Aluminum in lake water and organs of a fish Tribolodon hakonensis in strongly acidic lakes with a high aluminum concentration

Aluminum in lake water and in the organs of the fish Tribolodon hakonensis was investigated in Lake Usoriko (pH 3.6), Lake Inawashiroko (pH 5.0), and the Tenryu River (pH 7.7). The concentration of total soluble aluminum in the water was 0.51 mg l⁻¹ in Usoriko, 0.05 mg l⁻¹ in Inawashiroko, and less than 0.01 mg l⁻¹ in the Tenryu. The chemical forms of soluble aluminum in the acid water were characterized as Al³⁺, AlL²⁺, and AlL^≦1+. More than 90% of soluble aluminum in the water of Usoriko was Al³⁺, whereas AlL²⁺ was dominant in the water of Inawashiroko. The aluminum concentration in the organs of T. hakonensis in Usoriko was 42 μg g⁻¹ wet weight in gills, 4.2 μg g⁻¹ in muscle, 6.9 μg g⁻¹ in bone, 12.7 μg g⁻¹ in liver, 6.0 μg g⁻¹ in kidney, and 6.0 μg g⁻¹ in intestine, indicating accumulation of aluminum in the gills. The aluminum concentration in the organs of T. hakonensis living in Inawashiroko was approximately the same, in spite of the difference in water chemistry of the two acid lakes, especially for pH and aluminum. This suggests that aluminum accumulation might be controlled in the fish living in the acid lakes. In contrast, the aluminum concentration in the gills of T. hakonensis from the Tenryu was 2 μg g⁻¹. Received: May 20, 1999 / Accepted: December 10, 1999     

Pseudoxanthomonas bacteria that drive deposit formation of wood extractives can be flocculated by cationic polyelectrolytes

Runnability problems caused by suspended bacteria in water using industries, have, in contrast to biofilms, received little attention. We describe here that Pseudoxanthomonas taiwanensis, a wide-spread and abundant bacterium in paper machine water circuits, aggregated dispersions of wood extractives ("pitch") and resin acid, under conditions prevailing in machine water circuits (10⁹ cfu ml⁻¹, pH 8, 45°C). The aggregates were large enough (up to 50 μm) so that they could be expected to clog wires and felts and to reduce dewatering of the fiber web. The Pseudoxanthomonas bacteria were negatively charged over a pH range of 3.2–10. Cationic polyelectrolytes of the types used as retention aids or fixatives to flocculate "anionic trash" in paper machines were effective in flocculating the Pseudoxanthomonas bacteria. The polyelectrolyte most effective for this purpose was of high molecular weight (7–8 × 10⁶ g mol⁻¹) and low charge density (1 meq g⁻¹), whereas polyelectrolytes that effectively zeroed the electrophoretic mobility (i.e., neutralized the negative charge) of the bacterium were less effective in flocculating the bacteria. Based on the results, we concluded that the polyelectrolytes functioning by bridging mechanism, rather than by neutralization of the negative charge, may be useful as tools for reducing harmful deposits resulting from interaction of bacteria with wood extractives in warm water industry.     

Localization of ion-regulatory epithelia in embryos and hatchlings of two cephalopods

The tissue distribution and ontogeny of Na⁺/K⁺-ATPase has been examined as an indicator for ion-regulatory epithelia in whole animal sections of embryos and hatchlings of two cephalopod species: the squid Loligo vulgaris and the cuttlefish Sepia officinalis. This is the first report of the immunohistochemical localization of cephalopod Na⁺/K⁺-ATPase with the polyclonal antibody α (H-300) raised against the human α1-subunit of Na⁺/K⁺-ATPase. Na⁺/K⁺-ATPase immunoreactivity was observed in several tissues (gills, pancreatic appendages, nerves), exclusively located in baso-lateral membranes lining blood sinuses. Furthermore, large single cells in the gill of adult L. vulgaris specimens closely resembled Na⁺/K⁺-ATPase-rich cells described in fish. Immunohistochemical observations indicated that the amount and distribution of Na⁺/K⁺-ATPase in late cuttlefish embryos was similar to that found in juvenile and adult stages. The ion-regulatory epithelia (e.g., gills, excretory organs) of the squid embryos and paralarvae exhibited less differentiation than adults. Na⁺/K⁺-ATPase activities for whole animals were higher in hatchlings of S. officinalis (157.0 ± 32.4 μmol g_FM⁻¹ h⁻¹) than in those of L. vulgaris (31.8 ± 3.3 μmol g_FM⁻¹ h⁻¹). S. officinalis gills and pancreatic appendages achieved activities of 94.8 ± 18.5 and 421.8 ± 102.3 μmol_ATP g_FM⁻¹ h⁻¹, respectively. High concentrations of Na⁺/K⁺-ATPase in late cephalopod embryos might be important in coping with the challenging abiotic conditions (low pH, high pCO₂) that these organisms encounter inside their eggs. Our results also suggest a higher sensitivity of squid vs. cuttlefish embryos to environmental acid-base disturbances.     

