Coupling of fermentation and microfiltration for α-amylase production from Bacillus amyloliquefaciens

Abstract: A strain of Bacillus amyloliquefaciens secreting α-amylase was cultivated continuously in a fermentor coupled to a filtration unit. Under the tested operating conditions, the maximum flux was 91 m^-2 h^-1 during the first day and 61 m^-2 h^-1 during the 2nd day. The α-amylase retention was around 30%. Compared to a batch process, continuous cultivation with cell recycling led to lower α-amylase concentrations but to a doubling of volumetric productivities.     

Thermoalkalophilic lipase-producing Bacillus selected by continuous cultivation

Abstract Several lipase-producing thermophilic bacteria were isolated by continuous culture from samples collected near Bulgarian hot springs. Most of them had lipase activity of about 0.5 U ml⁻¹ when activated in shaken flasks. Three strains, Gram-positive sporeforming rods, possess higher enzyme activity in a Tween-80 containing medium. The highest lipase activity (2.0–3.0 U ml⁻¹) was observed in the newly-isolated thermoalkalophilic Bacillus sp. strain MC7.     

Continuous ethanol production by flocculating yeast in the fluidized bed bioreactor

Abstract: Continuous fermentation by a highly flocculant strain of the yeast Saccharomyces cerevisiae was carried out in a tower fluidized-bed bioreactor. The synthetic and molasses media with a total sugar concentration of 17% (w/v) were used for fermentation. Different dilution rates were tested. Stable cell densities of 50 kg m^-3(dry weight) were maintained for all dilution rates. The ethanol productivity was increasing linearly with dilution rates up to 15—20 kg m^-3 h^-1. Aeration of the culture stabilized flocculating activity and viability of yeast and also permitted long-term operation of the bioreactor.     

Production of oil-processing compounds by microorganisms from the Daqing oil field,China

Twenty pure cultures isolated from formation waters of the Daqing oil field were studied with respect to their capacity to produce surface-active compounds in media with individual hydrocarbons, lower alcohols, and fatty acids. Aerobic saprotrophic bacteria belonging to the genera Bacillus, Brevibacillus, Rhodococcus, Dietzia, Kocuria, Gordonia, Cellulomonas, Clavibacter, Pseudomonas, and Acinetobacter decreased the surface tension of cultivation media from 55-63 to 28-44 mN/m. Strains of Bacillus cereus, Rhodococcus ruber, and Bacillus licheniformis produced biosurfactants most actively. Bacteria of the genera Rhodococcus, Dietzia, Kocuria, and Gordonia produced exopolysaccharides in media with hydrocarbons. Culture liquids of the strains of R. ruber and B. licheniformis exhibited oil-releasing effect. Thus, the Daqing oil field is inhabited by aerobic bacteria capable of producing effective oil-releasing agents.     

Screening and production of rhamnolipids by Pseudomonas aeruginosa 47T2 NCIB 40044 from waste frying oils

World production of oils and fats is about 2.5 million tonnes, 75% of which are derived from plants. Most of them are used in the food industry for the manufacture of different products, or directly as salad oil. Great quantities of waste are generated by the oil and fat industries: residual oils, tallow, marine oils, soap stock, frying oils. It is well known that the disposal of wastes is a growing problem and new alternatives for the use of fatty wastes should be studied. Used frying oils, due to their composition, have great potential for microbial growth and transformation. The use of economic substrates such as hydrophobic wastes meets one of the requirements for a competitive process for biosurfactant production. In the Mediterranean countries, the most used vegetable oils are sunflower and olive oil. Here we present a screening process is described for the selection of micro-organism strains with the capacity to grow on these frying oils and accumulate surface-active compounds in the culture media. From the 36 strains screened, nine Pseudomonas strains decreased the surface tension of the medium to 34-36 mN/M; the emulsions with kerosene remained stable for three months. Two Bacillus strains accumulated lipopeptide and decreased the surface tension to 32-34 mN/m. Strain Ps. aeruginosa 47T2 was selected for further studies. The effect of nitrogen and a C/N of 8. 0 gave a final production of rhamnolipid of 2.7 g l-1 as rhamnose, and a production yield of 0.34 g g-1.     

