The role of nitrate reductase in the degradation of the explosive RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) by <Emphasis Type="Italic">Penicillium</Emphasis> sp. AK96151

In liquid culture on a defined growth medium, Penicillium sp. AK96151 efficiently degraded the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX, hexogen), causing > 80 % disappearance after 10 d. RDX degradation was reduced to a basal level (< 15 % degraded after 10 d) by the presence of > 150 μM ammonium ions or when the molybdenum component of the medium was replaced by sodium tungstate. An equivalent effect of ammonium, molybdenum and tungsten was observed in protoplasts of this fungus assayed for nitrate reductase activity. This enzyme was not inhibited by RDX itself. The involvement of a nitrate reductase in RDX degradation by Penicillium has practical implications for bioremediation strategies which are discussed.     

Coexisting <Emphasis Type="Italic">Curtobacterium</Emphasis> Bacterium Promotes Growth of White-Rot Fungus <Emphasis Type="Italic">Stereum</Emphasis> sp.

White-rot basidiomycetes are the main decomposers of woody biomass in forest ecosystems. Little is known, however, about the interactions between white-rot fungi and other microorganisms in decayed wood. A wood-rotting fungus, Stereum sp. strain TN4F, was isolated from a fruit body, and its coexisting cultivable bacteria were isolated from its substrate; natural white-rot decayed wood. The effects of bacteria on fungal growth were examined by confrontational assay in vitro. A growth-promoting bacterium for this Stereum strain was identified as Curtobacterium sp. TN4W-19, using 16SrRNA sequencing. A confrontational assay revealed that Curtobacterium sp. TN4W-19 significantly promoted the mycelial growth of Stereum sp. TN4F in the direction of the bacterial colony, without direct contact between the mycelium and bacterial cells. This is the first report of a positive interaction between a white-rot fungus and a coexisting bacterial strain in vitro.     

Potassium-induced inhibition of nitrogen and phosphorus metabolism as a strategy of controlling <Emphasis Type="Italic">Microcystis</Emphasis> blooms

To explore potassium toxicity in Microcystis sp., growth, chlorophyll a, carotenoid and phycocyanin content, uptake of nitrate, phosphate and ammonium and activities of the assimilatory enzymes nitrate reductase, alkaline phosphatase and glutamine synthetase (GS) were studied. Nitrate, phosphate, ammonium uptakes and chlorophyll a and phycocyanin contents decreased with increase in the concentration of potassium, but carotenoid content registered an increase at increasing potassium concentration. Alkaline phosphatase and GS activities followed the trend of inhibition of their respective nutrients, whereas nitrate and nitrate reductase showed negative correlation (p < 0.01). Potassium was found to inhibit the activities of all the assimilatory enzymes in a non-competitive manner. Inhibitions of these parameters support the view that potassium has the potential to regulate Microcystis blooms in an eco-friendly manner.     

Effects of root residues on growth ofPinus radiata seedlings and a mycorrhizal fungus

The height of Pinus radiata Don seedlings grown on soils incorporating different proportions of root tissues (25 and 50%) was shown to be reduced by 20–80%. Seedlings watered with aqueous extracts from soil/root mixtures also showed a significant reduction in growth. This growth-retarding effect was partially overcome by the addition of nutrients or by soil sterilisation. It is postulated that growth retardation was caused not only by nutrient deficiency resulting from the addition of organic matter with a high C/N ratio, but also by phytotoxic substances present in the root tissue. The effect of water extracts of roots from old P. radiata trees on the growth of young P. radiata seedlings and on a mycorrhizal fungus (Rhizopogon sp.) under aseptic conditions was also studied. Extracts from the inner bark of roots caused complete growth inhibition of the mycorrhizal fungus as well as root necrosis and wilting of P. radiata seedlings. A water-soluble substance or substances, toxic both to the roots of P. radiata and to its mycorrhizal fungus, is postulated as the causal agent.     

