Short-term Results of the Masking Effect of an Infiltrant Resin on Mild Molar Incisor Hypomineralization Lesions in Anterior Teeth.

This non-controlled clinical study evaluated the masking effect of an infiltrant resin on mild molar incisor hypomineralization (MIH) lesions. Thirty MIH-affected anterior teeth with creamy/ white opacities from 12 children aged 6-15 years received the application of an infiltrant resin (Icon- DMG). Standard photographs were taken before (T1), immediately after (T2), and 1 week after (T3) resin infiltration. Two calibrated examiners qualitatively analyzed the color match using the Fédération Dentaire Internationale (FDI) scale. The binomial distribution test analyzed the scores of the color match at T1 with T2 and T3, and McNemar's test analyzed the scores of the color match between T2 and T3 (α=0.05). There was a significant increase in color match between T1 and T2 (p=0.0005), between T1 and T3 (p=0.0005), and between T2 and T3 (p=0.0019). It was concluded that infiltrant resin was effective in improving the esthetic appearance of creamy/white opacities on MIH-affected anterior teeth.

Retention of Manually or CAD/CAM-customized Fiberglass Posts Luted to Enlarged Root Canals with Different Resin Cements.

The aim of this laboratory study was to evaluate the pull-out force of a prefabricated fiberglass post (PP), relined fiberglass post (RP), or milled fiberglass post (MP) luted with Multilink N (MN), RelyX Unicem 2 (RXU2) or RelyX Ultimate (RU) to enlarged root canals. The thickness of the resin cements and the presence of voids in the resin cement film were observed. The root canals of 90 bovine incisors were enlarged, endodontically treated, and randomly divided into 9 groups (n=10) according to the post type and resin cement. The specimens were scanned using micro-CT to analyze the thickness of the resin cement and the presence of voids. The specimens were submitted to mechanical cyclic loading (500,000 cycles at 50 N load) and subjected to pull-out force testing. Two-way ANOVA and Tukey's test analyzed the pull-out force and resin cement thickness data. Kruskal-Wallis and Bonferroni tests analyzed the void scores. The interaction between factors (post × resin cement) was significant (p=0.0001) for the pull-out force. Higher pull-out forces were obtained for RP and MP compared to PP. The post factor was significant (p=0.0001) for resin cement thickness, which was higher for PP (1054 µm), followed by MP (301 µm) and RP (194 µm). More void formation occurred for PP, being less for RP, differing significantly among the posts. Post customization (RP and MP) decreased resin cement thickness and void formation, favoring a higher pull-out force. Resin cements requiring an adhesive application (MN and RU) favored higher pull-out force than self-adhesive resin cement (RXU2).

Effect of pH Cycling Followed by Simulated Toothbrushing on the Surface Roughness and Bacterial Adhesion of Bulk-fill Composite Resins.

The aim was to evaluate, in vitro, quantitatively and qualitatively, the effect of pH cycling and simulated toothbrushing on surface roughness (Ra) and bacterial adhesion (Cn) of bulk-fill composite resins. Thirty specimens of each composite resin, 5 mm wide and 4 mm high, were obtained: group 1 (control): Filtek Z250 (Z250); group 2: Filtek Bulk-Fill (FTK); group 3: Tetric N-Ceram Bulk-Fill (TTC); and group 4: Aura Bulk-Fill (AUR). After 24 hours, the specimens were polished and then alternated with demineralization/remineralization solutions for 15 cycles of 24 hours each at 37°C. Then the specimens were submitted to simulated toothbrushing. The Ra and Cn measurements were quantitatively analyzed in three stages: after polishing (Ra0 and Cn0), after pH cycling (Ra1 and Cn1), and after simulated toothbrushing (Ra2 and Cn2). The Ra values were submitted to two-way analysis of variance, followed by the Tukey test (α=0.05). The Kruskal-Wallis test, followed by multiple comparisons, was applied for Cn analysis. Surface topography and bacterial adhesion were observed by scanning electron microscopy (SEM). Z250, FTK, and TTC showed no significant change in Ra regardless of the treatment performed; AUR obtained increased Ra at Ra2 (p<0.05). FTK differed from the others at Cn0 and Cn1 (p<0.05). At Cn2, there was no difference among the composite resins. SEM images showed the exposure of fillers and microcavities at Ra1 and Ra2. There was greater bacterial adhesion at Cn1 for Z250 and FTK. It was concluded that the pH cycling caused surface degradation of all composite resins, which was potentiated by simulated toothbrushing. However, only AUR presented an increased Ra. Bacterial adhesion occurred on all composite resins after pH cycling; however, after simulated toothbrushing, adhesion of dispersed bacteria was similar for all the composite resin groups.

