Accumulation of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid-co-4-hydroxyvaleric acid) by mutants and recombinant strains ofAlcaligenes eutrophus

Alcaligenes eutrophus accumulated a terpolyester of 3-hydroxybutyric acid (3HB), 3-hydroxyvaleric acid (3HV), and 4-hydroxyvaleric acid (4HV) during cultivation with 4HV as carbon and energy source under nitrogen starvation. The polyester accumulated by wild-type strains under these conditions contained 4HV at a molar fraction of approximately 5 mol% only. A catabolic pathway of 4HV was postulated, which included the activation of 4HV to 4HV-CoA and a conversion of 4HV-CoA to 3HV-CoA. Tn5::mob-induced mutants were isolated fromA. eutrophus HF39, which were affected in 4HV and/or valeric acid catabolism. Among 83 mutants were 27 4HV-negative or 4HV-leaky mutants; two mutants were identified which accumulated a terpolyester with a molar fraction of 10.1 to 22.7 mol% 4HV. In addition, a further increase in the molar fraction of 4HV in poly(3HB-co-3HV-co-4HV) and a two- to fourfold increase in the PHA synthase activity were monitored in these mutants or others and also in HF39, if the cells were complemented with the hybrid plasmid pHP1014::PP1, which contained the PHA biosynthesis genes ofA. eutrophus H16. Application of mutagenesis plus recombinant DNA techniques resulted in the accumulation of a terpolyester with up to 30 mol% 4HV and with approximately equimolar fractions of 3HB, 3HV, and 4HV.     

Photodynamic action of some sensitizers by photooxidation of luminol

We report the development of a novel simple experimental method which allows the comparison of new photosensitizers based on their production of reactive oxygen species. A high-performance liquid chromatography (HPLC) assay permits the monitoring of several substances (sensitizer, reactant and oxidized end product) simultaneously on a single chromatogram. Photoreactions were monitored throughout their course by the HPLC assay surveying the sensitizers' efficiency of singlet oxygen production by the oxidative decomposition of luminol. Several photosensitizers were tested: Rose Bengal, Methylene Blue, Protoporphyrin IX, Photosan III, Photofrin, Hypericin and Pseudohypericin. Additionally, photoreactions were monitored by a standard pO₂ detection system. The measurements of the two detection methods were strongly correlated. Rose Bengal proved to be the most efficient photosensitizer, clearly decreasing the luminol concentration and causing a corresponding increase in aminophthalic acid. Our experiments show that when factors necessary for photochemical reactions are absent or are blocked (antioxidants), no reaction can be detected.     

Effect of organic acid on Cd toxicityin tomato and bean growth

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Ultrastructural study and calcium and cadmium localization in the marine coccolithophoridCricosphaera elongata

Cells of the coccolithophoridCricosphaera elongata (Droop) Braarud (Prymnesiophyceae) were concentrated by means of tangential ultrafiltration. Ultrastructural features of control cells were typical of the genusCricosphaera. The flagellar apparatus consists of two flagella, a bulbous haptonema, an axosome and a basal helical band, as recently reported in the literature for another coccolithophorid. We observed a very-well developed Golgi apparatus, displaying all the features of the peculiar Golgi of the Prymnesiophytes. Observations with the light microscope did not reveal any difference between control algae and algae contaminated with 1, 10 and 20 µmol Cd 1^–1 (i.e., 112 µg 1^–1, 1.12 and 2.24 mg Cd 1^–1) for 48 h. However, algae treated with the highest cadmium concentration exhibited loss of motility after 6 h contact, but without apparent ultrastructural damage. Qualitative X-ray microanalysis of the dense granules present in Golgi vacuoles of control cells demonstrated the presence of calcium and chlorine and, at a lower level, sulphur and aluminium. The same elements were also identified in the dense part of the lysosomes. In situ,C. elongata produces coccoliths; this did not occur in culture. However, the coccolith-generating apparatus (Golgi complex and Ca²⁺) was present in culture, and the cells were able to synthesize unmineralized scales.     

