Fishery by-catches of marine mammals in Galician waters: results from on-board observations and an interview survey of fishermen

Rates of cetacean by-catch were investigated in Galician waters (NW Spain) using a combination of observer trips on fishing vessels, a carcase recovery scheme and an interview survey of fishermen, carried out over two years (1998-1999). All these data sources are suspected of underestimating by-catch due to the sample of co-operating fishermen being, necessarily, self-selecting. No by-catches were seen during observer trips, although not all sectors of the fishery could be covered. The carcass recovery scheme yielded seventeen cetacean carcases over two years, which compares to around 35 by-caught cetaceans recorded annually by the Galician strandings network. Analysis of interview data suggested that around 200 cetaceans might be caught annually in inshore waters and around 1500 in offshore waters. Confidence limits were wide for all estimates. The highest by-catch rates were estimated for gillnets and offshore trawling. The majority of by-catches are small dolphins, probably mainly Delphinus delphis. Smaller numbers of Tursiops truncatus and Globicephala melas are also reported. Comparing the interview estimates of by-catch rates with minimum estimates of population size, it is suggested that by-catches of D. delphis and T. truncatus may be unsustainably high and that routine monitoring of fishery by-catches in Galician fisheries is required.     

Linking colonies to fisheries: Spatio-temporal overlap between common murres (Uria aalge) from Tatoosh Island and coastal gillnet fisheries in the Pacific Northwest, USA

The population-level impact of seabird bycatch is difficult to assess because colony-of-origin is often unknown. As an alternative and complementary approach to ship-derived observations, we assessed the relative potential for bycatch of a known seabird population by quantifying spatio-temporal overlap with local fisheries. Common murres (Uria aalge) have been reported as the most abundant seabird inadvertently caught in Washington and British Columbia coastal gillnet fisheries. In 1999-2001, we tracked 48 common murres from Tatoosh Island, the closest colony to the fisheries, during post-breeding. Using capture-mark-recapture models, we estimated weekly murre movement probabilities to/from three strata (offshore of, centered around, and inshore of Tatoosh Island). Based on movement probabilities and population size, we projected strata- and week-specific murre abundance. We created an index of overlap by calculating the product of murre abundance × gillnet fishing effort as a function of strata and time. The majority of murres (80%) moved inshore, where fishing effort was consistently the highest, suggesting that up to 4000 Tatoosh murres were vulnerable to bycatch. Index scores in the inshore stratum were 5-25 times higher relative to the offshore and Tatoosh strata, respectively. Overlap was sensitive to phenology, as index scores increased by 50% when dispersal was shifted four weeks earlier, while a two weeks delay decreased scores by 20%. Until the long-term impact of cumulative mortality in gillnet fisheries is determined, we believe a precautionary approach is warranted in the inshore stratum where the potential for bycatch was highest. We advocate the use of visible netting in inshore fisheries, a proven solution that reduces murre bycatch while maintaining fishing efficiency.     

Behavioral responses of bottlenose dolphins, Tursiops truncatus, to gillnets and acoustic alarms

Along the east coast of the United States, by-catches of bottlenose dolphins, Tursiops truncatus, in gillnet fisheries exceed removal levels set under the US Marine Mammal Protection Act. One measure proposed to reduce this mortality is the use of acoustic alarms, or pingers, which have proven effective in reducing by-catches of other small cetaceans, but have not been tested with bottlenose dolphins. We examined the responses of bottlenose dolphins to a commercial gillnet equipped with functional (active) and non-functional (control) Dukane NetMark® 1000 alarms near Fort Macon, NC. Between 5 April and 10 May 2001 we used a theodolite to track 59 groups of dolphins around the net. Choice of treatment was random each day and the two shore-based observers were unaware of whether alarms were active (13 days) or controls (9 days). There were no significant differences in the number of groups observed (P=0.315; 1−β=0.835) or in the closest observed approach to the net (P=0.307; 1−β=0.828) between treatments. However, dolphins entered a circular buffer approximately 100 m around the net more frequently with control than active alarms (P=0.015). We conclude that some dolphins responded to the alarms by avoiding the net, but caution that the potential efficacy of alarms is confounded by dolphin behavior. Most dolphins were aware of the net, regardless of the status of alarms, and some dolphins fed on fish in the net or discarded by the fishing vessel. We believe that it would be unwise to use pingers in these fisheries because of the limited behavioral responses we observed in our experiment. Furthermore, the responses we observed are likely to diminish or change over time as dolphins habituate or sensitize to these alarms. Further research is required to understand the behavior responsible for entanglement.     

