Prodigiosin formation by Serratia marcescens in a chemostat

In a study of the control of metabolite formation, prodigiosin production by Serratia marcescens was used as a model. Specific production rates of prodigiosin formation were determined using batch culture technique. Sucrose as carbon source and NH₄NO₃ as nitrogen source resulted in a specific production rate of 0.476 mg prodigiosin (g cell dry weight)⁻¹ h⁻¹. Prodigiosin formation and productivity was inversely correlated to growth rate when the bacterium was grown under carbon limitation on a defined medium in a chemostat culture. The maximum specific growth rate (μ_max) was 0.54 h⁻¹ and prodigiosin was formed in amounts over 1 mg l⁻¹ up to a growth rate (μ) of 0.3 h⁻¹ at steady state conditions. At a dilution rate of 0.1 h⁻¹ growth at steady state with carbon and phosphate limitation supported prodigiosin formation giving a similar specific yield [1.17 mg prodigiosin (g cell dry weight)⁻¹ and 0.94 mg g⁻¹, respectively], however, cells grown with nitrogen limitation [(NH₄)₂SO₄] did not form prodigiosin. Productivity in batch culture was 1.33 mg l⁻¹ h⁻¹ as compared to 0.57 mg l⁻¹ h⁻¹ in the chemostat.     

Benthic organic carbon influences denitrification in streams with high nitrate concentration

1. Anthropogenic activities have increased reactive nitrogen availability, and now many streams carry large nitrate loads to coastal ecosystems. Denitrification is potentially an important nitrogen sink, but few studies have investigated the influence of benthic organic carbon on denitrification in nitrate‐rich streams. 2. Using the acetylene‐block assay, we measured denitrification rates associated with benthic substrata having different proportions of organic matter in agricultural streams in two states in the mid‐west of the U.S.A., Illinois and Michigan. 3. In Illinois, benthic organic matter varied little between seasons (5.9–7.0% of stream sediment), but nitrate concentrations were high in summer (>10 mg N L⁻¹) and low (<0.5 mg N L⁻¹) in autumn. Across all seasons and streams, the rate of denitrification ranged from 0.01 to 4.77 μg N g⁻¹ DM h⁻¹ and was positively related to stream‐water nitrate concentration. Within each stream, denitrification was positively related to benthic organic matter only when nitrate concentration exceeded published half‐saturation constants. 4. In Michigan, streams had high nitrate concentrations and diverse benthic substrata which varied from 0.7 to 72.7% organic matter. Denitrification rate ranged from 0.12 to 11.06 μg N g⁻¹ DM h⁻¹ and was positively related to the proportion of organic matter in each substratum. 5. Taken together, these results indicate that benthic organic carbon may play an important role in stream nitrogen cycling by stimulating denitrification when nitrate concentrations are high.     

Influence of the concentrations of d-xylose and yeast extract on ethanol production by Pachysolen tannophilus

To determine the most favorable conditions for the production of ethanol by Pachysolen tannophilus, this yeast was grown in batch cultures with various initial concentrations of two of the constituents of the culture medium: d-xylose (s_o), ranging from 1 g·l⁻¹ to 200 g·l⁻¹, and yeast extract (l_o), ranging from 0 g·l⁻¹ to 8 g·l⁻¹. The most favorable conditions proved to be initial concentrations of S_o=25 g·l⁻¹ and l_o=4 g·l⁻¹, which gave a maximum specific growth rate of 0.26 h⁻¹, biomass productivity of 0.023 g·l⁻¹·h⁻¹, overall biomass yield of 0.094 g·g⁻¹, specific xylose-uptake rate (q_s) of 0.3 g·g⁻¹·h⁻¹ (for t=50 h), specific ethanol-production rate (q_E) of 0.065 g·g⁻¹·h⁻¹ and overall ethanol yield of 0.34 g·g⁻¹; q_s values decreased after the exponential growth phase while q_E remained practically constant.     

In vitro Clonal Propagation and Organogenesis in Spilanthes acmella (L) Murray: A Herbal Pesticidal Plant of North-East India

Multiple shoots were regenerated in MS medium using different concentrations of BAP and Kn and different combinations of BAP with IAA, NAA and IBA. Highest multiplication of shoots was obtained with BAP (0.75 mg l^?1) with 28.4 shoots per explant after 60 days of culture. Shoots rooted best on IBA (0.5 mg l^?1), numbering 48.8 per explant. Organogenesis was maximum in callus cultured on MS medium supplemented with BAP (2.0 mg l^?1) and IAA (1.0 mg l^?1).     

