Enzymatic degradation of polyhydroxyalkanoate using lipase from <Emphasis Type="Italic">Bacillus subtilis</Emphasis>

Polyhydroxyalkanoates (PHAs) are an important class of biodegradable polymers synthesized by a few bacteria under nutrient-limiting conditions. In this study, the lipase-catalysed degradation of PHA synthesized by Enterobacter sp. was monitored. For this, the lipase-encoding gene from Bacillus subtilis DI2 was PCR-amplified, cloned into a T vector system and sequenced. It was expressed in Escherichia coli DH5α cells, the recombinant enzyme was purified 24.25-fold, and its molecular weight was determined to be around 28 kDa. When PHA biodegradation studies were carried out with this enzyme, gel permeation chromatography showed 21.3 and 28.3 % molecular weight decrease and weight loss, respectively. Further, scanning electron micrographs revealed alterations in polymer surface morphology. Changes in molecular vibrations were noticed in the FTIR spectra. When the chemical shifts in NMR spectra were studied, a steep reduction in area under the peak at 1.57 ppm was observed. In the heating range of 30–930 °C employed during thermogravimetry analysis, the degraded sample showed a total of 45.82 % weight loss, as against 18.89 % for the native sample. The melting temperature (T _m) of the polymer was also brought down from 126.22 to 118.18 °C, as inferred from differential scanning calorimetry. Lipase-catalysed chain scission reactions could thus be used to generate low molecular weight functional biopolymers with wide-ranging pharmaceutical applications, such as in sustained drug release.     

Toxicity effects of four typical nanomaterials on the growth of <Emphasis Type="Italic">Escherichia coli</Emphasis>, <Emphasis Type="Italic">Bacillus subtilis</Emphasis> and <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>

To offer an insight into the toxicity of nanomaterials (NM) on the growth of bacteria, Escherichia coli (E. coli), Bacillus subtilis (B. subtilis) and Agrobacterium tumefaciens (A. tumefaciens) were exposed to nano-Au, nano-Ag, nano-Fe and fullerene (C₆₀) in this study. As an effective bactericide, nano-Ag induced high toxicity on these three bacteria; C₆₀ could inhibit their growth; however, B. subtilis and E. coli could recover as exposure time extended. Nano-Au and nano-Fe had hardly any effect on three bacteria. A. tumefaciens showed the lowest resistance and slowest growth rate during exposure. Images obtained by scanning electron microscope (SEM) revealed that nano-Ag could cause damage to the cell structure of three bacteria at 1 μg/mL. Slight damage on E. coli was found when exposed to C₆₀, whereas no obvious physical damage was found after exposure to nano-Au or nano-Fe. It is assumed that surface activities of NM might be responsible for the different toxic effects on these bacteria.     

Toxinogenicity and cytotoxicity of <Emphasis Type="Italic">Alternaria,Aspergillus</Emphasis> and <Emphasis Type="Italic">Penicillium</Emphasis> moulds isolated from working environments

There is currently limited research available on the secondary metabolites of moulds in workplaces. The aim of this study was to determine the mould contamination in museums (N = 4), composting plants (N = 4) and tanneries (N = 4) and the secondary metabolite profiles of Alternaria, Aspergillus and Penicillium isolates from these workplaces. Alternaria, Aspergillus and Penicillium species were identified using the ITS1/2 sequence of the rDNA region. Mould metabolites were quantitatively analysed on standard laboratory medium and mineral medium containing materials specific to each workplace using liquid chromatography-mass spectrometry. We also examined the cytotoxicity of the moulds using MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) assays. Air microbiological contamination analyses showed a number of microorganisms, ranging from 2.4 × 10³ CFU m^?3 (composting plants) to 6.8 × 10⁴ CFU m^?3 (tanneries). We identified high percentages of Alternaria, Aspergillus and Penicillium moulds (air 57–59%, surfaces 10–65%) in all workplaces. The following moulds were the most cytotoxic (>90%): Alternaria alternata, A. limoniasperae, Aspergillus flavus, Penicillium biourgeianum, P. commune and P. spinulosum. The same mould species isolated from different working environments exhibited varying toxigenic and cytotoxic properties. Modifying the culture medium to simulate environmental conditions most often resulted in the inhibition of secondary metabolite production. Moulds isolated from the working environments produced the following mycotoxins (ng g^?1): chanoclavines (0.28–204), cyclopiazonic acid (27.1–1045), fumigaclavines (0.33–10,640,000), meleagrin (0.57–13,393), roquefortins (0.01–16,660), rugulovasines (112–220), viridicatin (0.12–957), viridicatol (4.23–2753) and quinocitrinines (0.07–1104), which may have a negative impact on human health.     