Subtropical plantations are large carbon sinks: Evidence from two monoculture plantations in South China

Quantifying the net carbon (C) storage of forest plantations is required to assess their potential to offset fossil fuel emissions. In this study, a biometric approach was used to estimate net ecosystem productivity (NEP) for two monoculture plantations in South China: Acacia crassicarpa and Eucalyptus urophylla. This approach was based on stand-level net primary productivity (NPP, based on direct biometric inventory) and heterotrophic respiration (R_h). In comparisons of R_h determination based on trenching vs. tree girdling, both trenching and tree girdling changed soil temperature and soil moisture relative to undisturbed control plots, and we assess the effects of corrections for disturbances of soil moisture and soil moisture on the estimation of soil CO₂ efflux partitioning. Soil microbial biomass and dissolved organic carbon were significantly lower in trenched plots than in tree girdled plots for both plantations. Annual soil CO₂ flux in trenched plots (R_h-t) was significantly lower than in tree-girdled plots (R_h-g) in both plantations. The estimates of R_h-t and R_h-g, expressed as a percentage of total soil respiration, were 58 ± 4% and 74 ± 6%, respectively, for A. crassicarpa, and 64 ± 3% and 78 ± 5%, respectively, for E. urophylla. By the end of experiment, the difference in soil CO₂ efflux between the trenched plots and tree-girdled plots had become small for both plantations. Annual R_h (mean of the annual R_h-t and R_h-g) and net primary production (NPP) were 470 ± 25 and 800 ± 118 g C m⁻² yr⁻¹, respectively, for A. crassicarpa, and 420 ± 35 and 2380 ± 187 g C m⁻² yr⁻², respectively, for E. urophylla. The two plantations in the developmental stage were large carbon sinks: NEP was 330 ± 76 C m⁻² yr⁻¹ for A. crassicarpa and 1960 ± 178 g C m⁻² yr⁻¹ for E. urophylla.     

Response of travelling bottlenose dolphins (Tursiops aduncus) to experimental approaches by a powerboat in Jervis Bay, New South Wales, Australia

Powerboats are potentially a significant source of disturbance to coastal cetaceans. Information is scarce, however, on the nature of interactions between powerboats and dolphins, particularly when both surface and acoustic behaviour are combined. The surface behaviour and acoustic response of travelling dolphins to approaches by a powerboat were assessed by a series of experimental trials between November 2001 and November 2003 in Jervis Bay, New South Wales, Australia. Dolphin behaviour was monitored continuously from an independent research boat before, during and after a powerboat approached (n = 12). Treatments were interspersed with control observations (n = 12). Changes in surface behaviour indicated differences between the treatment and control periods (z = 2.24, p = 0.025), with dolphins tending to alter their surface behaviour when exposed to the powerboat approach. Analysis also revealed a change in the direction of travel by dolphin groups when approached (z = 3.22, p = 0.001). Changes in surface behaviour occurred at vessel approach distances outside the minimum approach distance of 30 m for recreational and commercial vessels, as proposed by the New South Wales National Parks and Wildlife Service. In contrast, there were no changes in dolphin whistle rates (F_3,12 = 0.74, p = 0.54) or the duration of echolocation click bouts (F_3,12 = 0.76, p = 0.59) when approached. These findings indicate that powerboats do affect the surface behaviour and direction of travelling inshore bottlenose dolphins in Jervis Bay; however it appears that this impact is not reflected in their acoustic behaviour.     

Prey depletion caused by overfishing and the decline of marine megafauna in eastern Ionian Sea coastal waters (central Mediterranean)

Surveys primarily aimed at determining dolphin encounter rates were conducted from small inflatable craft in eastern Ionian Sea coastal waters between 1997 and 2004. During 633 surveys totalling 21,276 km of effort, observations of cetaceans and other marine species spotted in a study area of 480 km² were systematically recorded. Common dolphin encounter rates declined 25-fold across the study period, steadily decreasing from 2.18 encounters/100 km in 1997 to 0.09 encounters/100 km in 2004. Encounter rates of tuna also declined significantly. Swordfish encounter rates dropped from 1.03 encounters/100 km in 1997 to 0-0.12 in 1998-2004. Encounter rates of bottlenose dolphins did not show significant trends. The decline of high-order marine predators feeding on epipelagic prey was consistent with the hypothesis of prey depletion, likely resulting from intensive exploitation of local fish stocks, particularly anchovies and sardines. The catholic feeding habits and opportunistic behaviour of bottlenose dolphins may allow them to withstand the effects of overfishing at their present low density.     

