Simple and fast method for recognition of reducing and nonreducing neutral carbohydrates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Negative-ion mode matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry (TOF-MS) was used for the characterization of storage, neutral oligosaccharides extracted from Jerusalem artichoke, red onion, and wheat. The oligosaccharides from the real samples were analyzed with 2,4,6-trihydroxyacetophenone as the most convenient matrix that was selected in advance with the standard carbohydrate samples (inulin and maltooligosaccharides). The oligosaccharides from Jerusalem artichoke and red onion (similarly as inulin) produced [M - H](-) peaks as the main distribution, which reflects their nonreducing composition. On the contrary, the cross-ring fragmentations [M - H - 120](-) formed the main distribution in the mass spectra of hydrolyzed wheat starch similarly to reducing maltooligosaccharides and dextrans. The negative-ion mode MALDI-TOF MS is capable of recognizing reducing and nonreducing oligosaccharides. Such a simple differentiation of malto or inulin type of oligosaccharides is not possible in the positive-ion mode.     

How timing of stem girdling affects needle xylem structure in Scots pine

While needles represent a proportionally large fraction of whole-plant hydraulic resistance, no studies to date have investigated how source–sink disturbances affect needle xylem structure. In this study, we evaluated structural changes in xylem in current-year needles of Scots pine 227 and 411 days after stem girdling (hereafter referred to as DAG). Maximum and minimum tracheid lumen diameters and therefore also the size of tracheid lumen areas increased in needles 227 DAG compared to control needles. In contrast, tracheid dimensions were similar in needles 411 DAG as in the control needles, but smaller xylem area and lower number of tracheids resulted in the lower theoretical needle hydraulic conductivity of those needles. Several needle xylem parameters were intercorrelated in both control and girdled trees. These observed changes provide a new understanding of the processes that occur following a source–sink disturbance. Considering anatomical parameters such as the number of tracheids, tracheid dimension, or needle xylem area, which are rarely described in physiological studies, could be helpful, for example, in understanding to tree hydraulic systems or for modeling gas exchange. Finally, empirical equations were developed to calculate needle theoretical hydraulic conductivity and the number of tracheids in needles using an easily measurable parameter of needle xylem area.     

Incidence and control of the American giant liver fluke, Fascioloides magna, in a population of wild ungulates in the Danubian wetlands east of Vienna

Fascioloides magna,the giant liver fluke, is an introduced parasite imported to Europe with infected game which is endemic in the Austrian wetlands of the Danube from Vienna to the Slovakian border. Due to its pathogenicity (especially in roe deer) and its potential of transmission to domestic ruminants a project on the epidemiology and control of this parasite was carried out between 2000 and 2005. To assess distribution faecal droppings from red deer were collected in an area along the Danube and livers of red and roe deer shot or found dead were examined. Simultaneously, triclabendazole-medicated feed was offered repeatedly in several areas of known fluke presence from 2001. The average prevalence of infection as demonstrated by Fascioloides eggs in droppings (n = 145) was 6.3%, with > 50% in some areas. Infection rates decreased from 11.3% (01/2001) to 1.5% (04/2002) in the whole area. In Fischamend, a highly endemic area, prevalences in monthly samples (n = 25) decreased from 70% (01/2001) to less than 1% (01/2005). Similarly, the number of eggs per gram of faeces was significantly reduced. Of 457 red deer livers 15.8% were positive. In the Fischamend area, a reduction of positive livers was also found (2000:100%; 2005:13%), as well as a reduction of the numbers of flukes/liver. Control of F. magna in the Danubian wetlands by anthelmintic treatment proved to be successful although eradication was not achieved.     