Effect of Different Computer-aided Design/Computer-aided Manufacturing (CAD/CAM) Materials and Thicknesses on the Fracture Resistance of Occlusal Veneers.

The aim was to evaluate, in vitro, the influence of different computer-aided design/computer-aided manufacturing (CAD/CAM) materials (IPS e.max CAD, Vita Enamic, and Lava Ultimate) and thicknesses (0.6 mm and 1.5 mm) on the fracture resistance of occlusal veneers. Sixty human third molars were prepared to simulate advanced erosion of the occlusal surface, and the teeth were randomly divided into six experimental groups (n=10) according to the material and thickness used to build the veneers. Ten sound teeth formed the control group. The veneers were adhesively luted and submitted to mechanical cyclic loading (1 million cycles at 200-N load). The fracture resistance test was performed in a universal testing machine. The failures were classified as "reparable" and "irreparable." According to two-way analysis of variance and the Tukey test, the interaction (material × thickness) was significant ( p=0.013). The highest fracture resistance was obtained for IPS e.max CAD at a 1.5-mm thickness (4995 N) and was significantly higher compared to the other experimental groups ( p<0.05). The lowest fracture resistance was obtained for Vita Enamic at 0.6 mm (2973 N), although this resistance was not significantly different from those for IPS e.max CAD at 0.6 mm (3067 N), Lava Ultimate at 0.6 mm (3384 N), Vita Enamic at 1.5 mm (3540 N), and Lava Ultimate at 1.5 mm (3584 N) ( p>0.05). The experimental groups did not differ significantly from the sound teeth (3991 N) ( p>0.05). The failures were predominantly repairable. The occlusal veneers of IPS e.max CAD, Vita Enamic, and Lava Ultimate, with thicknesses of 0.6 mm and 1.5 mm, obtained fracture resistances similar to those associated with sound teeth.

Relined Fiberglass Post: Effect of Luting Length, Resin Cement, and Cyclic Loading on the Bond to Weakened Root Dentin.

This study evaluated the effects of luting length of the post, the resin cement, and cyclic loading on pull-out bond strength of fiberglass posts relined with composite resin in weakened roots. The canals of 80 bovine incisors were endodontically treated and weakened with diamond burs. The teeth were randomly divided into eight groups (n=10) according to the luting procedures of the relined fiberglass post (RFP): In groups 1, 2, 3, and 4, the RFPs were luted with RelyX ARC, and in groups 5, 6, 7, and 8 they were luted with RelyX U200. In groups 1, 3, 5, and 7, the RFPs were luted at a length of 5 mm, and in groups 2, 4, 6, and 8 they were luted at a length of 10 mm. Specimens from groups 3, 4, 7, and 8 were submitted to cyclic loading. Specimens were subjected to a pull-out bond strength test in a universal testing machine. The results (MPa) were analyzed by three-way analysis of variance and the Tukey post hoc test (α=0.05). Six human upper anterior teeth were used to analyze the bond interface by confocal laser scanning microscopy (CLSM). The pull-out bond strength of RFPs luted with RelyX U200 was statistically higher than that of RelyX ARC. Cyclic loading influenced the bond strength only for the luting length of 5 mm. CLSM analysis revealed the formation of resin cement tags for both materials. Luting length is an important factor in retaining RFPs in weakened roots when they are subjected to cyclic loading, and RelyX U200 resulted in greater bond strengths to the root canal in comparison with RelyX ARC.

Influence of immediate dentin sealing techniques on cuspal deflection and fracture resistance of teeth restored with composite resin inlays.