Assessing the uncertainty of estimated annual totals of net ecosystem productivity: A practical approach applied to a mid latitude temperate pine forest

Values for annual NEP of micrometeorological tower sites are usually published without an estimate of associated uncertainties. Few authors quantify total uncertainty of annual NEP. Moreover, different methods to assess total uncertainty are applied, usually addressing only one aspect of the uncertainty. This paper presents a robust and easy to apply method to quantify uncertainty of annual totals of Net Ecosystem Productivity (NEP), related to multiple factors involved therein. The method was applied to NEP observations for a Scots pine forest (Loobos) in the Netherlands. Total uncertainty of annual NEP for the Loobos site was on average ±32 g C m⁻² a⁻¹ (±8% of NEP), which is a quarter of the standard deviation of annual NEP (127 g C m⁻² a⁻¹).     

Nitrogenase activity and trehalose content of nodules of drought-stressed common beans infected with effective (Fix) and ineffective (Fix) rhizobia

Nitrogenase activity and trehalose accumulation were measured in nodulated and non-nodulated common beans (Phaseolus vulgaris) that were exposed to drought. Plants were infected with the Fix⁺Rhizobiumsp NGR234, or a Fix⁻ derivative (NGRΩfixF), or high trehalose-producing, native rhizobia. Trehalose content increased significantly while acetylene reduction activity (ARA) decreased in the nodules of plants exposed to drought. Nevertheless, ARA decreased at a slower rate in nodules with high trehalose levels. Under water stress, nodules infected with NGRΩfixF tended to accumulate more trehalose than nodules infected with wild-type NGR234 (9±0.1 vs 8±0.1 mg g⁻¹ dw, respectively). Highest trehalose accumulations were registered in nodules of plants infected with native rhizobia (average 16 mg g⁻¹ dw), and these plants also exhibited the highest relative water content (65%), while in plants infected with the NGRΩfixF RWC was significantly lower (56%). Our results suggest that nodule trehalose may protect bacterial nitrogenase activity under drought conditions, and that both trehalose and biological nitrogen fixation (BNF) contribute to drought tolerance.     

Genetic isolation and morphological divergence of Black Sea bottlenose dolphins

The Black Sea is a semi-enclosed body of water that differs from the adjacent Mediterranean Sea in terms of its biodiversity, oceanographical and ecological characteristics. There is growing international concern about pollution in the Black Sea and other anthropogenic threats to its fauna. The bottlenose dolphin (Tursiops truncatus) is one of three species of cetaceans living in the Azov-Black Sea basin. Despite considerable research on bottlenose dolphins elsewhere, the extent of human impacts on the Black Sea populations is unknown. Previous attempts to award special conservation status to Black Sea cetaceans have failed specifically because policy makers have viewed their ecological and evolutionary uniqueness as equivocal. This study assessed divergence between Black Sea, Mediterranean Sea and Atlantic Ocean bottlenose dolphins for 26 cranial measurements (n = 75 adult bottlenose dolphin skulls) and mitochondrial DNA (n = 99 individuals). Black Sea bottlenose dolphins are smaller than those in the Mediterranean, and possess a uniquely shaped skull. As in a previous study, we found the Black Sea population to be genetically distinct, with relatively low levels of mtDNA diversity. Population genetic models suggest that Black Sea bottlenose dolphins have so little gene flow with the Mediterranean due to historical isolation that they should be managed separately.     

Lead tolerance in Aporrectodea rosea earthworms from a clay pigeon shooting site

Lead tolerance in individuals of the earthworm species Aporrectodea rosea collected from a clay pigeon shooting site was investigated. Lead concentrations in the shooting site soil and the un-shot control site were 6410±2250 and 296±98 mg_Pb kg⁻¹ dry weight, respectively. Of these concentrations 1050±240 and 12±9 mg_Pb kg⁻¹ dry weight were suggested to be available, using ammonium acetate (1 M), respectively. With respect to earthworm body burdens of lead the shooting site earthworms had a body burden of 6.1±1.2 mg_Pb g⁻¹ dry weight while the uncontaminated site earthworms had almost a 1000-times lower body burden of 7.1±9.0 μg_Pb g⁻¹ dry weight. Lead tolerance was assessed in uncontaminated soil that had been augmented with lead, using lead nitrate solutions, to obtain lead concentrations in soil of 0.5, 5 and 50 mg_Pb kg⁻¹ dry weight. Earthworms were exposed for 28 days during which time a semi-qualitative assessment was made of their condition. Results showed no decrease in condition in the shooting site earthworms with increasing exposure time or concentration. In contrast, earthworms collected from an uncontaminated site showed a significant (p<0.05) decrease in condition when exposed to lead concentrations above, and including, a concentration of 5 mg kg⁻¹ dry weight soil. These results suggested lead tolerance in the shooting site earthworms.     