Toxicity of vegetable tannin extract from <Emphasis Type="Italic">Acacia mearnsii</Emphasis> in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Vegetable tannins are complex polyphenols, which occur widely in nature. Traditionally, natural tannins have been used for tanning leather. In Brazil, the main tannin-based products are obtained from Acacia mearnsii, which is a leguminous tree native to Southeastern of Australia, and the first seeds were brought to Brazil in 1928. The main cultivation of acacia was established in Rio Grande do Sul state due to the possibility of raw material for charcoal, adhesives to fuel and for tanning leather. The leather tanning processes based on plant tannins are thought to be less harmful than chromium-based tanning, and it has been used as a sustainable alternative. However, there is scarce information about the environmental impact of the leather tanning processes, with most studies reporting inhibitory effects against microorganisms at high doses and stimulatory and positive health effects at low concentrations. The aim of this study was to evaluate the toxicity of tannin preparations extracted from A. mearnsii in Saccharomyces cerevisiae. Vegetable tannin toxicity in yeast was tested using two tannin treatments in different concentrations. In general, the results showed toxicity of vegetable in yeast, BY4741 and gsh1Δ strains.     

New thermoelastic parameters of natural <Emphasis Type="Italic">C</Emphasis>2/<Emphasis Type="Italic">c</Emphasis> omphacite

The compressibility at room temperature and the thermal expansion at room pressure of two disordered crystals (space group C2/c) obtained by annealing a natural omphacite sample (space group P2/n) of composition close to Jd₅₆Di₄₄ and Jd₅₅Di₄₅, respectively, have been studied by single-crystal X-ray diffraction. Using a Birch–Murnaghan equation of state truncated at the third order [BM3-EoS], we have obtained the following coefficients: V ₀ = 421.04(7) Å³, K _T0 = 119(2) GPa, K′ = 5.7(6). A parameterized form of the BM3 EoS was used to determine the axial moduli of a, b and c. The anisotropy scheme is β _c  ≤ β _a  ≤ β _b , with an anisotropy ratio 1.05:1.00:1.07. A fitting of the lattice variation as a function of temperature, allowing for linear dependency of the thermal expansion coefficient on the temperature, yielded α_V(1bar,303K) = 2.64(2) × 10⁻⁵ K⁻¹ and an axial thermal expansion anisotropy of α _b  ≫ α _a  > α _c . Comparison of our results with available data on compressibility and thermal expansion shows that while a reasonable ideal behaviour can be proposed for the compressibility of clinopyroxenes in the jadeite–diopside binary join [K _T0 as a function of Jd molar %: K _T0 = 106(1) GPa + 0.28(2) × Jd_(mol%)], the available data have not sufficient quality to extract the behaviour of thermal expansion for the same binary join in terms of composition.     

<Emphasis Type="Italic">An </Emphasis><Emphasis Type="Italic">Inconvenient Truth</Emphasis>: a focus on its portrayal of the hydrologic cycle

An Inconvenient Truth (AIT) has earned Al Gore an Oscar and a share of the 2007 Nobel Peace Prize and has been widely acclaimed by the mass media. However, significant errors exist in the film, owing to alarmism and exaggeration. As this forum does not provide for a detailed examination of these errors, this paper will focus only on the portrayal of the hydrologic cycle by AIT—precipitation and floods, soil moisture and droughts, and storminess. AIT argues that precipitation and intense rainfalls, floods, droughts, and the total number, intensity, and duration of tropical cyclones have all increased due exclusively to anthropogenically-driven climate change; indeed, AIT paints a picture of near scientific certainty with an overwhelming bias toward catastrophe scenarios. A closer look at the science, however, reveals that the data do not support these claims and that the scientific community is divided as to what the impact of anthropogenic climate change on the hydrologic cycle will be. Thus, the film gives a false impression of both the current state of climate change and that ‘the science is settled’.

<Emphasis Type="Italic">BV I</Emphasis><Subscript><Emphasis Type="Italic">c</Emphasis></Subscript> photometry of RR Lyrae stars

We have obtained 26 372 CCD frames in the B, V, and I _c filters for 81 RR Lyrae stars in 2008–2010, using the 76-cm telescope of the South African Astronomical Observatory and the 40-cm telescope of the Cerro Armazones Observatory, North Catholic University (Chile) using an SBIG ST-10XME CCD camera. For 12 of these RR Lyrae stars, we also obtained 337 brightness measurements in the B and V bands in 2000–2001 using the 60-cm telescope of the High Altitude Mt. Maidanak Observatory (Republic of Uzbekistan). We present tables of observations, light curves, and improved light-curve elements for all these RR Lyrae stars. The Blazhko effect was detected for SU Hor.     

