The effect of recombinant human granulocyte/macrophage-colony-stimulating factor (rHu GM-CSF) and rHu G-CSF administration on neutrophil chemiluminescence assay in patients following cyclic chemotherapy

Secondary infections related to neutropenia and functional defects of phagocytes are common consequences in patients treated for cancer. The hematopoietic colony-stimulating factors (CSF) have been introduced into clinical practice as additional supportive measures that can reduce the incidence of infectious complications in patients with cancer and neutropenia. The aim of this study was to determine the role of␣granuolcyte/macrophage(GM)-CSF and granulocyte(G)-CSF in enhancing in vivo human neutrophil function. A luminol-dependent chemiluminescence assay was developed to evaluate whether the repair in neutropenia accompanies the ability of neutrophils to function. A dose of 5 μg G-CSF kg⁻¹ day⁻¹ [recombinant human (rHu) G-CSF; filgrastim] or 250 μg GM-CSF m⁻² day⁻¹ (rHu GM-CSF; molgramostim) was administered subcutaneously once daily to 12 metastatic cancer patients being treated with different cytotoxic regimens. All injections of CSF were given after the initiation of neutropenia and continued until the occurrence of an absolute neutrophil recovery. rHu GM-CSF and rHu G-CSF, administered once daily at the 250 μg m⁻² day⁻¹ and 5 μg kg⁻¹ day⁻¹ level, were effective in increasing the absolute neutrophil count and neutrophil function, as measured by an automated chemiluminescence system. Received: 26 February 1998 / Accepted: 21 May 1998     

Reclamation of an activated-sludge microbial consortium by selective biostimulation

Our previous study showed that an activated-sludge process broke down at the phenol-loading rate of 1.5 g l⁻¹ day⁻¹, when non-flocculating bacteria (called R6T and R10) overgrew the sludge, resulting in a sludge washout. In this study, we attempted to circumvent this breakdown problem by reclaiming the consortium structure. Activated sludge was fed phenol, and the phenol-loading rate was increased stepwise from 0.5 g l⁻¹ day⁻¹ to 1.0 g l⁻¹ day⁻¹ and then to 1.5 g l⁻¹ day⁻¹. Either galactose or glucose (at 0.5 g l⁻¹ day⁻¹) was also supplied to the activated sludge from the phenol-loading rate of 1.0 g l⁻¹ day⁻¹. Pure culture experiments have suggested galactose to be a preferential substrate for a floc-forming bacterium (R6F) that predominantly degrades phenol under low phenol-loading conditions. Supplying galactose allowed sustainment of the R6F population and suppression of the overgrowth of R6T and R10 at the phenol-loading rate of 1.5 g l⁻¹ day⁻¹. This measure allowed the activated-sludge process to treat phenol at a phenol-loading rate up to 1.5 g l⁻¹ day⁻¹, although it broke down at 2.0 g l⁻¹ day⁻¹. In contrast, supplying glucose reduced the R6F population and allowed the activated-sludge process to break down at the phenol-loading rate of 1.0 g l⁻¹ day⁻¹. This study demonstrated that reclamation of the activated-sludge consortium by selective biostimulation of the floc-forming population improved the phenol-treating ability of the process. Received: 13 January 2000 / Received revision: 10 March 2000 / Accepted: 7 April 2000     

Thermophilic acidification of dairy wastewater

Acidification of simulated dairy wastewater was conducted in an upflow reactor at 55 °C. Results showed that the degree of acidification decreased with the increase in chemical oxygen demand (COD) loading rate, from 60.8% at 4 g l⁻¹ day⁻¹ to 27.1% at 24 g l⁻¹ day⁻¹. Carbohydrate was readily degraded at all loading rates, but degradation of protein and lipid decreased with the increase in loading rate. Most carbohydrate degradation occurred at the reactor bottom, whereas protein was degraded mainly after the carbohydrate became depleted. The predominant acidification products were acetate, propionate, butyrate and ethanol, whereas formate, i-butyrate, valerate, i-valerate, caproate, lactate, methanol, propanol and butanol were present in lesser quantities. The increase in loading rate resulted in the increase of propionate and the decrease of acetate, but had little effect on ethanol and butyrate productions. Only 2.5–8.8% of influent COD was converted to hydrogen and methane. The biomass yield was 0.30–0.43 mg VSS mg⁻¹ COD. Received: 8 December 1999 / Received revision: 14 February 2000 / Accepted: 25 February 2000     