Comparison of California Mastitis Test (CMT), Somatic Cell Counts (SCC) and bacteriological examinations for detection of camel (Camelus dromedarius) mastitis in Ethiopia

A total of 956 quarter milk samples from 253 traditionally managed lactating camels were collected aseptically from Negele (Borena Region), Dire Dawa, and Gewane (Afar Region), Ethiopia, according to multi-stage sampling. The quarter milk samples were subjected to California Mastitis Test (CMT), Somatic Cell Counts (SCC) and bacteriological examinations. Five hundred and seventy one (59.7%) quarter milk samples had microorganisms. Of these, 428 (75.0%) had isolates that were identified as major pathogens (MAP) and 143 (25.0%) as minor pathogens (MIP). A positive correlation was found between CMT scores and bacteriological classes (MAP, MIP) (p-value = 0.00). Strong correlation (p-value = 0.00) between CMT scores and SCC was recorded. The differences among the median log SCC of bacteriological classes (MAP, MIP) were not significant (p-value = 0.24). Similarly, the application of the cut-off level of 2.5 x 10(5) ml(-1) indicated less agreement (p-value = 0.32) for bacteriological classes MAP and MIP.     

Effect of cutting frequency on species richness and dry matter yield of permanent grassland after grazing cessation

The effect of grazing cessation management has only rarely been investigated. In this study, the influence of cutting regime in the absence of fertilization was examined during 2005–2011 on a semi-natural grassland in Rapotin (Czech Republic) which had been grazed for more than 30 years before the start of the trial. This was conducted as a randomized plot design with five levels of treatment: 4 C – intensive (four cuts per year); 3 C – medium intensive (three cuts per year); 2 C – low intensive (two cuts per year); 1 C – extensive (one cut per year); 0 C – abandoned (control; without any management). Species richness was significantly lower in treatments 1 C and 0 C than in other treatments. Management and year had similar and significant influence on species composition and explained 23% of its variability. Dry matter (DM) yields were found to be lower in three-cut than in two-cut treatments (5.56 t ha^?1 and 6.22 t ha^?1, respectively). In the case of grazing cessation, a suitable compromise of three cuts per year can be recommended to guarantee maintenance of species richness, botanical composition, and appropriate grass forage yield under similar site conditions.     

Butterfly dispersal in inhospitable matrix: rare,risky, but long-distance

Metapopulation models typically assume that suitable habitats occupied by local populations and unsuitable matrix separating them form a ‘black-and-white’ landscape mosaic, in which dispersal is primarily determined by the spatial configuration of habitat patches. In reality, however, the matrix composition is also likely to influence dispersal. Using intensive mark-recapture surveys we investigated inter-patch movements in Maculinea (Phengaris) nausithous and M. teleius occurring sympatrically in six metapopulations. Three of these metapopulations had the matrix dominated by forest, an inhospitable environment for grassland butterflies, whereas in the remaining three the matrix was mostly composed of open environments. Dispersal parameters derived with the Virtual Migration model revealed significant differences between both groups of metapopulations. Both species had a lower propensity to emigrate from their natal habitat patches, and they suffered substantially higher dispersal mortality in the metapopulations with forest matrix. On the other hand, mean dispersal distances were roughly an order of magnitude longer in forest matrix as compared with open landscapes (ca. 500–1,500 vs. 100–200 m). Our results suggest that inhospitable forest matrix induces strong selection against dispersal, leading to a reduced emigration rate. At the same time, the selection may promote emigrants with good dispersal abilities, which are able to perform long-distance movements. Thus, while it is generally believed that a matrix structurally similar to the habitat of a species should improve the functional connectivity of habitat patches, our findings imply that this may not necessarily be the case.     

Vitrification of fully grown and growing porcine oocytes using germinal vesicle transfer