Phytoplankton Production at Two Stations in Lindåspollene,a Norwegian Land-locked Fjord,and Limiting Nutrients Studied by Two Kinds of Bio-assays

At two stations in a Norwegian land-locked fjord, one more and one less influenced by freshwater run-off, studies were made of the primary production, chlorophyll a concentrations, and the quantity and composition of phytoplankton. Samples were taken for analyses of phosphate, nitrate, and silicate, and temperature and salinity were measured. Two kinds of bio-assays were performed: combinations of phosphate, nitrate, and silicate were added to I, natural water incubated in situ, and II, natural water incubated in the laboratory. Lower salinities and higher concentrations of all nutrients were found at the station most influenced by fresh-water. Although the bio-assays indicated that nitrogen was the main limiting nutrient, the biological parameters showed no consistent differences between the two stations. The similarity is tentatively explained by the growth of nanoplankton algae, partly depending on nutrients other than those analysed.     

BRYOPHYTE LEAF EPIFLORA: AN SEM AND TEM STUDY OF POLYTRICHUM COMMUNE HEDW.

Epiphytes on leaves of Polytrichum commune Hedw. were observed and studied with SEM and TEM. A unicellular green alga, a fungus, and a rod-shaped bacterium were present on both leaf surfaces. Culturing experiments led to the identification of the alga as Chlorococcum sp. and the fungus as Trichoderma viride Lk. ex Fr. (Deuteromycetes). No attempt was made to culture and identify the bacterium. Terminal cells of the leaf lamellae form a ventral pseudo-epidermis which provides a microhabitat for the epiphytes. Fungal hyphae were seen intracellularly as well as penetrating cell walls between lamellar cells. The alga and bacterium were not observed intracellularly. Chlorococcum sp. displayed a distinctive wall morphology which consisted of short (0.3–0.5 μm), blunt papillae. The rod-shaped bacterium may be similar to the Agrobacterium-like isolates reported recently from Polytrichum commune. Host-epiphyte associations relative to potential beneficial and/or pathological aspects of growth require further investigation.     

Dracaena Leaf Proliferosis, a Newly Recorded Disease Affecting Dracaena sanderiana in Egypt

Dracaena leaf proliferosis is a newly reported disease affecting Dracaena sanderiana in Egypt. A cause and effect relationship between this disease and the fungus,Fusarium proliferatum var. minus has been established. In addition to D. sanderiana the fungus was found to be pathogenic, when tested in the laboratory, to several other members of the family Liliaceae. While the in vitro growth of the fungus is optimum at 25 °C, symptom expression is best at 30°C. Twelve fungicides were tested for their in vitro effect on fungal growth. Benlate, Rubigan, Saprol, Cercobin and Vitavax-200 came first on the list and inhibited growth at 2.5, 12.0, 55.0, 75.0, and 94.0 μg/ml, respectively. Although, Benlate was the most effective fungicide in this respect it failed to demonstrate similar effect on disease development when applied to plants artificially inoculated with the fungus. Fungal growth was completely inhibited on PDA medium by a bacterium belonging to Bacillus sp. but when the bacterium at a concentration of 1 × 10¹¹ cell/ml was applied 24 h before, at the same time with, or 24 h after inoculation no control of the disease was achieved. Naturally-infected plants could, however, be freed from infection when subjected to a hot air treatment at 35 ± 5 °C during day time and 25 ± 5° C at night for 3 months.     

Biodegradation of p-nitrophenol by methyl parathion-degrading Ochrobactrum sp. B2

Ochrobactrum sp. B2, a methyl parathion-degrading bacterium, was proved to be capable of using p-nitrophenol (PNP) as carbon and energy source. The effect of factors, such as temperature, pH value, and nutrition, on the growth of Ochrobactrum sp. B2 and its ability to degrade p-nitrophenol (PNP) at a higher concentration (100 mg l⁻¹) was investigated in this study.The greatest growth of B2 was observed at a temperature of 30 °C and alkaline pH (pH 9–10). pH condition was proved to be a crucial factor affecting PNP degradation. Enhanced growth of B2 or PNP degradation was consistent with the increase of pH in the minimal medium, and acidic pH (6.0) did not support PNP degradation. Addition of glucose (0.05%, 0.1%) decreased the rate of PNP degradation even if increased cell growth occurred. Addition of supplemental inorganic nitrogen (ammonium chloride or ammonium sulphate) inhibited PNP degradation, whereas organic nitrogen (peptone, yeast extract, urea) accelerated degradation.     