Plant secondary metabolites as mammalian feeding deterrents: separating the effects of the taste of salicin from its post-ingestive consequences in the common brushtail possum (Trichosurus vulpecula)

The effect of the phenolic glycoside, salicin, on food intake of the common brushtail possum (Trichosurus vulpecula) was studied in a series of feeding experiments. Increasing the concentration of salicin in a diet of fruits and cereals led to significant reductions of food intake in the short term (6 days). After prolonged (20 days) exposure to salicin, food intake (19 g kg^−0.75 day⁻¹) was still reduced relative to controls (31 g kg^−0.75 day⁻¹) but not reduced to the same extent as in the short-term experiments. Nonetheless, over these 20 days, common brushtail possums regulated their intake of salicin so as not to exceed a threshold limit of 1.9 ± 0.1 g kg^−0.75 day⁻¹. Manipulative experiments sought to determine whether this threshold intake was in response to pre-ingestive factors (taste) or the post-ingestive consequences of ingesting salicin. Dietary salicin (0.17–5.0% DM) had no significant effect on nitrogen balance or urea metabolism and injection of a specific serotonin receptor antagonist, ondansetron, did not lead to increases in salicin intake as has been found for some other plant secondary metabolites. Similarly, administration of 1.3 g salicin by gavage had no significant effect on the subsequent intake of salicin compared to controls that were gavaged with water. We concluded that pre-ingestive factors were responsible for common brushtail possums limiting their intake of salicin-rich diets rather than any measurable post-ingestive consequence of feeding. Accepted: 7 December 1999     

Effects of 3 days of carbohydrate supplementation on muscle glycogen content and utilisation during a 1-h cycling performance

This study compared the effects of supplementing the normal diets of six trained cyclists [maximal oxygen uptake O_2max) 4.5 (0.36)l · min⁻¹; values are mean (SD)] with additional carbohydrate (CHO) on muscle glycogen utilisation during a 1-h cycle time-trial (TT). Using a randomised crossover design, subjects consumed either their normal diet (NORM) for 3 days, which consisted of 426 (137) g · day⁻¹ CHO [5.9 (1.4) g · kg⁻¹ body mass (BM)], or additional CHO (SUPP) to increase their intake to 661 (76) g · day⁻¹ [9.3 (0.7) g · kg⁻¹ BM]. The SUPP diet elevated muscle glycogen content from 459 (83) to 565 (62) mmol · kg⁻¹ dry weight (d.w.) (P < 0.05). However, despite the increased pre-exercise muscle glycogen stores, there was no difference in the distance cycled during the TT [40.41 (1.44) vs 40.18 (1.76) km for NORM and SUPP, respectively]. With NORM, muscle glycogen declined from 459 (83) to 175 (64) mmol · kg⁻¹ d.w., whereas with SUPP the corresponding values were 565 (62) and 292 (113) mmol · kg⁻¹ d.w. Accordingly, both muscle glycogen utilisation [277 (64) vs 273 (114) mmol · kg⁻¹ d.w.] and total CHO oxidation [169 (20) vs 165 (30) g · h⁻¹ for NORM and SUPP, respectively] were similar. Neither were there any differences in plasma glucose or lactate concentrations during the two experimental trials. Plasma glucose concentration averaged 5.5 (0.5) and 5.6 (0.6) mmol · l⁻¹, while plasma lactate concentration averaged 4.4 (1.9) and 4.4 (2.3) mmol · l⁻¹ for NORM and SUPP, respectively. The results of this study show that when well-trained subjects increase the CHO content of their diet for 3 days from 6 to 9 g · kg⁻¹ BM there is only a modest increase in muscle glycogen content. Since supplementary CHO did not improve TT performance, we conclude that additional CHO provides no benefit to performance for athletes who compete in intense, continuous events lasting 1 h. Furthermore, the substantial muscle CHO reserves observed at the termination of exercise indicate that whole-muscle glycogen depletion does not determine fatigue at this exercise intensity and duration. Accepted: 25 November 1996     