Carob pulp as raw material for production of the biocontrol agent <Emphasis Type="Italic">P. agglomerans</Emphasis> PBC-1

Large-scale production has been the major obstacle to the success of many biopesticides. The spreading of microbial biocontrol agents against postharvest disease, as a safe and environmentally friendly alternative to synthetic fungicides, is quite dependent on their industrial mass production from low-cost raw materials. Considerable interest has been shown in using agricultural waste products and by-products from food industry as nitrogen and carbon sources. In this work, carob pulp aqueous extracts were used as carbon source in the production of the biocontrol agent Pantoea agglomerans PBC-1. Optimal sugar extraction was achieved at a solid/liquid ratio of 1:10 (w/v), at 25°C, for 1 h. Batch experiments were performed in shake flasks, at different concentrations and in stirred reactors at two initial inoculums concentrations, 10⁶ and 10⁷ cfu ml⁻¹. The initial sugar concentration of 5 g l⁻¹ allowed rapid growth (0.16 h⁻¹) and high biomass productivity (0.28 g l⁻¹ h⁻¹) and was chosen as the value for use in stirred reactor experiments. After 22 and 32 h of fermentation the viable population reached was 3.2 × 10⁹ and 6.2 × 10⁹ cfu ml⁻¹ in the fermenter inoculated at 10⁶ cfu ml⁻¹ and 2.7 × 10⁹ and 6.7 × 10⁹ cfu ml⁻¹ in the bioreactor inoculated at 10⁷ cfu ml⁻¹. A 78% reduction of the pathogen incidence was achieved with PBC-1 at 1 × 10⁸ cfu ml⁻¹, grown in medium with carob extracts, on artificially wounded apples stored after 7 days at 25°C against P. expansum.     

Aerobiological dynamics of potentially pathogenic fungi in a rice agroecosystem in La Habana, Cuba

The aerobiology can play a key role in protecting the rice crop since many fungi can cause serious damage to agricultural areas. In this way, the ideal time to implement different security measures can be identified. To determine the presence of potentially pathogenic fungi in the air of the rice agroecosystem, a weekly monitoring of viable fungi was carried out using a volumetric sampler. Collected fungi were quantified, isolated, and identified based on their morphological characteristics. The results obtained demonstrated that the annual average concentration of filamentous fungi in the atmosphere of rice agroecosystem studied was 1,225 cfu m⁻³ levels ranging between 115 cfu m⁻³ (April) and 2,865 cfu m⁻³ (August). Pyricularia grisea was detected in the air for 5 months, since the second week of June until the first week of October, and highest average concentration (25 cfu m⁻³) was observed in August. Of the meteorological factors evaluated, temperature and relative air humidity influence the concentration of propagules of P. grisea in the air. Besides, other fungi were detected such as Curvularia, Bipolaris, Alternaria, and Cercospora, all with relevance to rice cultivation. This is the first characterization of aeromycological biodiversity in the studied region.     