SUMMARY This research evaluated the influence of immediate dentin sealing (IDS) techniques on cuspal deflection and fracture resistance of teeth restored with composite resin inlays. Forty-eight maxillary premolars were divided into four groups: G1, sound teeth (control); G2, without IDS; G3, IDS with Clearfil SE Bond (CSE); and G4, IDS with CSE and Protect Liner F. The teeth from groups 2, 3, and 4 received mesio-distal-occlusal preparations. The impressions were made with vinyl polysiloxane, followed by provisional restoration and storage in water for seven days. The impressions were poured using type IV die stone, and inlays with Filtek Z250 composite resin were built over each cast. The inlays were luted with Panavia F. After storage in water for 72 hours, a 200-N load was applied on the occlusal surface using a metal sphere connected to a universal testing machine, and the cuspal deflection was measured with a micrometer. The specimens were then submitted to an axial load until failure. The following mean cuspal deflection (µm) and mean fracture resistance (N) followed by the same lowercase letter represent no statistical difference by analysis of variance and Tukey (p<0.05): cuspal deflection: G1, 3.1 ± 1.5(a); G2, 10.3 ± 4.6(b); G3, 5.5 ± 1.8(ac); and G4, 7.7 ± 5.1(bc); fracture resistance: G1, 1974 ± 708(a); G2, 1162 ± 474(b); G3, 700 ± 280(b); and G4, 810 ± 343(b). IDS with CSE allowed cuspal deflection comparable with that associated with sound teeth. The application of Protect Liner F did not contribute to a decrease in cuspal deflection. The IDS techniques did not influence the fracture resistance of teeth.

Up-dosing with bilastine results in improved effectiveness in cold contact urticaria.

BACKGROUND: Cold contact urticaria (CCU) is characterized by itchy wheal and flare responses due to the release of histamine and other pro-inflammatory mediators after exposure to cold. The treatment of choice is nonsedating antihistamines, dosages of which may be increased up to fourfold if standard doses are ineffective. Here, we assess the effects of a standard 20 mg dose and up-dosing to 40 and 80 mg of bilastine in reducing the symptoms of CCU and inflammatory mediator release following cold challenge. METHODS: Twenty patients with CCU were included in this randomized, crossover, double-blind, placebo-controlled 12-week study. They received placebo, 20, 40 or 80 mg of bilastine daily each for 7 days with 14-day washout periods. The primary readout was change in critical temperature thresholds (CTT). Secondary readouts were changes in pruritus, levels of histamine IL-6, IL-8 and TNF-α collected by skin microdialysis and safety and tolerability of bilastine. RESULTS: Bilastine 20 mg was highly effective (P < 0.0001) in reducing CTT. Up-dosing to 80 mg significantly (P < 0.04) increased its effectiveness. At this dose, 19 of 20 (95%) patients responded to treatment, with 12 of 20 (60%) becoming symptom free. Only one patient was refractory to treatment. Microdialysis levels of histamine, IL-6 and IL-8 assessed 1-3 h after cold challenge were significantly (P < 0.05) decreased following up-dosing with 80 mg bilastine. Bilastine treatment was well tolerated without evidence of increased sedation with dose escalation. CONCLUSIONS: Bilastine was effective in reducing the symptoms of patients with CCU. Increased efficacy of bilastine with fourfold up-dosing was without sedation and supports urticaria treatment guidelines.

In vitro marginal fit of three all-ceramic crown systems before and after cementation.

STATEMENT OF THE PROBLEM: Full-coverage all-ceramic restorations are widely used. The impact of various classifications of luting agent on marginal discrepancies is not well understood. PURPOSE: The purpose of this study was to evaluate the cervical fit of all ceramic crowns (IPS e.maxPress, Cergogold, and In Ceram) on bovine teeth with two luting agents before and after cementation. MATERIALS AND METHODS: Ninety bovine incisors were embedded in resin. The coronal portions of the teeth were prepared to receive full-coverage crowns. Thirty crowns of 7.0 ± 0.5 mm height, 8.0 mm cervical diameter, and 4.2 mm incisal diameter were fabricated for each ceramic system. The crowns were seated on the teeth, and the marginal discrepancy was measured using a measuring microscope. Then, 15 crowns of each ceramic system were luted on the teeth with resin cement (Variolink II) or resin-modified glass ionomer cement (Rely X luting), and the marginal discrepancy was measured. The results were submitted to analysis of variance, t test and Tukey's test (p<0.05). RESULTS: The three ceramic systems showed cervical fits after cementation statistically inferior to cervical fits before cementation for the two cements. The IPS e.maxPress showed values for cervical fit statistically superior to Cergogold before cementation. No statistically significant difference was found between IPS e.maxPress and In Ceram and In Ceram and Cergogold. After cementation, no statistically significant difference was found for the three ceramics systems when luted with resin or resin-modified glass ionomer luting agents. CONCLUSIONS: Within the limitations of this study, it can be concluded that both cements studied increase the marginal discrepancy between the crown and the preparation for the three ceramic systems evaluated.

