Air biocontamination in a variety of agricultural industry environments in Egypt: a pilot study

Inhalation of airborne microorganisms and organic dust is an occupational concern among workers in agricultural industries. Airborne microorganisms and particulate matter samples were collected from poultry house, flourmill, textile, and food industry sites by use of liquid impinger and gravimetric samplers. Particulate matter concentrations were recorded at median concentrations of 1.56, 1.92, 4.39, and 0.7 mg/m³ in the occupied poultry house, textile, flourmill, and food indoor working environments, respectively. The highest median particulate matter concentration (27.9 mg/m³) was detected at the flourmill’s stack site. The highest median indoor concentration of culturable airborne bacteria (6.23 × 10⁵ CFU/m³) was found at the occupied poultry-house site and the lowest concentration (4.6 × 10³ CFU/m³) was found at the food industry site. The highest median indoor concentration of culturable airborne fungi (3.15 × 10⁴ CFU/m³) was found at the flourmill site whereas the lowest (1.24 × 10³ CFU/m³) was found at the textile industry site. Bacillus and Staphylococcus were the predominant Gram-positive bacteria whereas Acinetobacter and Klebsiella were the predominant Gram-negative bacteria. Escherichia coli and Salmonella were only detected in the indoor air at the poultry house site. Aspergillus flavus, Aspergillus niger, Penicillium, and yeast were the predominant fungal types at flourmill, textile, food industry, and poultry house, respectively. Workers were continuously exposed to airborne microorganisms at a median value of 10⁴ CFU/m³ in all the industries studied.     

Patterns of litter production across a salinity gradient in a Pterocarpus officinalis tropical wetland

Historically, Pterocarpus officinalisJacq. (Leguminoseae) dominated freshwater wetlands in the coastal plains of Puerto Rico, but deforestation has reduced its distribution to small patches adjacent to mangrove forests in areas of higher salinity. The objective of this study was to determine how a gradient in soil salinity affected litter, flower, and fruit production in a Pterocarpus officinalis.Three 100 m² plots were established in each of three sites along a salinity gradient: pasture/Pterocarpus edge (low salinity, mean salinity at 60 cm–9.7 g Kg^–1), Pterocarpus forest (intermediate salinity, 11.5 g Kg^–1) and a Pterocarpus/mangrove ecotone (high salinity, 15.0 g Kg^–1). Across this gradient, P. officinalis accounted for 100% of the relative basal area in the low and intermediate sites and 43% in the high salinity site which was domimated by Laguncularia racemosa. The basal area of P. officinalis decreased along the gradient from 73.5 m² ha^–1 in the low salinity site to 42.0 m² ha^–1 in the high salinity site. Litterfall was sampled on average every 23 days in 45 0.25 m² traps (5 traps per plot) for two years. Annual litterfall for the forest was 11.9 Mg ha^–1 yr^–1. Peaks in litterfall were associated with high precipitation in May 1995 and tropical storms in September 1995. Leaf fall of P. officinalis was significantly higher in the low salinity site (4.8 Mg ha^–1 yr^–1) than the high salinity site (1.8 Mg ha^–1 yr^–1), but total stand litterfall was greatest in the area of high salinity due to the greater contribution of L. racemosa. Pterocarpus flower and fruit production was approximately 10 times greater in low and intermediate salinity sites in comparison with the high salinity site. An increase in global temperature, will lead to higher sea level and higher soil salinity in costal wetlands. To conserve this wetland forest type it is critical to expand the distribution into areas of lower salinity where this species occurred historically.     