Effects of Rhamnolipids from Pseudomonas aeruginosa DS10-129 on Luminescent Bacteria: Toxicity and Modulation of Cadmium Bioavailability

In this study, the mixture of mono- and di-rhamnolipids produced by Pseudomonas aeruginosa DS10-129 was characterized for its toxicity and modulatory effects on Cd availability to different bacteria. Gram-negative naturally bioluminescent Vibrio fischeri and recombinant bioluminescent Pseudomonas fluorescens, P. aeruginosa, Escherichia coli, and Gram-positive Bacillus subtilis were used as model organisms. Rhamnolipids reduced the bioluminescence of these bacteria in less than a second of exposure even in relatively low concentrations (30-min EC₅₀ 45–167 mg l⁻¹). Toxicity of Cd to Gram-negative bacteria (30-min EC₅₀ values 0.16 mg l⁻¹ for E. coli, 0.96 mg l⁻¹ for P. fluorescens, and 4.4 mg l⁻¹ for V. fischeri) was remarkably (up to 10-fold) reduced in the presence of 50 mg l⁻¹ rhamnolipids. Interestingly, the toxicity of Cd to Gram-positive B. subtilis (30-min EC₅₀ value 0.49 mg l⁻¹) was not affected by rhamnolipids. Rhamnolipids had an effect on desorption of Cd from soil: 40 mg l⁻¹ rhamnolipids increased the water-extracted fraction of Cd twice compared with untreated control. However, this additionally desorbed fraction of Cd remained bound with rhamnolipids and was not available to bacteria. Hence, in carefully chosen concentrations (still effectively complexing heavy metals but not yet toxic to soil bacteria), rhamnolipids could be applied in remediation of polluted areas.     

Solid state bioreactor production of transglutaminase by Amazonian Bacillus circulans BL32 strain

In this work, we investigated the production of transglutaminase (TGase) by an Amazonian isolated strain of Bacillus circulans by solid-state cultivation (SSC). Several agro-industrial residues, such as untreated corn grits, milled brewers rice, industrial fibrous soy residue, soy hull, and malt bagasse, were used as substrates for microbial growth and enzyme production. Growth on industrial fibrous soy residue, which is rich in protein and hemicellulose, produced the highest TGase activity (0.74 U g⁻¹ of dried substrate after 48 h of incubation). A 2³ central composite design was applied to determine the optimal conditions of aeration, cultivation temperature and inoculum cell concentration to TGase production. The best culture conditions were determined as being 0.6 L air min⁻¹, 33 °C and 10 log ₁₀ CFU g⁻¹ of dried substrate, respectively. Under the proposed optimized conditions, the model predicted an enzyme production of 1.16 U g⁻¹ of dried substrate, closely matching the experimental activity of 1.25 U g⁻¹. Results presented in this work point to the use of this newly isolated B. circulans strain as a potential alternative of microbial source for TGase production by SSC, using inexpensive culture media.     

The influence of cadmium on life-history parameters and gut microflora of <Emphasis Type="Italic">Archegozetes longisetosus</Emphasis> (Acari: Oribatida) under laboratory conditions

We tested the effect of cadmium (25, 130 μg Cd g⁻¹), administered via Chinese cabbage (Brassica chinensis L.) as food on life-history parameters and gut microflora of tritonymphs and adults of the oribatid mite, Archegozetes longisetosus Aoki. Both concentrations of Cd had an adverse effect on offspring mortality, and the higher concentration also reduced female fecundity, as well as the number of bacteria, fungi and actinomycetes, and it changed the community structure of bacteria; the proportion of gram-negative bacteria increased while that of gram-positive bacteria declined. Interestingly, at the lower Cd concentration microflora was more abundant and diverse than in the control group, especially in the tritonymphs, although the mean activity of gut microflora was reduced. The higher Cd concentration reduced microflora activity both in the tritonymphs and adults.     

Measurement of culturable airborne staphylococci downwind from a naturally ventilated broiler house

The objective of this study was to estimate the possible travel distance of airborne bacteria emitted from a naturally ventilated broiler house by using staphylococci as indicator organisms. Air samples were taken during the second half of three fattening periods with Impinger (AGI-30) in the barn and simultaneously upwind and downwind from the building. Staphylococci concentrations varied between 1 × 10⁶ and 1 × 10⁷ cfu m⁻³ in the barn. No Staphylococci were detected in air samples at the upwind side. A strong exponential decrease of these bacteria was observed at three sampling heights (1.5, 4.0 and 9.5 m) in the main wind direction downwind of the barn. Staphylococci concentrations up to 5.9 × 10³ cfu m⁻³ were detected at the farthest sampling point (333 m) downwind. Identification to the species level by means of a 16S–23S ITS PCR confirmed that Staphylococcus spp. from downwind samples originated from the barn. Staphylococci served as an useful indicator to demonstrate the travel distance of bacterial emissions originating from a naturally ventilated broiler house. These findings indicate that airborne transmission of viable bacteria from this type of housing system to adjacent residential dwellings or animal houses several hundred metres away is possible.     

