Partial sequencing and expression of genes involved in glucose metabolism in adipose tissues and skeletal muscle of healthy cats

Impaired insulin sensitivity is increasingly recognised in cats, but sequences of genes involved in insulin-signalling are largely undetermined in this species. In this study, extended feline mRNA sequences were determined for the adiponectin, glucose transporter-1 (GLUT1), GLUT4, peroxisome proliferative activated receptor-gamma1 (PPARgamma1), PPARgamma2, plasminogen activator inhibitor-1 (PAI-1), monocyte chemoattractant protein-1 (MCP-1) and insulin receptor genes. Conserved dog-specific primers identified from human-dog mRNA alignments were used to amplify feline cDNA in the polymerase chain reaction (PCR). The feline sequences determined by this method were used to design feline-specific primers suitable for real-time PCR for quantification of gene expression in insulin sensitive tissues of healthy cats. Partial sequences of feline mRNAs had 86-95% identity with dog and human genes. Expression of adiponectin, GLUT1, GLUT4, PPARgamma1, PPARgamma2, PAI-1 and insulin receptor mRNA was detected and quantified in subcutaneous and visceral fat and skeletal muscle, whereas MCP-1 mRNA was detected in adipose tissue but not in skeletal muscle. Further characterisation of genes related to glucose metabolism in cats will provide additional insights into insulin-signalling mechanisms in this species.     

Stable carbon isotopes accurately predict diet selection by sheep fed mixtures of C3 annual pastures and saltbush or C4 perennial grasses

The stable carbon isotope technique has been widely used to infer the dietary ecology of a range of animal species; however calibration of the technique with animals fed known diets is essential for accurate back-calculation of dietary preferences. The aim of this study was to identify suitable samples and back-calculation methods to predict short-term (2 to 3 week) dietary selection by sheep among plants with C₃ and C₄ photosynthetic pathways. Variation in integration time of dietary carbon into plasma and faeces; diet-tissue discrimination of carbon isotopes (fractionation) and the importance of accounting for the digestible or indigestible components of the diet was investigated. The results indicate that faecal and rumen samples provided the most accurate prediction of short term dietary changes in sheep selecting between C₃ and C₄ plants. The most accurate back-calculation method for these samples used δ¹³C of the C₃ and C₄ plants and accounted for both diet-tissue discrimination and differences in the indigestibility between the C₃ and C₄ forage. For faecal samples, the organic matter content of the diet originating from C₄ plants could be predicted with a mean error as low as 2.7%. Wool and plasma samples were not conducive to predicting proportion of C4 forage in the diet within 18 days after a change in diet; however plasma could be used to discriminate between animals fed 100% C₃ and C₄ diets after 3 days. The δ¹³C technique provides a valuable tool for researchers when designing pastures for dual environmental and production purposes. An understanding of what sheep select allows for development of appropriate grazing management strategies to optimise productivity and/or persistence of target species.     

A method to generate soilscapes from soil maps

Digital soil mapping for large areas is challenging if mapping resolution should be as high as possible and sampling should be as sparse as possible. Generally, the more complex the soil associations in a landscape, the more samples are required to systematically cover the entire feature space. Moreover, regions should be modeled separately if the patterns of spatial variation vary on subregion level. A systematic segmentation of a landscape into soilscapes is additionally important for a feasible application of soil‐sensing approaches. In this paper, we introduce a semiautomated approach to segment nominal spatial datasets based on the local spatial frequency distribution of the mapping units. The aim is to provide homogeneous and nonfragmented segments with smoothed boundaries. The methodological framework for the segmentation comprises different spatial and nonspatial techniques and focuses mainly on a moving‐window analysis of the local frequency distribution and a k‐means cluster analysis. Based on an existing soil map (1:50,000), we derived six segments for the Nidda catchment (Central Hesse, Germany), comprising 1600 km². As segmentation is based on a soil map, soilscapes are derived. In terms of the feature space, these soilscapes show a higher homogeneity compared to the entire landscape. Advantages compared to an existing map of landscape units are discussed. Segmenting a landscape as introduced in this study might also be of importance for other disciplines and can be used as a first step in biodiversity analysis or setting up environmental‐monitoring sites.     

Removal of short‐range‐order minerals prior to grain‐size analysis of volcanic ash soils

Incomplete dispersion of aggregates is a well‐known difficulty when analyzing soil texture of Andosols, particularly if no fresh soil material is available. To facilitate such investigations, several dispersion procedures were carried out on air‐dried samples rich in short‐range‐order minerals, originating from selected Andosol profiles (S Mexico). As a result, we propose an improved method based on a sequential chemical treatment with K oxalate, NH₄ oxalate, and oxalic acid that reliably removes cementing agents and allows for a stable dispersion of soil particles after addition of Na pyrophosphate. This method was proved to be well reproducible and highly reliable for routine grain‐size determinations. The potential error caused by the inherent risk of dissolving primary minerals or fresh volcanic glass remains far beyond the analytical error resulting from the incomplete and irreproducible dispersion of highly resistant aggregates by other commonly used techniques.     