The survival rate of vitrified germinal vesicle (GV) stage porcine oocytes is very low, and it is not known if the vitrification damages the nucleus, cytoplasm or both. We have evaluated the eventual GV or cytoplasmic damage in fully grown (FG) and growing vitrified oocytes. Fifty-five percent of nonvitrified FG cumulus-denuded oocytes reached the metaphase II (MII) stage in culture. When growing oocytes from preantral (PA) and early antral (EA) follicles were matured in vitro, almost all oocytes were arrested at the GV stage (GV stage: PA 88.9 and EA 79.5%, respectively). When fresh GVs from FG, PA and EA oocytes were transferred into fresh enucleated FG oocytes and matured in vitro, some of them reached the MII stage (MII stage: FG/FG 57.5%, PA/FG 9.3% and EA/FG 35.3%, respectively). The maturation rate of vitrified FG oocytes was only 6.1% but increased dramatically when vitrified GVs from FG, PA and EA oocytes were transferred into fresh enucleated FG oocytes (MII stage: VitFG/FG 43.9%, VitPA/FG 7.1% and VitEA/FG 26.3%, respectively). These results were not significantly different from those for the nonvitrified groups (MII stage: FG/FG 57.5%, PA/FG 9.3% and EA/FG 35.3%, respectively). We activated the reconstructed oocytes that received fresh or vitrified GVs (FG/FG, EA/FG, VitFG/FG and VitEA/FG) and examined their embryonic development. Cleaved embryos (nonvitrified groups 13.0-61.8%, vitrified groups 33.3-40.0%) and blastocysts (nonvitrified groups 0.0-18.2%, vitrified groups 0.0-2.9%) were obtained after activation. These results demonstrate that vitrified porcine GVs maintain maturational and developmental competence and that vitrification predominantly damages the cytoplasm.     

Determination of ergosterol levels in barley and malt varieties in the Czech Republic via HPLC

Ergosterol is considered to be a suitable indicator of mold infestation in barley and malt. In this study ergosterol levels in different varieties of barley and malt produced in the Czech Republic were determined. A modified high-performance liquid chromatography (HPLC) method was statistically processed, validated (Effivalidation program), and applied to 124 samples of barley and malt. Ergosterol was isolated by extraction and saponification, and the quantification was performed using HPLC with diode array detection. The content of ergosterol ranged between the limit of detection (LOD) and 36.3 mg/kg in barley and between the LOD and 131.1 mg/kg in malt. Ergosterol is presumably connected with metabolites generated when barley grain is attacked by pathogens, and such barley often shows a high overfoaming (gushing) value. However, it was found that the content of ergosterol does not correlate with the degree of beer gushing.     

Plasma cholesterol levels and short-term adaptations of metabolism and milk production during feed restriction in early lactating dairy cows on pasture

Low plasma total cholesterol (TC) concentrations are characteristic during the negative energy balance in early lactating dairy cows. The objective was to investigate short-term effects of different TC concentrations during an aggravated energy deficiency through a 1-week concentrate withdrawal on adaptations of metabolism and milk production. Multiparous Holstein cows (n = 15) were investigated during 3 week beginning at 24 ± 7 DIM (mean ± SD). Cows were kept on pasture and received additional concentrate in experimental week 1 and 3, while in week 2, concentrate was withdrawn. Blood was sampled once and milk twice daily. Based on their average TC concentration during week 1 (prior to concentrate withdrawal), cows were retrospectively assigned into a high (H-Chol; n = 8, TC ≥ 3.36 mmol/L) and a low TC groups (L-Chol; n = 7, TC < 3.36 mmol/L). Concentrations of phospholipids and lipoproteins were higher in H-Chol compared to L-Chol throughout the study (p < 0.05). During concentrate withdrawal, milk yield, glucose and insulin concentrations decreased similarly in both groups, while milk fat, milk acetone and plasma BHB were higher in H-Chol compared to L-Chol (p < 0.05). Compared to initial values, plasma NEFA, TAG and VLDL increased in both groups within 2 days after concentrate withdrawal (p < 0.05). Concentrations of NEFA during week 2 were greater in L-Chol compared to H-Chol (p < 0.05). Despite reintroduction of concentrate, milk yield in H-Chol remained lower for two more days compared to week 1 (p < .05), whereas milk yield recovered immediately in L-Chol. Activity of aspartate aminotransferase was higher in H-Chol compared to L-Chol in week 2 (p < 0.05). Greater plasma TC concentrations were associated with a reduced increase of NEFA. Further research is warranted if TC concentrations are related to adipose tissue mobilization and fatty acid turnover.