Kinetic and crystallographic studies of a redesigned manganese-binding site in cytochrome c peroxidase

Manganese peroxidase (MnP) from the white rot fungus Phanerochaete chrysosporium contains a manganese-binding site that plays a critical role in its function. Previously, a Mn^II-binding site was designed into cytochrome c peroxidase (CcP) based on sequence homology (Yeung et al. in Chem. Biol. 4:215–222, 1997; Gengenbach et al. in Biochemistry 38:11425–11432, 1999). Here, we report a redesign of this site based on X-ray structural comparison of MnP and CcP. The variant, CcP(D37E, V45E, H181E), displays 2.5-fold higher catalytic efficiency (k _cat/K _M) than the variant in the original design, mostly due to a stronger K _M of 1.9 mM (vs. 4.1 mM). High-resolution X-ray crystal structures of a metal-free form and a form with Co^II at the designed Mn^II site were also obtained. The metal ion in the engineered metal-binding site overlays well with Mn^II bound in MnP, suggesting that this variant is the closest structural model of the Mn^II-binding site in MnP for which a crystal structure exists. A major difference arises in the distances of the ligands to the metal; the metal–ligand interactions in the CcP variant are much weaker than the corresponding interactions in MnP, probably owing to partial occupancy of metal ion at the designed site, difference in the identity of metal ions (Co^II rather than Mn^II) and other interactions in the second coordination sphere. These results indicate that the metal ion, the ligands, and the environment around the metal-binding site play important roles in tuning the structure and function of metalloenzymes. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Time spent in the drift by downstream-dispersing invertebrates in a Lake District stream

1. The objective was to determine the time spent in the drift by different taxa of stream invertebrates. Most data were obtained from an earlier experimental study to determine the distances travelled by drifting invertebrates of 16 taxa in Wilfin Beck. Experiments were performed at two sites: ‘site 4’ in a stony, fast‐flowing, section of stream, ‘site 3’ in a deeper stream section where macrophytes were abundant. 2. The significant relationship between the mean distance x (m) travelled in the drift and modal water velocity V (m s^?1) was described by a power function in the earlier study but, as the power was close to one, a linear relationship has now been found to provide a satisfactory model. The rate of increase in x (m) with increasing V varied considerably between taxa. The mean time [mean t (s)] spent in the drift was estimated by dividing each x (m) by the appropriate V. Mean t (s) for each taxon was usually very constant over a wide range of V at each site (0.10–0.60 m s^?1 at site 4, 0.15–0.53 m s^?1 at site 3). A simple model estimated the time spent in the drift by different percentages (e.g. 75, 50, 10 and 1%) of the drifting invertebrates. 3. The experimental taxa at site 4 were divided into three groups according to the mean time spent in the drift. Mean t (s) for the five taxa in group 1 (32.8 s) was not significantly different from that obtained in control experiments with a mixed group of dead invertebrates. A similar time (33.0 s) was obtained for the five taxa in group 2, except at water velocities less than 0.2 m s^?1 when the mean t (s) decreased to 15–21 s. Mean t (s) was constant for each of the six taxa in group 3, and significantly less than that for groups 1 and 2. Mean values ranged from 28.8 s for Ephemerella ignita to only 9.4 s for Baetis rhodani and Gammarus pulex. All mean values were lower at site 3, presumably because of the dense stands of macrophytes, with mean values of 12.9 s for the five taxa in group 1 (equalling the value for dead invertebrates). Mean values for the six remaining taxa varied from 6.4 s for Simulium spp. to only 4.9 s for Baetis rhodani and 4.8 s for Gammarus pulex. It was concluded from a discussion of this study that the time spent in the drift may provide a useful measure for comparing the downstream dispersal of invertebrates in different streams, and may be a useful addition to models for the drift feeding of salmonids.     

Assessing the Potential of Phytoremediation at a Site in the U.K. Contaminated With 137Cs