Groundwater dependent ecosystems. Part II. Ecosystem services and management in Europe under risk of climate change and land use intensification

Groundwater in sufficient amounts and of suitable quality is essential for potable water supplies, crop irrigation and healthy habitats for plant and animal biocenoses. The groundwater resource is currently under severe pressure from land use and pollution and there is evidence of dramatic changes in aquifer resources in Europe and elsewhere, despite numerous policy measures on sustainable use and protection of groundwater. Little is known about how such changes affect groundwater dependent ecosystems (GDEs), which include various aquatic and terrestrial ecosystems above ground and inside the aquifer. Future management must take this uncertainty into account. This paper focuses on multiple aspects of groundwater science, policy and sustainable management. Examples of current management methods and practices are presented for selected aquifers in Europe and an assessment is made of the effectiveness of existing policies such as the European Water Framework Directive and the Habitat Directive in practice and of how groundwaters and GDEs are managed in various conditions. The paper highlights a number of issues that should be considered in an integrated and holistic approach to future management of groundwater and its dependent ecosystems.     

Torpor use during gestation and lactation in a primate

Torpor is an energy-saving mechanism that allows endotherms to overcome energetic challenges. Torpor should be avoided during reproduction because of potential incompatibility with offspring growth. To test if torpor can be used during gestation and lactation to compensate for food shortage, we exposed reproductive female grey mouse lemurs (Microcebus murinus), a heterothermic primate, to different levels of food availability. Torpor use was characterised by daily skin temperature profiles, and its energetic outcome was assessed from changes in body mass. Food shortage triggered torpor during the end of the gestation period (n = 1), ranging from shallow in response to 40% food restriction to deep daily torpor in response to 80% restriction. During the early period of lactation, females fed ad libitum (n = 2) or exposed to a 40% restriction (n = 4) remained normothermic; but 80% food restricted females (n = 5) gave priority to energy saving, increasing the frequency and depth of torpor bouts. The use of torpor was insufficient to compensate for 80% energetic shortage during lactation resulting in loss of mass from the mother and delayed growth in the pups. This study provides the first evidence that a heterothermic primate can use torpor to compensate for food shortages even during reproduction. This physiological flexibility likely evolved as a response to climate-driven fluctuations in food availability in Madagascar.     

Biodiversity,carbon stocks and sequestration potential in aboveground biomass in smallholder farming systems of western Kenya

While Carbon (C) sequestration on farmlands may contribute to mitigate CO₂ concentrations in the atmosphere, greater agro-biodiversity may ensure longer term stability of C storage in fluctuating environments. This study was conducted in the highlands of western Kenya, a region with high potential for agroforestry, with the objectives of assessing current biodiversity and aboveground C stocks in perennial vegetation growing on farmland, and estimating C sequestration potential in aboveground C pools. Allometric models were developed to estimate aboveground biomass of trees and hedgerows, and an inventory of perennial vegetation was conducted in 35 farms in Vihiga and Siaya districts. Values of the Shannon index (H), used to evaluate biodiversity, ranged from 0.01 in woodlots through 0.4–0.6 in food crop plots, to 1.3–1.6 in homegardens. Eucalyptus saligna was the most frequent tree species found as individual trees (20%), in windrows (47%), and in woodlots (99%) in Vihiga and the most frequent in woodlots (96%) in Siaya. Trees represented the most important C pool in aboveground biomass of perennial plants growing on-farm, contributing to 81 and 55% of total aboveground farm C in Vihiga and Siaya, respectively, followed by hedgerows (13 and 39%, respectively) and permanent crop stands (5 and 6%, respectively). Most of the tree C was located in woodlots in Vihiga (61%) and in individual trees growing in or around food crop plots in Siaya (57%). The homegardens represented the second C pool in importance, with 25 and 33% of C stocks in Vihiga and Siaya, respectively. Considering the mean total aboveground C stocks observed, and taking the average farm sizes of Vihiga (0.6 ha) and Siaya (1.4 ha), an average farm would store 6.5 ± 0.1 Mg C farm^?1 in Vihiga and 12.4 ± 0.1 Mg C farm^?1 in Siaya. At both sites, the C sequestration potential in perennial aboveground biomass was estimated at ca. 16 Mg C ha^?1. With the current market price for carbon, the implementation of Clean Development Mechanism Afforestation/Reforestation (CDM A/R) projects seems unfeasible, due to the large number of small farms (between 140 and 300) necessary to achieve a critical land area able to compensate the concomitant minimum transaction costs. Higher financial compensation for C sequestration projects that encourage biodiversity would allow clearer win–win scenarios for smallholder farmers. Thus, a better valuation of ecosystem services should encourage C sequestration together with on-farm biodiversity when promoting CDM A/R projects.     

