Purification and Properties of Extracellular Poly(3-hydroxybutyrate) Depolymerase Produced by <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> 202

An extracellular poly(3-hydroxybutyrate) (PHB) depolymerase produced by a thermotolerant fungal soil isolate, Aspergillus fumigatus 202, was purified and characterized. Maximum PHB depolymerase production was obtained at the end of 48 h with initial medium pH 7.0 and 45 °C in Bushnell Haas Minerals medium containing PHB as sole source of carbon. The PHB depolymerase was purified using size exclusion chromatography to a fold purification of 20.62 and 61.62% yield. SDS-PAGE and isoelectric focusing revealed the molecular weight and pI of the purified enzyme as 63,744 Da and 4.2, respectively. N-terminal amino acid sequence of purified enzyme was HAXDAYLVK. This non-glycosylated enzyme was most active at pH 9.0 and 45 °C. Purified enzyme was inactivated by N-bromosuccinimide and dithiothreitol suggesting the involvement of tryptophan residues and disulfide bonds at its active site. Nonionic detergents like Tween 20, Tween 80 and Triton X-100 inhibited the enzyme activity. Ions like Ca⁺² and Mg⁺² (5 mM) increased the enzyme activity 1.5 times. Fe⁺² effectively inhibited the enzyme activity to 88% whereas Hg⁺² completely inhibited the enzyme.     

Local Responses to Participatory Conservation in Annapurna Conservation Area, Nepal

Biodiversity conservation has undergone a profound change in philosophy, policies and management approaches over the last forty years. The traditional top–down approach to nature protection has been widely criticized for failing to include critical social elements in management practices, and is being gradually replaced by a slew of participatory strategies under the rubric of bottom-up conservation. The new approach recognizes local communities as key partners in wildlife management and seeks their participation in social development and biodiversity conservation. However, every social context is different in its structure and functions, and in the way social groups respond to calls for participation. In order to gain a better understanding of the approach and the barriers encountered in its implementation, a questionnaire survey of 188 households was employed in the communities of the Upper Mustang extension of Annapurna Conservation Area (ACA) in Nepal. The study provides a comparative analysis of community participation and its barriers between Non-Tourist (NT) and Tourist (TV) villages. The results revealed important differences between the two groups in terms of their participation in community programs, barriers to participation, and perception of benefits from participation. Owing to their distinct spatial, demographic and attitudinal differences, the two village groups have their own sets of needs, values and motivation factors which cannot be generalized and treated as such. The research clearly identifies the need for the conservation agency to be creative in devising strategies and initiatives appropriate to specific social groups so as to optimize their input in participatory conservation.     

Denitrification in anaerobic lagoons used to treat swine wastewater

Anaerobic lagoons are commonly used for the treatment of swine wastewater. Although these lagoons were once thought to be relatively simple, their physical, chemical, and biological processes are very complex. This study of anaerobic lagoons had two objectives: (i) to quantify denitrification enzyme activity (DEA) and (ii) to evaluate the influence of lagoon characteristics on the DEA. The DEA was measured by the acetylene inhibition method. Wastewater samples and physical and chemical measurements were taken from the wastewater column of nine anaerobic swine lagoons from May 2006 to May 2009. These lagoons were typical for anaerobic swine lagoons in the Carolinas relative to their size, operation, and chemical and physical characteristics. Their mean value for DEA was 87 mg N2O-N m(-3) d(-1). In a lagoon with 2-m depth, this rate of DEA would be compatible with 1.74 kg N ha(-1) d(-1) When nonlimiting nitrate was added, the highest DEA was compatible with 4.38 kg N ha(-1) d(-1) loss. Using stepwise regression for this treatment, the lagoon characteristics (i.e., soluble organic carbon, total nitrogen, temperature, and NO3-N) provided a final step model R2 of 0.69. Nitrous oxide from incomplete denitrification was not a significant part of the system nitrogen balance. Although alternate pathways of denitrification may exist within or beneath the wastewater column, this paper documents the lack of sufficient denitrification enzyme activity within the wastewater column of these anaerobic lagoons to support large N2 gas losses via classical nitrification and denitrification.     