The Intertidal Burrowing Crab <Emphasis Type="Italic">Neohelice</Emphasis> (=<Emphasis Type="Italic">Chasmagnathus</Emphasis>) <Emphasis Type="Italic">granulata</Emphasis> Positively Affects Foraging of Rodents in South Western Atlantic Salt Marshes

The role of positive and indirect interactions is often crucial in communities with intense abiotic stress such as salt marshes. The burrowing crab, Neohelice (=Chasmagnathus) granulata, is the dominant benthic macroinvertebrate of southwest Atlantic marshes (southern Brazil to Northern Argentinean Patagonia), having strong direct and indirect effects on marsh soil and, in consequence, on marsh vegetation and primary consumers. In this work, we investigate if this crab indirectly modifies habitat use by the granivorous rodents, Akodon azarae and Oligoryzomys flavescens, by increasing nutrient availability and thus enhancing seed production by the marsh plant Spartina densiflora. The study was conducted at the Mar Chiquita Coastal Lagoon, Argentina (37°32′ S). Rodent frequencies in S. densiflora were positively correlated with crab densities throughout the low and middle marsh. Additionally, the highest quality of S. densiflora and inflorescence density was recorded at the highest crab densities. Experimental manipulation of crab densities shows that N. granulata indirectly enhances the performance of S. densiflora (e.g., decreased fiber content and C/N ratios) and increases density of seeds. Moreover, N. granulata also facilitates S. densiflora seed availability to rodents by concentrating them in sediment mound at their burrows entrances. Experimental rodent exclusions showed that rodent species used S. densiflora seeds, a variable positively related to crab burrow density. Thus, our results show that N. granulata drives the granivorous rodent distribution and the intensity of seeds–rodent interaction trough facilitative and indirect interactions in marsh community.     

Feeding Behavior of the Native Mussel <Emphasis Type="Italic">Ischadium recurvum</Emphasis> and the Invasive Mussels <Emphasis Type="Italic">Mytella charruana</Emphasis> and <Emphasis Type="Italic">Perna viridis</Emphasis> in FL,USA, Across a Salinity Gradient

The feeding behavior of three species of mussels, the native Ischadium recurvum and the invasives Mytella charruana and Perna viridis, was studied in an invaded ecosystem in Florida (USA). In situ feeding experiments using the biodeposition method were performed along a salinity gradient in four different locations along the St. Johns River. Water characteristics, such as salinity, temperature, dissolved oxygen, and seston loads, were recorded to identify relationships between these variables and the feeding behavior of the mussels. Feeding behavior of the species varied by study site. Clearance, filtration, organic ingestion, and absorption rates of I. recurvum were negatively affected by salinity. For the invasive mussel, M. charruana, rejection was positively related to salinity while total ingestion, organic ingestion, and absorption rates were positively related to the percentage of organic matter in the seston. For P. viridis, total and organic ingestion rates were negatively affected by salinity but positively affected by total particulate matter. Condition indices for P. viridis and M. charruana were 13.16?±?0.64 and 6.63?±?0.43, respectively, compared to 4.82?±?0.41 for the native species I. recurvum, indicating that these mussels are well adapted to the environmental conditions in the area. This study indicates that the three species have different preferred habitats because of their specific responses to water characteristics. Thus, the invasive mussels will not totally occupy the niche of the native mussel in Florida despite overlapping zones. These data may help identify potential invaded areas and understand the extent of the invasion.     

Ecophysiology of the Olympia Oyster, <Emphasis Type="Italic">Ostrea lurida</Emphasis>, and Pacific Oyster, <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

The native Olympia oyster, Ostrea lurida, was once abundant in many US Pacific Northwest (PNW) estuaries, but was decimated by human activity in the late nineteenth early to twentieth centuries. Having been the subject of only few modern, detailed studies, a dearth of basic physiological information surrounded O. lurida and how it compared to the now dominant, non-native Pacific oyster, Crassostrea gigas. Utilizing laboratory and in situ studies in Yaquina Bay, OR, we explored the clearance rates of both species across a wide range of conditions. Pacific oysters not only had greater size-specific clearance rates than Olympia oysters, but also had a lower optimum temperature. Clearance rates for both species were reduced at lower salinity, at lower organic content, and at higher turbidity. Clearance rate models were constructed for each species using three approaches: (1) a single mechanistic model that incorporated feeding response functions of each species to the effects of temperature, salinity, turbidity, and seston organic content based on laboratory studies; (2) another additive model in which the number and type of response functions from laboratory studies were allowed to vary; and (3) a statistical model that utilized environmental data collected during in situ feeding trials. Clearance rate models that correlated feeding activity with in situ environmental data were found to often better predict oyster clearance rates (based on Adj R ²) for both species in Yaquina Bay, OR, than mechanistic, additive models based on laboratory feeding response functions; however, in situ correlative models varied in accuracy by species and season. This work represents important first steps towards better understanding the physiological ecology of the native Olympia oyster and how it differs from introduced and now dominant Pacific oyster.     