Airborne concentrations of bacteria and fungi in Thailand border market

Thailand border market is where the local Thais, Cambodians, Laotians, and Burmeses exchange their goods and culture at the border checkpoints. It is considered to be the source of aerial disease transmission especially for foreigners because it is always very crowded with people from all walks of life. Unhealthy air quality makes this area high risk of spread of airborne diseases. This study assessed airborne concentrations of bacteria and fungi in a border market to improve exposure estimates and develop efficient control strategies to reduce health risk. The density and distribution of airborne bacteria and fungi were investigated in the Chong Chom border market in Surin Province, Thailand. Eighteen air sampling sites were taken from outdoors and various work environments including indoor footpaths, wooden handicraft shops, electronic shops, the secondhand clothing shops, and fruit market areas. Exposed Petri plate method and liquid impinger sampler were used for sampling at the breathing zone, 1.5 m above the floor level, during weekend and holiday. Meteorological factors such as relative humidity, temperature, and light intensity were collected by portable data logger. The relative humidity was 67–73%, and temperature 29–33°C, and light varied between 18 and 270 Lux m⁻². Gram-positive and Gram-negative bacteria were found at a mean value of 10⁴ CFU m⁻³, and airborne fungi of 10³ CFU m⁻³ were recorded. The highest concentration of culturable airborne microorganisms was found along the indoor footpath (9.62 × 10⁴ CFU m⁻³ and 750.00 CFU/plate/h for impingement and sedimentation methods, respectively), the fruit market area (7.86 × 10⁴ CFU m⁻³ and 592.42 CFU/plate/h for impingement and sedimentation methods, respectively), and the secondhand clothing shop (4.59 × 10³ CFU m⁻³ and 335.42 CFU/plate/h for impingement and sedimentation methods, respectively) for Gram-positive bacteria, Gram-negative bacteria, and fungi, respectively. The lowest concentration of Gram-positive bacteria, Gram-negative bacteria, and fungi was found only at the outdoor area at 1.53 × 10⁴ CFU m⁻³, 0.93 × 10⁴ CFU m⁻³ and 0.80 × 10³ CFU m⁻³ by means of impingement method and 136.67 CFU/plate/h, 69.25 CFU/plate/h, and 62.00 CFU/plate/h by means of sedimentation methods for Gram-positive bacteria, Gram-negative bacteria, and fungi, respectively. The most frequently present airborne bacteria were identified as Bacillus, Corynebacteria, Diplococcus, Micrococcus, Acinetobacter, Alcaligenes, Enterobacter, and spore former rods. Acremonium, Aspergillus, Cladosporium, Penicillium, and Sporotrichum were the most frequently found aerosol fungi genera. The distribution of airborne microorganisms correlated with relative humidity and light factors based on principal component analysis. In conclusion, the border market is a potential source of aerial disease transmission and a various hazards of bioaerosols for workers, consumers, sellers, and tourists. The bioaerosol concentration exceeded the standard of occupational exposure limit. Many major indicators of allergenic and toxigenic airborne bacteria and fungi, Acinetobacter, Enterobacter, Pseudomonas, Cladosporium, Alternaria, Aspergillus, and Penicillium, were found in the various market environments.     

Aerobic heterotrophic bacteria and petroleum-utilizing bacteria from cow dung and poultry manure

In this study, enumeration and identification of total aerobic heterotrophic bacteria and petroleum-utilizing bacteria as well as the degradative potential of petroleum-utilizing bacterial isolates were carried out. The average counts of total aerobic heterotrophic bacteria in cow dung and poultry manure were 74.25 × 10⁵ c.f.u. g⁻¹ and 138.75 × 10⁵ c.f.u. g⁻¹ respectively. Acinetobacter sp, Bacillus sp, Pseudomonas sp, and Serratia spp. occurred as aerobic heterotrophs in both cow dung and poultry manure. However, Alcaligenes spp. occurred only in cow dung while, Flavobacterium sp, Klebsiella sp, Micrococcus sp, and Nocardia spp. occurred only in poultry manure as aerobic heterotrophs. The average counts of petroleum-utilizing bacteria in cow dung and poultry manure were 9.25 × 10⁵ c.f.u. g⁻¹ and 17.25 × 10⁵ c.f.u. g⁻¹ respectively. Pseudomonas spp. occurred as petroleum utilizer in both cow dung and poultry manure. However, Bacillus spp. occurred only in cow dung while Acinetobacter spp. and Micrococcus spp. occurred only in poultry manure as petroleum utilizers. Relative abundance of petroleum utilizers in total aerobic heterotrophs ranged from 6.38% to 20.00% for cow dung and from 9.38% to 17.29% for poultry manure. Introduction of pure cultures of petroleum-utilizing bacteria from cow dung and poultry manure into sterile oil-polluted soil revealed oil degradation in one week period.     

