Enumeration and characterization of the soil microflora from hydrocarbon-contaminated soil sites able to mineralize polycyclic aromatic hydrocarbons (PAH)

The use of a plate screening technique allowed the direct isolation and quantification of polycylic aromatic hydrocarbon (PAH)-degrading bacteria from different soil sites. Bacteria that were able to grow on anthracene, phenanthrene, fluoranthene or pyrene as a sole carbon source were found with numbers between 10³ and 10⁵ colony-forming units (cfu)/g of soil dry weight, but only in samples that originated from PAH-contaminated sites. No isolates were found that could grow on perylene, triphenylene, benzo(a)pyrene or chrysene as sole carbon source. Bacteria that had been selected on the same PAH substrate showed a related degradation pattern for both other PAH and oil compounds and carbohydrate substrates even if they had been collected at distant soil sites. Based on these findings the isolates could be clustered into four different catabolic and taxonomic similarity groups. Taxonomic determination of representative isolates suggested that nocardioform actinomycetes of the genera Mycobacterium, Rhodococcus and Gordona represented a major part of the soil microflora able to mineralize PAH. Three new isolates able to grow on anthracene, pyrene or fluoranthene as the sole carbon source, respectively, have been isolated and identified (Sphingomonas paucimobilis BA2, Gordona sp. BP9, Mycobacterium sp. VF1). The ubiquitous presence of a potent and versatile mineralizing microflora in PAH-contaminated soils indicated that the microflora is not the limiting factor for the degradation of PAH with up to four rings.     

Nutrient balances and phytoplankton dynamics in two agriculturally loaded shallow lakes

The impact of agriculture was estimated on two shallow, eutrophic lakes, Lake Kotojärvi and Lake Villikkalanjärvi in southern Finland. The main emphasis was on phosphorus and nitrogen budgets and on the phytoplankton dynamics. Special attention was paid to internal P loading and blue-green algal blooms. The mean Tot-P load from agricultural land was 1.2 kg ha^-1 a^-1 in both basins and Tot-N loads were 19 kg ha^-1 a^-1 in L. Villikkalanjärvi and 12 kg ha^-1 a^-1 in L. Kotojärvi. The Tot-P input to L. Kotojärvi was on an average 0.62 g m^-2 a^-1 (per lake surface area), and the Tot-N input 9.1 g m^-2 a^-1. The corresponding inputs to L. Villikkalanjärvi were 3.1 and 57 g m^-2 a^-1, respectively. The annual variation followed the runoff volumes. About half of the Tot-P and one third of the Tot-N load was retained in L. Kotojärvi. In L. Villikkalanjärvi the retention was only 24% for Tot-P and 19% for Tot-N. The difference was very probably due to a longer theoretical retention time in L. Kotojärvi. In L. Villikkalanjärvi the mean concentration of Tot-P was 120 µg 1^-1 and that of Tot-N 1700 µg 1^-1 and the corresponding figures in L. Kotojärvi 67 and 990 µg 1^-1, respectively. The mean chlorophyll a concentration was, however, higher in L. Kotojärvi (26 µg 1^-1) than in L. Villikkalanjärvi (20 µg 1^-1). This was probably due to an internal P load in L. Kotojärvi: in 1988 the internal load of dissolved P was estimated to be as much as twofold the external load. In L. Villikkalanjärvi the internal dissolved P load was only up to 50% of the external input. In L. Kotojärvi the high internal P load coupled with a low DIN:DIP ratio resulted in a strong blue-green algal bloom in the summer of 1988. In L. Villikkalanjärvi blue-green algae were observed only in small amounts. Even in August 1990, when the DIN:DIP ratio was low enough to favor the occurrence of blue-green algae, they contributed only up to 10–15% of the total phytoplankton biomass.     