Effects of silver nanoparticles on the microbiota and enzyme activity in soil

To assess the impact of silver nanoparticles (SNP) on soil microbial biomass, microbial activity, and enzyme activities, a medium‐term experiment over four months was performed in which soil was applied with increasing SNP‐application rates compared to a control. The treatments included a single SNP‐application dose analogous to 3.2 (SNP‐1), 32 (SNP‐10), and 320 (SNP‐100) μg Ag (kg dry soil)^–1 and a control without SNP application, respectively. At the end of the experiment, clear evidence was found that microbial biomass was significantly decreased with increasing SNP‐application rate, while basal respiration was increased in this direction. In addition, metabolic quotients were increased in the SNP treatments compared to the control. This is at least circumstantial evidence that the efficiency of substrate use was lowered in SNP‐treated soils. Another suggestion might be that after four months microbial‐community composition was changed due to SNP. No treatment effects were found for microbial biomass N, fluorimetric enzymes, and the abiotic soil parameters pH and soil organic C.     

Mixed forests and a flexible harvest policy: a problem for conventional risk analysis?

The famous “Faustmann” equation, which allows for identifying the most profitable tree species on a given unstocked piece of land, assumes constant timber prices. In reality, timber prices may fluctuate dramatically. Several authors have proven for monocultures that waiting for an acceptable timber price (reservation price) before harvesting (flexible harvest policy) increases the net present value of forest management. The first part of this paper investigates how efficient a flexible harvest strategy may be applied in mixed forests and whether the optimal species mixture is changed under such harvest policy. Mixtures of the conifer Norway spruce [Picea abies (L.) Karst] and the broadleaf European beech (Fagus sylvatica L.) were investigated. In order to evaluate mixed forests, the risks and the correlation of risks between tree species as well as the attitude towards risk of the decision-maker (risk-aversion is assumed) were considered according to the classical theory of optimal portfolio selection. In the second part we took up a recent critique on modern financial theory by Mandelbrot. Whether or not the assumption of normally distributed financial flows, which are supposed to occur under risk, would be appropriate to evaluate the risk of forest management was investigated. Market and hazard risks as well as their correlation were integrated in the evaluation of mixed forests by means of Monte-Carlo simulations (MCS). The risk of the timber price fluctuation was combined with the natural hazard risk, caused mainly by insects, snow and wind. Applying the μ-σ-rule, the mean net present value (NPV) from 1,000 simulations and their standard deviation were used for the optimisation. Given a low-return, risk-free interest rate to assess potential species mixtures of the Norway spruce and European beech, optimal proportions of European beech increased according to the theory of optimum portfolio selection with growing risk aversion from 0 (ignorance of risk) to 60% (great risk-aversion). In relation to a fixed harvest policy, the net present value of both, Norway spruce and European beech, could be increased significantly. Since the hazard risks of European beech were substantially lower compared with the Norway spruce (relation of susceptibility 1:4) beech benefited more from the flexible harvest policy. A comparison of simulated frequency distributions of the NPV with the expected density functions under the assumption of a normal distribution revealed significant differences. Only in the case of European beech was the general shape of the simulated frequency distribution similar to a normal distribution (bell-shaped curve). However, the density of NPV close to the mean was much greater than expected under the assumption of a normal distribution. Consequently, the frequency of a negative NPV for a European beech forest was greatly overestimated when applying the normal distribution. Though the shape of the simulated frequency distribution was rather different from a normal distribution for Norway spruce the simulated part of negative NPV was quite well approximated by the normal distribution. Therefore the simulated and expected frequencies of negative NPV were similar in case of Norway spruce; only a slight underestimation was seen in the assumption of a normal distribution. It can be concluded that actually simulated frequencies of negative NPV seem to be better measures for risk than computed probabilities of negative NPV, which assume normal distribution. As the risk for European beech was greatly overestimated by the conventional assumption of a normal distribution, the optimal proportions of European beech were surely rather underestimated according to the theory of portfolio. MCS on optimum mixtures derived by the classical portfolio theory seems necessary to test the robustness of such mixtures.