EFFICIENCY OF GROWTH AND NUTRIENT UPTAKE FROM WASTEWATER BY HETEROTROPHIC,AUTOTROPHIC, AND MIXOTROPHIC CULTIVATION OF CHLORELLA VULGARIS IMMOBILIZED WITH AZOSPIRILLUM BRASILENSE1

Heterotrophic growth of microalgae presents significant economic advantages over the more common autotrophic cultivation. The efficiency of growth and nitrogen, phosphorus, and glucose uptake from synthetic wastewater was compared under heterotrophic, autotrophic, and mixotrophic regimes of Chlorella vulgaris Beij. immobilized in alginate beads, either alone or with the bacterium Azospirillum brasilense. Heterotrophic cultivation of C. vulgaris growing alone was superior to autotrophic cultivation. The added bacteria enhanced growth only under autotrophic and mixotrophic cultivations. Uptake of ammonium by the culture, yield of cells per ammonium unit, and total volumetric productivity of the culture were the highest under heterotrophic conditions when the microalga grew without the bacterium. Uptake of phosphate was higher under autotrophic conditions and similar under the other two regimes. Positive influence of the addition of A. brasilense was found only when light was supplied (autotrophic and mixotrophic), where affinity to phosphate and yield per phosphate unit were the highest under heterotrophic conditions. The pH of the culture was significantly reduced in all regimes where glucose was consumed, similarly in heterotrophic and mixotrophic cultures. It was concluded that the heterotrophic regime, using glucose, is superior to autotrophic and mixotrophic regimes for the uptake of ammonium and phosphate. Addition of A. brasilense positively affects the nutrient uptake only in the two regimes supplied with light.     

Production of the glycopeptide antibiotic A40926 by<Emphasis Type="Italic"> Nonomuraea</Emphasis> sp. ATCC 39727: influence of medium composition in batch fermentation

Nonomuraea sp. ATCC 39727 is a novel actinomycete species and the producer of A40926, a glycopeptide antibiotic structurally similar to teichoplanin. In the present study, a defined minimal medium was designed for Nonomuraea fermentation. The influence of initial phosphate, glucose and ammonium concentrations on antibiotic productivity was investigated in batch fermentation and the effect of glucose limitation was studied in fed-batch fermentation. It was found that low initial concentrations of phosphate and ammonium are beneficial for A40926 production and that productivity is not enhanced during glucose limitation. Furthermore, the initiation of A40926 production was not governed by residual ammonium and phosphate concentrations, although the level of these nutrients strongly influenced A40926 production rates and final titers. Electronic Publication     

Utilization of Hydrocarbons and Vitamin B12 Production by Bacillus badius

The accumulation of vitamin B₁₂ by Bacillus badins grown on hydrocarbon was investigated. The bacterium could assimilate n-alkanes of C₁₁–C₁₈, ethanol, fumarate, α-ketoglutarate and malate. n-Alkanes of C₁₆–C₁₈, were the best for vitamin B₁₂ production. The bacterium utilized well all of the nitrogen sources tested. Above all, ammonium dihydrogen phosphate was the best for the bacteria] growth and vitamin B₁₂ production. Addition of organic nutrients such as malt extract and meat extract, and addition of metal ions such as ferrous and cobalt promoted the growth and vitamin B₁₂ production. Interestingly, vitamin B₁₂ was produced mostly in the supernatant. The cyanoform of the corrinoid predominantly formed in the supernatant would confirm the identity with cobalamin.     

