. FTIR also showed that the addition of glycerol as co‐polyol increases crosslinking within the polymer backbone hence enhancing the bulk mechanical properties of the polymer. XPS data showed that glycerol incorporation leads to an enrichment of oxidized organic species on the surface of the polymers. Preliminary investigation into in vitro biocompatibility showed that serum protein adsorption can be controlled by varying the glycerol content with polymer backbone. An alamar blue assay looking at the metabolic activity of the cells indicated that castor oil based PU and its variants containing glycerol are non‐toxic to the cells. This study opens an avenue for using low cost bio‐polyurethane based on castor oil and glycerol for biomedical applications.     

Effect of functional oils on the immune response of broilers challenged with Eimeria spp.

Infection with Eimeria sp. results in the activation of multiple facets of the host immune system; the use of phytogenics can modulate the inflammatory response and improve the performance of the challenged animal. The aim of this study was to evaluate the effect of a commercial blend of cashew nut shell liquid (CNSL) and castor oil on the immune response of broilers challenged with coccidiosis. A total of 864 one-day-old male chicks (Cobb 500) were randomly distributed into six treatment groups (8 pens/treatment and 18 chicks/pen) in a three-by-two factorial design with three additives: control (non-additive), 100 ppm of monensin or 0.15% CNSL–castor oil. Challenge status was determined twice at 14 days of age. Unchallenged birds were inoculated by gavage with oocysts sporulated with Eimeria tenella, Eimeria acervulina and Eimeria maxima. Although the positive control (non-additive and challenged) and CNSL–castor oil treatment groups exhibited similar variation in weight gain (ΔBWG) compared to unchallenged birds fed without additives, the variation observed in birds fed diets containing CNSL–castor oil was associated with a higher maintenance requirement and not feed efficiency. In the second week after infection, ΔBWG of the CNSL–castor oil treatment group did not significantly change compared to the other treatment groups. At days 7 and 14 post-challenge, there was a higher excretion of oocysts in the control group, whereas the CNSL–castor oil and monensin groups did not differ. The CNSL–castor oil group exhibited increased gene expression of interferon (IFN), interleukin 6 (IL-6) and tumor necrosis factor (TNF), while the control group exhibited increased expression of cyclooxygenase (COX) and IL-1. The heterophils/lymphocyte ratio was low for the monensin treatment group. The unchallenged birds that received monensin treatment presented higher gene expression of IFN, COX and IL-1 compared to the other treatments, while the CNSL–castor oil group exhibited reduced gene expression, except for TNF. The commercial blend of cashew nut liquid and castor oil modulated the inflammatory response against Eimeria spp. In the absence of the parasite, there was no stimulation of genes involved in the inflammatory response, demonstrating that the blend is an effective tool in specifically modulating the immune system of birds afflicted with coccidiosis.     

Phases dispersion and oxygen transfer in a simulated fermentation broth containing castor oil and proteins

The sizes of air bubbles and castor oil drops were studied by image analysis as a function of the concentration of soluble protein (bovine serum albumin [BSA] and lipase, as model proteins) in a three-phase system using a simulated fermentation medium (aqueous salt solution, castor oil, and air). Small amounts of proteins (<0.02 g/L) caused an important decrease in oil drops and bubbles sizes, together with a pronounced decrease in surface tension. The extent and profiles of this decrease seem to be determined by the conformation of the protein at the interface. The kLa value increased considerably for increasing concentration (up to 0.02 g/L) of the two proteins but was very different (2-fold higher for the lipase) at the highest concentrations tested (0.5 g/L), a phenomenon that can be caused by the extent to which bubbles are trapped within oil drops.     

Schistosoma mansoni: experimental chemoprophylaxis in mice using oral anti-penetration agents

The ability to prevent schistosomiasis by using an oral chemoprophylactic agent, which acts by preventing cercarial penetration, has been unexplored. We initially examined the effect of praziquantel (PZQ) as such an agent and found that it was moderately effective in blocking cercarial penetration, but that this effect was dependent on the vehicle used to administer the drug. ICR mice were given a total of 200 mg/kg PZQ per os over an 8-hr period in a divided dose of 50 mg/kg/2 hr. At time periods ranging from 0 to 92 hr after the last dose, mice were exposed to approximately 75 75Se-labeled cercariae via the tail. Twenty-four hours later, mice were sacrificed, their tails were removed and subjected to autoradiography, and the percentage of penetration was calculated. Cremophor El, 50% PEG 200, 50% propylene glycol, vegetable oil, and cod liver oil were used as PZQ vehicles. When Cremophor El (ethoxylated castor oil) was used to administer PZQ, a 93% reduction in cercarial penetration was seen at 0 hr and a 98%+ reduction rate was seen from 4 to 24 hr postexposure. However, Cremophor El alone had an essentially equivalent effect on cercarial penetration from 8 to 92 hr after administration. These unexpected results led us to investigate both castor oil and ricinoleic acid (castor oil is 87% ricinoleate as triglyceride) as oral anti-penetration agents. Mice were given the lipids orally by gavage for 7 days. On Day 8, each group of 12 mice was exposed to approximately 75 75Se-radiolabeled cercariae. Castor oil gave protection rate ranging from 90 to almost 100% at an optimal concentration of 0.3 ml/day x 3 or 7 days (approximately 9.8 g/kg/day). These observations suggest that chemoprophylaxis may be possible by dietary supplementation with lipids having anti-penetration activity or by molecules that resemble these lipids.     

Oil and air dispersion in a simulated fermentation broth as a function of mycelial morphology

The culture conditions of a multiphase fermentation involving morphologically complex mycelia were simulated in order to investigate the influence of mycelial morphology (Trichoderma harzianum) on castor oil and air dispersion. Measurements of oil drops and air bubbles were obtained using an image analysis system coupled to a mixing tank. Complex interactions of the phases involved could be clearly observed. The Sauter diameter and the size distributions of drops and bubbles were affected by the morphological type of biomass (pellets or dispersed mycelia) added to the system. Larger oil drop sizes were obtained with dispersed mycelia than with pellets, as a result of the high apparent viscosity of the broth, which caused a drop in the power drawn, reducing oil drop break-up. Unexpectedly, bubble sizes observed with dispersed mycelia were smaller than with pellets, a phenomenon which can be explained by the segregation occurring at high biomass concentrations with the dispersed mycelia. Very complex oil drops were produced, containing air bubbles and a high number of structures likely consisting of small water droplets. Bubble location was influenced by biomass morphology. The percentage (in volume) of oil-trapped bubbles increased (from 32 to 80%) as dispersed mycelia concentration increased. A practically constant (32%) percentage of oil-trapped bubbles was observed with pelleted morphology at all biomass concentrations. The results evidenced the high complexity of phases interactions and the importance of mycelial morphology in such processes.