Spinacia oleracea L. cv. ‘Bloomsdale’, Beta vulgaris L. cv. ‘Flavescens’, Brassica juncea L. ‘OB825’, and Helianthus annuus L. cv. ‘Oranges and Lemons’ were grown for 8 weeks at a site contaminated with ¹³⁷Cs at Bradwell Nuclear Power Station, UK. The site was a trench approximately 1.5 m deep, 2 m wide, and 100 m long in ‘made ground’ consisting of alluvium with traces of illites, kaolinites, and smectites. ¹³⁷Cs activity concentration was measured in individual plants after 8 weeks growth and the soil in which they grew. The biomass produced and total ¹³⁷Cs removed to shoots differed significantly between species but ¹³⁷Cs activity concentrations and Transfer Factors (TFs) did not. B. vulgaris produced the most biomass and removed the greatest amount of ¹³⁷Cs. For all plants, and within each taxon, plants growing at low soil ¹³⁷Cs activity concentrations had significantly greater TFs than those growing at high soil ¹³⁷Cs activity concentrations. It is concluded that selecting plant taxa suited to a particular site can be an effective way of improving phytoremediation rates, that there is much scope for adjusting harvesting intervals to 8 weeks or less without affecting TFs, and that estimates of time taken for ¹³⁷Cs removal by phytoremediation should consider that TFs may increase as soil concentrations decrease. With refinements in methodology, phytoremediation has the potential to contribute significantly to decontamination of the site at Bradwell.     

Spatial and seasonal variation in nutrient excretion by benthic invertebrates in a eutrophic reservoir

1. Nitrogen (N) and phosphorus (P) fluxes via excretion by benthic invertebrates were quantified in a eutrophic reservoir (Acton Lake, Ohio, U.S.A.). We quantified variation in nutrient fluxes seasonally (June until November 1997), spatially (three sites) and among taxa (chironomids, tubificid oligochaetes and Chaoborus). 2. The three taxa differed in spatial distribution and contribution to nutrient fluxes. Tubificids were the most abundant taxon at two oxic sites (1.5 and 4 m depth), and were exceedingly rare at an anoxic, hypolimnetic site (8 m). Chironomids were abundant only at the shallowest oxic site. Chaoborus was the only abundant taxon at the anoxic site. Total benthic invertebrate biomass was greatest at the shallowest site and lowest at the anoxic, hypolimnetic site. 3. Mass‐specific excretion rate [μmol NH₄–N or soluble reactive P (SRP) excreted mg dry mass^–1 h^–1] varied among experiments and was influenced by temperature. Differences among taxa were not significant. Thus, nutrient flux through benthic invertebrates was affected more by total invertebrate biomass and temperature than by species composition. 4. Fluxes of N and P via benthic invertebrate excretion (μmol NH₄–N or SRP m^–2 day^–1) were greatest at the oxic sites, where fluxes were dominated by the excretion of tubificids and chironomids. The N and P fluxes at the anoxic site were much lower, and were dominated by excretion by Chaoborus. The ratio of N and P excreted by the benthic invertebrate assemblage varied seasonally and was lowest at the anoxic site. 5. Comparison with other measured inputs shows that excretion by benthic invertebrates could be an important source of nutrients, especially of P. However, the relative importance of nutrient excretion by the benthos varies greatly spatially and temporally.     

Preliminary results on sperm motility and testicular histology of two feral fish species, Oreochromis mossambicus and Clarias gariepinus, from a currently DDT-sprayed area, South Africa

The effects of DDT and its metabolites on the reproductive health of two fish species (Clarias gariepinus and Oreochromis mossambicus) were studied using the gonadosomatic index (GSI), gonadal histology and computer assisted sperm analysis (CASA). DDT and its metabolites, DDE and DDD are endocrine disrupting chemicals posing estrogenic and anti‐androgenic properties, which have detrimental effects on growth and reproduction. Although DDT was banned internationally, it is still used for Malaria vector control in areas of South Africa. Both species were sampled at a reference site, Albasini dam (AD), (outside the DDT‐sprayed area, <0.01 μg L^?1 of DDT) and at an exposed site, Xikundu weir (XW), in the same river ± 70 km within the DDT‐sprayed area, <0.01 μg L^?1 of DDT. Gill nets were used to acquire a sample size of 10 male fish per site for each species. Both testes were dissected out and the GSI calculated. Uncontaminated milt was collected and analyzed using a novel CASA system based on open source software for characterization of sperm motility parameters. Testis tissue sampled for histology was fixed in Bouins and processed according to standard methods. The sample size obtained at the reference site (AD) was O. mossambicus n = 6, C. gariepinus n = 9; and at the exposed site (XW) was O. mossambicus n = 10, C. gariepinus n = 3. The GSI results showed a lower value for O. mossambicus at XW (0.06) (n = 13) than AD (0.12) (n = 6); U = 16.0, P = 0.044. However the opposite was true for C. gariepinus. Exposure to various environmental toxicants can result in gonadal changes such as decreased GSI, morphological alterations or both. Histological assessment showed histopathological alterations to testes tissue including intersex (only in O. mossambicus) and detachment of basal membranes. The CASA results showed a decrease in parameters from the reference site (AD) to the exposed site (XW) for both species. The decrease in percent motility (% MOT) for O. mossambicus from the reference site (AD) (n = 6) to the exposed site (XW) (n = 13) was statistically significant; U = 13.0, P = 0.023. C. gariepinus from the exposed site (XW) was most affected in terms of percent motility (10.00%) and velocity (67.94 μm s^?1). Detailed assessment of the environmental effects of DDT in this area seems crucial before population impacts become evident.     

