Hydrolysis of Fucoidan by Fucoidanase Isolated from the Marine Bacterium,Formosa algae

Intracellular fucoidanase was isolated from the marine bacterium, Formosa algae strain KMM 3553. The first appearance of fucoidan enzymatic hydrolysis products in a cell-free extract was detected after 4 h of bacterial growth, and maximal fucoidanase activity was observed after 12 h of growth. The fucoidanase displayed maximal activity in a wide range of pH values, from 6.5 to 9.1. The presence of Mg²⁺, Ca²⁺ and Ba²⁺ cations strongly activated the enzyme; however, Cu²⁺ and Zn²⁺ cations had inhibitory effects on the enzymatic activity. The enzymatic activity of fucoidanase was considerably reduced after prolonged (about 60 min) incubation of the enzyme solution at 45 °C. The fucoidanase catalyzed the hydrolysis of fucoidans from Fucus evanescens and Fucus vesiculosus, but not from Saccharina cichorioides. The fucoidanase also did not hydrolyze carrageenan. Desulfated fucoidan from F. evanescens was hydrolysed very weakly in contrast to deacetylated fucoidan, which was hydrolysed more actively compared to the native fucoidan from F. evanescens. Analysis of the structure of the enzymatic products showed that the marine bacteria, F. algae, synthesized an α-l-fucanase with an endo-type action that is specific for 1→4-bonds in a polysaccharide molecule built up of alternating three- and four-linked α-l-fucopyranose residues sulfated mainly at position 2.     

Reproductive development response to high daytime temperature in field pea

Flowering plants are highly sensitive to heat stress during reproductive phase, which covers development from floral initiation to seed maturity. The objectives of this study were to diagnose high temperature effects on pollen production and morphology, production of reactive oxygen species (ROS) in pollen grains and ovules in pea cultivar “CDC Golden.” This study also investigated timing and duration of heat exposure at specific developmental stages of floral buds, open flowers and early set pods on flower and pod abortion, seed development and seed yield in “CDC Golden” and a second cultivar “CDC Sage.” The experiments were conducted in growth chambers with two temperature regimes (24/18°C and 35/18°C day/night temperature for 4–14 days) during reproductive development. Heat stress reduced the number of pollen grains per anther, induced smaller pollen grains and increased ROS production in pollen grains, but it did not affect ROS accumulation in ovules and ovule number per ovary. Heat exposure when young floral buds were visible at the first reproductive node was more detrimental to flower retention, seed set, pod development and seed yield compared to heat exposure started later when flowers at the second reproductive node were fully open.     

Characterization and plant expression of a glyphosate-tolerant enolpyruvylshikimate phosphate synthase

BACKGROUND: Glyphosate tolerance is a dominant trait in modern biotech crops. RESULTS: A gene encoding a glyphosate-tolerant EPSP synthase (aroA(1398)) from bacterial strain ATX1398 was cloned and characterized. The protein is initiated at a GTG translational start codon to produce a protein that provides robust glyphosate resistance in Escherichia coli (Mig) Cast & Chalm. The aroA(1398) protein was expressed and purified from E. coli, and key kinetic values were determined (K(i) = 161 microM; K(m)(PEP) = 11.3 microM; k(cat) = 28.3 s(-1)). The full-length enzyme is 800-fold more resistant to glyphosate than the maize EPSP synthase while retaining high affinity for the substrate phosphoenol pyruvate. To evaluate further the potential of aroA(1398), transgenic maize events expressing the aroA(1398) protein were generated. T(0) plants were screened for tolerance to glyphosate sprays at 1.3x commercial spray rates, and T(1) plants were selected that completely resisted glyphosate sprays at 1x, 2x and 4x recommended spray rates in field trials. CONCLUSION: These data suggest that aroA(1398) is a suitable candidate for conferring glyphosate tolerance in transgenic crop plants.     