Effects of light and temperature on the cell size and some biochemical components in two freshwater cryptophytes

The effects of light and temperature on cell size and cellular composition (chlorophyll, protein, carbohydrate) of two freshwater cryptophytes were studied with batch cultures. Neither of the species had a constant cell size but the size varied with growth conditions. At each temperature the smallest cells were recorded at the lowest experimental photon flux density. The smallest cells of Cryptomonas 979/67 had an average volume of 232 μm³ and the largest ones 1 020 μm³. In Cryptomonas 979/62 the smallest and largest cells measured 4 306 μm³ and 12 450 μm³. Both species increased their cellular chlorophyll content when PFB dropped below 110–120 μmol m^-2 s^-1. The highest and lowest chlorophyll contents of 979/67 were 7.45 fg μm^-3 and 0.55 fg μm^-2 respectively. For 979/62 the corresponding values were 10.23 fg μm^-3 and 0.93 fg μm^-3. In both species the protein content remained stable at PFDs higher than 110–120 μmol m^-2 S^-1. The highest content of protein measured in 979/67 was 638 fg μm^-3 and the lowest 147 fg μm^-3. For 979/62 these values were 1 036 fg μm^-3 and 148 fg μm^-3 respectively. The carbohydrate results were less clear and no pattern either in response to photon flux density or temperature was obvious. The lowest and highest contents recorded for 979/67 were 62 fg μm^-3 and 409 fg μ^-3 and for 979162, 36 fg μm^-3 and 329 fg μm^-3     

Chlorophyll fluorescence measured using the Fraunhofer line-depth principle and relationship to photosynthetic rate in the field

Abstract. A field study was conducted to determine the relationship of solar-excited chlorophyll a fluorescence to net CO₂ assimilation rate in attached leaves. The Fraunhofer line-depth principle was used to measure fluorescence at 656.3 nm wavelength while leaves remained exposed to full sunlight and normal atmospheric pressures of CO₂ and O₂. Fluorescence induction kinetics were observed when leaves were exposed to sunlight after 10 min in darkness. Subsequently, fluorescence varied inversely with assimilation rate. In the C₄ Zea mays , fluorescence decreased from 2.5 to 0.8 mW m^-2 nm^-1 as CO₂ assimilation rate increased from 1 to 8 μmol m^-2 s^-1 (r²= 0.520). In the C₃ Liquidambar styraciflua and Pinus taeda , fluorescence decreased from 6 to 2 mW m^-2 nm^-1 as assimilation rate increased from 2 to 5 or 0 to 2 μmol m^-2 s^-1 (r²= 0.44 and 0.45. respectively). The Fraunhofer line-depth principle enables the simultaneous measurement of solar-excited fluorescence and CO₂ assimilation rate in individual leaves, but also at larger scales. Thus, it may contribute significantly to field studies of the relationship of fluorescence to photosynthesis.     

Enrichment of linear alkylbenzenesulphonate (LAS) degrading bacteria in continuous culture

Enrichment experiments were carried out in continuous-flow units using a mineral medium with commercial linear alkylbenzenesulphonate (LAS) as the limiting carbon- and energy-source. The mixed bacterial culture originating from the waste water of a detergent plant consisted of five strains belonging to the genus Pseudomonas and two strains each of the genera Achromobacter and Acinetobacter. The cultivation conditions corresponding to dilution rates of 0.025-0.1 h^-1 and LAS concentrations of 20–50 mg/1 were examined. During the experiments the composition of mixed cultures and the kinetics of LAS biodegradation were followed. Continuous-flow enrichment experiments resulted in the selection of six bacterial cultures with different compositions of individual species and capability to utilize LAS. From the original seven strains at lower dilution rates (0.025 and 0.05 h^-1) six were selected, excluding Pseudomonas sp. 3, while at the highest dilution rate (0.1/h^-1) five strains were selected after eliminating Pseudomonas sp. 5 and Achromobacter sp. 1. All enriched mixed cultures were more efficient in primary than in ultimate LAS degradation. Two of the culture strains were able to achieve primary LAS degradation ( Pseudomonas sp. 1 in mineral medium with LAS as the sole carbon- and energy-source and Acinetobacter sp. 3 in medium supplemented by yeast extract and nutrient broth).
None of the strains could degrade LAS completely, which indicates that many types of interactions based on combined metabolic attack as well as those based on provision of specific nutrients, may exist between culture members during the complete LAS bio-oxidation.     