Assessing the above‐ground biomass of a complex tropical rainforest using a canopy crane

Abstract Current estimates of the total biomass in tropical rainforests vary considerably; this is due in large part to the different approaches that are used to calculate biomass. In this study we have used a canopy crane to measure the tree architectures in a 1 ha plot of complex mesophyll vine forest at Cape Tribulation, Australia. Methods were developed to measure and calculate the crown and stem biomass of six major species of tree and palm (Alstonia scholaris (Apocynaceae), Cleistanthus myrianthus (Euphorbiaceae), Endiandra microneura (Lauraceae), Myristica insipida (Myristicaceae), Acmena graveolens (Myrtaceae), Normanbya normanbyi (Arecaceae)) using the unique access provided by the crane. This has allowed the first non‐destructive biomass estimate to be carried out for a forest of this type. Allometric equations which relate tree biomass to the measured variable ‘diameter at breast height’ were developed for the six species, and a general equation was also developed for trees on the plot. The general equation was similar in form to equations developed for tropical rainforests in Brazil and New Guinea. The species equations were applied at the level of families, the generalized equation was applied to the remaining species which allowed the biomass of a total of 680 trees to be calculated. This has provided a current estimate of 270 t ha⁻¹ above‐ground biomass at the Australian Canopy Crane site; a value comparable to lowland rainforests in Panama and French Guiana. Using the same tree database seven alternative allometric equations (literature equations for tropical rainforests) were used to calculate the site biomass, the range was large (252–446 t ha⁻¹) with only three equations providing estimates within 34 t ha⁻¹ (12.5%) of the site value. Our use of multiple species‐specific allometric equations has provided a site estimate only slightly larger (1%) than that obtained using allometric equations developed specifically for tropical wet rainforests. We have demonstrated that it is possible to non‐destructively measure the biomass in a complex forest using an on‐site canopy crane. In conjunction the development of crown maps and a detailed tree architecture database allows changes in forest structure to be followed quantitatively.     

Feasibility of Using Phytoextraction to Remediate a Compost-Based Soil Contaminated with Cadmium

Greenhouse and in-situ field experiments were used to determine the potential for phytoextraction to remediate soil contaminated with Cd from municipal solid waste (MSW) and sewage sludge (SS) compost application at a Peterborough (Canada) site. For the greenhouse experiment, one native (Chenopodium album) and three naturalized (Poa compressa, Brassica juncea, Helianthus annuus) plant species were planted in soil containing no detectable Cd (<1.0 μg·g^?1), and soil from the site containing low (5.0 ± 0.3 μg·g^?1 Cd), and high (16.5 ± 1.2 μg?g^?1 Cd) Cd concentrations. Plant uptake was low (root BAFs ≤0.5) for all species except P. compressa in the low Cd treatment (BAF 1.0). Only B. juncea accumulated Cd in its shoots, though uptake was low (BAF ≤0.3). For the field experiment, B. juncea was planted in-situ in areas of low and high Cd concentrations. Brassica juncea Cd uptake was low (root and shoot BAFs <0.2) in both treatments. Sequential extraction analysis indicated that Cd is retained primarily by low bioavailability soil fractions, and phytoextraction is therefore not feasible at this site. Though low Cd bioavailability has negative implications for Cd phytoextraction from MSW/SS compost-based soils, it may limit receptor exposure to Cd sufficiently to eliminate the potential for risk at this site.     