Effect of nitrogen source concentration on curdlan production by Agrobacterium sp. ATCC 31749 grown on prairie cordgrass hydrolysates

The effect of nitrogen source concentration on the production of the polysaccharide curdlan by the bacterium Agrobacterium sp. ATCC 31749 from hydrolysates of prairie cordgrass was examined. The highest curdlan concentrations were produced by ATCC 31749 when grown on a medium containing a solids-only hydrolysate and the nitrogen source ammonium phosphate (2.2 mM) or on a medium containing a complete hydrolysate and 3.3 mM ammonium phosphate. The latter medium sustained a higher level of bacterial curdlan production than the former medium after 144 hr. Biomass production by ATCC 31749 was highest after 144 hr when grown on a medium containing a solids-only hydrolysate and 2.2 or 8.7 mM ammonium phosphate. On the medium containing the complete hydrolysate, biomass production by ATCC 31749 was highest after 144 hr when 3.3 mM ammonium phosphate was present. Bacterial biomass production after 144 hr was greater on the complete hydrolysate medium compared to the solids-only hydrolysate medium. Curdlan yield produced by ATCC 31749 after 144 hr from the complete hydrolysate medium containing 3.3 mM ammonium phosphate was higher than from the solids-only hydrolysate medium containing 2.2 mM ammonium phosphate.     

GROWTH DYNAMICS OF ULVA ROTUNDATA (CHLOROPHYTA) IN A FISH FARM: IMPLICATIONS FOR BIOMITIGATION AT A LARGE SCALE1

Changes in biomass of several macroalgae [Ulva rotundata Bliding; Gracilariopsis longissima (S. G. Gmel.) Steentoft, L. M. Irvine et Farnham; Ulva intestinalis L.; and Cladophora sp.] and marine plants (Zostera noltii and Ruppia cirrhosa) growing naturally in earthen ponds of a fish farm (Acuinova, San Fernando, Southern Spain) were recorded during a year. The farm is mainly devoted to the culture of gilthered seabream (Sparus aurata). The most conspicuous algal species thriving in the ponds was U. rotundata, which reached densities up to 600 g dry mass · m⁻² and produced up to 20.45 g C · m⁻² · d⁻¹. Dissolved nutrients (phosphate and ammonium), tissue nutrient content, and growth rates of this species were estimated during 2001 and 2002. Evidence of natural biomitigation by U. rotundata when water circulates throughout the fish farm is presented. Due to the fish cultivation, both phosphate and ammonium increased as water circulated from the preculture ponds to the postculture ponds. As a consequence, U. rotundata tissue nitrogen (N) and phosphorus (P) increased from algae growing in preculture ponds to algae growing in the outflow channel, so that mean C:N:P ratio varied from 773:57:1 in preculture ponds to 567:64:1 in the outflow channel. Phosphorus limited growth of U. rotundata during the spring. As growth rates increased as a function of tissue P, data were fitted to the Droop equation. From this equation, the estimated maximal growth rate was 0.295 ± 0.041 d⁻¹, the subsistence quota was 0.05 ± 0.01% P of dry mass, and the critical quota was 0.215% P of dry mass. The results suggest that management of the fish farm based on a large-scale integrated mariculture system of fish and macroalgae may increase the total ecological and economic benefits, both for the farm and for the environment.     

Effect of wheat resistance, the parasitoid Aphidius rhopalosiphi, and the entomopathogenic fungus Pandora neoaphidis, on population dynamics of the cereal aphid Sitobion avenae

The influence of wheat (Triticum aestivumL.) resistance, the parasitoid Aphidius rhopalosiphiDe Stephani-Perez (Hymenoptera: Braconidae) and the entomopathogenic fungus Pandora neoaphidis(Remaudière et Hennebert) Humber (Zygomycetes: Entomophthorales) on the density and population growth rate of the cereal aphid Sitobion avenae(F.) (Hemiptera: Aphididae) was studied under laboratory conditions. Partial wheat resistance was based on hydroxamic acids, a family of secondary metabolites characteristic of several cultivated cereals. The partial resistance of wheat cultivar Naofén, the action of the parasitoid and the joint action of the parasitoid and fungus, reduced aphid density. The lowest aphid densities were obtained with the combination of the parasitoid and the fungus, but wheat resistance under these circumstances did not improve aphid control. Significant reductions of population growth rate (PGR) of aphids were obtained with the joint action of wheat resistance and natural enemies. In particular, the combined effects of parasitoids and fungi showed significantly lower PGR than the control without natural enemies in both wheat cultivars. Our results support the hypothesis that wheat resistance and the utilization of biological control agents could be complementary strategies in an integrated pest management program against cereal aphids.     