Comparative analysis of the digestive efficiency and nitrogen and energy requirements of the phyllostomid fruit-bat (Artibeus jamaicensis) and the pteropodid fruit-bat (Rousettus aegyptiacus)

Nitrogen (N) and energy (E) requirements of the phyllostomid fruit bat, Artibeus jamaicensis, and the pteropodid fruit bat Rousettus aegyptiacus, were measured in adults that were fed on four experimental diets. Mean daily food intake by A. jamaicensis and R. aegyptiacus ranged from 1.1–1.6 times body mass and 0.8–1.0 times body mass, respectively. Dry matter digestibility and metabolizable E coefficient were high (81.1% and 82.4%, respectively) for A. jamaicensis and (77.5% and 78.0%, respectively) for R. aegyptiacus. Across the four diets, bats maintained constant body mass with mean metabolizable E intakes ranging from 1357.3 kJ · kg^−0.75 · day⁻¹ to 1767.3 kJ · kg^−0.75 · day⁻¹ for A. jamaicensis and 1282.6–1545.2 kJ · kg^−0.75 · day⁻¹ for R. aegyptiacus. Maintenance E costs were high, in the order of 3.6–5.4 times the basal metabolic rate (BMR). It is unlikely that the E intakes that we observed represent a true measure of maintenance E requirements. All evidence seems to indicate that fruit bats are E maximizers, ingesting more E than required and regulating storage by adjusting metabolic output. We suggest that true maintenance E requirements are substantially lower than what we observed. If it follows the eutherian norm of two times the BMR, fruit bats must necessarily over-ingest E on low-N fruit diet. Dietary E content did affect N metabolism of A. jamaicensis. On respective low- and high-E diets, metabolic fecal N were 0.492 mg N · g⁻¹ and 0.756 mg N · g⁻¹ dry matter intake and endogenous urinary N losses were 163.31 mg N · kg^−0.75 · day⁻¹ and 71.54 mg N · kg^−0.75 · day⁻¹. A. jamaicensis required 332.3 mg · kg^−0.75 · day⁻¹ and 885.3 mg · kg^−0.75 · day⁻¹ of total N on high- and low-E diets, respectively, and 213.7 mg · kg^−0.75 · day⁻¹ of truly digestible N to achieve N balance. True N digestibilities were low (29% and 49%) for low- and high-E diets, respectively. For R. aegyptiacus, metabolic fecal N and endogenous urinary N losses were 1.27 mg N · g⁻¹ dry matter intake and 96.0 mg N · kg^−0.75 · day⁻¹, respectively, and bats required 529.8 mg · kg^−0.75 · day⁻¹ (total N) or 284.0 mg · kg^−0.75 · day⁻¹ (truly digestible N). True N digestibility was relatively low (50%). Based on direct comparison, we found no evidence that R. aegyptiacus exhibits a greater degree of specialization in digestive function and N retention than A. jamaicensis. When combined with results from previous studies, our results indicate that all fruit bats appear to be specialized in their ability to retain N when faced with low N diet. Accepted: 24 November 1998     

Effect of ammonia on the anaerobic degradation of protein by a mesophilic and thermophilic biowaste population