Landscape reclamation at a central Florida phosphate mine

A 13-ha parcel of phosphate-mined land at the Gardinier Mine near Ft. Meade, Florida, USA, was reclaimed to test several design principles, establish planting and seed-dispersal trails, and monitor groundwater and surface water. The site was designed as a complete hydrologic unit with perched, basin, and lake-fringe wetland communities in upland communities. The design and placement of ecological communities resulted from the reclaimed topography and site hydrology. The percent growth for 11 wetland and mesic tree species averaged 127% (weighted average) in 3 years; some species averaged more than 160%. Percent survival ranged from 0% (Cephalanthus occidentalis) to 100% for several mesic forest tree species (Fraxinus pennsylvanica, Liquidambar styraciflua, Magnolia grandiflora). The percent survival of wetland species (weighted average) was 94.6%. An adjacent forested, floodplain wetland was evaluated as a source of windblown and bird-dispersed seeds to the site. Windblown seeds decreased in densities as distance from forest edge increased. Densities were from 125/m² to 380/m² within the forest, 50/m² to 120/m² at the forest edge and decreased exponentially as distance increased. Bird-dispersed seed densities at the base of constructed perches and tree “snags” ranged from 100/m² to more than 300/m². Rainfall and surface-water inflow along the edge of the site were the largest inputs to the reclaimed landscape's hydrologic budget. Groundwater flow and seepage were more important to growth and survival of planted species because of the maintenance of higher soil moisture during periods of drought.     

Relationship between small-scale structural variability and simulated vegetation productivity across a regional moisture gradient in southern Africa

The observed variability in vegetation structure within landscapes was used as the basis for model estimates of the range of annual productivity of landscape patches at four sites along a moisture gradient in southern Africa ranging from 879 to 365 mm mean annual rainfall. Principal components of patch‐scale variability in leaf area, woody biomass and vertical leaf profiles were derived from intensive characterization of the small‐scale spatial structure of woody vegetation at each site. For each site, the mean and extremes of the principal component distribution parameterized an ecophysiology model of vegetation productivity. Vegetation was most heterogeneous at intermediate locations along the rainfall gradient. Variability in vegetation structure led to a range of annual productivity within one site (600 mm) that accounted for 68% of the total range in mean productivity across all sites. Patch‐scale estimates of tree productivity were found to be primarily correlated to annual rainfall (r²=0.66, P=0.001) and not woody leaf area (r²=0.01, P=0.75), while grass productivity was found to be related to values of woody leaf area (r²=0.77, P<0.001) and not annual rainfall (r²=0.11, P=0.29). This result indicates that life‐form interactions have a significant role in controlling vegetation productivity across the rainfall gradient. The findings of this study emphasize the importance of considering heterogeneity rather than mean structure when modeling productivity, particularly when considering dynamic vegetation structure, where differences between landscape patches may not be well represented in the mean structure.     

Long-term impact of a stand-replacing fire on ecosystem CO2 exchange of a ponderosa pine forest

Ponderosa pine (Pinus ponderosa) forests of the southwestern United States are a mosaic of stands where undisturbed forests are carbon sinks, and stands recovering from wildfires may be sources of carbon to the atmosphere for decades after the fire. However, the relative magnitude of these sinks and sources has never been directly measured in this region, limiting our understanding of the role of fire in regional and US carbon budgets. We used the eddy covariance technique to measure the CO₂ exchange of two forest sites, one burned by fire in 1996, and an unburned forest. The fire was a high‐intensity stand‐replacing burn that killed all trees. Ten years after the fire, the burned site was still a source of CO₂ to the atmosphere [109±6 (SEM) g C m^?2 yr^?1], whereas the unburned site was a sink (?164±23 g C m^?2 yr^?1). The fire reduced total carbon storage and shifted ecosystem carbon allocation from the forest floor and living biomass to necromass. Annual ecosystem respiration was lower at the burned site (480±5 g C m^?2 yr^?1) than at the unburned site (710±54 g C m^?2 yr^?1), but the difference in gross primary production was even larger (372±13 g C m^?2 yr^?1 at the burned site and 858±37 g C m^?2 yr^?1at the unburned site). Water availability controlled carbon flux in the warm season at both sites, and the burned site was a source of carbon in all months, even during the summer, when wet and warm conditions favored respiration more than photosynthesis. Our study shows that carbon losses following stand‐replacing fires in ponderosa pine forests can persist for decades due to slow recovery of the gross primary production. Because fire exclusion is becoming increasingly difficult in dry western forests, a large US forest carbon sink could shift to a decadal‐scale carbon source.     