Abundance and size distribution dynamics of abyssal epibenthic megafauna in the northeast Pacific

The importance of interannual variation in deep-sea abundances is now becoming recognized. There is, however, relatively little known about what processes dominate the observed fluctuations. The abundance and size distribution of the megabenthos have been examined here using a towed camera system at a deep-sea station in the northeast Pacific (Station M) from 1989 to 2004. This 16-year study included 52 roughly seasonal transects averaging 1.2 km in length with over 35600 photographic frames analyzed. Mobile epibenthic megafauna at 4100 m depth have exhibited interannual scale changes in abundance from one to three orders of magnitude. Increases in abundance have now been significantly linked to decreases in mean body size, suggesting that accruals in abundance probably result from the recruitment of young individuals. Examinations of size-frequency histograms indicate several possible recruitment events. Shifts in size-frequency distributions were also used to make basic estimations of individual growth rates from 1 to 6 mm/month, depending on the taxon. Regional intensification in reproduction followed by recruitment within the study area could explain the majority of observed accruals in abundance. Although some adult migration is certainly probable in accounting for local variation in abundances, the slow movements of benthic life stages restrict regional migrations for most taxa. Negative competitive interactions and survivorship may explain the precipitous declines of some taxa. This and other studies have shown that abundances from protozoans to large benthic invertebrates and fishes all have undergone significant fluctuations in abundance at Station M over periods of weeks to years.     

Iteroparity in the variable environment of the salamander Ambystoma tigrinum

Simultaneous estimation of survival, reproduction, and movement is essential to understanding how species maximize lifetime reproduction in environments that vary across space and time. We conducted a four-year, capture-recapture study of three populations of eastern tiger salamanders (Ambystoma tigrinum tigrinum) and used multistate mark-recapture statistical methods to estimate the manner in which movement, survival, and breeding probabilities vary under different environmental conditions across years and among populations and habitats. We inferred how individuals may mitigate risks of mortality and reproductive failure by deferring breeding or by moving among populations. Movement probabilities among populations were extremely low despite high spatiotemporal variation in reproductive success and survival, suggesting possible costs to movements among breeding ponds. Breeding probabilities varied between wet and dry years and according to whether or not breeding was attempted in the previous year. Estimates of survival in the nonbreeding, forest habitat varied among populations but were consistent across time. Survival in breeding ponds was generally high in years with average or high precipitation, except for males in an especially ephemeral pond. A drought year incurred severe survival costs in all ponds to animals that attempted breeding. Female salamanders appear to defer these episodic survival costs of breeding by choosing not to breed in years when the risk of adult mortality is high. Using stochastic simulations of survival and breeding under historical climate conditions, we found that an interaction between breeding probabilities and mortality limits the probability of multiple breeding attempts differently between the sexes and among populations.     

Estimates of minimum patch size depend on the method of estimation and the condition of the habitat

Minimum patch size for a viable population can be estimated in several ways. The density-area method estimates minimum patch size as the smallest area in which no new individuals are encountered as one extends the arbitrary boundaries of a study area outward. The density-area method eliminates the assumption of no variation in density with size of habitat area that accompanies other methods, but it is untested in situations in which habitat loss has confined populations to small areas. We used a variant of the density area method to study the minimum patch size for the gopher tortoise (Gopherus polyphemus) in Florida, USA, where this keystone species is being confined to ever smaller habitat fragments. The variant was based on the premise that individuals within populations are likely to occur at unusually high densities when confined to small areas, and it estimated minimum patch size as the smallest area beyond which density plateaus. The data for our study came from detailed surveys of 38 populations of the tortoise. For all 38 populations, the areas occupied were determined empirically, and for 19 of them, duplicate surveys were undertaken about a decade apart. We found that a consistent inverse density area relationship was present over smaller areas. The minimum patch size estimated from the density-area relationship was at least 100 ha, which is substantially larger than previous estimates. The relative abundance of juveniles was inversely related to population density for sites with relatively poor habitat quality, indicating that the estimated minimum patch size could represent an extinction threshold. We concluded that a negative density area relationship may be an inevitable consequence of excessive habitat loss. We also concluded that any detrimental effects of an inverse density area relationship may be exacerbated by the deterioration in habitat quality that often accompanies habitat loss. Finally, we concluded that the value of any estimate of minimum patch size as a conservation tool is compromised by excessive habitat loss.