Threshold responses of songbirds to forest loss and fragmentation across the Marcellus-Utica shale gas region of central Appalachia,USA

Landscape Ecology - Since 2005, unconventional gas development has rapidly altered forests across the Marcellus-Utica shale basin in the central Appalachian region of the eastern United States, an...     

Multi-residue analysis of PAH, PCB, and OCP optimized for organic matter of forest soil

Purpose

Analyzing organic pollutants in forest soil is challenging because they are strongly physical and chemical bound to soil organic matter (SOM). Within the framework of a forest soil inventory, an analytical protocol for the determination of polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB), and organochlorine pesticides (OCP) should be established and validated using one and the same extraction and cleanup procedure. The protocol should be applicable for reliable analysis of a high number of samples in a short timeframe.

Materials and methods

Two different soil samples representative for the humic layer from a typical mixed and coniferous forest soil had been used for the analysis. Three solvents of different polarity, namely cyclohexane (CH), ethylacetate (EA)/CH (1/1, v/v), and acetone (AC)/CH (2/1, v/v), and the six standard extraction techniques (pressurized liquid extraction (PLE), soxhlet extraction, fluidized bed extraction, sonication, shaking, and one-step extraction recommended for analyzing agricultural soil in Germany (VDLUFA 2008)) were compared concerning their extraction efficiency. For additional matrix separation, two different cleanup procedures (gel permeation chromatography (GPC) and solid-phase extraction (SPE) with different sorbents) were tested. Quantification was carried out using gas chromatography combined with mass spectrometry (GC–MS) and two different injection systems (split/splitless injection and programmable temperature vaporizer (PTV) injection). Labeled internal standards, added prior to extraction, were used for method evaluation.

Results and discussion

For the simultaneous extraction of PAH, PCB, and OCP from organic forest soil PLE with acetone/cyclohexane (2/1, v/v) provided the highest extraction efficiency. A two-step cleanup procedure consisting of GPC followed by SPE with silica gel was entirely sufficient for the separation of humic substances without discrimination of analytes. Recovery rates for the different extraction and cleanup steps ranged between 89% and 106%. For quantification, a GC–MS method was developed using two different injection systems and two capillary columns of different selectivity.

Conclusions

By comparing six standard extraction techniques for PAH, PCB, and OCP from forest soil, we obtained the highest extraction efficiency when using PLE with AC/CH (2/1). For sample injection, we achieved best results using an optimized PTV injection system as it highly reduced the breakdown of thermolabile pesticides. Using this combination of technical equipment, it is possible to determine a concentration of the analytes in the trace level range of 1–2 μg kg^?1 in humic soil.     

Is CO2 evolution in saline soils affected by an osmotic effect and calcium carbonate?

Salt-affected soils are widespread, particularly in arid climates, but information on nutrient dynamics and carbon dioxide (CO₂) efflux from salt-affected soils is scarce. Four laboratory incubation experiments were conducted with three soils. To determine the influence of calcium carbonate (CaCO₃) on respiration in saline and non-saline soils, a loamy sand (6.3% clay) was left unamended or amended with NaCl to obtain an electrical conductivity (EC) of 1.0 dS?m^?1 in a 1:5 soil/water extract. Powdered CaCO₃ at rates of 0%, 0.5%, 1.0%, 2.5%, 5.0% and 10.0% (w/w) and 0.25-2 mm mature wheat residue at 0% and 2% (w/w) were then added. Cumulative CO₂-C emission from the salt amended and unamended soils was not affected by CaCO₃ addition. To investigate the effect of EC on microbial activity, soil respiration was measured after amending a sandy loam (18.8% clay) and a silt loam (22.5% clay) with varying amount of NaCl to obtain an EC_1:5 of 1.0–8.0 dS?m^?1 and 2.5 g glucose C?kg^?1 soil. Soil respiration was reduced by more than 50% at EC_1:5?≥?5.0 dS?m^?1. In a further experiment, salinity up to an EC_1:5 of 5.0 dS?m^?1 was developed in the silt loam with NaCl or CaCl₂. No differences in respiration at a given EC were obtained between the two salts, indicating that Na and Ca did not differ in toxicity to microbial activity. The effect of different addition rates (0.25–2.0%) of mature wheat residue on the response of respiration to salinity was investigated by adding NaCl to the silt loam to obtain an EC_1:5 of 2.0 and 4.0 dS?m^?1. The clearest difference between salinity levels was with 2% residue rate. At a given salinity level, the modelled decomposition constant ‘k’ increased with increasing residue addition rate up to 1% and then remained constant. Particulate organic carbon left after decomposition from the added wheat residues was negatively correlated with cumulative respiration but positively correlated with EC. Inorganic N (NH ₄ ⁺ -N and NO ₃ ^? -N) and resin P significantly decreased with increasing salinity. Resin P was significantly decreased by addition of CaCl₂ and CaCO₃.     

Eosinophilic keratoconjunctivitis in two rabbits

The purpose of this case report is to describe the clinical course and cytologic findings, treatment, and outcome of eosinophilic keratoconjunctivitis in two rabbits. Ophthalmic examination revealed ocular discharge, dacryocystitis, blepharitis, conjunctivitis, white conjunctival and corneal plaques, corneal vascularization, and stromal infiltration with different degrees of severity in each case. In case 2 there was also ulcerative disease of the cornea. Computerized tomography scan of the head, corneal biopsy for histopathologic examination with additional Luna and Giemsa stain were performed in case 2 and conjunctival as well as corneal specimens were obtained for bacteriologic culture and cytologic examination in case 1. Based on test results, a diagnosis of eosinophilic keratoconjunctivitis was made in case 2 and a tentative diagnosis of eosinophilic keratoconjunctivitis was made in case 1. Response to treatment with a topical steroid and topical cyclosporin was supportive of the diagnosis in both cases and shared many similarities with the response to treatment previously described in cats. Eosinophilic keratitis should be considered as part of a differential diagnosis list in rabbits with a history of keratitis.     