Uptake,accumulation and metabolism of auxins in tobacco leaf protoplasts

Uptake and metabolism of exogenous naphthalene-1-acetic acid (NAA) and indole-3-acetic acid (IAA) have been studied in tobacco (Nicotiana tabacum L. cv. Xanthi) mesophyll protoplasts. Both auxins entered protoplasts by diffusion under the action of the transmembrane pH gradient without any detectable participation of an influx carrier. Molecules were accumulated by an anion-trapping mechanism and most of them were metabolized within hours, essentially as glucose-ester and amino-acid conjugates. Protoplasts were equipped with a functional auxin-efflux carrier as evidenced by the inhibitory effect of naphthylphtalamic acid on IAA efflux. Basically, similar mechanisms of NAA and IAA uptake occurred in protoplasts. However, the two auxins differed in their levels of accumulation, due to different membrane-transport characteristics, and the nature of the metabolites produced. This shows the need to estimate the accumulation and the metabolism of auxins when analyzing their effects in a given cell system. The internal auxin concentration could be modulated by changing the transmembrane pH gradient, giving an interesting perspective for discriminating between the effects of intra- and extracellular auxin on physiological processes.Abbreviations BA benzoic acid - C_i/C_e accumulation ratio of auxin - IAAasp N-[3-indolylacetyl]-dl-aspartic acid - NAA naphthalene-1-acetic acid - NAAasp N-[1-naphthylacetyl]-l-aspartic acid - NPA N-1-naphthylphthalamic acid The authors thank Dr. M. Caboche (I.N.R.A, Versailles, France) for his generous gifts of some amide derivatives of 1-NAA, Mr. P. Varennes and Dr. B. Das (I.C.S.N., C.N.R.S., Gif-sur-Yvette, France) for recording and interpreting the mass spectra of NAA glucose ester, and Prof. P. Manigault (Institut des Sciences Végétales, Gif-sur-Yvette) for microscopy measurements of protoplast dimensions. This work was supported by funds from the C.N.R.S, I.N.R.A, and E.E.C.     

Effects of energy deprivation on the fly pupil mechanism: evidence for a rigor state

The energy dependence of the pupil pigment-migrations in the fly Musca domestica was studied in live animals, using optical techniques and nitrogen-gas induced anoxia. The results obtained can be summarized in 3 points:

1. Energy deficiency can make the pupil mechanism stop in any state, extreme or intermediate.
2. Anoxia induced during intermittent stimulation makes the pupil stop in the closed state (aggregated pigment granules).
3. During long-term anoxia the pupil very slowly opens (dispersal of pigment granules), irrespective of ambient intensity.

The slow anoxic opening (point 3) is more than 1000 times slower than that predicted for free diffusion of pigment granules in water. Assuming realistic values of cytoplasm viscosity, this implies that anoxia causes the pigment granules to attach to rigid structures in the cells, in analogy with the rigor state in anoxic muscles. The rigor phenomenon in the pupil mechanism prevents experimental discrimination between active and passive processes of pigment migration. Normal pupil opening has a time course which agrees reasonably with a passive diffusion process, but it is argued that an active transportation of granules away from the rhabdom is more likely in the dark adapted eye.     

Optimization of transverse alternating field electrophoresis for strain identification of Leuconostoc oenos

Genomic DNA of several strains oof oenological lactic bacteria belonging to the species Lactobacillus plantarum, Leuconostoc oenos and Pediococcus pentosaceus was digested by the rare-cutting endonucleases ApaI and SmaI. The restriction products were separated by transverse alternating field electrophoresis (TAFE). The size of the genome of L. oenos estimated by adding the molecular size of the ApaI fragments was on average 1320 kb. A strong polymorphism was observed between the strains, which could be easily differentiated except for two industrial strains of L. oenos. A simple modification of the TAFE apparatus is proposed to improve the separation of the DNA fragments. Correspondence to: J.-N. Hallet     

Enzymatic hydrolosis of isolated and fibre-bound galactoglucomannans from pine-wood and pine kraft pulp