A comparison of water and carbon dioxide exchange at a windy alpine tundra and subalpine forest site near Niwot Ridge,Colorado

Eddy covariance measurements of the surface energy balance and carbon dioxide exchange above high-elevation (3,480 m above sea level) alpine tundra located near Niwot Ridge, Colorado, were compared to simultaneous measurements made over an adjacent subalpine forest over two summers and one winter, from June 9, 2007 to July 3, 2008. The surface energy balance closure at the alpine site averaged 71 and 91%, winter and summer, respectively, due to the high wind speeds, short turbulent flux footprint, and relatively flat ridge-top location of the measurement site. Throughout the year, the alpine site was cooler with higher relative humidity, and had a higher horizontal wind speed, especially in winter, compared to the forest site. Wind direction was persistently downslope at the alpine site (summer and winter, day and night), whereas upslope winds were common at the forest site during summer daytime periods. The latent and sensible heat fluxes were consistently larger in magnitude at the forest site, with the largest differences during summer. The horizontal advective flux of CO₂ at the alpine site averaged 6% of the net ecosystem exchange (NEE) during summer nights (5% during summer daytime), and was small in relation to the high wind speeds, relatively flat site, and weak sources of CO₂ upwind of the site. The magnitudes and diurnal behavior of the alpine NEE calculated using three methods; eddy-covariance, friction velocity filter, and with advection and storage calculations, gave similar results. The period of net CO₂ uptake (negative NEE) was 100 days at the alpine site with a net uptake of 16 g C m⁻², compared to 208 days at the forest site with a net uptake of 108 g C m⁻², with initiation of net uptake coinciding with air temperatures reaching +10°C. Winter respiration loss at the alpine site was 164 g C m⁻² over 271 days, compared to 52 g C m⁻² over 175 days at the forest site, with the initiation of net loss coinciding with air temperatures reaching −10°C at each site.     

Soil organic matter as a potential net nitrogen sink in a fertilized cornfield,South Deerfield,Massachusetts, USA

Net nitrogen (N) mineralization in situ and N mineralization potential (N₀) over one complete year (1986–1987) were examined for a conventionally managed silage cornfield that received at least 235 kg fertilizer N ha^-1. Net N mineralization at the site, measured by sequential in situ polyethylene-bag incubations, totaled –54 kg N ha^-1 yr^-1, and –31 kg N ha^-1 over the May-to-August growing season. Nitrogen mineralization potential of the soil organic matter (SOM), measured by laboratory anaerobic incubations, was positive uniformly and varied with month of sample collection. The soil gained 72 kg inorganic N ha^-1 from April to October, principally because of a fall manuring, only 7 kg N ha^-1 from April to September. The in situ incubations, likely more representative of the balance between N mineralization and immobilization under N-fertilized conditions, suggest that SOM at the site is accumulating N.Contribution from the Department of Forestry and Wildlife Management, University of Massachusetts, Amherst, MA 01003, USA.Contribution from the Department of Forestry and Wildlife Management, University of Massachusetts, Amherst, MA 01003, USA.     