Fracture resistance of root filled molar teeth restored with glass fibre bundles.

AIM: To evaluate the effect of unidirectional or woven glass fibre tapes inserted into MOD cavity preparations on the fracture resistance of root filled molar teeth. METHODOLOGY: Extracted human molar teeth were randomly divided into six groups (n = 15) : G1 - sound teeth, control; G2 - MOD cavity preparation; G3 - MOD + root canal treatment (Endo); G4 - MOD + Endo + composite resin restoration (Resin); G5 - MOD + Endo + unidirectional fibre (UF) + Resin; G6 - MOD + ;Endo + woven fibre (WF) + Resin. The teeth were subjected to a compressive fracture test in a universal testing machine. After testing, two failure modes were classified: pulp chamber floor or cusp. RESULTS: The highest and the lowest mean fracture strengths were found in sound teeth (G1) (4960N) and MOD + root canal treatment (G3) (612.84N), respectively, with significant differences from the other groups (P < 0.05). The remaining groups had statistically similar means. In G5 and G6, there was a tendency for fracture to occur in the pulp chamber floor compromising tooth integrity. CONCLUSIONS: The insertion of glass fibres into MOD cavity preparations and restoring them with composite resin was not different than molar teeth filled with composite resin only in terms of fracture resistance. Fibres placed into MOD cavities do not reinforce teeth.

The effect of surface treatments on the micro-shear bond strength of a resin luting agent and four all-ceramic systems.

OBJECTIVE: The current study evaluated the micro-shear bond strength between a resin luting agent and four strengthened all-ceramic systems under different surface treatments. METHODS: Rectangular specimens of IPS Empress 2 (Ivoclar-Vivadent), Cergogold (DeguDent), In Ceram Alumina (Vita) and Cercon (DeguDent) ceramics were fabricated and randomly divided into three groups: 1-no treatment; 2-etched with 9.5% hydrofluoric acid and 3-airborne-particle abraded with 50 microm aluminum oxide particles. The ceramic surfaces of the specimens were coated with a silane agent (Clearfil Porcelain Bond, Kuraray), then bonded with a resin-luting agent (Panavia F, Kuraray). A micro-shear bond test was carried out to measure the bond strength. Moreover, each ceramic surface was observed morphologically by scanning electron microscopy. The results were submitted to analysis of variance and Tukey's post-hoc analysis (p<0.05). RESULTS: The bond strength of all ceramic systems evaluated was affected by the surface treatments (p<0.05). The highest values for bond strength of IPS Empress 2 were found when the surface treatment used was hydrofluoric acid etching, followed by airborne particle abrasion treatment. On the other hand, airborne particle abrasion treatment and acid etching were not different for Cergogold and In Ceram Alumina ceramics, but they were higher when compared to the control (p<0.05). The highest bond strength to Cercon was found when it was treated with airborne particle abrasion with aluminum oxide. The SEM photographs showed that the hydrofluoric acid etching treatment affected the surface of IPS Empress 2 and Cergogold; however, Cercon and In Ceram surface morphology were not changed by the hydrofluoric acid etching. The airborne particle abrasion treatment altered the Cercon ceramic morphology but it did not change the other ceramic's surface.

Tensile bond strength of four adhesive systems to dentin.