The effect of mixing ground leaf litters to soil on the development of pitch pine ectomycorrhizal and soil arthropod communities in natural soil microcosm systems

The addition of leaf litter to soil influences both the nutrients and polyphenols of soil. It is likely that contrasting nutrient and polyphenolic composition of different plant litters may affect plant growth, mycorrhizal and soil arthropod communities. We report results from a microcosm experiment of effects of incorporation of three single leaf litter species and a mixture of all three on pitch pine seedling growth, their ectomycorrhizal community and soil arthropod community. The three litter species (pine, oak and huckleberry) represent co-dominant species within the New Jersey pine barrens ecosystem. We show that the leaf litters have different composition of nutrients and polyphenols, with rooting matrix containing pine litter having lower inorganic nitrogen content (1.6 μg g⁻¹) than oak (19.9 μg g⁻¹) and huckleberry (4.4 μg g⁻¹), but oak litter having the highest extractable phosphorus (13.3 cf. 0-0.08 μg g⁻¹) and total phenol content and lowest condensed tannin content. These differences were imparted to rooting matrix of homogenized humic (Oa) layer of pine barrens soil to which milled leaf litter was added and used in the microcosms. Pitch pine seedlings grew significantly better in un-amended rooting matrix (0.33±0.02 g) than any of the litter treatments (0.15±0.02-0.17±0.01 g) and tissue P concentrations tracked phosphate concentrations in the rooting matrix. Total P accumulation into plant tissue was higher in oak than control, attributable to a significantly higher (P<0.05) accumulation in roots (3.3±0.19 mg g⁻¹) compared to other species (1.1±0.04-2.3±0.08 mg g⁻¹). No relationship was seen between tissue N concentration and soil N, but seedlings growing in huckleberry litter amended soil accumulated less N than control. The effect of leaf litters on the ectomycorrhizal community composition were determined by PCA (first two axes accounted for 81% of the variance) and stepwise multiple regression analysis. These analyses showed that huckleberry leaf litter had a significant impact on mycorrhizal community composition with morphotypes Cg and DB being more abundant in the presence of huckleberry litter (178±13 cf. 68±15-106±15 for Cg and 141±11 cf. 88±23-111±18 for DB) and its influence of elevating nitrate nitrogen, organic nitrogen, total phenols and protein precipitation content of the rooting matrix. Mycorrhizal morphotypes BS and SB were significantly more abundant in the community where these soil factors were low in the absence of leaf litter addition. Total ectomycorrhizal abundance was negatively related to hydrolysable tannin concentration in the rooting matrix (r²=0.132, P<0.05). There was no influence of leaf litter type on mite density (dominated by non-burrowing phthiracarids), but collembolan density (dominated by Folsomia spp) showed a greater than threefold reduction in population density in the presence of leaf litter (F=6.47, P<0.05). Collembolan density was positively correlated with mycorrhizal morphotypes GS and SB (P<0.05) and negatively related to morphotypes DB (P<0.05) and soil extractable NH₄-N (P<0.05), suggesting a possible selection of fungal species in their diet and a relationship between collembola and nitrification.     

Distribution of monoterpenes between organic resources in upper soil horizons under monocultures of Picea abies, Picea sitchensis and Pinus sylvestris