Spreading of West Nile virus infection in Croatia

West Nile virus (WNV) is an emerging zoonotic pathogen with rapid global expansion. The virus circulation is confirmed in many countries of Mediterranean Basin and Southern and Central Europe. In our study detection of specific WNV antibodies was performed in horses and cattle sera samples collected from October 2010 to April 2011. Serum samples were randomly taken from different parts of Croatia and tested by IgG and IgM ELISA. Positive serological results were confirmed by virus neutralization assay (VN-assay) and plaque reduction neutralization test (PRNT). Results showed that WNV antibodies were present in 72 out of 2098 horse sera (3.43%) and 3 of 2695 cattle sera (0.11%). The highest seroprevalence was found in Eastern Croatia in counties next to Hungarian, Serbian and Bosnia and Herzegovinian state borders. In Adriatic part of Croatia positive animals were found only in the westernmost county, near Slovenian and Italian borders. Geographic distribution and number of positive horses indicated that WNV is highly present in Croatia and spreading from East to West. However, positive horses in westernmost part of country indicate possible second origin of spreading. Location of serological positive cattle supports the hypothesis that seropositive cattle could be indicators of high WNV activity in the respective geographic regions.     

Microbial decomposition of soil organic matter is mediated by quality and quantity of crop residues: mechanisms and thresholds

Crop residue quality and quantity have contrasting effects on soil organic matter (SOM) decomposition, but the mechanisms explaining such priming effect (PE) are still elusive. To reveal the role of residue quality and quantity in SOM priming, we applied two rates (5.4–10.8 g kg^?1) of ¹³C-labeled wheat residues (separately: leaves, stems, roots) to soil and incubated for 120 days. To distinguish PE mechanisms, labeled C was traced in CO₂ efflux and in microbial biomass and enzyme activities (involved in C, N, and P cycles) were measured during the incubation period. Regardless of residue type, PE intensity declined with increasing C additions. Roots were least mineralized but caused up to 60% higher PE compared to leaves or stems. During intensive residue mineralization (first 2–3 weeks), the low or negative PE resulted from pool substitution. Thereafter (15–60 days), a large decline in microbial biomass along with increased enzyme activity suggested that microbial necromass served as SOM primer. Finally, incorporation of SOM-derived C into remaining microbial biomass corresponded to increased enzyme activity, which is indicative of SOM cometabolism. Both PE and enzyme activities were primarily correlated with residue-metabolizing soil microorganisms. A unifying model demonstrated that PE was a function of residue mineralization, with thresholds for strong PE increase of up to 20% root, 44% stem, and 51% leaf mineralization. Thus, root mineralization has the lowest threshold for a strong PE increase. Our study emphasizes the role of residue-feeding microorganisms as active players in the PE, which are mediated by quality and quantity of crop residue additions.     

Heat‐inactivated Mycobacterium bovis protects zebrafish against mycobacteriosis

Control of mycobacterial infection constitutes a priority for human and animal health worldwide. However, effective vaccines are needed for the control of human and animal tuberculosis (TB). Adult zebrafish have become a useful model for studying the pathophysiology of mycobacterial infection and for the development of novel interventions for TB control and prevention. Recently, parenteral and oral immunization with the heat‐inactivated Mycobacterium bovis vaccine (M. bovis IV) protected wild boar against TB. The objectives of this study were to provide additional support for the role of M. bovis IV in TB control using the zebrafish model and to conduct the first trial with this vaccine for the control of fish mycobacteriosis. The results showed that M. bovis IV protected zebrafish against mycobacteriosis caused by low and high infection doses of Mycobacterium marinum and provided evidence suggesting that the protective mechanism elicited by M. bovis IV in zebrafish as in other species is based on the activation of the innate immune response through the C3 pathway, with a role for the regulatory protein Akr2 in this process. These results encourage the use of M. bovis IV for TB control in different species.     