Evaluation of the gene expression changes in Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) as affected by the bio-removal of toxic textile dyes from aqueous solution in small-scale bioreactor

A laboratory-scale bioremediation unit was designed, built and tested for the bio-removal of several Direct textile dyes. Four experiments were carried out to assess the efficiency of the bioremediation unit using Aspergillus niger fungal strain. Three commonly used Direct dyes and textile dyes mixture (simulated effluent: Direct brown, Direct violet, Direct green) were tested in this study. The strain of A. niger was efficient in the removal of the three Direct dyes. The decolorization percentages of the dyes after 24 h of incubation were 56.2, 51.7, and 95.4% for Direct brown, Direct green, Direct violet dyes, respectively. The percentages increased up to 79.4, 86.4, and 96.7% after 72 h of incubation for the same dyes, respectively. The results also showed that the fungal strain reduced the chemical oxygen demand values of simulated dye effluents from 165 to 564 mg/l with most of the dyes. The assessment of bioremediation products on biomodel was conducted using a fresh water fish. The liver and brain of Nile tilapia were tested to evaluate the expression of genes coding for several proteins related to stress such as metallothioneins (MTs), cytochrome P450 (CYP450), and heat shock proteins (HSPs). To assess the alterations in the gene expression, ten animals from each group were killed after 4 weeks of treatment. The results revealed significant increases in the brain and hepatic mRNA levels of all stress protein genes MT, CYP450, Hsp70a, b, and Hsp47 in the fish groups treated with industrial Direct violet, green, and brown dye water. Exposure of tilapia to bioremediation products after treatment with A. niger fungi reduced the over-expression of the stress protein genes in the brain and liver tissues.     

Carbon Storage in Soil Size Fractions Under Two Cacao Agroforestry Systems in Bahia, Brazil

Shaded perennial agroforestry systems contain relatively high quantities of soil carbon (C) resulting from continuous deposition of plant residues; however, the extent to which the C is sequestered in soil will depend on the extent of physical protection of soil organic C (SOC). The main objective of this study was to characterize SOC storage in relation to soil fraction-size classes in cacao (Theobroma cacao L.) agroforestry systems (AFSs). Two shaded cacao systems and an adjacent natural forest in reddish-yellow Oxisols in Bahia, Brazil were selected. Soil samples were collected from four depth classes to 1 m depth and separated by wet-sieving into three fraction-size classes (>250 μm, 250–53 μm, and <53 μm)—corresponding to macroaggregate, microaggregate, and silt-and-clay size fractions—and analyzed for C content. The total SOC stock did not vary among systems (mean: 302 Mg/ha). On average, 72% of SOC was in macroaggregate-size, 20% in microaggregate-size, and 8% in silt-and-clay size fractions in soil. Sonication of aggregates showed that occlusion of C in soil aggregates could be a major mechanism of C protection in these soils. Considering the low level of soil disturbances in cacao AFSs, the C contained in the macroaggregate fraction might become stabilized in the soil. The study shows the role of cacao AFSs in mitigating greenhouse gas (GHG) emission through accumulation and retention of high amounts of organic C in the soils and suggests the potential benefit of this environmental service to the nearly 6 million cacao farmers worldwide.     