The aim of this study was to compare the monoterpene content and distribution in litters and roots of three conifer species: Picea abies (L.) Karst, Picea sitchensis (Bong.) Carr. and Pinus sylvestris (L.). We analysed the monoterpene content of green needles, needle litter, F (fermentation) layer material and roots collected from monoculture plots. The rate of loss of monoterpenes from freshly fallen litter in the field was also studied at two monthly intervals over 10 months, to assess the length of time that monoterpenes entering the litter layer remain. Monoterpene analysis was carried out by extracting homogenised samples in hexane and identifying and quantifying the resulting monoterpenes using gas chromatography with flame ionisation detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Mean total monoterpene concentrations varied significantly between the three species examined (e.g. in freshly fallen litter 1531 ± 96, 100 ± 5 and 1175 ± 122 μg g⁻¹ d. wt for P. abies, P. sitchensis and P. sylvestris); each species had distinctive and consistent monoterpene profiles associated with each type of tissue, and total monoterpene concentrations in green needles varied between individual trees of the same species, particularly for P. sitchensis. A substantial proportion of the monoterpene content of green needles remained in the needles after litter fall for P. abies (42%), P. sitchensis (11%) and P. sylvestris (30%). Although rates of monoterpene loss from needle litters varied initially (P. sylvestris > P. abies > P. sitchensis), the majority of the monoterpene content was lost after 4-6 months. Maximum monoterpene emission rates from decaying litter were calculated of 39 (P. abies), 1.7 (P. sitchensis) and 39 μg m⁻² h⁻¹ (P. sylvestris). Monoterpene concentrations in F layer material were very low (<10 μg g⁻¹ d. wt). Roots, particularly in P. sylvestris, represented a significant pool of monoterpenes (185 ± 16, P. abies; 258 ± 54, P. sitchensis; 2133 ± 200 μg g⁻¹ d. wt, P. sylvestris). The monoterpene profile was similar between roots and litter of P. sylvestris (α-pinene most abundant), and for P. sitchensis, (limonene and α-pinene most abundant), although a different pattern was observed between needle litter (most abundant β-pinene) and roots (most abundant myrcene) of P. abies. The relatively high concentrations and different profiles of monoterpenes characterised in upper organic soil horizons here emphasise the need for their influence on soil ecological processes to be assessed.     

Nitrate leaching in soil: Tracing the NO3 sources with the help of stable N and O isotopes

Legumes increase the plant-available N pool in soil, but might also increase NO₃⁻ leaching to groundwater. To minimize NO₃⁻ leaching, N-release processes and the contribution of legumes to NO₃⁻ concentrations in soil must be known. Our objectives were (1) to quantify NO₃⁻-N export to >0.3 m soil depth from three legume monocultures (Medicago x varia Martyn, Onobrychis viciifolia Scop., Lathyrus pratensis L.) and from three bare ground plots. Furthermore, we (2) tested if it is possible to apply a mixing model for NO₃⁻ in soil solution based on its dual isotope signals, and (3) estimated the contribution of legume mineralization to NO₃⁻ concentrations in soil solution under field conditions. We collected rainfall and soil solution at 0.3 m soil depth during 1 year, and determined NO₃⁻ concentrations and δ¹⁵N and δ¹⁸O of NO₃⁻ for >11.5 mg NO₃⁻-N l⁻¹. We incubated soil samples to assess potential N release by mineralization and determined δ¹⁵N and δ¹⁸O signals of NO₃⁻ derived from mineralization of non-leguminous and leguminous organic matter.Mean annual N export to >0.3 m soil depth was highest in bare ground plots (9.7 g NO₃⁻-N m⁻²; the SD reflects the spatial variation) followed by Medicago x varia monoculture (6.0 g NO₃⁻-N m⁻²). The O. viciifolia and L. pratensis monocultures had a much lower mean annual N export (0.5 and 0.3 g NO₃⁻-N m⁻²). The averaged NO₃⁻-N leaching during 70 days was not significantly different between field estimates and incubation for the Medicago x varia Martyn monoculture.The δ¹⁵N and δ¹⁸O values in NO₃⁻ of rainfall (δ¹⁵N: 3.3±0.8‰; δ¹⁸O: 30.8±4.7‰), mineralization of non-leguminous SOM (9.3±0.9‰; 6.7±0.8‰), and mineralization of leguminous SOM (1.5±0.6‰; 5.1±0.9‰) were markedly different. Applying a linear mixing model based on these three sources to δ¹⁵N and δ¹⁸O values in NO₃⁻ of soil solution during winter 2003, we calculated 18-41% to originate from rainfall, 38-57% from mineralization of non-leguminous SOM, and 18-40% from mineralization of leguminous SOM.Our results demonstrate that (1) even under legumes NO₃⁻-N leaching was reduced compared to bare ground, (2) the application of a three-end-member mixing model for NO₃⁻ based on its dual isotope signals produced plausible results and suggests that under particular circumstances such models can be used to estimate the contributions of different NO₃⁻ sources in soil solution, and (3) in the 2nd year after establishment of legumes, they contributed approximately one-fourth to NO₃⁻-N loss.     