Beneficial microorganism survival on seed, roots and in rhizosphere soil following application to seed during drum priming

Priming is a technique used to improve seedling establishment of direct-seeded crops such as onion and carrot, resulting in a quick and uniform emergence. This work investigated the application of four selected beneficial microorganisms (Pseudomonas chlororaphis MA342, Pseudomonas fluorescens CHA0, Clonostachys rosea IK726d11 and Trichoderma harzianum T22) to onion and carrot seed during drum priming, and their subsequent survival and establishment in the rhizosphere once the seed was planted. Different application rates of fungi (7 log₁₀ cfu g⁻¹ dry seed) and bacteria (6 log₁₀ cfu g⁻¹ dry seed) were required on onion to achieve the end target of 5 log₁₀ cfu g⁻¹ dry seed, whereas a lower rate (5 log₁₀ cfu g⁻¹ dry seed for both bacteria and fungi) was successful on carrot. Microorganism-treated seed was planted in soil in the glasshouse and root and rhizosphere soil samples were taken at 2, 4 and 8 weeks post-planting. All seed-applied microorganisms were recovered throughout the experiment, although differences in the survival patterns were seen. The bacterial isolates declined in number over time, with P. fluorescens CHA0 showing better overall survival than P. chlororaphis MA342, particularly on the roots and in the rhizosphere soil of carrot. In contrast to the bacteria, the fungal isolate C. rosea IK726d11 showed good survival on both onion and carrot, and increased significantly in number throughout the 8-week period. Trichoderma harzianum T22 remained relatively constant in number throughout the experiment, but showed better survival on carrot than onion roots. Similar results were found in three different soil-types.     

Changes in intestinal morphology and microbiota caused by dietary administration of inulin and Bacillus subtilis in gilthead sea bream (Sparus aurata L.) specimens

Changes produced in gilthead sea bream (Sparus aurata L.) intestinal morphology and microbiota caused by dietary administration of inulin and Bacillus subtilis have been studied. Gilthead sea bream specimens were fed diets containing 0 (control), inulin (10 g kg^?1), B. subtilis (10⁷ cfu g^?1), or B. subtilis + inulin (10⁷ cfu g^?1 + 10 g kg^?1) for four weeks. Curiously, fish fed the experimental diets (inulin, B. subtilis, or B. subtilis + inulin) showed the same morphological alterations when studied by light and electron microscopy, while significant differences in the signs of intestinal damage were detected by the morphometric study. All of the observed alterations were present only in the gut mucosa, and intestinal morphometric study revealed no effect of inulin or B. subtilis on the intestinal absorptive area. Furthermore, experimental diets cause important alterations in the intestinal microbiota by significantly decreasing bacterial diversity, as demonstrated by the specific richness, Shannon, and range-weighted richness indices. The observed alterations demonstrate that fish fed experimental diets had different signs of gut oedema and inflammation that could compromise their body homeostasis, which is mainly maintained by the epithelial lining of the gastrointestinal tract. To our knowledge, this is the first in vivo study regarding the implications of the use of synbiotics (conjunction of probiotics and prebiotics) on fish gut morphology and microbiota.     