LIGHT-DEPENDENT GROWTH, DARK SURVIVAL, AND GLUCOSE UPTAKE BY CRYPTOPHYTES ISOLATED FROM A FRESHWATER CHEMOCLINE

Clones of Cryptomonas phaseolus Skuja , Cryptomonas rostratiformis ( Skuja ) Skuja in Huber-Pestalozi, and Cryptomonas undulata Gervais were isolated from the deep chlorophyll maximum near the oxic/anoxic boundary layer of the mesoeutrophic lake Schlachtensee, Germany. Different autecological features of these species were studied in batch culture experiments . Cryptomonas cf . ovata Ehrenberg and Chroomonas sp. that never dominated in the deep chlorophyll layer were also isolated from Schlachtensee to study their light-dependent growth in comparison to the deep-living species . Cryptomonas undulata, C. cf . ovata, and C. phaseolus had a very low light compensation point ( 5–7 μmol.m^-2.s^-1 ), whereas the growth rate of Chroomonas sp. and C. rostratiformis was positive above 16 and 24 μmol.m^-2.s^-1 . Cryptomonas phaseclus and Chroomonas sp. became photoinhibited above photon flux densities of 92 and 116 μmol.m^-2.s^-1 . Cryptomonas rostratiformis, C. cf . ovata, and C. undulata reached a maximum growth rate at a considerably higher photon flux density (198–250 μmol.m^-2.s^-1 ). Cryptomonas phaseolus grew fastest under light-limiting conditions . Chyptomonas phaseolus and C. undulata were best able to suruive prolonged periods of darkness . Cryptomonas phaseolus, C. rostratiformis, and C. undulata did not show any uptake of fluorescent latex beads. When labeled glucose was provided in naturally occurring concentrations, carbon uptake by C. phaseolus, C. rostratiformis, and C. undulata was negligibly small in comparison to cellular carbon content. I suggest that the adaptation to a low-light environment is an important preadaptation for the dominance of C. phaseolus and C. undulata near the freshwater chemocline .     

Production of Oil-Releasing Compounds by Microorganisms from the Daqing Oil Field,China

Twenty pure cultures isolated from formation waters of the Daqing oil field were studied with respect to their capacity to produce surface-active compounds in media with individual hydrocarbons, lower alcohols, and fatty acids. Aerobic saprotrophic bacteria belonging to the genera Bacillus, Brevibacillus, Rhodococcus, Dietzia, Kocuria, Gordonia, Cellulomonas, Clavibacter, Pseudomonas, and Acinetobacter decreased the surface tension of cultivation media from 55–63 to 28–44 mN/m. Strains of Bacillus cereus, Rhodococcus ruber, andBacillus licheniformis produced biosurfactants most actively. Bacteria of the genera Rhodococcus, Dietzia, Kocuria, and Gordonia produced exopolysaccharides in media with hydrocarbons. Culture liquids of the strains of R. ruberand B. licheniformis exhibited an oil-releasing effect. Thus, the Daqing oil field is inhabited by aerobic bacteria capable of producing effective oil-releasing agents.     

A modified drop-net method for sampling mobile epifauna on marine shallow sandy bottoms

The seasonal variations in numbers and biomass of the mobile epifauna of a shallow, sandy bay in Gullmar Fjord (Sweden) was investigated during 1976–1977 using the drop-net technique. Juvenile plaice, Pleuronectes platessa L., sand goby, Pomatoschistus minutus (Pallas), and brown shrimp Crangon crangon L. are the dominant epifaunal species. A net enclosing an area of 100 m² was used, and the organisms captured were collected with a small trawl. Three stations with a water depth≤ 1 m were regularly sampled with a total number of 73 drops. Maximum abundance was for plaice in June (6 ind m^-2), for the shrimp in July-August (61 ind m^-2), and for sand goby in September (2 ind m^-2). Standing stocks (dry wt) were 0.2, 1.5 and 0.2 g m^-2 respectively. Production estimates were based on seasonal changes in size frequency and information on specific growth rates. The production rate was for plaice (0-group) 0.3 g m^-2 yr^-1, for the shrimp 1.5, and for sand goby 0.2 g m^-2 yr^-1. Reliable estimates of the absolute abundance can be derived from the mean catch per unit area only when the efficiency of the fishing gear is known. After 6 hauls with the trawl in the drop-net enclosure more than 85% of the shrimps were caught and after the fifteenth haul less than 1% were still left. The fishing efficiency of a push-net in relation to the drop-net was for the shrimp 50%, for plaice 35% and for sand goby 17%.     