Microbial activity and functional composition among northern peatland ecosystems

The extent to which complex interrelationships between plants and microorganisms influence organic matter dynamics is critical to our understanding of global C cycles in changing environments. We examined the hypothesis that patterns of soil microbial activity and functional composition differ among vegetation types in northern peatland ecosystems. Microbial characteristics were compared among peatlands differing in plant growth form (tree, shrub/moss, sedge) in two regions (New York State and West Virginia). Microbial activity (basal respiration) was greater in surface (0-15 cm) than subsurface (15-30 cm) peat and from sites dominated by shrubs and Sphagnum moss (3.9±0.65 μg C g⁻¹ h⁻¹) compared to forested (1.8±0.20 μg C g⁻¹ h⁻¹) or sedge-dominated sites (1.9±0.38 μg C g⁻¹ h⁻¹). Microbial activity was not related to decomposability of peat organic matter among vegetation types, and activity was unexpectedly higher in sites with lower peat pH and higher water table level. Substrate-induced respiration (SIR) did not show a clear pattern among vegetation types, but was greater in surface than subsurface peat. Microbial responsiveness to added glucose was very low. The ratio of basal respiration to SIR varied between 0.39 and 0.72 and, like activity, was highest in shrub/Sphagnum sites. Microbial substrate utilization patterns (assayed with BIOLOG^® GN plates) also differed between shrub/Sphagnum sites and forest or sedge sites, suggesting that C fluxes were mediated by different assemblages of microorganisms in shrub/Sphagnum peatlands. Principal component (PC) scores indicated more utilization of N-containing compounds and carboxylic acids, and less utilization of carbohydrates by microbial communities in shrub/Sphagnum sites. PC scores were much more variable both within and among vegetation types for sites in West Virginia than in New York State, and a greater diversity of C sources were utilized in WV (57±3) than NYS (47±2) peat. Our results suggest a link between microbial respiratory activity and microbial functional composition as they vary among these peatland vegetation types.     

Soil CO2 efflux in a temperate deciduous forest: Environmental drivers and component contributions

Soil CO₂ efflux is a large component of total respiration in many ecosystems. It is important to understand the environmental controls on soil CO₂ efflux, in order to evaluate potential responses of ecosystems to climate change. This study investigated the relationship between total soil CO₂ efflux and soil temperature, soil moisture and solar radiation on an interannual basis for a plot of temperate deciduous ancient semi-natural woodland at Wytham Woods in central southern England. We also aimed to quantify the contribution of soil organic matter decomposition (SOM), root-and-rhizosphere respiration, and mycorrhizal respiration components to total soil CO₂ efflux, and determine their environmental correlates. Total soil CO₂ efflux was measured regularly from April 2006 to December 2008 and found to average 4.1 Mg C ha⁻¹ yr⁻¹ in both 2007 and 2008. In addition, we applied a recently developed approach to partition the efflux into SOM, root-and-rhizosphere, and mycorrhizal components in situ using mesh bags. SOM decomposition, root-and-rhizosphere, and mycorrhizal respiration were estimated to contribute 70 ± 6%, 22 ± 6% and 8 ± 3% of total soil CO₂ efflux respectively, equating to 3.0 ± 0.3, 0.9 ± 0.2 and 0.3 ± 0.1 Mg C ha⁻¹ yr⁻¹. In order to avoid the effect of temporal correlation between variables caused by seasonality, we investigated interannual variability by examining the relationship between CO₂ flux anomalies and anomalies in environmental variables. Variation in soil temperature explained 50% of the interannual variance in soil CO₂ efflux, and soil moisture a further 18% of the residual variance. Solar radiation, as a proxy for plant photosynthesis, had no significant effect on total soil CO₂ efflux, but was positively correlated with root-and-rhizosphere respiration, and mycorrhizal respiration. The relationship between anomalies in soil CO₂ efflux and soil temperature was highly significant, with a sensitivity of 0.164 ± 0.023 μmol CO₂ m⁻² s⁻¹ °C⁻¹. For mean peak summer efflux rates (2.03 μmol CO₂ m² s⁻¹), this is equivalent to 8% per °C, or a Q₁₀ temperature sensitivity of 2.2 ± 0.2. We demonstrate the utility of an anomaly analysis approach and conclude that soil temperature is the key driver of total soil CO₂ efflux primarily through its positive relationship with SOM-decomposition rate.     