Plasmid cloning vectors that can be nicked at a unique site

Summary We describe ColE1-type plasmids, with relaxed DNA replication, based on pMB9, and carrying the Cm^R determinant of R1, in addition to the Tc^R determinant of pMB9. One of the plasmids, pPH207, has unique sites for EcoRI, HindIII, BamI, SalI and Hpal. Insertion of foreign DNA into all but the last of these inactivates cither the Cm^R or the Tc^R determinant.The original Cm^R Tc^R plasmid (pCM2) contains a copy of IS1 which produces deletions to left and to right. Most of these inactivate either the Cm^R or the Tc^R determinant. An internal 280 bp deletion of IS1 DNA in pPH207 greatly reduces the frequency at which deletions are observed.The main feature of these plasmids is a site that is cleaved by some preparations of EcoRI in only one strand of the DNA duplex (the EcoRIⁿ site). This site facilitates strand separation of sequences inserted at the HindIII, BamI and SalI sites of the Tc^R gene, and also of any inserted at the true EcoRI site by a method that destroys that site. Since the oricntation of the EcoRIⁿ site is known, the orientation of sequences inserted at the neighbouring sites can be easily determined.Plasmid pPH207 is not mobilised by a Hfr, but its mobilisation is promoted by ColE1. It is therefore Mob ^- bom ⁺. Experiments with minicells show that it directs the copious synthesis of chloramphenicol transacetylase.     

Cloning Vectors for Lactococci Based on a Plasmid Encoding Resistance to Cadmium

An 8.8-kb plasmid (pND302) was identified in Lactococcus lactis spp lactis M71 which encodes cadmium resistance (Cd^R). Most of the commercial lactococcal strains tested were sensitive to cadmium. Therefore, Cd^R should provide a useful selectable marker for constructing cloning vectors in lactococci. pND302 was mapped with a number of restriction enzymes and found to contain a unique EcoRI site suitable for cloning. Two E. coli/L. lactis shuttle cloning vectors, pND304 and pND624, were constructed by subcloning of the E. coli plasmids pBR322 and pGEM-7Zf(+) containing a 1.6-kb gene encoding nisin resistance (Nis^R) of lactococcal origin into the EcoRI site of pND302, separately. The E. coli DNA component of pND624 was removed and the resulting plasmid, pND625, consisted of only lactococcal DNA, expressing Nis^R and Cd^R, with two synthetic polylinkers that contain multiple restriction sites for versatile cloning. Both pND302 and pND625 can be transformed by electroporation into L. lactis LMO230 at 10³/μg DNA and maintained stably in LMO230. The results indicated that pND302 and pND625 are potential food-grade cloning vectors for lactococci. Received: 27 November 1995 / Accepted: 29 December 1995     

Carbon storage and fluxes in ponderosa pine forests at different developmental stages

We compared carbon storage and fluxes in young and old ponderosa pine stands in Oregon, including plant and soil storage, net primary productivity, respiration fluxes, eddy flux estimates of net ecosystem exchange (NEE), and Biome‐BGC simulations of fluxes. The young forest (Y site) was previously an old‐growth ponderosa pine forest that had been clearcut in 1978, and the old forest (O site), which has never been logged, consists of two primary age classes (50 and 250 years old). Total ecosystem carbon content (vegetation, detritus and soil) of the O forest was about twice that of the Y site (21 vs. 10 kg C m^?2 ground), and significantly more of the total is stored in living vegetation at the O site (61% vs. 15%). Ecosystem respiration (R_e) was higher at the O site (1014 vs. 835 g C m^?2 year^?1), and it was largely from soils at both sites (77% of R_e). The biological data show that above‐ground net primary productivity (ANPP), NPP and net ecosystem production (NEP) were greater at the O site than the Y site. Monte Carlo estimates of NEP show that the young site is a source of CO₂ to the atmosphere, and is significantly lower than NEP(O) by c. 100 g C m^?2 year^?1. Eddy covariance measurements also show that the O site was a stronger sink for CO₂ than the Y site. Across a 15‐km swath in the region, ANPP ranged from 76 g C m^?2 year^?1 at the Y site to 236 g C m^?2 year^?1 (overall mean 158 ± 14 g C m^?2 year^?1). The lowest ANPP values were for the youngest and oldest stands, but there was a large range of ANPP for mature stands. Carbon, water and nitrogen cycle simulations with the Biome‐BGC model suggest that disturbance type and frequency, time since disturbance, age‐dependent changes in below‐ground allocation, and increasing atmospheric concentration of CO₂ all exert significant control on the net ecosystem exchange of carbon at the two sites. Model estimates of major carbon flux components agree with budget‐based observations to within ± 20%, with larger differences for NEP and for several storage terms. Simulations showed the period of regrowth required to replace carbon lost during and after a stand‐replacing fire (O) or a clearcut (Y) to be between 50 and 100 years. In both cases, simulations showed a shift from net carbon source to net sink (on an annual basis) 10–20 years after disturbance. These results suggest that the net ecosystem production of young stands may be low because heterotrophic respiration, particularly from soils, is higher than the NPP of the regrowth. The amount of carbon stored in long‐term pools (biomass and soils) in addition to short‐term fluxes has important implications for management of forests in the Pacific North‐west for carbon sequestration.     