Effects of feed sugar concentration on continuous ethanol fermentation of cheese whey powder solution (CWP)

Cheese whey powder (CWP) solution with different CWP or sugar concentrations was fermented to ethanol in a continuous fermenter using pure culture of Kluyveromyces marxianus (DSMZ 7239). Sugar concentration of the feed CWP solution varied between 55 and 200 g l⁻¹ while the hydraulic residence time (HRT) was kept constant at 54 h. Ethanol formation, sugar utilization and biomass formation were investigated as functions of the feed sugar concentration. Percent sugar utilization and biomass concentrations decreased and the effluent sugar concentration increased with increasing feed sugar concentrations especially for the feed sugar contents above 100 g l⁻¹. Ethanol concentration and productivity (DP) increased with increasing feed sugar up to 100 g l⁻¹ and then decreased with further increases in the feed sugar content. The highest ethanol concentration (3.7%, v v⁻¹) and productivity (0.54 gE l⁻¹ h⁻¹) were obtained with the feed sugar content of 100 g l⁻¹ or 125 g l⁻¹. The ethanol yield coefficient (Y_P/S) was also maximum (0.49 gE gS⁻¹) when the feed sugar was between 100 and 125 g l⁻¹. The growth yield coefficient (Y_X/S) decreased steadily from 0.123 to 0.063 gX gS⁻¹ when the feed sugar increased from 55 to 200 g l⁻¹ due to adverse effects of high sugar contents on yeast growth. The optimal feed sugar concentration maximizing the ethanol productivity and sugar utilization was between 100 and 125 g l⁻¹ under the specified experimental conditions.     

SODIUM: SOME EFFECTS ON BLUEGREEN ALGAL GROWTH1

The growth of heterocystous bluegreen algae in various concentrations of sodium, was examined in axenic culture as well as in situ studies. Anabaena cylindrica Lemm. with no Na⁺ added, suffered from decreased rates of acetylene reduction, ¹⁴C, assimilation, excretion of organic C as well as lower concentrations of chlorophyll a and particulate organic C compared to cultures supplied with 5, 10, and 50 mg Na⁺·l⁻¹ Sodium deficient algae released, extracellularly a higher percentage of previously fixed C as organic C. No differences in any parameter measured were demonstrable among cultures grown with 5, 10, and 50 mg Na⁺·l⁻¹ High nitrate concentrations (20 mg NO₃⁻·l⁻¹) resulted in decreased rates of acetylene reduction and heterocyst numbers in. Na sufficient, and Na deficient cultures: however, decreased, cellular Na content at high NO₃⁻ levels occurred only in N deficient, cultures. Higher percentages of excreted organic C occurred with increasing NO₃⁻ concentrations in Na deficient cultures. Sodium enrichment of natural bluegreen populations with the addition of 50, 100, and 200 mg Na⁺·l⁻¹ elicited neither a stimulatory nor an inhibitory response in photosynthetic C fixation. In contrast, the addition of small amounts of Na⁺ (5 mg·l⁻) resulted in increased C fixation. However, since the Na. concentration of the lake water, at ca. 5 mg Na⁺·l⁻¹, was sufficient for growth of the bluegreens present, sodium, is not assumed to be limiting under most natural conditions. No increase in in situ acetylene reduction rates occurred with additions of sodium.     

Primary Production and Effects of Enrichment with Nitrate and Phosphate on Phytoplankton in the Barra Bonita Reservoir (State of São Paulo,Brazil)

Primary productivity of the phytoplankton was evaluated by the ¹⁴C and dissolved oxygen methods in December 1981 at the Barra Bonita Reservoir (São Paulo State, Brazil). The primary production varied between 0.17 to 14.51 mg C m⁻³h⁻¹ at 4 and 0 m depth, respectively. About 57 to 94% of the photosynthetic activity was due to phytoplankton > 50 μm. The highest value of assimilation rate (3.36 mg C mg Chl⁻¹h⁻¹) was found in the surface water. Dissolved nutrient concentrations were very high and consisted mainly of nitrate. Light penetration was low, the aphotic zone accounting for about 90% of the water column. Enrichment with nitrate and phosphate showed that both N and P stimulated the production of biomass (chlorophyll a), mainly due to the addition of phosphate. The enrichment experiment also indicated that phosphate addition has a significant stimulatory effect on the growth of Melosira sp. The limiting effect of light penetration on photosynthetic activity is more severe than that of nutrients.     