Bioassay for phosphate demand in phytoplankton from acidified lakes: Lake Njupfatet,an example of phosphate deficiency induced by liming

A bioassay was developed, involving steady-state ATP level determinations, for estimation of phosphate demand and deficiency in natural phytoplankton communities. The studies were performed on phytoplankton from the moderately acidified Lake Njupfatet in central Sweden before and after liming. Phytoplankton samples from in situ enclosure experiments with low-dose enrichments of nitrate and phosphate and removal of large (> 100 µm) zooplankton and from the lake water were collected. The phytoplankton were concentrated by through-flow centrifugation and post-cultured in the laboratory with or without the addition of phosphate. A relative increase in the ATP:chlorophyll a ratio after the phosphate treatment as compared to samples without phosphate enrichment was found to be a highly reproducible indicator of phosphate deficiency in the natural phytoplankton population. In contrast, the absolute ATP:chlorophyll a ratio varied substantially between different sampling occasions. No phosphate deficiency was detected in phytoplankton from the acidic lake or from fertilized in situ enclosures. However, phytoplankton from in situ enclosures without added nutrients showed evidence of phosphate limitation after 21 days incubation. Also, the phytoplankton community developed a significant phosphate deficiency the summer after lake liming. The results from the ATP analyses are compared with chemical data of the lake water, phytoplankton community structure and phosphatase activities in the lake before and after liming. The average total biomass of phytoplankton and the average Tot-P measured during May to September decreased with appr. 30% after liming while Tot-N was essentially unaffected and the phosphatase activities increased by 1000–2000%.     

The Effect of Chitinase on Didymella applanata, the Causal Agent of Raspberry Cane Spur light

In vitro and in vivo studies were conducted to assess the efficacy of the two microbial chitinases Chi I (from Streptomyces sp.) and Chi II (from Serratia marcescens) on Didymella applanata (Niessl.) Sacc., the fungus which causes spur blight of raspberry (Rubus idaeus L.). D. applanata was isolated from canes of diseased raspberries in a plantation in Novosibirsk, Russia. In vitro, the effective concentration of Chi I that reduced the growth of D. applanata was 0.4 U/ml (p = 0.05), but Chi II had no influence on the growth of the fungus in medium. In inoculation experiments on raspberry canes, both chitinases at the rate 0.5 U/ml reduced fungal development. In plantation where canes were inoculated after spraying with chitinase, fruiting bodies of fungus failed to form in all enzyme treatments, whereas a significant number of these fungal fruiting bodies (12.8 per cm²) developed in control treatments lacking chitinases spraying. The chitinases reduced the size of lesions and limited the infection of internal tissues of canes. Field testing of Chi I under natural conditions showed a significant suppression of the independent spur blight. These studies form the basis for further evaluation of ecologically benign control measures for raspberry spur blight.     

Effects of photon flux density and agricultural fertilizers on the development of <Emphasis Type="Italic">Sarcothalia crispata</Emphasis> tetraspores (Rhodophyta,Gigartinales) from the Strait of Magellan,Chile

Tetraspores of Sarcothalia crispata from San Juan Bay, Strait of Magellan, Chile, were cultivated under different combinations of photon flux densities and agricultural fertilizers in the laboratory. In the experiment, the S. crispata specimens were cultured in combinations of different photon flux densities (50, 100, 150 μmol photons m^-2 s^-1) and enriched seawater solutions (sodium nitrate + monocalcium phosphate, urea + monocalcium phosphate, ammonium nitrate + monocalcium phosphate), always adjusting the N and P concentrations to 10 and 3 mg L^-1, and in sea water as control. After 45 days, the tetrasporeling plants were found to be larger at photon flux densities of 50 and 100 μmol photons m^-2 s^-1 in the nutrient enrichment experiments; growth was greatest in the sea water enriched with ammonium nitrate and urea. An analysis of the combined effect of the photon flux density and nutrients revealed that the best combination for sporeling growth was the ammonium nitrate and urea solution at 50–100 μmol photons m^-2 s^-1.     