Establishment of a novel sensitive method for detecting methylation modification on DNA of escherichia coli cell

This study focused on finding a novel sensitive method to determine the methylation modification at DNA dam (GATC) sites in Escherichia coli. A new plasmid which contained three GATC sites recognized by restriction enzyme BclI and one GAATTC site recognized by EcoRI was transformed into E. coli stains AB1157(dam ⁺) and GM2929(dam ⁻) respectively. Then the plasmid DNA was digested by restriction enzyme BclI(T*GATCA), which was sensitive to methylation. The results showed that the plasmid derived from AB1157 was not digested while that from GM2929 was, for the methylation level of the former was high while the latter was low. So by detecting the methylation of plasmid transferred into the strain, we could determine whether methylaion existed at DNA dam (GATC) site in E. coli. This method was effective and rapid; moreover, the digested fragments were not dispersive. It also made a basis for the detection of whether methylation occurred in mode beings by low-energy ion beam. The article is published in the original.     

Paired-tower measurements of carbon and energy fluxes following disturbance in the boreal forest

Disturbances by fire and harvesting are thought to regulate the carbon balance of the Canadian boreal forest over scales of several decades. However, there are few direct measurements of carbon fluxes following disturbances to provide data needed to refine mathematical models. The eddy covariance technique was used with paired towers to measure fluxes simultaneously at disturbed and undisturbed sites over periods of about one week during the growing season in 1998 and 1999. Comparisons were conducted at three sites: a 1‐y‐old burned jackpine stand subjected to an intense crown fire at the International Crown Fire Modelling Experiment site near Fort Providence, North‐west Territories; a 1‐y‐old clearcut aspen area at the EMEND project near Peace River, Alberta; and a 10‐y‐old burned, mixed forest near Prince Albert National Park, Saskatchewan. Nearby mature forest stands of the same types were also measured as controls. The harvested site had lower net radiation (R_n), sensible (H) and latent (LE) heat fluxes, and greater ground heat fluxes (G) than the mature forest. Daytime CO₂ fluxes were much reduced, but night‐time CO₂ fluxes were identical to that of the mature aspen forest. It is hypothesized that the aspen roots remained alive following harvesting, and dominated soil respiration. The overall effect was that the harvested site was a carbon source of about 1.6 gC m^?2 day^?1, while the mature site was a sink of about ?3.8 gC m^?2 day^?1. The one‐year‐old burn had lower R_n, H and LE than the mature jackpine forest, and had a continuous CO₂ efflux of about 0.8 gC m^–2 day^?1 compared to the mature forest sink of ? 0.5 g C m^?2 day^?1. The carbon source was likely caused by decomposition of fire‐killed vegetation. The 10‐y‐old burned site had similar H, LE, and G to the mature mixed forest site. Although the diurnal amplitude of the CO₂ fluxes were slightly lower at the 10‐y‐old site, there was no significant difference between the daily integrals (? 1.3 gC m^?2 day^?1 at both sites). It appears that most of the change in carbon flux occurs within the first 10 years following disturbance, but more data are needed on other forest and disturbance types for the first 20 years following the disturbance event.