Denitrification by Alcaligenes denitrificans in a closed rotating biological contactor

Biological denitrification using a pure culture of Alcaligenes denitrificans was investigated in a closed rotating biological contactor, which operated with a hydraulic retention time of 2 h, a carbon/nitrogen ratio of 2:1, with a dissolved O₂ concentration below 6 mg l^–1 and under three different phosphate concentrations. Alcaligenes denitrificans was not repressed by O₂ limitation and the removal of nitrate was about 30% more efficient at the intermediate phosphate concentration (20 mg P l^–1).     

Assessment of two techniques for measuring nitrification rates in aerated slurries of seagrass-bed sediments

Two methods were assessed to measure rates of nitrification in aerated slurries of seagras-bed sediments; (1) monitoring ammonium concentrations in samples with and without nitrapyrin ((2-chloro-6-(trichloromethyl) pyridine) treatment and (2) monitoring the accumulation of nitrate plus nitrite. Nitrifying bacteria were inhibited completely after 14 days exposure to nitrapyrin at 5 mg⁻¹, but the effect was poor over shorter periods, probably owing to limited diffusion of the inhibitor. Hence, nitrapyrin was not a satisfactory inhibitor to use in these sediments. Nitrification rates could not be measured by monitoring the concentrations of nitrate plus nitrite, because nitrate reduction occurred simultaneously with nitrification, even though the slurries contained from 3.5 to 8.9 mg O₂ l⁻¹.     

Successional changes in habitat caused by sedimentation in navigation pools

Effects of Cu and Zn through food as well as through the water on the growth and emergence of the young larvae of Epeorus latifolium (Ephemeroptera), were investigated using an indoor model stream. The critical lowest concentrations of Cu which have significant effects on the growth of the larvae were between 10 and 15 µg l^-1 Cu. Growth and emergence of the larvae fed on the algae (diatoms) which accumulated more than 1000 µg Cu g^-1 (dry wt.) were significantly impaired. Growth of the mayfly larvae exposed to 100 or 300 µg l^-1 Zn ceased after the second week, and all died before emergence. At 30 µg l^-1 Zn, the growth rate decreased gradually and many larvae died before emergence. The molt interval also nearly doubled that of the control at these concentrations. Growth and emergence of the mayfly fed the algae which accumulated more than 2000 µg g^-1 of Zn were significantly affected.     

Cadmium induction of metallothionein synthesis in Mytilus galloprovincialis

1. The accumulation and subcellular distribution of cadmium and resulting induction of metallothionein has been studied in mussels, Mytilus galloprovincialis, exposed to 400 μg Cd 1⁻¹.2. Cadmium accumulated by mussels was primarily associated with metallothionein: a maximum four-fold increase in concentrations of the metal-binding protein (3 mg g⁻¹ to 12 mg g⁻¹) was observed after 30 days.3. Results for M. galloprovincialis confirm the importance of metallothionein in the detoxification of intracellular cadmium and indicate that induction rates are identical to those observed in the closely-related M. edulis.4. Intercomposition of data between these species may therefore be valid when employing metallothionein assays to determine sub-lethal response to cadmium contamination.     

The effect of exogenous prolactin on lactation performance of first-litter sows given protein-deficient diets during the first pregnancy