Screening of bacteria to produce polyhydroxyalkanoates from xylose

Although xylose is a major constituent of lignocellulosic feedstock and the second most abundant sugar in nature, only 22% of 3,152 screened bacterial isolates showed significant growth in xylose in 24 h. Of those 684, only 24% accumulated polyhydroxyalkanoates after 72 h. A mangrove isolate, identified as Bacillus sp. MA3.3, yielded the best results in literature thus far for Gram-positive strains in experiments with glucose and xylose as the sole carbon source. When glucose or xylose were supplied, poly-3-hydroxybutyrate (PHB) contents of cell dry weight were, respectively, 62 and 64%, PHB yield 0.25 and 0.24 g g⁻¹ and PHB productivity (P_PHB) 0.10 and 0.06 g l⁻¹ h⁻¹. This 40% P_PHB difference may be related to the theoretical ATP production per 3-hydroxybutyrate (3HB) monomer calculated as 3 mol mol⁻¹ for xylose, less than half of the ATP/3HB produced from glucose (7 mol mol⁻¹). In PHB production using sugar mixtures, all parameters were strongly reduced due to carbon catabolite repression. PHB production using Gram-positive strains is particularly interesting for medical applications because these bacteria do not produce lipopolysaccharide endotoxins which can induce immunogenic reactions. Moreover, the combination of inexpensive substrates and products of more value may lead to the economical sustainability of industrial PHB production.     

Low Concentration of Copper Inhibits Colonization of Soil by the Arbuscular Mycorrhizal Fungus <Emphasis Type="Italic">Glomus intraradices</Emphasis> and Changes the Microbial Community Structure

Common agricultural practices result in accumulation of copper in agricultural soils worldwide. The effect of bioavailable copper ([Cu]_bio) on colonization of soil by the AM fungus Glomus intraradices and other soil microorganisms was investigated in microcosms containing copper-amended soil. To avoid indirect effects through the plant, copper was only added to root-free microcosm compartments. [Cu]_bio was measured using a Pseudomonas fluorescens biosensor strain. In the range of 0–1.5 μg g⁻¹ [Cu]_bio, a log–log linear relationship between added copper and [Cu]_bio was found. Microbial colonization of the root-free compartment was evaluated by whole-cell fatty acid analysis (WCFA) and amplified rDNA restriction analysis (ARDRA). The WCFA analysis showed that the AM fungus soil colonization was severely inhibited by Cu with a 50% reduction of mycorrhizal growth at 0.26 μg g⁻¹ [Cu]_bio. The growth of other main microbial groups was not significantly affected by copper. However, ARDRA analysis showed a very strong effect of copper on the bacterial community composition probably caused by an increased proportion of Cu-resistant bacteria. Our results suggest that problems with plant yield may arise when converting slightly copper-contaminated soils to land uses such as low-input and sustainable agriculture that are dependent on AM fungal symbiosis.     

A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA

An improved method for the electrotransformation of wild-type Corynebacterium glutamicum (ATCC 13032) is described. The two crucial alterations to previously developed methods are: cultivation of cells used for electrotransformation at 18 °C instead of 30 °C, and application of a heat shock immediately following electrotransformation. Cells cultivated at sub optimal temperature have a 100-fold improved transformation efficiency (10⁸ cfu μg⁻¹) for syngeneic DNA (DNA isolated from the same species). A heat shock applied to these cells following electroporation improved the transformation efficiency for xenogeneic DNA (DNA isolated from a different species). In combination, low cultivation temperature and heat shock act synergistically and increased the transformation efficiency by four orders of magnitude to 2.5 × 10⁶ cfu μg⁻¹ xenogeneic DNA. The method was used to generate gene disruptions in C. glutamicum. Received: 26 March 1999 / Received revision: 9 June 1999 / Accepted: 11 June 1999     

The viability to a wall shear stress and propagation of <Emphasis Type="Italic">Bifidobacterium longum</Emphasis> in the intensive membrane bioreactor

Bifidobacterium longum grew at 65 L pilot scale of the membrane bioreactor (MBR), externally fitted with ceramic membrane (0.7 m²). Cell mass at the MBR reached 22.18 g L⁻¹ as dry cell weight in 12 h, which is 8.44 times higher than cell mass attained at the vial culture. The growth rate in the vial culture was μ = 0.385 h⁻ and at the batch culture was μ = 1.13 h⁻ in the exponential period and μ = 0.31 h⁻¹ in the stationary period. In the fed-batch mode was μ = 1.102 h⁻¹ for 6 h with inoculation and declined to μ = 0.456 h⁻¹ with feeding of feed medium. The growth rate at the MBR was μ = 0.134 h⁻¹. The number of viable cells was 6.01 × 10¹² cfu L⁻¹ at the batch culture, but increased to 1.15 × 10¹⁴ cfu L⁻¹ at the MBR culture. The specific growth rate of viable cell number (colony-forming units per liter, per hour) improved by 6.01 times from the batch to the MBR culture. The wall shear stress mainly generated by the pump, and the membrane incorporated into the MBR was controlled during the cultivation at the MBR. The viability of B. longum declined to under 10% in the first 2 weeks of the 4-week stability test (40°C) as B. longum was exposed to over wall shear stress 713 Pa, but the viability improved to 30–40% in wall shear stress of 260 Pa or STR culture. The loss in the cell viability can be saved by managing with wall shear stress during the cultivation at the MBR.     

