Search for hydrophilic marine fungal metabolites: a rational approach for their production and extraction in a bioactivity screening context

In the search for bioactive natural products, our lab screens hydrophobic extracts from marine fungal strains. While hydrophilic active substances were recently identified from marine macro-organisms, there was a lack of reported metabolites in the marine fungi area. As such, we decided to develop a general procedure for screening of hydrophobic metabolites. The aim of this study was to compare different processes of fermentation and extraction, using six representative marine fungal strains, in order to define the optimized method for production. The parameters studied were (a) which polar solvent to select, (b) which fermentation method to choose between solid and liquid cultures, (c) which raw material, the mycelium or its medium, to extract and (d) which extraction process to apply. The biochemical analysis and biological evaluations of obtained extracts led to the conclusion that the culture of marine fungi by agar surface fermentation followed by the separate extraction of the mycelium and its medium by a cryo-crushing and an enzymatic digestion with agarase, respectively, was the best procedure when screening for hydrophilic bioactive metabolites. During this development, several bioactivities were detected, confirming the potential of hydrophilic crude extracts in the search for bioactive natural products.     

Breeding perennial ryegrass ( Lolium perenne L.) for turf usage: an assessment of genetic improvements in cultivars released in Europe, 1974–2004

An assessment of genetic improvement in turf‐type perennial ryegrass was performed at a network of six locations. A comparison was made of the turf performances of five natural populations, five forage‐type cultivars used for turf seeding until the 1980s and 31 turf‐type cultivars released from 1974 to 2004. Populations and cultivars were also compared in two spaced‐plant experiments and in two seed‐yield trials. Trait regressions on registration year of turf‐type cultivars showed that breeding had been highly successful in improving the turf aesthetic merit (from +8·8 to +12·5% per decade according to seasons), wear tolerance (+5·4% per decade) and crown‐rust resistance (+8·9% per decade) and in lessening the turf height increase rate (?0·43 mm day^?1 per decade). Turf winter greenness had been marginally improved, whereas summer greenness and seed yield had not been significantly changed. A multivariate analysis provided evidence that turf density and fineness played a major role in the visual assessment of turf aesthetic merit and that wear tolerance was closely associated with turf density. Conflicting trait associations may have precluded improvements in turf ground‐cover 3 months after sowing, turf winter greenness and turf persistency.     

Lack of correlation between the levels of soluble cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) and the CT-60 genotypes

Background

Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) plays a critical role in downregulation of antigen-activated immune response and polymorphisms at the CTLA-4 gene have been shown to be associated with several autoimmune diseases including type-1 diabetes (T1D). The etiological mutation was mapped to the CT60-A/G single nucleotide polymorphism (SNP) that is believed to control the processing and production of soluble CTLA-4 (sCTLA-4).

Methods

We therefore determined sCTLA-4 protein levels in the sera from 82 T1D patients and 19 autoantibody positive (AbP) subjects and 117 autoantibody negative (AbN) controls using ELISA. The CT-60 SNP was genotyped for these samples by using PCR and restriction enzyme digestion of a 268 bp DNA segment containing the SNP. Genotyping of CT-60 SNP was confirmed by dye terminating sequencing reaction.

Results

Higher levels of sCTLA-4 were observed in T1D (2.24 ng/ml) and AbP (mean = 2.17 ng/ml) subjects compared to AbN controls (mean = 1.69 ng/ml) with the differences between these subjects becoming significant with age (p = 0.02). However, we found no correlation between sCTLA-4 levels and the CTLA-4 CT-60 SNP genotypes.

Conclusion

Consistent with the higher serum sCTLA-4 levels observed in other autoimmune diseases, our results suggest that sCTLA-4 may be a risk factor for T1D. However, our results do not support the conclusion that the CT-60 SNP controls the expression of sCTLA-4.

     

Stop me if you can: quantification of the effect of interfaces between plots on the dispersal of Cosmopolites sordidus

BACKGROUND

Cosmopolites sordidus is one of the most damaging pests of banana worldwide. To date, most studies have addressed the control of this pest at the plot level, without considering the landscape scale, whereas between plots dispersion could be important. The aim of this study was to investigate the ability of C. sordidus to cross contrasted field interfaces. The 10 following interfaces were investigated: forests, hedgerows, field tracks, grassy areas, finely and coarsely tilled soil zones, ditches with and without water, vegetable gardening zones, and pheromone trap lines. Individually marked weevils were released on one side of the interface and recovered daily on the other side, allowing the estimation of the velocity and the crossing success of C. sordidus for each interface.

RESULTS

Highest permeabilities (with a crossing success above 70%) were obtained for vegetable gardening zones, finely tilled soil zones, forests, and coarsely tilled soil zones. Intermediate permeabilities were measured for hedgerows, field tracks, grassy areas, and ditches without water. Only the line of pheromone traps and submerged ditches had a strong barrier effect on C. sordidus, with a crossing success below 12%. Wind strength, percentage of sky, and vegetation height were negatively correlated to the C. sordidus crossing success.

CONCLUSION

Overall, our results showed that only ditches with water and lines of pheromone traps were efficient in stopping the dispersal of C. sordidus. The next step will be to conduct research to incorporate the use of these two interfaces at farm and landscape scale into integrated pest management strategies. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Cortisol levels in skimmed milk during the first 22?weeks of lactation and response to short-term metabolic stress and lameness in dairy cows

Background

Cortisol is secreted into blood in reaction to acute stress, but also in phases of diminished feed intake and changed animal behavior. As cows do not always show clear signs of discomfort, reliable diagnostic markers could be used to provide information regarding individual cows’ distress. The objective of this study was to establish an ether free immunoassay for the detection of cortisol and to determine values during the first 22 weeks of lactation. Furthermore, the response in milk cortisol levels was assessed during times of metabolic stress and pain associated symptoms of lameness.

Methods

Milk yield and composition, blood serum glucose, NEFA and BHBA as well as milk cortisol were determined in 24 multiparous Holstein-Friesian cows over the course of the first 22 weeks of lactation. Animals were further checked for signs of clinical diseases on a daily basis. Two feed restrictions over three days (FR; 70 % of precious ad libitum intake) were performed during the 4^th wk and the 21^st wk, respectively. An ELISA for cortisol measurement in easily accessible bovine skimmed milk was established and applied.

Results

On the last day of FR in early lactation, a reduction in milk yield and changes in serum metabolites compared to respective previous values were detected. The FR in mid-lactation resulted in no changes in milk production and serum metabolites. Milk cortisol was highest during first wk of lactation and remained on comparable levels thereafter. Milk yield and composition were not influenced by FR. Lameness resulted in enhanced milk cortisol levels.

Conclusion

Milk cortisol could be used as an indicator of painful symptoms such as lameness. Higher values of milk cortisol levels during first wk of lactation should be taken into account for interpretation.     

The effect of temperature on pollen production in carnations

Summary For breeding carnations the supply of good pollen may be a problem. For this reason the effect of temperature on pollen production in carnations was studied.In phytotron experiments a temperature of 23°C proved to be the optimum.     

Observing pollen tubes by means of fluorescence

Summary There appears to be interest, also on the part of private breeders, in studying pollen tubes with the aid of fluorescence.The method described, which is applied in research on self- and cross-incompatibility, makes a rapid and correct interpretation possible.