PURPOSE: To assess, in vitro, the tensile bond strength to dentin of the following adhesive systems: Single Bond (3M), One Coat Bond (Coltène), Clearfil Liner Bond 2 V (Kuraray) and Etch & Prime 3.0 (Degussa). MATERIALS AND METHODS: Thirty third molars, extracted according to therapeutic indication, were used. The roots were removed at the cement-enamel junction level and the crowns were sectioned in a mesial-distal direction, obtaining buccal and lingual halves. The dental portions were included in self-cured acrylic resin, abraded on a lathe under water spray and polished to 400 and 600 grit. The adhesive systems were applied on dentin according to manufacturers' instructions and resin-based composite (Z100) cones were bonded. For each adhesive system, 15 specimens were prepared, stored in distilled water at 37 degrees C during 24 hrs and submitted to tensile bond strength tests on a universal testing machine (EMIC DL-2000) at a crosshead speed of 0.5 mm/min. RESULTS: The mean tensile bond strength to dentin (MPa +/- SD) were statistically different (ANOVA, Tukey's tests, P < or = 0.01, shown by asterisks) from each other: (1) Clearfil Liner Bond 2 V* = 24.8 +/- 3.3; (2) One Coat Bond** = 20.4 +/- 2.0; (3) Single Bond** = 18.1 +/- 2.4; (4) Etch & Prime 3.0*** = 5.8 +/- 2.4.

Effect of refrigeration on tensile bond strength of three adhesive systems.

The purpose of this study was to investigate the tensile bond strengths of three adhesive systems applied to dentin at refrigerated and room temperatures. Ninety bovine incisor teeth were obtained, embedded in self-cured acrylic resin, abraded on a lathe under water spray and polished to 400 and 600 grit to form standardized dentin surfaces before randomly assigning to six groups (n = 15). The adhesive systems Scotchbond Multi-Purpose, Single Bond and Prime & Bond NT were applied to dentin according to the manufacturers' instructions at refrigerated temperature (4 degrees C) and at room temperature (23 degrees C), before bonding resin-based composite (Z 100). The specimens were stored in distilled water at 37 degrees C for 24 hours and submitted to tensile bond strength tests on a universal testing machine (EMIC DL-2000) at a crosshead speed of 0.5 mm/min. The resulting data were statistically analyzed using analysis of variance and Tukey's test. No statistical differences were found when the adhesive systems were applied at refrigerated and room temperatures. Scotchbond Multi-Purpose and Single Bond had significantly stronger tensile bond strengths than Prime & Bond NT at room and refrigerated temperatures (p < 0.01). Scotchbond Multi-Purpose and Single Bond were statistically similar. No adverse effects upon tensile bond strength were observed when adhesive systems were taken directly from refrigerated storage.

alpha-Amylase production in high cell density submerged cultivation of Aspergillus oryzae and A. nidulans.

The effect of biomass concentration on the formation of Aspergillus oryzae alpha-amylase during submerged cultivation with A. oryzae and recombinant A. nidulans strains has been investigated. It was found that the specific rate of alpha-amylase formation in chemostats decreased significantly with increasing biomass concentration in the range of approx. 2-12 g dry weight kg(-1). When using a recombinant A. nidulans strain in which the gene responsible for carbon catabolite repression of the A. oryzae alpha-amylase gene (creA) was deleted, no significant decrease in the specific rate of alpha-amylase formation was observed. On the basis of the experimental results, it is suggested that the low value of the specific alpha-amylase productivity observed at high biomass concentration is caused by slow mixing of the concentrated feed solution in the viscous fermentation medium.

Role of substrate concentration in mitosis and hyphal extension of Aspergillus.

The filamentous fungi Aspergillus oryzae and A. niger grow by apical extension of multinucleate hyphae that are subdivided into compartments by cross-walls called septa. Submerged cultivation, image analysis, and fluorescence microscopy were used to study the role of the carbon source on mitosis and hyphal extension in these fungi. In the two species of Aspergillus, the length of the apical compartment, the number of nuclei in the apical compartment, and the hyphal diameter were regulated in response to the surrounding glucose concentration. A long apical compartment with many nuclei was the result of a high glucose concentration, whereas a short apical compartment with few nuclei was the result of a low glucose concentration. This is the first study of the influence of glucose concentration on nuclear mitosis and septation in filamentous fungi grown submerged. In addition, this is the first time a model of the duplication cycle in higher filamentous fungi has been simulated.