Gulf Ribbed Mussels (<Emphasis Type="Italic">Geukensia granosissima</Emphasis>) Increase Methane Emissions from a Coastal <Emphasis Type="Italic">Spartina alterniflora</Emphasis> Marsh

Understanding methane emissions from natural sources is becoming increasingly important with future climactic uncertainty. Wetlands are the single largest natural source of methane; however, little attention has been given to how biota and interactions between aboveground and belowground communities may affect methane emission rates in these systems. To investigate the effects of vegetative disturbance and belowground biogeochemical alterations induced by biota on methane emissions in situ, we manipulated densities of Littoraria irrorata (marsh periwinkle snails) and Geukensia granosissima (gulf ribbed mussels) inside fenced enclosures within a Spartina alterniflora salt marsh and measured methane emissions and sediment extracellular enzyme activity (phosphatase, β-glucosidase, cellobiohydrolase, N-acetyl-β-D-glucosaminidase, peroxidase, and phenol oxidase) over the course of a year. Changes in snail density did not have an effect on methane emission; however, increased densities of ribbed mussels significantly increased the emission of methane. Sediment extracellular enzyme activities for phosphatase, cellobiohydrolase, N-acetyl-β-D-glucosaminidase, and phenol oxidase were correlated to methane emission, and none of the enzymes assayed were affected by the snail and mussel density treatments. While methane emissions from salt marsh ecosystems are lower than those from freshwater systems, the high degree of variability in emission rates and the potential for interactions with naturally occurring biota that increase emissions warrant further investigations into salt marsh methane dynamics.     

Evaluation of <Emphasis Type="Italic">smtA</Emphasis> expression and <Emphasis Type="Italic">E. coli</Emphasis> survival against cadmium ions

Cadmium and other heavy metals lead to environmental danger, and these heavy metals are a great threat to human and other animal’s health. Investigation of the relationship between survival of E. coli and metallothionein smtA gene expression against cadmium ion is the goal of this research. Survival of recombinant bacteria containing smtA gene was analyzed against various concentrations of cadmium chloride salt using optical density (OD). At the resistive range, recombinant bacteria were subjected to different treatments. At the logarithmic phase of bacterial growth, sampling, RNA extraction and cDNA synthesis were performed and smtA gene expression was then analyzed by real-time PCR using designed primers for smtA gene and Amp resistance (as the calibrator gene). Relative gene expression was calculated using the ??C_t method. The resistive range against cadmium chloride was 0.5–0.7 mM (minimum inhibitory concentration (MIC = 0.5 mM)). Survival and gene expression analysis showed that in induced bacteria, smtA expression was increased significantly that in turn conferred resistance to cadmium chloride prominently. There was a direct relationship between increased smtA gene expression and survival of the recombinant bacteria. Therefore, our result may help to confront to cadmium metal environmental pollution using overexpression of smtA gene expression in recombinant bacteria.     

<Emphasis Type="Italic">Rhizedra lutosa</Emphasis>, a natural enemy of<Emphasis Type="Italic">Phragmites australis</Emphasis> in North America

Rhizedra lutosa (Hübner) (Lepidoptera: Noctuidae), native to Europe, was first found in North America in 1988 in southern New Jersey. The insect is reported to cause serious damage toPhragmites australis in the Netherlands, but it now appears to be relatively unimportant throughout central Europe. We are investigating this insect as part of an effort to characterize existing natural enemies ofP. australis in North America toward a goal of biological control of this invasive plant. We trapped adults ofR. lutosa with blacklights in Rhode Island, Massachusetts, Connecticut, New York, Delaware, and Maryland; we also found it in Ohio and Pennsylvania.R. lutosa larvae feed in stems and rhizomes ofP. australis growing in dry sites. Based uponR. lutosa damage surveys andP. australis biomass measurements, we have determined that this insect does cause a reduction of plant growth in some sites, however, because of low moth densities, this effect is small. Despite spreading rapidly,R. lutosa densities do not appear to be increasing at sites we have investigated, and this insect does not currently appear to have much of an impact uponP. australis in North America.     