Identification of fatty acid esters of pectenotoxin-2 seco acid in blue mussels (Mytilus edulis) from Ireland

Pectenotoxins from marine dinoflagellates of the genus Dinophysis are rapidly hydrolyzed by many shellfish to give pectenotoxin-2 seco acid, which isomerizes to 7-epi-pectenotoxin-2 seco acid. Three series of fatty acid esters of pectenotoxin-2 seco acid (PTX-2 seco acid) and 7-epi-PTX-2 seco acid were detected by LC-MS analysis of extracts from blue mussels (Mytilus edulis) from Ireland. The locations of the fatty acid ester linkages were identified by a combination of LC-MSn in positive- and negative-ion modes, LC-MS analysis of the products from reaction of the esters with sodium periodate, and NMR analysis of purified samples of the two most abundant ester derivatives. The 37-O-acyl esters of PTX-2 seco acid were the most abundant, followed by the corresponding 11-O-acyl esters, accompanied by low levels of the 33-O-acyl esters. The most abundant fatty acid esters in the fractionated sample were, in order, the 16:0, 22:6, 14:0, 16:1, 18:4, and 20:5 fatty acids, although a wide array of other PTX-2 seco acid fatty acid esters were also present at low levels.     

Direct aqueous injection liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry analysis of water for atrazine, simazine, and their chlorotriazine metabolites

A method is reported for the determination of atrazine, simazine, and their respective dealkylated chlorotriazine metabolites in ground, surface, and finished drinking water. Water samples are diluted 1:4 in an injection vial prior to analysis using liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry (LC/ESI-MS/MS). The lower limit of method validation is 0.10 microg/L (ppb) for 2-chloro-4-(ethylamino)-6-isopropylamino)-s-triazine (atrazine, G-30027), 2-chloro-4, 6-(diethylamino)-s-triazine (simazine, G-27692), 2-amino-4-chloro-6-(isopropylamino)-s-triazine (deethylatrazine, DEA, or G-30033), 2-amino-4-chloro-6-(ethylamino)-s-triazine (deisopropylatrazine, DIA, or G-28279), and 2,4-diamino-6-chloro-s-triazine (didealkylatrazine, DDA, or G-28273). The overall mean procedural recoveries (and % relative standard deviations) for atrazine, simazine, DEA, DIA, and DDA are 98 (4.4), 102 (3.6), 99 (4.8), 103 (4.0), and 109% (4.8%), respectively, in finished drinking water; 108 (2.7), 104 (5.4), 113 (4.5), 111 (5.2), and 105% (5.3%), respectively, in groundwater; and 96 (6.9), 103 (4.2), 102 (4.4), 102 (5.2), and 102% (8.2%), respectively, in surface water. The method validation was conducted under U.S. EPA FIFRA Good Laboratory Practice Guidelines 40 CFR 160.     

Evidence of breed-dependent differences in susceptibility to porcine circovirus type-2-associated disease and lesions

Porcine circovirus type 2 (PCV2) has been confirmed as the primary cause of postweaning multisystemic wasting syndrome (PMWS). However, in the field, PMWS is seen only in a small percentage of pigs infected with PCV2. The overall objective of the study reported here was to determine whether host genetic differences in the susceptibility to PCV2-associated disease exist among selected breeds of pigs. This study included Duroc (n = 23), Landrace (n = 19), and Large White (n = 21) pigs. The pigs were infected intranasally and intramuscularly at 5-7 weeks of age with PCV2. A portion of the pigs (31/63; 30.2%) had low passively acquired PCV2 antibodies at the time of infection. There were no differences in mean weight gain, rectal temperature, or respiratory score. Clinical disease compatible with PMWS was observed only in the Landrace pigs. Most of the PCV2-infected pigs had enlarged lymph nodes, and individual Duroc and Landrace pigs had mottled tan lungs. PCV2-associated lymphoid depletion and granulomatous inflammation were observed in pigs of all breeds. Three of 19 Landrace pigs and none of the Duroc or Large White pigs developed severe lymphoid lesions associated with large amounts of intralesional PCV2 antigen typical of PMWS. Compared with seronegative Landrace pigs, Landrace pigs that had low maternal antibodies at the time of PCV2 inoculation had significantly (P < 0.05) less-severe PCV2-associated lesions. The results suggest a predisposition of the Landrace pigs of this study to PCV2-induced disease and lesions, and that low levels of passively acquired antibodies are protective.     

Cytochrome oxidase gene sequencing reveals channel catfish ovary cell line is contaminated with brown bullhead cells

The channel catfish (Ictalurus punctatus, Rafinesque) ovary (CCO) cell line is the standard cell line used for channel catfish diagnostics. Next‐gen sequencing studies of a virus cultured in the CCO cells revealed mitochondrial sequences matching those of brown bullhead (Ameiurus nebulosus, Lesueur). Therefore, we systematically performed partial cytochrome oxidase 1 gene sequencing of several sources of the CCO cell line and all matched the brown bullhead and not the channel catfish.