The influence of ammonia on the anaerobic degradation of peptone by mesophilic and thermophilic populations of biowaste was investigated. For peptone concentrations from 5 g l⁻¹ to 20 g l⁻¹ the mesophilic population revealed a higher rate of deamination than the thermophilic population, e.g. 552 mg l⁻¹ day⁻¹ compared to 320 mg l⁻¹ day⁻¹ at 10 g l⁻¹ peptone. The final degree of deamination of the thermophilic population was, however, higher: 102 compared to 87 mg NH₃/g peptone in the mesophilic cultures. If 0.5–6.5 g l⁻¹ ammonia was added to the mesophilic biowaste cultures, deamination of peptone, degradation of its chemical oxygen demand (COD) and formation of biogas were increasingly inhibited, but no hydrogen was formed. The thermophilic biowaste cultures were most active if around 1 g ammonia l⁻¹ was present. Deamination, COD degradation and biogas production decreased at lower and higher ammonia concentrations and hydrogen was formed in addition to methane. Studies of the inhibition by ammonia of peptone deamination, COD degradation and methane formation revealed a K _i (50%) for NH₃ of 92, 95 and 88 mg l⁻¹ at 37 °C and 251, 274 and 297 mg l⁻¹ at 55 °C respectively. This indicated that the thermophilic flora tolerated significantly more NH₃ than the mesophilic flora. In the mesophilic reactor effluent 4.6 × 10⁸ peptone-degrading colony-forming units (cfu)/ml were culturable, whereas in the thermophilic reactor effluent growth of only 5.6 × 10⁷ cfu/ml was observed. Received: 24 April 1998 / Received revision: 26 June 1998 / Accepted: 27 June 1998     

Production of Spirulina sp. in sea water supplemented with anaerobic effluents in outdoor raceways under temperate climatic conditions

The use of untreated sea water supplemented with anaerobic effluents from digested pig waste and sodium bicarbonate was evaluated as a low-cost medium for semi-continuous cultivation of a mixed culture of two Spirulina strains in outdoor raceways under temperate climatic conditions (pond temperature in the range 21–26 °C and light intensity in the range 225–957␣μE m⁻² s⁻¹). The mixed culture had a predominant population (86.6 ± 3.9%) of an atypical Spirulina strain consisting of straight filaments, which appeared spontaneously after the strain with helicoidal trichomes had been subcultured. Morphological studies for the identification of the type and size of trichomes of the two strains (HF and SF) were carried out. The proportions of the two strains were observed to be stable during the monitoring period (30 days). Three different sets of semicontinuous cultures were carried out. Sets 1 and 2 were operated under regime 1 (a single addition of anaerobic effluents at time zero and no pH control) during the same season (June and July) of different years. Set 3 was operated under regime 2 (semi-continuous addition of anaerobic effluents and pH control) during the autumn. A minimum productivity of 3.6 g m⁻² day⁻¹ was obtained at one of the lowest temperatures (22.1 °C) and light intensities (245 μE m⁻² s⁻¹) and a maximum productivity of 10.9 g m⁻² day⁻¹ was observed at the highest temperature (25 °C) and highest average light intensity (618 μE m⁻² s⁻¹) registered for sets 1 and 2. The protein content in the Spirulina biomass harvested from these two sets varied from 17% to 65.6%. In set 3, a maximum productivity of 9.0 g m⁻² day⁻¹ was recorded at an average temperature of 24.4 °C and at an average light intensity of 668 μE m⁻² s⁻¹. The protein content in this set under regime 2 varied within a narrower range than in set 1 and set 2 (from 34.8% to 49.1%), apparently because of a continuous availability of ammonia nitrogen at a level of 30–50 mg l⁻¹. However, in terms of the removal of ammonia nitrogen and chemical oxygen demand, regime 1 was more efficient than regime␣2. Received: 3 September 1996 / Received revision: 19 February 1997 / Accepted: 7 March 1997     

Hydrodynamic aspects and Arthrospira growth in two outdoor tubular undulating row photobioreactors

Two tubular undulating row photobioreactors (TURPs) with a very high illuminated surface/volume ratio (400 m⁻¹) were designed and constructed for the growth of photosynthetic micro-organisms. Experiments were conducted under outdoor conditions; and Arthrospira recycling was performed with airlifts (one for each row). The rows in each reactor faced east-west and consisted of a flexible polyvinyl chloride pipe (22 m long, 0.01 m bore) arranged in a sinusoidal shape. We studied the hydraulic performance of the sine-shaped photobioreactor rows during culture recycling in the TURPs at a very high Reynolds number (4200), when Arthrospira showed Newtonian fluid behavior. The sinusoidal pipe arrangement imposed a sine waveform on the culture, which led to better light utilization. During summer, a volumetric productivity of 2.2 g l⁻¹ day⁻¹ was reached in the TURP-5r (5 rows m⁻²), whereas an area productivity of 35 g m⁻² day⁻¹ was obtained in the TURP-10r (10 rows m⁻²). This was due to more light being available in the TURP-5r, because its rows were more spaced out and the photic ratio (R _f) was low (3.0). In the TURP-10r, the closer rows caused a dilution of the sunlight, but gave a better light distribution inside the Arthrospira culture and improved the light utilization. This was attributed to the high R _f (6.0) of this reactor. Received: 8 October 1999 / Received revision: 20 January 2000 / Accepted: 23 January 2000     