Use of breath hydrogen testing to detect experimentally induced disaccharide malabsorption in healthy adult dogs.

OBJECTIVE: To develop a noninvasive method to detect disaccharide malabsorption in dogs by measuring hydrogen concentration ([H2]) in exhaled breath before and after experimentally induced disaccharide malabsorption. ANIMALS: 8 healthy mixed-breed dogs. PROCEDURE: [H2] was measured every 30 minutes for 8 hours after administration of disaccharide solutions (lactose, 0.5 g/kg of body weight; lactose, 1.0 g/kg; sucrose, 2.0 g/kg; maltose, 1.5 g/kg; and lactose [0.5 g/kg] and sucrose [2.0 g/kg]) to determine reference ranges of [H2] for each solution, which were compared with [H2] in dogs with experimentally induced disaccharide malabsorption. To induce disaccharide malabsorption, dogs were given a mild overdose of lactose (1.5 g/kg) or a disaccharidase inhibitor. In the latter experiment, acarbose (10 mg/kg, PO) was given with the combination of lactose (0.5 g/kg) and sucrose (2 g/kg), and with maltose (1.5 g/kg). RESULTS: Overdosing with lactose resulted in [H2] persistently outside the reference range for lactose in 5 of 8 dogs. Acarbose administration resulted in [H2] persistently outside the reference range in 7 of 8 dogs that received a combination of sucrose and lactose but did not consistently affect [H2] after administration of maltose. CONCLUSIONS: Disaccharide malabsorption resulted in [H2] outside the reference ranges in most of the adult dogs studied, suggesting that the technique may be useful in detecting naturally occurring disaccharidase deficiency.

On-line study of growth kinetics of single hyphae of Aspergillus oryzae in a flow-through cell.

Using image analysis the growth kinetics of the single hyphae of the filamentous fungus Aspergillus oryzae has been determined on-line in a flow-through cell at different glucose concentrations in the range from 26 mg L-1 to 20 g L-1. The tip extension rate of the individual hyphae can be described with saturation type kinetics with respect to the length of the hyphae. The maximum tip extension rate is constant for all hyphae measured at the same glucose concentration, whereas the saturation constant for the hyphae varies significantly between the hyphae even within the same hyphal element. When apical branching occurs, it is observed that the tip extension rate decreases temporarily. The number of branches formed on a hypha is proportional to the length of the hypha that exceeds a certain minimum length required to support the growth of a new branch. The observed kinetics has been used to simulate the outgrowth of a hyphal element from a single spore using a Monte Carlo simulation technique. The simulations shows that the observed kinetics for the individual hyphae result in an experimentally verified growth pattern with exponential growth in both total hyphal length and number of tips.

Reproducibility of breath hydrogen concentration measurements in dogs after change of diet.

OBJECTIVE: To assess reproducibility of breath hydrogen (H2) concentration measurements in dogs on 3 consecutive days after introduction of a new diet. ANIMALS: 10 healthy colony-source dogs. Procedure Dogs were fed a commercial pet food for a minimum of 10 days before introduction of a high-carbohydrate commercial diet. Expired breath samples were collected at baseline and every 90 minutes for 18 hours on the first 3 days after introduction of the new diet. The area under the curve (AUC) for breath H2 concentration versus time on days 1, 2, and 3 were compared by use of repeated-measures ANOVA. RESULTS: The (AUC) did not differ significantly among test days 1, 2, and 3. CONCLUSIONS: Expired breath H2 concentration in dogs indicated good reproducibility during the first 3 days after introduction of a new diet. CLINICAL RELEVANCE: It is not necessary to provide a period for adaptation to a test diet prior to performing breath H2 tests in dogs.

Growth and product formation of Aspergillus oryzae during submerged cultivations: verification of a morphologically structured model using fluorescent probes.