A Generic Bio-Economic Farm Model for Environmental and Economic Assessment of Agricultural Systems

Bio-economic farm models are tools to evaluate ex-post or to assess ex-ante the impact of policy and technology change on agriculture, economics and environment. Recently, various BEFMs have been developed, often for one purpose or location, but hardly any of these models are re-used later for other purposes or locations. The Farm System Simulator (FSSIM) provides a generic framework enabling the application of BEFMs under various situations and for different purposes (generating supply response functions and detailed regional or farm type assessments). FSSIM is set up as a component-based framework with components representing farmer objectives, risk, calibration, policies, current activities, alternative activities and different types of activities (e.g., annual and perennial cropping and livestock). The generic nature of FSSIM is evaluated using five criteria by examining its applications. FSSIM has been applied for different climate zones and soil types (criterion 1) and to a range of different farm types (criterion 2) with different specializations, intensities and sizes. In most applications FSSIM has been used to assess the effects of policy changes and in two applications to assess the impact of technological innovations (criterion 3). In the various applications, different data sources, level of detail (e.g., criterion 4) and model configurations have been used. FSSIM has been linked to an economic and several biophysical models (criterion 5). The model is available for applications to other conditions and research issues, and it is open to be further tested and to be extended with new components, indicators or linkages to other models.     

A contextual framework for understanding good practice in integrated catchment management

Principles of good practice for collaborative resource management were derived from the literature and their use studied in a range of integrated catchment management processes. Desk-based reviews and interviews with participants allowed the principles to be refined and described within a framework that illustrates the interrelationships between core principles, enabling principles, precursors to a project and the influence of external factors on such collaborative processes. The findings illustrate the importance of these relationships in understanding how success is defined and under what conditions successful outcomes can be achieved. Understanding how these procedural aspects influence outcomes contributes to the wider literature on collaborative resource management that often treats processes separately from their context.     

Benzyl-penicillin (Penicillin G) transformation in aqueous solution at low temperature under controlled laboratory conditions

Antibiotics are released into the environment in a variety of ways: via wastewater effluent as a result of incomplete metabolism in the body after use in human therapy, as runoff after use in agriculture, through improper disposal by private households or hospitals or through insufficient removal by water treatment plants. Unlike in most European countries, in Arctic regions effluents are not suitably treated prior to their release into the aquatic environment. Also, many of the scattered human settlements in remote regions of the Arctic do not possess sewage treatment facilities and pharmaceutical residues therefore enter the aqueous environment untreated. Only limited data are available on the biodegradation of antibiotics under Arctic conditions. However, such information is needed to estimate the potential harm of antibiotics for the environment. Pen-G is used in this study since it is a widely prescribed antibiotic compound whose environmental properties have not yet been investigated in detail. Thus, for a very first assessment, the OECD approved biodegradation Zahn-Wellens test (ZWT, OECD 302 B) was used to study biodegradation and non-biotic elimination of the antibiotic Benzyl-penicillin (Pen-G) at different temperatures (5°C, 12.5°C and 20°C). The testing period was extended from the OECD standard of 28-42d. In addition to dissolved organic carbon (DOC), Pen-G levels and major transformation products were recorded continuously by LC-ion-trap-MS/MS. DOC monitoring revealed considerable temperature dependence for the degradation process of Pen-G. DOC loss was slowest at 5°C and considerably faster at 12.5°C and 20°C. In the initial step of degradation it was found that Pen-G was hydrolyzed. This hydrolyzed Pen-G was subsequently further degraded by decarboxylation, the result of which was 2-(5,5-dimethyl-1,3-thiazolidin-2-yl)-2-(2-phenylacetamido)acetic acid. Furthermore, direct elimination of 2-phenyl-acetaldehyde from the hydrolyzed and decarboxylated Pen-G also led to the formation of 2-[amino(carboxy)methyl]-5,5-dimethyl-1,3-thiazolidone-4-carboxylic acid. Since biodegradation slows down considerably at a low temperature, the resulting transformation products had considerably longer residence times at 5°C compared to higher temperature conditions within the 42-d experiment. The results presented here clearly demonstrate that a risk assessment for pharmaceuticals present in low ambient temperature environments (i.e. the Arctic) cannot be based on test results obtained under standard laboratory conditions (i.e. 20°C ambient temperatures).