Laccase and peroxidase isoenzymes during leaf litter decomposition of Quercus ilex in a Mediterranean ecosystem

The dynamics of leaf litter decomposition of Quercus ilex (L.) were investigated over a 2 year period by determining the activities and isoenzyme distribution of laccases and peroxidases. The analysis of isoenzymes was performed by isoelectric focusing on high stability pH gradients with high resolving power. The preparation of zymograms was carried out using the leaf litter extract without previous concentration. During litter decomposition, laccase and peroxidase activities changed as well as the type and number of enzyme isoforms. The activities of both enzymes were low (≤0.017 and ≤0.031 mmol o-tolidine oxidized h⁻¹ g⁻¹ d.w. for laccase and peroxidase, respectively) in first year and increased in October-January of the second year of litter decay. The highest activities measured after 15-18 months of litter exposure (0.37±0.03 and 0.19±0.02 mmol o-tolidine oxidized h⁻¹ g⁻¹ d.w. for laccase and peroxidase, respectively), showed that litter chemical composition affected the growth of ligninolytic microbial community. The activation energy for laccase and peroxidase reactions also changed during decomposition: the highest values (55±6 kJ mol⁻¹ for laccase and 60±6 kJ mol⁻¹ for peroxidase) occurred in autumn-winter, even if spatial changes were evidenced. Some enzyme isoforms (pI=5.3 and 5.5 for laccase and pI=5.0 and 5.1 for peroxidase, respectively), contributed more than others to the overall laccase and peroxidase activity, suggesting that some ligninolytic species bloomed in particular seasons of the year, even if other species with similar functional activities colonized the litter.     

Variation in NH4 mineralization and microbial communities with stand age in lodgepole pine (Pinus contorta) forests, Yellowstone National Park (USA)

Soils and vegetation were analyzed in 20 lodgepole pine (Pinus contorta) forest stands, varying in age from 50 to 350 years, that had initiated following stand-replacing fire. Our goal was to determine how nitrogen availability (NH₄⁺-N) and microbial community composition varied with stand age-class and to determine whether differences could be explained by canopy, soil, or understory characteristics. Gross NH₄⁺ mineralization was measured using laboratory isotopic pool dilution, and microbial community composition was evaluated using microbial membrane lipids. The microbial community composition of stands in the 300-350 age class was distinct from stands in younger age classes. Microbial community composition among sites varied with pH, % organic matter, and phosphorus. Gross NH₄⁺ mineralization rates averaged 1.45±0.07 mg NH₄⁺ kg soil⁻¹ d⁻¹ while consumption averaged 1.37±0.20 mg NH₄⁺ kg soil⁻¹ d⁻¹, resulting in low net NH₄⁺ mineralization rates (0.08±0.18 mg NH₄⁺ kg soil⁻¹ d⁻¹), but rates were not significantly different with stand age-class at p<0.05. At p<0.10, net NH₄⁺ mineralization was significantly higher in the 300-350 age class compared to the 125-175 age class. None of the measured variables significantly explained NH₄⁺ consumption and net mineralization patterns. However, gross NH₄⁺ mineralization rates were best explained by information on microbial community structure (i.e. lipids). Variation among stands within a given age-classes was high, indicating that patterns of N cycling across landscapes reflect substantial heterogeneity among mature stands.     

Microbial community response to nitrogen deposition in northern forest ecosystems

The productivity of temperate forests is often limited by soil N availability, suggesting that elevated atmospheric N deposition could increase ecosystem C storage. However, the magnitude of this increase is dependent on rates of soil organic matter formation as well as rates of plant production. Nonetheless, we have a limited understanding of the potential for atmospheric N deposition to alter microbial activity in soil, and hence rates of soil organic matter formation. Because high levels of inorganic N suppress lignin oxidation by white rot basidiomycetes and generally enhance cellulose hydrolysis, we hypothesized that atmospheric N deposition would alter microbial decomposition in a manner that was consistent with changes in enzyme activity and shift decomposition from fungi to less efficient bacteria. To test our idea, we experimentally manipulated atmospheric N deposition (0, 30 and 80 kg NO₃⁻-N) in three northern temperate forests (black oak/white oak (BOWO), sugar maple/red oak (SMRO), and sugar maple/basswood (SMBW)). After one year, we measured the activity of ligninolytic and cellulolytic soil enzymes, and traced the fate of lignin and cellulose breakdown products (¹³C-vanillin, catechol and cellobiose).In the BOWO ecosystem, the highest level of N deposition tended to reduce phenol oxidase activity (131±13 versus 104±5 μmol h⁻¹ g⁻¹) and peroxidase activity (210±26 versus 190±21 μmol h⁻¹ g⁻¹) and it reduced ¹³C-vanillin and ¹³C-catechol degradation and the incorporation of ¹³C into fungal phospholipids (p<0.05). Conversely, in the SMRO and SMBW ecosystems, N deposition tended to increase phenol oxidase and peroxidase activities and increased vanillin and catechol degradation and the incorporation of isotope into fungal phospholipids (p<0.05). We observed no effect of experimental N deposition on the degradation of ¹³C-cellulose, although cellulase activity showed a small and marginally significant increase (p<0.10). The ecosystem-specific response of microbial activity and soil C cycling to experimental N addition indicates that accurate prediction of soil C storage requires a better understanding of the physiological response of microbial communities to atmospheric N deposition.     