Production of B-group vitamins by two Rhizobium strains in chemically defined media

Twenty-eight strains of Rhizobium spp. were tested for their ability to grow in chemically-defined medium lacking growth factors. Two strains, R. meliloti GR4B and Rhizobium spp. ( Acacia ) GRH28, were selected, on the basis of their good growth under the conditions imposed, for further quantification of the production of water-soluble vitamins (thiamine, niacin, riboflavin, pantothenic acid and biotin) in chemically defined media amended with different compounds (mannitol, glucose or sodium succinate) as sole carbon sources. Qualitative and quantitative production of vitamins in chemically-defined media was significantly affected by the use of C sources of a different nature and the age of the cultures. Strain GRH28 produced all the vitamins analysed, and high biological levels of biotin (14 ng ml^–1 culture) were detected after 6 d of culture in mineral medium amended with mannitol. Pantothenic acid was the vitamin detected in the highest amounts (up to 1 μg ml^–1 of culture) in culture supernatant fluids of strain GR4B grown for 6 d with succinate as sole carbon source.     

一株石油烃降解菌的细胞疏水性及其乳化性质

【目的】从新疆油田石油污染土壤中分离到一株在25 °C条件下利用烃类产生生物表面活性剂的菌株红球菌(Rhodococcus sp.) HL-6, 对其菌体细胞疏水性及所产表面活性剂进行研究。【方法】通过细胞粘附性、表面张力及乳化活性测定对菌株所产表面活性剂进行性质研究。【结果】菌株HL-6在亲水性和疏水性基质中均能产生生物表面活性剂, 在疏水性基质中可以将培养液表面张力由初始的62.487 mN/m降到30.667 mN/m, 培养液在pH 6?9及NaCl浓度1%?5%范围内乳化效果良好, 在4 °C到55 °C范围内乳化效果均为100%, 菌株对柴油的耐受能力很高, 在30%柴油浓度下依然生长良好并且有44%的乳化活性。【结论】HL-6菌株的细胞表面具有很强的疏水性, 这有助于菌体细胞对烃类的摄取。该菌株能够利用烃类基质生产生物表面活性剂, 可以明显降低培养液表面张力并且对石油烃具有良好的乳化作用。说明菌株HL-6能够适应海洋滩涂石油污染的环境, 并可用于严重石油污染区域的生物修复。     

Habitat selection by grayling-II. Preliminary results on larval and juvenile daytime habitats

Larval grayling were found along the shoreline at velocities <20 cm s^-1 depths <40 cm, shear stress <2 dyn m^-2 and over sand and silt. Juveniles were found in the river channel at currents of 20-40 cm s^-1 depths of 40-60 cm and shear stresses of 2-4 dyn m^-2, over gravel and pebbles.     

Bacillus tianmuensis sp. nov., isolated from soil in Tianmu Mountain national natural reserve, Hangzhou, China

In a project aiming to isolate strains with the ability to produce nonribosomal peptides, a Gram-negative, endospore-forming, rod-shaped strain, designated B5^T, was isolated from a soil sample collected from Tianmu Mountain national natural reserve in Hangzhou, China. Strain B5^T contained meso -diaminopimelic acid in the cell wall peptidoglycan. The major cellular fatty acids were anteiso-C_15:0 and iso-C_15:0. The DNA G+C content was 42.5 mol%. The phylogenetic analysis based on 16S rRNA gene sequence indicated that strain B5^T fell within the genus Bacillus , with highest sequence similarity values to Bacillus barbaricus DSM 14730^T (96.4%) and Bacillus macauensis JCM 13285^T (95.5%). The isolate, however, could be distinguished from Bacillus strains with validly published names by low 16S rRNA gene sequence similarity values, distinct phenotypic and chemotaxonomic characteristics. On the basis of these polyphasic evidences, it is demonstrated that the isolate B5^T represents a novel species of the genus Bacillus , for which the name Bacillus tianmuensis sp. nov. is proposed. The type strain is B5^T (=DSM 22111^T=CGMCC 1.8879^T).     

