A novel extracellular peroxidase and nucleases from a milky sap of Chelidonium majus

Using affinity chromatography, SDS-PAGE, peroxidase activity assay and mass spectrometry data, a new extracellular peroxidase (CMP) from Chelidonium majus milky sap was isolated and characterized. The protein has a molecular weight of about 40 kDa and belongs to secretory class III plant peroxidases. The peroxidase activity is also accompanied by DN-ase activities. A novel CMP combined with other proteins is probably involved in development and differentiation of the plant and defence against different pathogens. It suggests that the biological activity of C. majus whole plants and extracts may depend not only on its alkaloidal content but also on the presence of biologically active proteins.     

Digital radiography (DR) and imaging analysis for evaluating the penetration and distribution of organic substances used in wood conservation

Consolidation treatments are commonly employed in museum conservation studios for treating biologically deteriorated wooden cultural heritage objects. Impregnation using either a solvent/resin or a polymerizing system is an example of such an intervention, which is often difficult to describe in terms of its behavior within the object’s structure. A new simple method has been devised to evaluate the effectiveness of these impregnation treatments in terms of spatial distribution of consolidant solution. A combination of digital radiography and imaging analysis has permitted to evaluate the degree of penetration of a consolidant and to determine its location within the artifact by studying the X-ray images taken before, during, and after treatment. The adequacy of polymer solutions or their effectiveness as wood consolidants is properties which are often difficult to investigate due to limited visual access to the interior of the object. The proposed method provides an alternative way of studying these parameters through analysis of X-ray attenuation recorded on two images: taken before and after the conservation treatment. The resulting image is then converted into a two-dimensional map of consolidation agent within the object using a straightforward calibration procedure.     

Effects of thermal-time artificial scarification on the germination dynamics of black locust (<Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> L.) seeds

The aim of this study was to determine the efficiency of methods for the artificial scarification of black locust seeds based on the influence of the air and liquid nitrogen temperatures. Three populations were selected for the analyses: a managed tree stand, a selected seed stand and a seed orchard. Samples of 200 seeds (4 × 50) were randomly selected. Before scarification, the seeds were stored at ?70 °C for 24 h. Nine different scarification treatments were evaluated. To calculate the germination capacity and the dynamics of germination, observations were made on days 3, 5, 7, 10 and 14 after treatment. For statistical analyses, we used repeated measures analysis of variance. The obtained results indicated that the highest scarification efficiency for black locust seeds occurred in the treatment in liquid nitrogen and in an air temperature range of 85–90 °C. The provenance of seeds had a significant effect on the dynamics of germination. Thermal methods and the use of liquid nitrogen can be alternatives to the labour intensive methods of cutting the seeds or using measures that require specialised equipment.     

Metal Concentrations in Organs of the Clam Amiantis umbonella and Their Use in Monitoring Metal Contamination of Coastal Sediments

The aim of this study was to evaluate the use of metal concentrations in clam organs to monitor metal contamination in coastal sediments. The concentrations of Cd, Cr, Cu, Hg, Ni, Pb, V, and Zn were measured in the kidneys, gonads, mantles, gills, digestive gland, and hearts of the infaunal clam Amiantis umbonella collected from a contaminated site near desalination and power plant discharges, and a reference site in Kuwait Bay. Metal concentrations in sediment and sediment pore water were also measured at the collection sites of individual clams at the contaminated site. The concentrations of all metals in all organs (except Zn in the digestive gland) were significantly higher in clams from the contaminated site than from the reference site. Metal concentrations in several organs in A. umbonella from the contaminated site were correlated with those in the sediments and pore waters to which they were exposed. However, fresh weights of gonads, gills, and mantles were significantly lower in clams from the contaminated site compared to the reference site, indicating that the observed elevated concentrations of metals in the organs of clams from the contaminated site largely reflect lower organ weights, rather than higher metal loads, and that these organs in A. umbonella and perhaps other clams are not appropriate for use as biomonitors of metal contamination. Metal concentrations in clam kidneys showed a wide dynamic range with respect to environmental contamination and kidney weight was not variable. Therefore, metal concentrations in clam kidneys provide a reliable biomonitor of contaminant metals in coastal marine sediments.     

Preliminary assessment of serum clusterin as a potential biomarker for canine lymphoma

Clusterin (CLU), also known as apolipoprotein J, is a widely expressed, heterodimeric, glycoprotein, important in tumourigenesis, apoptosis and immunoregulation. In humans, CLU expression has been associated with anaplastic large cell and Hodgkin's lymphoma. In this study, serum CLU levels in dogs with multicentric lymphoma (MLSA) were compared with healthy control dogs, using both western blot and enzyme‐linked immunosorbent assay (ELISA). Western blot confirmed the presence of CLU in dog sera at the predicted molecular weight and the relative levels detected correlated with the levels detected by ELISA. CLU level analysis by ELISA found treatment naïve dogs with MLSA had a significantly (P < .001) lower serum CLU level compared with healthy controls. However, there was no significant difference between MLSA dogs prior to treatment and in complete remission. The wide variation in serum CLU levels may limit its potential as a single candidate biomarker for MLSA, although any prognostic predictive value of serum CLU concentrations has yet to be assessed.     