Use of 13C and 15N natural abundance techniques to characterize carbon and nitrogen dynamics in composting and in compost-amended soils

Isotope fractionation during composting may produce organic materials with a more homogenous δ¹³C and δ¹⁵N signature allowing study of their fate in soil. To verify this, C, N, δ¹³C and δ¹⁵N content were monitored during nine months covered (thermophilic; >40 °C) composting of corn silage (CSC). The C concentration reduced from 10.34 to 1.73 g C (g ash)⁻¹, or 83.3%, during composting. Nitrogen losses comprised 28.4% of initial N content. Compost δ¹³C values became slightly depleted and increasingly uniform (from −12.8±0.6‰ to −14.1±0.0‰) with composting. Compost δ¹⁵N values (0.3±1.3 to 8.2±0.4‰) increased with a similar reduced isotope variability.The fate of C and N of diverse composts in soil was subsequently examined. C, N, δ¹³C, δ¹⁵N content of whole soil (0-5 cm), light (<1.7 g cm⁻³) and heavy (>1.7 g cm⁻³) fraction, and (250-2000 μm; 53-250 μm and <53 μm) size separates, were characterized. Measurements took place one and two years following surface application of CSC, dairy manure compost (DMC), sewage sludge compost (SSLC), and liquid dairy manure (DM) to a temperate (C₃) grassland soil. The δ¹³C values and total C applied (Mg C ha⁻¹) were DM (−27.3‰; 2.9); DMC (−26.6‰; 10.0); SSLC (−25.9‰; 10.9) and CSC (−14.0‰; 4.6 and 9.2). The δ¹³C of un-amended soil exhibited low spatial (−28.0‰±0.2; n=96) and temporal (±0.1‰) variability. All C₄ (CSC) and C₃ (DMC; SSLC) composts, except C₃ manure (DM), significantly modified bulk soil δ¹³C and δ¹⁵N. Estimates of retention of compost C in soil by carbon balance were less sensitive than those calculated by C isotope techniques. One and two years after application, 95 and 89% (CSC), 75 and 63% (SSLC) and 88 and 42% (DMC) of applied compost C remained in the soil, with the majority (80-90%) found in particulate (>53 μm) and light fractions. However, C₄ compost (CSC) was readily detectable (12% of compost C remaining) in mineral (<53 μm) fractions. The δ¹⁵N-enriched N of compost supported interpretation of δ¹³C data. We can conclude that composts are highly recalcitrant with prolonged C storage in non-mineral soil fractions. The sensitivity of the natural abundance tracer technique to characterize their fate in soil improves during composting, as a more homogeneous C isotope signature develops, in addition to the relatively large amounts of stable C applied in composts.     

Modification and application of a soil ATP determination method

Accurate estimation of microbial adenosine 5′-triphosphate (ATP) is a pre-requisite to quantify the impact of varying environments on microbial activity of soil. We investigated the effectiveness of a high efficiency soil ATP determination method (PA) [Webster, J.J., Hampton, G.J., Leach, F.R., 1984. ATP in soil: a new extractant and extraction procedure. Soil Biology & Biochemistry 4, 335-342] in 10 Ontario (Canada) soils collected along a 100 m transect and spanning a textural class gradient ranging from a sandy loam to clay loam with increasing organic matter. Modifications of the method involved using an extract of autoclaved soil to make the standard curve, as it was found that the light emitted by ATP luciferin-luciferase bioluminescence reaction in the pure extractant was different from that in the extracts. Replacing Tricine with Tris buffer in the assay significantly improved the light emission. On an average, the internal standard calibration method (ISM) measured a smaller amount of extracted ATP (1199 ng ATP g⁻¹ soil) and a lower recovery of ATP spike (82.4±7.2%) than did the standard curve method (SCM) (1246 ng ATP g⁻¹ soil and 91.2±4.5%, respectively) (P<0.05 for both comparisons). However, the average total estimated ATP was higher with ISM (1474±102 ng ATP g⁻¹ soil) than with SCM (1373±88 ng ATP g⁻¹ soil) (P<0.07). While the recovery rates determined using SCM were consistent among the soils tested, the rates measured using ISM was negatively correlated with soil clay and organic matter content, implying that the latter assay was affected by the soil properties. Our results confirmed that the recovery rates obtained by the PA method were the highest among those reported, when only SCM was used.