Potential of Bacillus sp. to produce polyhydroxybutyrate from biowaste

Aim:  To test the Bacillus strains for their abilities to produce polyhydroxybutyrate (PHB) from different sugars and biowaste (Pea-shells).
Methods and Results:  Six Bacillus strains were checked for their ability to produce PHB from GM2 medium supplemented with different sugars at the rate of 1% (w/v) and from biowaste and GM2 (BW : M) combinations (3 : 7, 1 : 1, 7 : 3). Glucose supplemented GM2 medium resulted in maximum PHB production of 435 mg l⁻¹ constituting 31–62% w/w of the total cell dry mass. Substituting GM2 medium to the extent of 50% with biowaste (pea-shell slurry) resulted in 945–1205 mg l⁻¹ PHB (55–65% w/w). Optimization for additional nitrogen supplementation, inoculum size resulted in a final PHB production of 3010–3370 mg l⁻¹ equivalent to 300 g kg⁻¹ biowaste (dry wt).
Conclusion:  The Bacillus strains were able to produce PHB from biowaste (Pea-shells) as cheap source of substrate.
Significance and Impact of the Study:  This is the first report on usage of pea-shells as feed for PHB production, opening new possibilities for its use for production of PHB and Bacillus as potential candidate for the purpose.     

Peat respiration and decomposition in Antarctic terrestrial moss communities

Oxygen uptake by the peat of two Antarctic bryophyte communities (a moss turf and a moss carpet) is converted to organic matter loss and used to derive the rate of decomposition. The decay rates obtained in this way are evaluated in two mathematical models which simulate the accumulation of dead organic matter (DOM) in the communities from the litter production and decomposition rate. Litter production, the extent of DOM accumulations at present on the sites and mean decomposition rates (i.e. fraction of standing crop lost per year) were 409 g m^-2 year^-1, 33.5 kg m^-2 and 0.017 g g^-1 year^-1 in the moss turf and 392 g m^-2 year^-1, 29.6 kg m-² and 0.010 g g^-1 year^-1 in the moss carpet respectively (all weights expressed as dry weight). Aerobic decomposition rate declined with depth in both communities. From the model's predictions it is suggested that the observed decay rate was too high in the moss turf and too low in the carpet. Possible reasons for this are discussed and suggestions made for future work.     

The growth rate of eel in tributaries of the lower River Severn, England, and its relationship with stock size

The growth rate of 1980 eel Anguilla anguilla from 15 sites in the Severn system varied between 16·4 and 27·9 mm year^-1, density from 0·12 to 1·14 m^-2 and biomass from 2·56 to 25·24 g m^-2. There was no significant relationship between growth rate and density or biomass ( P > 0·05).     

Growth and production of five species of Ephemeroptera larvae from an experimental recirculating stream

SUMMARY. The production and growth of the larvae of five species of Ephemeroptera in an experimental recirculating stream-channel are described. The most abundant species were Baetis rhodani , which achieved a maximum population density of c. 29,000 m^-2 in August, and Ephemerella ignita which reached a density of c. 13,000 m^-2 in the spring. Three other common speeies, Paraleptophlebia submarginata, Centroptilum luteolum and Caenis rivulorum were present at lower densities (< 1000 m^-2). B. rhodani had five cohorts in a year whilst C. luteolum had two or possibly three. E. ignita, C. rivulorum and P. submarginata were univoltine. Mean speeifie growth rates were calculated for each species, the maximum being 3.26±0.49% length day^-1 for E. ignita (3 April—15 May). Production of each of these five species of Ephemeroptera was calculated giving a total annual production of 10.2 g dry wt m^-2 (computational method).     

Observations on the growth and production of chub Leuciscus cephalus and dace Leuciscus leuciscus in a small lowland river in southeast England

SUMMARY. The River Eden (Kent, England) holds a mixed coarse fish population in which minnows ( Phoxinus phoxinus ), gudgeon ( Gobio gobio ), chub ( Leuciscus cephalus ) and dace ( Leuciscus leuciscus ) are numerically predominant. Chub and dace provide the major interest to anglers and their growth and production were studied. Observed growth rates of both species were marginally below recorded averages from other British habitats. Back-calculations showed that year-class strength and relative growth rates varied between years. Instantaneous mortality rate Z was 0.147 for chub aged 1–10, 0.438 for chub aged 10 and over, and 0.172 for dace. Exclusive of the 0-group, fish numbers, biomass (wet wt) and production were found to be 0.1305 m^-2, 19.12 gm^-2 and 5.38 gm^-2 year^-1, respectively, for chub and 0.0968 m^-2, 4.33 gm^-2 and 1.69 gm^-2 year^-1 for dace. Smoothing of data produced theoretical production figures of 6.69 gm^-2 year^-1 for chub and 1.15 gm^-2 year^-1 for dace.