Simultaneous bioconversion of glucose and xylose to ethanol by Saccharomyces cerevisiae in the presence of xylose isomerase

Simultaneous isomerisation and fermentation (SIF) of xylose and simultaneous isomerisation and cofermentation (SICF) of a glucose/xylose mixture was carried out by Saccharomyces cerevisiae in the presence of xylose isomerase. The SIF of 50 g l⁻¹ xylose gave an ethanol concentration and metabolic yield of 7.5 g l⁻¹ and 0.36 g (g xylose consumed)⁻¹. These parameters improved to 13.4 g l⁻¹ and 0.40 respectively, when borate was added to the medium. The SICF of a mixture of 50 g l⁻¹ glucose and 50 g l⁻¹ xylose gave an ethanol concentration and metabolic yield of 29.8 g l⁻¹ and 0.42 respectively, in the presence of borate. Temperature modulation from 30 °C to 35 °C during fermentation further enhanced the above parameters to 39 g l⁻¹ and 0.45 respectively. The approach was extended to the bioconversion of sugars present in a real lignocellulose hydrolysate (peanut-shell hydrolysate) to ethanol, with a fairly good yield. Received: 14 May 1999 / Received revision: 27 September 1999 / Accepted: 2 October 1999     

Repeated batch cultivation of the microalga Spirulina platensis

Summary The cultivation of photosynthetic microorganisms such as the microalga Spirulina platensis can provide an alternative source of food. The water in Mangueira Lagoon (Rio Grande do Sul state, southern Brazil) has several required nutrients for the growth of Spirulina and could be added to culture medium to reduce the cost of producing S. platensis. Although little studied, repeated batch cultivation is a very useful technique because it has a better cost–benefit ratio than other cultivation methods. In a series of runs, we studied the influence of cell concentration, renewal rate and strain on the specific growth rate and biomass productivity of S. platensis during repeated batch cultivation, the runs taking place in 2-l Erlenmeyer flasks for 2160 h at 30 °C and a light intensity of 2500 lux under a 12 h photoperiod. The three factors studied had a significant (P < 0.05) effect on the results (specific growth rate and productivity). Using Zarrouk’s medium, the highest specific growth rate (μ_X) was 0.111 day⁻¹ while the biomass productivity (P _X) was 0.0423 g l⁻¹ day⁻¹, while Mangueira Lagoon water supplemented with 10% Zarrouk’s medium gave μ_X = 0.113 day⁻¹ and a productivity P _X = 0.0467 g l⁻¹ day⁻¹. These values were two to three times higher than the results obtained in batch cultivation, indicating that the repeated batch cultivation of S. platensis is attractive and convenient.     

Batch and continuous cultivation of Anaerobiospirillum succiniciproducens for the production of succinic acid from whey

Batch and continuous cultivation of Anaerobiospirillum succiniciproducens were systematically studied for the production of succinic acid from whey. Addition of 2.5 g l⁻¹ yeast extract and 2.5 g l⁻¹ polypeptone per 10 g l⁻¹ whey was most effective for succinic acid production from both treated and nontreated whey. When 20 g l⁻¹ nontreated whey and 7 g l⁻¹ glucose were used as cosubstrates, the yield and productivity of succinic acid reached at the end of fermentation were 95% and 0.46 g (l h)⁻¹, respectively. These values were higher than those obtained using nontreated whey alone [93% and 0.24 g (l h)⁻¹ for 20 g l⁻¹ whey]. Continuous fermentation of A. succiniciproducens at an optimal dilution rate resulted in the production of succinic acid with high productivity [1.35 g (l h)⁻¹], high conversion yield (93%), and higher ratio of succinic acid to acetic acid (5.1:1) from nontreated whey. Received: 23 July 1999 / Received revision: 17 November 1999 / Accepted: 24 December 1999     