Plasma free amino acid kinetics in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) using a bolus injection of <Superscript>15</Superscript>N-labeled amino acids

To gain insight into the metabolic design of the amino acid carrier systems in fish, we injected a bolus of ¹⁵N amino acids into the dorsal aorta in mature rainbow trout (Oncorhynchus mykiss). The plasma kinetic parameters including concentration, pool size, rate of disappearance (R _d), half-life and turnover rate were determined for 15 amino acids. When corrected for metabolic rate, the R _d values obtained for trout for most amino acids were largely comparable to human values, with the exception of glutamine (which was lower) and threonine (which was higher). R _d values ranged from 0.9 μmol 100 g⁻¹ h⁻¹ (lysine) to 22.1 μmol 100 g⁻¹ h⁻¹ (threonine) with most values falling between 2 and 6 μmol 100 g⁻¹ h⁻¹. There was a significant correlation between R _d and the molar proportion of amino acids in rainbow trout whole body protein hydrolysate. Other kinetic parameters did not correlate significantly with whole body amino acid composition. This indicates that an important design feature of the plasma-free amino acids system involves proportional delivery of amino acids to tissues for protein synthesis.     

Effects of spironolactone and RU486 on gene expression and cell proliferation after freshwater transfer in the euryhaline killifish

We have explored the possible mechanisms by which mineralocorticoid (MR) and glucocorticoid (GR) receptors regulate the response to freshwater transfer in the gills of the euryhaline killifish Fundulus heteroclitus. Killifish were implanted with RU486 (GR antagonist) or spironolactone (MR antagonist) at doses of 0.1–1.0 mg g⁻¹, and subsequently transferred from 10‰ brackish water to freshwater. Compared to brackish water sham fish, mRNA expression of CFTR and NKCC1 decreased in the gills of sham fish transferred to freshwater, whereas Na⁺,K⁺–ATPase α_1a mRNA expression and α protein abundance, as well as cell proliferation (detected using BrdU) increased. Spironolactone inhibited the normal increase in cell proliferation and Na⁺,K⁺-ATPase expression after freshwater transfer. RU486 increased plasma cortisol levels and may have slightly inhibited Na⁺,K⁺–ATPase activity, but did not change α _1a expression. RU486 had no effect on cell proliferation in the non-lamellar region of the gills, but increased proliferation in the lamellar region. Neither antagonist inhibited the suppression of CFTR or NKCC1 expression after freshwater transfer. Glucocorticoid receptor expression was reduced in all sham and antagonist treatments compared to untreated controls, but no other consistent differences were observed. The effects of spironolactone suggest that MR is important for regulating ion transport in killifish gills after freshwater transfer.     

Hydrophobic response of the fungus <Emphasis Type="Italic">Rhinocladiella similis</Emphasis> in the biofiltration with volatile organic compounds with different polarity

Rhinocladiella similis biodegraded volatile organic compounds (VOCs) of different polarity in gas-phase biofilters. Elimination capacities, (EC) of 74 g_hexane m⁻³ h⁻¹, 230 g_ethanol m⁻³ h⁻¹, 85 g_toluene m⁻³ h⁻¹ and 30 g_phenol m⁻³ h⁻¹ were obtained. EC values correlated with the solubility of the VOCs. R. similis grown with n-hexane or ethanol in biofilters packed with Perlite showed that the surface hydrophobicity was higher with n-hexane than ethanol. The hydrophobin-like proteins extracted from the mycelium produced with n-hexane (15 kDa) were different from those in the ethanol biofilter (8.5 kDa and 7 kDa).