Characteristics of capercaillie (Tetrao urogallus) semen analysed with flow cytometry combined with fertility results

In order to increase the reproductive indices of capercaillie kept in closed breeding facilities, it is necessary to constantly expand the methods of better understanding the characteristics of sperm and their fertilizing potency. The aim of the study was to analyse selected features of capercaillie sperm using flow cytometry and their connection with fertility results. The study included five males, three of which were kept in a family group with eight females and two were kept alone. For sperm viability, acrosome integrity, mitochondrial potential and DNA defragmentation were assessed. Paternity analyses were performed in order to confirm the paternity of the individual and to link the evaluated semen traits with reproductive success. Analyses carried out in the flow cytometer showed any significant differences between males in sperm characteristics. In the semen of male No. 101, the father of all chicks from the analysed family group, 91.3% of live sperm, 91.5% with intact acrosome, 83.6% with active mitochondria and 2.0% with DNA defragmentation were observed. The average fertility rate was 71.0%, and chick hatchability was 100%. Using flow cytometry in the analysis of capercaillie semen and its connection with the results of natural mating, we were able to obtain deeper knowledge about new sperm characteristics that were not examined before and which in the future may be helpful in selecting males for the reproductive flocks and developing assisted reproduction techniques.     

The effect of human activity on birds at a coastal bay

I examined the direct and indirect effects of human activity on birds at a coastal bay refuge along the Atlantic Coast. Over the year, human activity varied at different sample sites on the refuge, but people were present on part of the refuge every day, although activity was concentrated on designated paths around a freshwater pond and at a fishing pier. On the refuge (exclusive of the ponds) people were present at the sample sites 17% of the time, birds were present 42% of the time when people were present, but birds were present 72% of the time when people were absent. Human activities, such as jogging or grass mowing, which involved rapud movement or close proximity to roosting birds, usually caused them to flush. Slow-walking bird watchers and clammers did not usually cause birds to flush. Gulls and terns were least affected as they usually relanded where they had been, ducks usually flushed and flew to the centre of the pond, and herons, egrets and shorebirds were most disturbed and flushed to distant marshes. These results suggest that if management objectives include providing roosting areas for migrating shorebirds then some areas must be protected from close and fast-moving human activities.     

The effect of a piperonyl butoxide/tau-fluvalinate mixture on pollen beetle (Meligethes aeneus) and honey bees (Apis mellifera)

BACKGROUND: Previous work has characterised pyrethroid resistance in pollen beetle (Meligethes aeneus F.) as principally an oxidative mechanism. Piperonyl butoxide (PBO) can synergise this resistance in the field, but its effects on the honey bee are thought to be unacceptable. RESULTS: A field trial in Poland was conducted to show that a mixture of PBO and tau‐fluvalinate at the registered rate gave increased and longer‐lasting control of resistant pollen beetle. Four days after spraying with tau‐fluvalinate, only 20% of pollen beetles were controlled, compared with 70% if the tau‐fluvalinate/PBO mixture was used. No detriment to honey bee health was observed using the same mixture. CONCLUSIONS: PBO, if used in conjunction with a pyrethroid of relatively low bee toxicity, can successfully overcome pyrethroid resistance in pollen beetle without incurring an increased loss of honey bees, even if they are present at the time of spraying. Copyright © 2012 Society of Chemical Industry     

Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis

Background

The blaZ gene encoding penicillin resistance can be located either chromosomally or on plasmids. The aim of this study was to investigate the genetic relationships and to determine the location of the blaZ gene in S. aureus isolated in bovine mastitis in Finland and Sweden.

Methods

Seventy-eight β-lactamase positive S. aureus isolates from bovine mastitis (34 from Finland and 44 from Sweden) were included in the study. The localization of blaZ gene was determined by Southern blotting. The blaZ genes of the isolates were sequenced and the sequences were translated to beta-lactamase proteins and further grouped as different protein signatures. The isolates and, as control, 33 Swedish and 36 Finnish beta-lactamase negative isolates were typed with pulsed-field gel electrophoresis (PFGE).

Results

In 26 out of 34 Finnish isolates (76.5%) and in 25 out of 44 Swedish isolates (56.8%) the blaZ gene was localized on a plasmid. Six different protein signatures were found. One signature was found only in four Swedish isolates, but all other signatures were found both in Finnish and Swedish isolates. The PFGE results revealed a diversity of S. aureus clones. The protein signatures were not clearly associated with certain pulsotypes.

Conclusions

The plasmid location of the blaZ gene was not statistically significantly more common in Finland than in Sweden, and hence does not explain the higher proportion of penicillin-resistant isolates of S. aureus causing bovine mastitis in Finland compared to Sweden.