Twenty-four sows were used to study the effects of dietary protein restriction during pregnancy and exogenous porcine prolactin (pPRL) during late pregnancy and throughout lactation on lactation performance. Eight sows were given a protein-adequate diet containing 179 g crude protein (CP)kg⁻¹ during their first pregnancy while the remaining 16 sows received the same amount of a diet containing 80 g CP kg⁻¹. Eight of the sows given 80 g CP kg⁻¹ during pregnancy were injected with 15 mg pPRL i.m. twice daily at 08:00 and 20:00 between day (d) 102.1 (±0.3) of pregnancy and weaning after their first lactation. Pregnant sows offered the low protein diet gained significantly less body weight during gestation and tended to eat less in the subsequent lactation than sows given the protein-adequate diet. Dietary protein had no significant effect on birth weight, milk yield, milk composition or growth rate of the litter during lactation. Neither dietary protein intake during pregnancy nor exogenous prolactin affected the concentrations of plasma glucose, serum insulin, urea or non-esterified fatty acid (NEFA) during lactation. The concentration of lactose in plasma during lactation was unaffected by treatment, but at d 105 of pregnancy, plasma lactose levels were greater in sows which had received exogenous prolactin (32.4 vs. 6.2 mg l⁻¹, P < 0.05). The concentrations of RNA and DNA in mammary tissue biopsies were unaffected by either dietary protein or pPRL. The concentration of RNA and DNA increased between d 70 and 90 from 0.66 to 2.77 mg g⁻¹ and from 0.54 to 1.19 mg g⁻¹, respectively. Thereafter, RNA increased to 4.40 mg g⁻¹ at d 14 of lactation whilst DNA concentration remained at a similar level of 0.90 mg g⁻¹.Milk yield of sows between d 5 and 8 and between d 19 and 22 of lactation was reduced from 8.36 to 7.00 kg day⁻¹ and from 10.74 to 8.22 kg day⁻¹, respectively, in sows given pPRL. The protein content of colostrum from sows treated with pPRL was reduced from 164 to 104 g kg⁻¹ whereas the fat content increased from 47 to 127 g kg⁻¹. These results indicate that the administration of exogenous pPRL during late pregnancy and throughout lactation initiated lactogenesis prematurely and reduced subsequent milk yield during established lactation.     

Enhancement of polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by optimization of medium compositions and feeding of sucrose

A strategy that optimization of medium compositions for maximum biomass followed by feeding of sucrose for maximum polysaccharide synthesis was developed for enhancing polysaccharide production in suspension culture of protocorm-like bodies (PLBs) of Dendrobium huoshanense C.Z. Tang et S.J. Cheng. In growth stage, the original half-strength MS medium was optimized with carbon sources, nitrogen sources and metal ion combinations. The effects of different carbon sources on PLBs growth were remarkable and sucrose at 35 g l⁻¹ was the most suitable. Sole nitrate nitrogen of 30 mmol l⁻¹ was the best for PLBs growth. Metal ions (Ca²⁺, Fe²⁺, Mn²⁺ and Zn²⁺) showed different influences on PLBs growth. The optimal concentration of Ca²⁺, Fe²⁺, Mn²⁺ and Zn²⁺ was 4.5 mmol l⁻¹, 0.1 mmol l⁻¹, 0.5 mmol l⁻¹ and 0.06 mmol l⁻¹, respectively. In the optimized medium (sucrose, nitrate, Ca²⁺, Fe²⁺, Mn²⁺ and Zn²⁺ concentration as described above, the other component concentration seen in half-strength MS), 33.9 g DW l⁻¹ PLBs were harvested after 30 days of culture and biomass increase was improved 245% as compared with that in the original medium. In production stage, polysaccharide synthesis was significantly improved by the feeding sucrose. The maximum polysaccharide production (22 g l⁻¹) was obtained in the case of 50 g l⁻¹ sucrose feeding at day 30 of culture, which was about 109-fold higher than that in the original medium without feeding of sucrose.     

Food consumption of Sarotherodon mossambicus (Trewaves) exposed to sublethal concentration of diammonium phosphate

S. mossambicus was exposed to toxic and sublethal concentrations of the fertilizer diammonium phosphate (0.2 to 1.0 g l^–1). Mortality, food utilization and growth were studied. At a concentration of 0.6 g l^–1 DAP, 100% mortality was observed within 96 h; no mortality occurred at 0.5 g l^–1; LC50 was 0.55 g l^–1. Rearing the fish in increasing sublethal concentrations of DAP, it was found that the feeding rate decreased from 25.4 mg g^–1 fish^–1 d^–1 (fish reared in DAP-free water) to 10.1 mg g^–1 d^–1 at the highest sublethal concentration (0.5 g l^–1). Growth rate was drastically reduced. At high sublethal concentrations of DAP, the fish lost reserve energy, in addition to the energy obtained from food intake for survival, as a result of increased swimming activity and opercular beats.     