Maintenance nitrogen requirement of an obligate nectarivore, the honey possum, Tarsipes rostratus

A nitrogen balance feeding trial was carried out with the marsupial honey possum, Tarsipes rostratus, using four pollen-honey diets varying in nitrogen content from 9.4 mg · g⁻¹ to 2.3 mg · g⁻¹ dry matter. The dietary maintenance nitrogen requirement (MNR) was determined by regression analysis as 89 ± 21 mg N · kg^−0.75 · day⁻¹ and the truly digestible MNR was 79 mg N · kg^−0.75 · day^−1.. Regressing nitrogen balance on daily nitrogen intake separately for ten males and seven females revealed that the slopes of the fitted lines did not vary significantly, but the difference in the intercepts approached significance. This suggests that the MNR for females may be lower than that of males. The nitrogen digestibility of the diet was 76% and the biological value (BV) was 58%. A comparison of the MNR of the honey possum with that of other marsupials shows that it is indeed much lower than that of herbivorous macropodid marsupials and is close to that of the sap- and gum-feeding sugar glider, Petarurus breviceps. The endogenous urinary nitrogen excretion (EUN) of the honey possum was 42 mg N · kg^−0.75 · day⁻¹ and a regression analysis with other published data showed that the EUN per unit basal heat production is significantly lower than that of eutherian mammals. Measurements of the actual feeding rates of animals in the field, taken together with the low MNR, do not lend support to the hypothesis that the honey possum exhibits a reduced rate of reproduction due to a deficiency in dietary nitrogen. It is possible that the quality of nitrogen provided by pollen, as reflected in its composition of essential amino acids, may be a limiting factor. Accepted: 15 September 2000     

Energetics during nursing and early postweaning fasting in hooded seal (Cystophora cristata ) pups from the Gulf of St Lawrence, Canada

In this study we measured growth and milk intake and calculated energy intake and its allocation into metabolism and stored tissue for hooded seal (Cystophora cristata) pups. In addition, we measured mass loss, change in body composition and metabolic rate during the first days of the postweaning fast. The mean body mass of the hooded seal pups (n = 5) at the start of the experiments, when they were new-born, was 24.3 ± 1.3 kg (SD). They gained an average of 5.9 ± 1.1. kg · day⁻¹ of which 19% was water, 76% fat and 5% protein. This corresponds to an average daily energy deposition of 179.8 ± 16.0 MJ. The pups were weaned at an average body mass of 42.5 ± 1.0 kg 3.1 days after the experiment was initiated. During the first days of the postweaning fast the pups lost an average of 1.3 ± 0.5␣kg of body mass daily, of which 56% was water, 16% fat and 28% protein. During the nursing period the average daily water influx for the pups was 124.6 ± 25.8 ml · kg⁻¹. The average CO₂ production during this period was 1.10 ± 0.20 ml · g⁻¹ · h⁻¹, which corresponds to a field metabolic rate of 714 ± 130 kJ ·  kg⁻¹ · day⁻¹, or 5.8 ± 1.1 times the predicted basal metabolic rate according to Kleiber (1975). During the postweaning fast the average daily water influx was reduced to 16.1 ± 6.6 ml · kg⁻¹. The average CO₂ production in␣this period was 0.58 ± 0.17 ml · g⁻¹ · h⁻¹ which corresponds to a field metabolic rate of 375 ± 108 kJ · kg⁻¹ · day⁻¹ or 3.2 ± 0.9 times the predicted basal metabolic rate. Average values for milk composition were 33.5% water, 58.6% fat and 6.2% protein. The pups drank an average of 10.4 ± 1.8␣kg of milk daily, which represents an energy intake of 248.9 ± 39.1 MJ · day⁻¹. The pups were able to store 73.2 ± 7.7% of this energy as body tissue. Accepted: 15 August 1996     