Fibre-bound and isolated galactoglumanans from pine-wood and pine kraft pulp were hydrolysed with purified mannanases from Trichoderma reesei and Bacillus subtilis. The isolated galactoglucomannans from both wood and pulp could be hydrolysed fairly extensively with both enzymes. In addition to mixed oligomers, the fungal mannase produced mannobiose as the main hydrolysis product whereas the bacterial mannanase produced mannobiose, mannotriose and mannotetraose. Both enzymes hydrolysed the native galactoglucomannan in finely ground pinewood, whereas galactoglucomannan in pine kraft pulp was only hydrolysed by the T. ressei mannanase. Thus, mannanases exhibit different specificities on fibre-bound, modified substrates. In spite of the high enzyme loading, the degree of hydrolysis of fibre-bound substrates did not exceed 10% of the theoretical, probably due to poor accessibility of the substrates. Correspondence to: M. Rättö     

The Effect of Transport Stress on Plasma Levels of Catecholamines,Cortisol, Corticosteroid-Binding Globulin,Blood Cell Count,and Lymphocyte Proliferation in Pigs

The effect of transport stress on the plasma levels of catecholamines, Cortisol, and corticosteroid-binding globulin were studied in 6 gilts. To assess the effect on immune status, white blood cells were also counted and the cell-mediated immunity was estimated. The adrenaline level increased significantly during transport, from a basal mean level of 0.03 ng/ml to a plateau level of 0.11 to 0.12 ng/ml. The noradrenaline level fluctuated, but not constantly, during transport. The mean plasma Cortisol level before loading was approximately 40 nmol/1 and rose immediately after the start of transport to 70 nmol/1 (p< 0.05) and to 87 nmol/1 (p< 0.01) within 10 and 30 min, respectively. After unloading the Cortisol level rapidly decreased and a minimum level was seen 4 h after the transport, whereafter the diurnal rhythm was resumed. The plasma corticosteroid-binding globulin level increased nonsignificantly during the day of transport, from 25 nmol/1 to a level of 34 nmol/1, and it continued to increase until a plateau level was reached on the second day after transportation. The total white blood cell number increased significantly (from 13.7 to 15.5 × 109 cells/1), the number of lymphocytes decreased significantly (from 8.4 to 7.0 × 109 cells/1), and the number of polymorphonuclear neutrophils increased significantly (from 4.3 to 7.2 × 109 cells/1) during transport. No significant variation in the proliferation response was seen in the whole blood cell cultures. The main results were the significant signs of simultaneous activity of both the adrenal cortex and the adrenal medulla during transport.     

Omphalina foliacea, a new basidiolichen from America

A relatively large Leptogium -like lichen overgrowing mosses in wet paramos has been discovered in the northern Andes and Costa Rica. It has, however, a green photobiont and basidiomycete hyphae. No fruitbodies have been observed, only asexual reproduction by fragmentation of the fragile thallus and by goniocysts formed on the ecorticate lower surface. Some aspects of the taxonomy of Omphalina are briefly discussed.     

The braincase and palate of the tetrapodomorph sarcopterygian mandageria fairfaxi: morphological variability near the fish–tetrapod transition

The braincase of the Late Devonian tristichopterid sarcopterygian Mandageria fairfaxi , from Canowindra, NSW, Australia, differs radically from the conservative pattern present in other 'osteolepiforms' (stem–group tetrapodomorph fishes) and non–dipnoan sarcopterygian fishes in general. The basioccipital region is short, displaced anteriorly, and either unossified or loosely articulated to the exoccipital, leaving most or all of the notochordal tunnel open ventrally. The exoccipital complex, which is developed into a large saddle that would have rested on top of the notochord, carries large, triangular articular facets on its posterior face and appears to have formed part of a functional neck joint, a synovial articulation between the skull and vertebral column that allows the former to rotate against the latter. Such a joint is characteristic of post–Devonian tetrapods, but unknown in other sarcopterygians. We infer that the ventrally open notochordal tunnel allowed gentle flexion of the cranial notochord during (predominantly vertical) rotational movement at the occiput; this is a mechanically unique solution to the problem of creating a mobile neck. Other unusual features of Mandageria include a posteriorly located lateral commissure, and structures on the entopterygoid and lateral commissure that may have been associated with an elaborate spiracular tract.