A morphologically structured model is well suited for obtaining a good description of growth and product formation of filamentous fungi for use in a process model, for example. This article describes a new morphologically structured model and its application to an alpha-amylase producing strain of Aspergillus oryzae. The model is based on a division of the fungal hyphae into three different regions: an extension zone, representing the tips of the hyphae; an active region, which is responsible for growth and product formation; and an inactive hyphal region. Two metamorphosis reactions describing branching and inactivation are included in the model, and the kinetics of branching and tip extension are based on known experimentally verified models of fungal microscopic morphology. To verify the structure of the model a double-staining method, based on a combination of fluorescence microscopy and automated image analysis, has been developed for measuring the fraction of active cells. The method employs the fluorescent dye 3, 3'-dihexyloxocarbocyanin to stain organelles inside the hyphae and Calcoflour White to stain the cell wall. The ratio between the projected areas of the organelles and of the entire hyphal element is then taken to be proportional to the fraction of active cells. When applied to chemostat and fed-batch experiments, the double-staining method seemed to confirm the basic morphological structure of the model. The model is able to produce accurate simulations of steady-state and transient conditions in chemostats, of batch cultivations, and even the formation of a single hyphal element from a spore, all with the same values of the model parameters.

On-line study of fungal morphology during submerged growth in a small flow-through cell.

A flow-through cell is designed to measure the growth kinetics of hyphae of Aspergillus oryzae grown submerged in a well controlled environment. The different stages of the growth process are characterized, from the spore to the fully developed hyphal element with up to 60 branches and a total length lt up to 10,000 micrometer. Spore swelling is found to occur without change in the form of the spore (circularity index constant at about 1.06) and the spore volume probably increases exponentially. The germ tube appears after about 4 h. The branching frequency and the rate of germ tube extension is determined. After about 10 h growth at a glucose concentration of 250 mg L-1, 6-7 branches have been set, and both the total hyphal length lt and the number of tips increase exponentially with time. The specific growth rate of the hyphae is 0. 33 h-1 while the average rate of the extension of the growing tips approaches 55 micrometer h-1. The growth kinetics for all the branches on the main hypha have also been found. The main hypha and all the branches grow at a rate which can be modeled by saturation kinetics with respect to the branch length and with nearly equal final tip speeds (160 micrometer h-1). Branches set near the apical tip of the main hypha attain their final tip speed in the shortest time, i.e., the value of the saturation parameter is small. Finally, the influence of substrate (glucose) concentration cs on the values of the morphological parameters has been determined. It is found that saturation type kinetics can be used to describe the influence of cs on the growth. Experiments with recirculation of effluent from the cell back to the inlet strongly suggest that the fungus secretes an inducer for growth and branching.

Morphology and physiology of an alpha-amylase producing strain of Aspergillus oryzae during batch cultivations.

The microscopic morphology, that is, total hyphal length and total number of tips, has been characterized during batch cultivations of Aspergillus oryzae. The specific growth rate estimated by measuring the total hyphal length (mu(h)) corresponds well with the specific growth rate estimated from dry weight measurements during cultures grown as free hyphal elements. The average tip extension rate can be described with a saturation type kinetics with respect to the average total hyphal length, and the branching frequency is closely related to the total hyphal length. For the applied strain of A. oryzae, pellet formation occurs by coagulation of spores. The agglomeration process is pH dependent and pellets are formed at pH values higher than 5, whereas low pH (<3.5) results in growth as freely dispersed hyphal elements. The maximum specific growth rate has a broad pH optimum between 3 and 7, whereas the alpha-amylase production has a sharper maximum at about pH 6. During batch cultivation with pellets the growth is described well by the cube-root law when pellet fragmentation can be neglected. The kinetic parameter k in the cube-root law is derived from the growth kinetics with no mass transfer limitation, k = mu(h)/3. Based on an oxygen balance, the active growth layer in the pellet is estimated to be 200 to 325 mum and, consequently, up to 50% of the biomass is limited by oxygen for large pellets. Ethanol production (up to 1 g L(-1)) was observed during batch cultivations with pellets, suggesting that ethanol is produced in the oxygen limited part of the biomass. A constitutive, low alpha-amylase production was observed at high glucose concentration. The specific alpha-amylase production was significantly higher for filamentous growth than for pellets and oxygen appears to be necessary for production of alpha-amylase. (c) 1996 John Wiley & Sons, Inc.