Different nitrogen sources and growth responses of Spirulina platensis in microenvironments

Spirulina platensis was cultivated, in comparative studies, using several sources of nitrogen. The standard source used (sodium nitrate) was the same as that used in the synthetic medium Zarrouk, whereas the alternative nitrogen sources consisted of ammonium nitrate, urea, ammonium chloride, ammonium sulphate or acid ammonium phosphate. The initial nitrogen concentrations tested were 0.01, 0.03 and 0.05 M in an aerated photobioreactor at 30 °C, with an illuminance of 1900 lux, and 12 h-light/12 h-dark photoperiod over a period of 672 h. Maximum biomass was produced in medium containing sodium nitrate (0.01–0.03–0.05 M), followed by ammonium nitrate (0.01 M) and urea (0.01 M). The final biomass concentrations were 1.992 g l^–1 (0.03 M sodium nitrate), 1.628 g l^–1 (0.05 M sodium nitrate), 1.559 g l^–1 (0.01 M sodium nitrate), 0.993 g l^–1 (0.01 M ammonium nitrate) and 0.910 g l^–1 (0.01 M urea). This suggested that it is possible to utilize nitrogen sources other than sodium nitrate for growing S. platensis, in order to decrease the production costs of scaled up projects.     

The gills as an important uptake route for the essential nutrient iron in freshwater rainbow trout Oncorhynchus mykiss

Short-term (3h) acquisition of iron (16 nmol ⁵⁹FeCl₃ l⁻¹) from oxic, alkaline fresh water was assessed in rainbow trout Oncorhynchus mykiss in the presence or absence of a range of iron chelators, all of which had differing binding affinities for ferric iron [100 μmol l⁻¹ of desferrioxamine (DFO), Log₁₀K₁ 32·5; citric acid Log₁₀K₁ 11·9; nitrilotriacetic acid (NTA) Log₁₀K₁ 15·9, CP20 and CP94 (Log₁₀K₁ > 30), as well as humic acid (HA), Log₁₀K₁ 5·04, 5 mg l⁻¹]. In the absence of chelators (control conditions) O. mykiss acquired iron from the water under laboratory lights (wavelength range of the lights 440–650 nm, peak intensity 548–626 nm) via the gill. In these conditions iron uptake onto the gill had a maximum transport capacity (J_max) of 11·2 pmol Fe g⁻¹ h⁻¹ (gill organ mass) and a K_m of 21·3 nmol Fe l⁻¹ h⁻¹. Furthermore, there were two components to iron accumulation into the carcass of these fish, a slow rate of aqueous iron uptake at low concentrations (6–24 nmol Fe l⁻¹), followed by a faster rate of uptake at higher iron concentrations (48–96 nmol Fe l⁻¹), suggesting that the rate-limiting step of iron uptake at low iron concentrations is the apical entry step. O. mykiss also acquired iron in the presence of HA, although the majority of the other chelators prevented iron uptake. Ultraviolet light (354 nm) treatment of Fe-DFO increased iron bioavailability. Results suggest that rainbow trout are able to access either the predicted very low concentrations (picomolar) of ferrous iron present in fresh water or the ferric oxide complexes present in oxic environments. The iron uptake rate measured (0·75 pmol g⁻¹ h⁻¹) would be sufficient to provide a substantial proportion (c. 85%) of the daily iron requirements of growing salmonid fry.     

Evaluation of distant carbon sources in biosurfactant production by a gamma ray-induced Pseudomonas putida mutant

Pseudomonas putida 33 wild strain, subjected to gamma ray mutagenesis and designated as P. putida 300-B mutant was used as microbial rhamnolipid-producer by using distant carbon sources (viz. hydrocarbons, waste frying oils ‘WFOs’, vegetable oil refinery wastes and molasses) in the minimal media under shake flask conditions. The behavior of glucose as co-substrate and growth initiator was examined. The 300-B mutant strain showed its ability to grow on all the substrates tested and produced rhamnolipid surfactants to different extents however; soybean and corn WFOs were observed to be preferred carbon sources followed by kerosene and paraffin oils, respectively. The best cell biomass (3.5 g l⁻¹) and rhamnolipids yield (4.1 g l⁻¹) were obtained with soybean WFO as carbon source and glucose as growth initiator under fed-batch cultivation showing an optimum specific growth rate (μ) of 0.272 h⁻¹, specific product yield (q_p) of 0.318 g g⁻¹ h and volumetric productivity (P_V) of 0.024 g l⁻¹ h. The critical micelle concentration of its culture supernatant was observed to be 91 mg rhamnolipids l⁻¹ and surface tension as 31.2 mN m⁻¹.     