Irreversible sorption of humic substances causes a decrease in wettability of clay surfaces as measured by a sessile drop contact angle method

Purpose

The objective of the study was to obtain quantitative assessments of the hydrophobic impact of irreversible sorption of humic substances (HSs) onto clay mineral surfaces using a sessile drop contact angle method.

Materials and methods

Two clays (kaolin and montmorillonite) were modified with four humic materials: (1) sod podzolic soil, (2) chernozem, (3) peat, and (4) coal (leonardite). The humic materials were characterized using elemental analysis, size exclusion chromatography, and ¹³C NMR spectroscopy. Both clay samples were saturated with Ca²⁺ prior to modification with HS using a sorption isotherm technique. Contact angles (CAs) of the obtained HS-clay complexes were determined using a static sessile drop method after drying the obtained HS-clay complexes in the form of a thin film.

Results and discussion

HS modification rendered both clays under study—kaolin and montmorillonite—more hydrophobic. In case of Ca-kaolin, the CA values increased from 27° (Ca-kaolin) up to 31°–32° (all HS-kaolin complexes) with no significant difference among the HS types used for modification. In the case of Ca-montmorillonite, the CA values increased from 41° (Ca-montmorillonite) up to 51°–83° with the following ascending trend for the humic types investigated: chernozem HS < coal HS < peat HS < sod-podzolic HS. This trend is in reverse to the degree of aromaticity of the HS, expressed as the content of aromatic carbon, and it is directly proportional to the molecular weight of each HS.

Conclusions

Application of a sessile drop method showed increased surface hydrophobicity of HS-modified clays. Much more substantial hydrophobization was observed for montmorillonite as compared to kaolin, which was explained by the differences in the sorption mechanism.

     

Pre‐slaughter mortality among turkeys related to their transport

Overall mortality among turkeys transported for slaughter in the Czech Republic from 2009 to 2014 was 0.147%. The lowest mortality (0.023%) was found in turkeys transported for distances up to 50 km; longer distances were associated with increasing death rates, with the highest losses (0.543%) recorded for distances from 201 to 300 km. Differences were also found when comparing transport‐related mortality rates according to the season of the year. The highest mortality (0.228%) was associated with transports carried out in summer, whereas the lowest death rates were found in turkeys transported for slaughter in winter (0.105%) and autumn (0.113%). According to the ambient temperatures in which transports associated with the highest death rates among turkeys were carried out (14 to 21°C), it appears that heat stress can be a major factor in the birds’ mortality. However, low ambient temperatures (i.e. below ?2°C) also appear to increase transport‐related mortality, as shown by our results. In addition to shortening the lengths of transport distances, appropriate consideration of prevailing climatic conditions and the sensitivity of turkeys to high and low temperatures during transport may also lead to a reduction in the number of turkeys dying during transport for slaughter.     

Ionic thiocyanate (SCN(-)) production, fate, and phytotoxicity in soil amended with Brassicaceae seed meals

Brassicaceae seed meals produce ionic thiocyanate (SCN (-)), a bioherbicidal compound. This study determined the fate of SCN (-) in a field soil amended with seed meals of Sinapis alba, Brassica juncea, and Brassica napus and quantified crop phytotoxicity by monitoring carrot ( Daucus carota) emergence. Meals were applied at 1 or 2 t ha (-1), and soils were sampled to 35 cm for SCN (-). Maximum SCN (-) (211 micromol kg (-1) of soil) was measured at 5 days in 0-5 cm samples from plots amended with S. alba meal at 2 t ha (-1). Less than 30 micromol of SCN (-) kg (-1) of soil was measured at soil depths below 15 cm. At 44 days, SCN (-) was <15 micromol kg (-1) of soil in all treatments. Emergence inhibition of carrots seeded 15-36 days after meal amendment was found only in S. alba treatments. The rapid decrease of SCN (-) concentrations in Brassicaceae meal-amended soil indicates limited potential for off-site environmental impacts.