Inhibition of Clostridium butyricum by 1,3-propanediol and diols during glycerol fermentation

1,3-Propanediol inhibition during glycerol fermentation to 1,3-propanediol by Clostridium butyricum CNCM 1211 has been studied. The initial concentration of the 1,3-propanediol affected the growth of the bacterium more than the glycerol fermentation. μ _max was inversely proportional to the initial concentration of 1,3-propanediol (0–65 g l⁻¹). For glycerol at 20 g l⁻¹, the growth and fermentation were completely stopped at an initial 1,3-propanediol concentration of 65 g l⁻¹. However, for an initial 1,3-propanediol concentration of 50 g l⁻¹ and glycerol at 70 g l⁻¹, the final concentration (initial and produced) of 1,3-propanediol reached 83.7 g l⁻¹(1.1 M), with complete consumption of the glycerol. Therefore, during the fermentation, the strain tolerated a 1,3-propanediol concentration higher than the initial inhibitory concentration (65 g l⁻¹). The addition of 1,2-propanediol or 2,3-butanediol (50 g l⁻¹) in the presence of glycerol (50–100 g l⁻¹), showed that 2-diols reduced the μ _max in a similar way to 1,3-propanediol. The measurement of the osmotic pressure of glycerol solutions, diols and diol/glycerol mixtures did not indicate any differences between these compounds. The hypothesis of diol inhibition was discussed. Taking into account the strain tolerance of highly concentrated 1,3-propanediol during fermentation, the fermentation processes for optimising production were considered. Received: 15 November 1999 / Revision received: 1 February 2000 / Accepted: 4 February 2000     

Production of xylitol by Candida peltata

Candida peltata NRRL Y-6888 to ferment xylose to xylitol was evaluated under different fermentation conditions such as pH, temperature, aeration, substrate concentration and in the presence of glucose, arabinose, ethanol, methanol and organic acids. Maximum xylitol yield of 0.56 g g⁻¹ xylose was obtained when the yeast was cultivated at pH 6.0, 28°C and 200 rpm on 50 g L⁻¹ xylose. The yeast produced ethanol (0.41 g g⁻¹ in 40 h) from glucose (50 g L⁻¹) and arabitol (0.55 g g⁻¹ in 87 h) from arabinose (50 g L⁻¹). It preferentially utilized glucose > xylose > arabinose from mixed substrates. Glucose (10 g L⁻¹), ethanol (7.5 g L⁻¹) and acetate (5 g L⁻¹) inhibited xylitol production by 61, 84 and 68%, respectively. Arabinose (10 g L⁻¹) had no inhibitory effect on xylitol production. Received 24 December 1998/ Accepted in revised form 18 March 1999     

Treatment of cyanide-containing wastewater from the food industry in a laboratory-scale fixed-bed methanogenic reactor

During the process of producing cassava starch from Manihot esculenta roots, large amounts of cyanoglycosides were released, which rapidly decayed to CN⁻ following enzymatic hydrolysis. Depending on the varying cyanoglycoside content of the cassava varieties, the cyanide concentration in the wastewater was as high as 200 mg/l. To simulate anaerobic stabilization, a wastewater with a chemical oxygen demand (COD) of about 20 g/l was prepared from cassava roots and was fermented in a fixed-bed methanogenic reactor. The start-up phase for a 99% degradation of low concentrations of cyanide (10 mg/l) required about 6 months. After establishment of the biofilm, a cyanide concentration of up to 150 mg CN⁻/l in the fresh wastewater was degraded during anaerobic treatment at a hydraulic retention time of 3 days. All nitrogen from the degraded cyanide was converted to organic nitrogen by the biomass of the effluent. The cyanide-degrading biocoenosis of the anaerobic reactor could tolerate shock concentrations of cyanide up to 240 mg CN⁻/l for a short time. Up to 5 mmol/l NH₄Cl (i.e. 70 mg N/l = 265 mg NH₄Cl/l) in the fresh wastewater did not affect cyanide degradation. The bleaching agent sulphite, however, had a negative effect on COD and cyanide removal. For anaerobic treatment, the maximum COD space loading was 12 g l⁻¹ day⁻¹, equivalent to a hydraulic retention time of 1.8 days. The COD removal efficiency was around 90%. The maximum permanent cyanide space loading was 50 mg CN⁻ l⁻¹ day⁻¹, with tolerable shock loadings up to 75 mg CN⁻ l⁻¹day⁻¹. Under steady-state conditions, the cyanide concentration of the effluent was lower than 0.5 mg/l. Received: 15 August 1997 / Received revision: 10 October 1997 / Accepted: 14 October 1997     