An efficient micropropagation protocol for Citrus jambhiri Lush. and assessment of clonal fidelity employing anatomical studies and RAPD markers

Citrus jambhiri (rough lemon) is considered a major rootstock source for a number of Citrus species. A simple method for micropropagation from nodal segments is reported. Nodal segments of C. jambhiri were inoculated on MS medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP), kinetin, and N6-(2-isopentenyl) adenosine (2iP). Maximum multiple shoot regeneration response (75?%) was observed with BAP at 3?mg?l^?1. Shoots were multiplied for 30?d on fresh medium with similar composition. A total of 67?% of the cultures showed multiplication with the optimum number of shoots (4.02) and height of shoots (1.81?cm) with BAP (3?mg?l^?1) alone. Maximum rooting response (87?%) was observed with naphthaleneacetic acid at 0.5?mg?l^?1. Transverse sections of shoot stems obtained in vivo (sampled from seedlings) and in vitro (regenerated from nodal segments), showed similar anatomies. Randomly amplified polymorphic DNA analysis confirmed that all the regenerated plants were genetically identical to their donor plant, suggesting absence of detectable genetic variation in the regenerated plantlets.     

Optimising the foliar uptake of zinc oxide nanoparticles: Do leaf surface properties and particle coating affect absorption?

Foliar absorption of zinc (Zn) is limited by several barriers, the first of which is the leaf cuticle. In this study, we investigated the absorption of Zn from Zn oxide nanoparticles (ZnO-NPs) in wheat (Triticum aestivum cv Gladius) and sunflower (Helianthus annuus cv Hyoleic 41) to determine the importance of NP surface coating for Zn absorption. Fourier transform infrared (FTIR) spectroscopy showed a higher polysaccharide content in the wheat cuticle than sunflower, indicated by a more pronounced glycosidic bond at 1020 cm⁻¹, but wax and cutin content were similar. Scanning electron microscopy (SEM) revealed that t richome density was twice as high in wheat (3600 ± 900 cm⁻²) as in sunflower (1600 cm⁻²) and stomatal density four times higher in sunflower (6400 ± 800 cm⁻² in wheat and 22 900 cm⁻² in sunflower). Suspensions of ZnO-NPs with coatings of different hydrophobicity were applied to leaves to compare Zn absorption using X-ray fluorescence microscopy (XFM) and inductively coupled plasma mass spectroscopy (ICP-MS). Absorption of Zn was similar between wheat and sunflower when Zn was applied at 1000 mg Zn l⁻¹, but much less Zn was absorbed from all ZnO products than from soluble Zn fertiliser. Particle coating did not affect Zn absorption, but it may facilitate particle adhesion to leaves, providing a longer-term source of resupply of Zn ions to the leaves. Differences in leaf surface characteristics did not affect Zn absorption, indicating that the cuticle is the main pathway of absorption under these conditions.     

Some Physical and Chemical Properties of Water from an Undisturbed Tropical Catchment Area (Guma Dam,Sierra Leone)

The water had a very low content of total suspended solids (<1 mg l⁻¹), but its transparency was high (5–8 Hazen units). The concentration of Mg⁺⁺ and Ca⁺⁺ were below 1 mg l⁻¹, but that of SiO₂ was 5.3 mg l⁻¹. Free CO₂ was abundant (15.3 mg l⁻¹) and the pH was low (<6.5). These conditions were accompanied by low concentrations of PO₄-P, total iron and NO₃-N. The DO content of the water was always below 80 %, but the chloride concentration (8.4 mg l⁻¹) seems relatively high for this type of freshwaters. It is, however, much lower than that known for arid areas. It is concluded that the geology and vegetation dominate the chemistry of the water from the Guma Dam catchment area. The influence of man is acknowledged only in as far as he has protected the catchment area.