Dense fouling in acid transfer pipelines by an acidophilic rubber degrading fungus

An unique case of dense fouling by an acidophilic, hard rubber (polymerized rubber) degrading fungus in the acid transfer pipelines of a boron enrichment plant located at Kalpakkam, India is reported. In spite of a highly adverse environment for survival (pH 1.5, no dissolved nutrients), the fungus thrived and clogged the pipeline used for transferring 0.1N hydrochloric acid (HCl). Detailed investigations were carried out to isolate and identify the fungus and examine the nutrient source for such profuse growth inside the system. Microscopic observation showed the presence of a thick filamentous fungal biomass. Molecular characterization by 18S rRNA gene sequencing showed 98% similarity of the isolate with the acidophilic fungus Bispora sp. In laboratory studies the fungus showed luxuriant growth (specific growth rate of 13 mg day^?1) when scrapings of the hard rubber were used as the sole source of carbon. Scanning electron microscopy revealed extensive incursion of the fungus into the hard rubber matrix. In the laboratory, fungal growth was completely inhibited by the antifungal agent sodium omadine. The study illustrates an interesting example of biofouling under extreme conditions and demonstrates that organisms canphysiologically adapt to grow under unfavourable conditions, provided that a nutrient source is available and competition is low. The use of this fungal strain in biodegradation and in development of environmentally compatible processes for disposal of rubber wastes is envisaged.     

Nutrient interactions between phytoplankton and bacterioplankton under different carbon dioxide regimes

Light, nutrients, temperature, pH, and salinity are important factors in controlling the growth of phytoplankton and bacterioplankton. Supply of key nutrients to these communities can result in mutualistic or competitive relationships between bacterioplankton and phytoplankton. In this study, we investigated growth and uptake of nutrients by the marine prasinophyte flagellate Tetraselmis chui (strain PLY429) in the presence and absence of a community of bacterioplankton at two pH levels. Growth of PLY429 and total nutrient uptake were calculated for each treatment. The addition of bacterioplankton resulted in lower growth rates of PLY429, but the removal of ammonium was greater in those cultures with bacterioplankton present. The division rate of PLY429 was affected by pH; however, pH changes did not result in different uptake rates of nitrate, ammonium, or phosphate by the mixed algal and bacterial assemblage. These findings suggest that bacterioplankton and phytoplankton were competing for ammonium and that a lower pH resulted in more rapid algal growth. Mention of a trade name does not imply endorsement by the National Marine Fisheries Service.     

Biological control of alligatorweed (Alternanthera philoxeroides) with a Fusarium sp.

Alligatorweed (Alternanthera philoxeroidesG.) has become a serious weed in different crops in China. A fungal pathogen was found in Chongqing and Sichuan Provinces and was identified as a species in the Fusarium genus. The fungus produced macroconidia and chlamydospores abundantly on potato sucrose agar (PSA) plates. The bestconidial production and germination and colonygrowth of Fusarium sp. were at 23–31°C and pH 6.7–7.0. Light period and flooding did not affect fungal growth and conidium formation. The herbicides, glyphosate and paraquat, inhibited the fungal development in vitro. The fungus did not affect seed germination and seedling growth of paddy rice, wheat, maize, oilseed rape and broad bean inlaboratory or greenhouse trials. Inoculum density and wetness duration influenced the efficiency of Fusarium sp. to control alligatorweed; a concentration of 1.0 × 10⁵ spores^–1 ml and 12 h of high humidity duration after inoculation produced goodinfections on the weed at 23°C in the laboratory. When the fungus was applied to alligatorweed grown in greenhouse and in the field, good biocontrol efficiency was obtained: the plants started to wilt after four to five (greenhouse) or six days (field), and were killed 9–10 (greenhouse) or 13–14 (field) days after spraying the fungal inoculum. This was similar to the control efficiency resulting from glyphosate treatment. Therefore, this Fusarium sp. appeared to be a good candidatefor further studies and a promising biocontrol agent to manage alligatorweed in some terrestrial and aquatic crops.