Discovery of <Emphasis Type="Italic">Elephas</Emphasis> cf. <Emphasis Type="Italic">namadicus</Emphasis> from the late Pleistocene strata of Marginal Ganga Plain

We describe an elephant skull recovered from a cliff section of Dhasan river of Marginal Ganga Plain. The dental morphology and cranial features of the skull have been compared with the known species of Elephas from the Indian subcontinent. Although it shows very near resemblance to Elephas namadicus, but being an isolated specimen its specific identity cannot be proclaimed with certainty. As such, the specimen is provisionally referred as E. cf. namadicus. The Optically Stimulated Luminescence ages place this find at ~56 ka BP. This is the first chronologically well constrained report of E. cf. namadicus from the Ganga Plain.     

Physiological Constraints in Juvenile <Emphasis Type="Italic">Ostrea chilensis</Emphasis> Fed the Toxic Dinoflagellate <Emphasis Type="Italic">Alexandrium catenella</Emphasis>

Frequent blooms of the dinoflagellate Alexandrium catenella in southern Chile encouraged undertaking the present study which uses the oyster Ostrea chilensis as a model for evaluating the feeding, growth, lipid storage and mortality responses to diets containing paralytic shellfish poisoning (PSP) produced by A. catenella. Medium-term (30 days) physiological responses of two groups of juvenile oysters were measured every 10 days. Five replicates were exposed to diets containing A. catenella and other five replicates were fed with a diet containing the non-toxic algae Isochrysis galbana. Diets were continuously supplied at a concentration of 2 mg L^?1, in which the feeding and metabolic activity was measured, and the scope for growth calculated. Lipids storage, actual growth and mortality were also measured every 10 days. The results showed that the toxic diet has dramatic negative effects on feeding and metabolism of the juvenile individuals of O. chilensis, with high reduction of the lipid storage and growth. Mortality was also increased in individuals fed with the contaminated diet. This study supports the conclusion that the toxic dinoflagellate A. catenella restricts the energy acquisition in the juvenile O. chilensis, an important fishery and aquaculture resource in southern Chile.     

Isolation of <Emphasis Type="Italic">Bacillus cereus</Emphasis> from botanical soil and subsequent biodegradation of waste engine oil

Waste engine oil causes a vital environmental pollution when it spill during change and transportation and products of waste engine oil causes lethal effects to the living systems. Thus, abiotic and biotic approaches are being extensively used for removal of waste engine oil pollution. Therefore in present study, waste engine oil degradation was accomplished by a new bacterial culture, isolated from the soil by an enrichment technique. Morphological, biochemical and gene sequence analysis revealed that isolate was Bacillus cereus. Subsequently, biodegradation potential of B. cereus for waste engine oil was studied. Experimental variables, such as pH, substrate concentration, inoculum size, temperature and time on the biodegradation, were checked in mineral salt medium. The biodegradation efficiency of B. cereus was determined by gravimetry, UV–visible spectrophotometry and gas chromatography. In addition, waste engine oil was also characterized by GC–MS and FTIR for its major constituents, which showed total 38 components in waste engine oil, including hopanes, benzopyrene, long-chain aliphatic hydrocarbons, dibenzothiophenes, biphenyl and their derivatives. Results of successive biodegradation indicated that B. cereus was capable to degrade 1% of waste engine oil with 98.6% degradation potential at pH 7 within 20 days. Hence, B. cereus presents an innovative tool for removing the engine oil from the contaminated area.     

Estimation of aseismic crustal-strain using radon precursors of the 2003 <Emphasis Type="Italic">M</Emphasis> 6.8, 2006 <Emphasis Type="Italic">M</Emphasis> 6.1, and 2008 <Emphasis Type="Italic">M</Emphasis> 5.0 earthquakes in eastern Taiwan

Aseismic crustal-strain signals prior to the 2003 Mw 6.8 Chengkung, 2006 Mw 6.1 Taitung, and 2008 Mw 5.0 Antung earthquakes with epicenters located 20, 55 and 11 km, respectively, from the Antung radon-monitoring station have been calculated using the radon anomalies recorded. Specifically, radon decreased from background levels of 791 ± 46, 762 ± 57, and 735 ± 48 pCi/L to minima of 326 ± 9, 371 ± 9, and 480 ± 43 pCi/L prior to the 2003, 2006, and 2008 earthquakes, respectively. The estimated aseismic crustal-strain maxima at the Antung hot spring during the rock dilation stage were 3.6, 2.7, and 1.3 ppm, respectively. The v-shaped radon pattern recognized in all three anomalies is valuable for detecting the aseismic strain precursory to disastrous earthquakes in the Antung hot spring which is situated in a brittle fractured aquifer of limited recharge surrounded by ductile mudstone.