Different immune response of pigs to Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis infection

Mycobacterium avium subsp. avium (MAA) and Mycobacterium avium subsp. hominissuis (MAH) are the most common mycobacterial species isolated from granulomatous lesions in swine in countries with controlled bovine tuberculosis. This study is focused on the immunological aspect of MAA and MAH infection in pigs. We detected induction of humoral and cell-mediated immunity in experimentally infected pigs. Specific antibodies were analyzed in serum by ELISA and the IFN-γ release assay was used for evaluation of cell-mediated immunity. While MAA induced a significant increase of both types of immune responses, MAH-infected pigs had an unvarying level of specific antibodies and showed low cell-mediated immunity with high individual variability. The subsequent in vitro experiment confirmed the lower immunogenicity of the MAH strain in comparison to MAA. MAH-infected porcine monocyte-derived macrophages showed a weaker induction of pro-inflammatory mediators in comparison to MAA, which included mRNA for IL-1β, TNF-α, IL-23p19, IL-18 and chemokines CCL-3, CCL-5, CXCL-8 and CXCL-10. Additionally, qualitative proteomic analysis revealed 28 proteins exclusively in MAA and 7 proteins unique to MAH. In conclusion, closely related M. avium subspecies MAA and MAH showed different capacities to stimulate the porcine immune system. From a diagnostic point of view, the IFN-γ release assay showed higher sensitivity than the detection of specific antibodies by ELISA and seems to be an effective tool for discrimination of MAA-infected pigs. In the case of MAH infection, the IFN-γ release assay could fail because of the low immunogenic capacity of the MAH strain.     

Effects of sanitation processes on survival of Synchytrium endobioticum and Globodera rostochiensis

The two quarantine pests Synchytrium endobioticum, the causal agent of potato wart disease and Globodera rostochiensis, the yellow potato cyst nematode are currently present in Germany. Winter sporangia of Synchytrium endobioticum and cysts of Globodera rostochiensis can be spread with waste from potato processing industries, if infected tubers are processed. The German Biowaste Ordinance prescribes sanitation of organic waste before it can be used on arable land as fertilizer or filling material. Sanitation parameters prescribed by the German Biowaste Ordinance include composting for 7 days at 65°C or 14 days at 55°C or pasteurisation for 60 min at 70°C. The effect of composting and pasteurisation processes on winter sporangia of Synchytrium endobioticum and cysts of Globodera rostochiensis was tested with varying time-temperature relations. Cysts of Globodera rostochiensis were killed by composting for 7 days at 50–55°C and by pasteurisation for 30 min at 70°C. In contrast, viable winter sporangia of Synchytrium endobioticum could be extracted from sample material after composting for 70 days at 30–45°C, composting for 21 days at 50–55°C and after composting for 12 days at 60–65°C. Likewise viable winter sporangia could be extracted after pasteurisation for 90 min at 70°C and heating in a water bath at 80°C and in a dry oven at 90°C for 8 h. The parameters prescribed in the German Biowaste Ordinance are sufficient to kill cysts of Globodera rostochiensis but not sufficient to kill winter sporangia of Synchytrium endobioticum in organic waste.     

Postnatal functional maturation of blood phagocytes in pig

Developmental changes of functional ability of peripheral blood phagocytes from days 1 to 100 of life were investigated. Luminol enhanced chemiluminiscence was used to establish the ability of phagocytes to produce reactive oxygen species (ROS). Simple superoxide anion production was determined by spectrophotometrical measurement of cytochrome c. Activity of surface aminopeptidase N was assessed by spectrophotometrical measurement of l-alanine-p-nitroanilide. Flow cytometric measurements of CD18 and CD45 expression were performed. The ROS production per 0.5microl of blood did not show any trend; however, the values recalculated per 500 granulocytes had a decreasing course. The most noteworthy increase in production of superoxide anion occurred between days 17 and 26. Activity of aminopeptidase N decreased during the first 4 weeks. Expression of CD18 and CD45 intensively increased from days 1 to 14 with gradual decrease by day 100. Natural immunity develops during the early postnatal life and seems to be influenced by exposure of the organism to environmental antigens.     

Solvent-free lipase-catalyzed preparation of long-chain alkyl phenylpropanoates and phenylpropyl alkanoates

An enzymatic method was developed for the preparation of medium- or long-chain alkyl 3-phenylpropenoates (alkyl cinnamates), particularly alkyl hydroxy- and methoxy-substituted cinnamates such as oleyl p-coumarate and oleyl ferulate. The various alkyl cinnamates were formed in high to moderate yield by lipase-catalyzed esterification of cinnamic acid and its analogues with fatty alcohols in vacuo at moderate temperatures in the absence of drying agents and solvents. Immobilized Candida antarctica lipase B was the most effective biocatalyst for the various esterification reactions. The relative esterification activities were of the following order: dihydrocinnamic > cinnamic > 3-methoxycinnamic > dihydrocaffeic approximately 3-hydroxycinnamic > 4-methoxycinnamic > 2-methoxycinnamic > 4-hydroxycinnamic > ferulic approximately 3,4-dimethoxycinnamic > 2-hydroxycinnamic acid. With respect to the position of the substituents at the phenyl moiety, the esterification activity increased in the order meta > para > ortho. Rhizomucor miehei lipase demonstrated moderate esterification activity. Compounds with inverse chemical structure, that is, 3-phenylpropyl alkanoates such as 3-(4-hydroxyphenyl)propyl oleate, were also obtained in high yield by esterification of fatty acids with the corresponding 3-phenylpropan-1-ols.