Candida bombicola : production of novel alkyl glycosides based on glucose/2-dodecanol

Candida bombicola produces glycolipids containing sophorose and a glycosidically/esterically bound ω- or (ω−1)-hydroxy C₁₆₍₁₈₎ acid. Here we describe novel glycolipids from this source. Glucose and 2-dodecanol were used for the cultivation of the yeast, one part of the racemic secondary alcohol being connected directly with a glucose or a sophorose unit. A relatively high content of yeast extract, up to 4 g l⁻¹, and subsequently higher biomass concentrations favoured the production of novel products. The provision of 150 g l⁻¹ glucose and 15 g l⁻¹ 2-dodecanol resulted in maximum production of 22 g l⁻¹ novel alkyl glycosides (more than 90% novel products). The molecular structures were analysed by gas chromatography, fast atom bombardment/mass spectrometry, ¹H- and ¹³C-nuclear magnetic resonance and optical rotation studies. Sophorose and glucose were detected as carbohydrate moieties, (S)-(+)-2-dodecanol (88%) was found to be the major lipid moiety. The new glycolipids are suitable biosurfactants, reducing the surface tension of water from 72 mN m⁻¹ to 32–38 mN m⁻¹. Received: 8 December 1997 / Received revision: 19 March 1998 / Accepted: 20 March 1998     

Production of lipids in 10 strains of Chlorella and Parachlorella, and enhanced lipid productivity in Chlorella vulgaris

We tested 10 different Chlorella and Parachlorella strains under lipid induction growth conditions in autotrophic laboratory cultures. Between tested strains, substantial differences in both biomass and lipid productivity as well as in the final content of lipids were found. The most productive strain (Chlorella vulgaris CCALA 256) was subsequently studied in detail. The availability of nitrates and/or phosphates strongly influenced growth and accumulation of lipids in cells by affecting cell division. Nutrient limitation substantially enhanced lipid productivity up to a maximal value of 1.5 g l⁻¹ day⁻¹. We also demonstrated the production of lipids through large-scale cultivation of C. vulgaris in a thin layer photobioreactor, even under suboptimal conditions. After 8 days of cultivation, maximal lipid productivity was 0.33 g l⁻¹ day⁻¹, biomass density was 5.7 g l⁻¹ dry weight and total lipid content was more than 30% dry weight. C. vulgaris lipids comprise fatty acids with a relatively high degree of saturation compared with canola oil offering a possible alternative to the use of higher plant oils.     

Enantioselective reduction of 3,4-methylene-dioxyphenylacetone using Candida famata and Zygosaccharomyces rouxii

In an effort to prepare 3,4-methylene-dioxyphenyl-(S)-isopropanol from 3,4-methylene-dioxyphenylacetone, an initial screen of microbes indicated that Candida famata could catalyze this reaction efficiently at low substrate concentration. A dilute, large-scale process was developed to provide experimental material for the chemical synthesis to be explored. However, the productivity number of this process [0.134 g product (g␣wet␣weight cells)⁻¹ day⁻¹ was too low to be practical. C.␣famata was also extremely sensitive to concentrations of both the ketone and the alcohol greater than 2 g/l. A more extensive screen of yeast and fungi revealed that Zygosaccharomyces rouxii was more tolerant to higher substrate concentrations and had a higher productivity number [0.8 g (g wet weight cells)⁻¹ day⁻¹]. These characteristics suggested that Z. rouxii could be used in a large-scale process at high substrate concentrations. Received: 8 July 1996 / Received revision: 9 September 1996 / Accepted: 18 September 1996