<Emphasis Type="Italic">Pichia anomala</Emphasis> in biocontrol for apples: 20 years of fundamental research and practical applications

Fungal pathogens such as Botrytis cinerea, Penicillium expansum and the Gloeosporioides group are mainly responsible for important economical losses of post-harvest apples. Application of biological control agents (BCAs) is an emerging alternative to synthetic fungicides. However, before becoming an economically feasible alternative to chemical control, BCAs have to satisfy different requirements related to biological, technological and toxicological properties. The different steps for a successful strategy of disease control (selection, production and formulation, study of mechanisms of action, ecological characterization, molecular monitoring, pilot efficacy trials, registration) are reviewed in this paper considering the antagonistic yeast Pichia anomala strain K. This strain was selected for its high and reliable antagonistic activity against B. cinerea and P. expansum on apples. The studies of mode of action and ecological fitness are emphasized because they can lead to a better efficacy of strain K. Recently advanced molecular techniques have contributed to improving knowledge on the modes of action. Thanks to the identification of genes involved in biocontrol properties, the genetic basis of action mechanisms can be understood. That approach was adopted for P. anomala strain K and led to the identification of genes coding for exo-β-1,3-glucanases implicated in the efficacy. Based on that identification, a formulation involving β-1,3-glucans was developed and applied with higher efficacy in controlled conditions. The importance of ecological characterisation is also highlighted in the context of pre-harvest application of P. anomala strain K. UV light, temperature and humidity were identified as major factors influencing the strain K population. A model taking into consideration temperature and humidity was developed and could be useful in deciding whether pre-harvest treatment is sufficient to allow fast colonization of wounds prior to the arrival of wound pathogens, or whether it is wise to apply further post-harvest treatment to increase the yeast population density. This summary presenting 20 years of work also paid attention to practical application of strain K and its integration with other methods of control.     

Development of cyanobacterium-based biofilms and their in vitro evaluation for agriculturally useful traits

The ability of cyanobacteria to be useful as matrices for agriculturally important bacteria was evaluated. Biofilms were generated with the selected strain Anabaena torulosa after co-culturing with Azotobacter chroococcum, Pseudomonas striata, Serratia marcescens, and Mesorhizobium ciceri. The biochemical attributes were compared with individual bacterial and cyanobacterial cultures. The biofilms were characterized in terms of proteins, chlorophyll, IAA production, acetylene-reducing activity, phosphate solubilization, and antagonism towards selected phytopathogenic fungi. An enhancement in the population counts was recorded in A. torulosa–S. marcescens and A. torulosa–P. striata biofilms. The A. torulosa–A. chroococcum and A. torulosa–M. ciceri biofilms were also able to utilize new saccharides as compared to the individual cultures. Such novel biofilms with agriculturally useful traits can provide additional advantages including the broader spectrum of activity and the presence or formation of biologically active compounds; they also suggest the way to effective inoculants for sustainable and environment friendly agriculture.     

Isolation and phylogenetic classification of culturable psychrophilic prokaryotes from the Collins glacier in the Antarctica

Culturable psychrophilic prokaryotes were obtained of samples of glacier sediment, seaside mud, glacier melted ice, and Deschampsia antarctica rhizosphere from Collins glacier, Antarctica. The taxonomic classification was done by a culture-dependent molecular approach involving the Amplified Ribosomal DNA Restriction Analysis. Two hundred sixty colonies were successfully isolated and sub-cultivated under laboratory conditions. The analysis showed a bacterial profile dominated by Beta-proteobacteria (35.2%) followed by Gamma-proteobacteria (18.5%), Alpha-proteobacteria (16.6%), Gram-positive with high GC content (13%), Cytophaga–Flavobacterium–Bacteroides (13%) and Gram-positive with low GC content (3.7%). Eleven of the isolates have been reported previously and the others microorganisms remain uncharacterized. The isolated microorganisms here could be a potential source for biotechnological products, such as cold-active enzymes and secondary metabolites.     

A novel β-glucosidase with lipolytic activity from a soil metagenome

Moonlighting proteins have two different functions within a single polypeptide chain. Exploring moonlighting enzymes from the environment using the metagenomic approach is interesting. In the present study, a novel β-glucosidase gene, designated as bgl1D, with lipolytic activity (renamed Lip1C) was cloned through function-based screening of a metagenomic library from uncultured soil microorganisms. The deduced amino acid sequence comparison and phylogenetic analysis also indicated that Lip1C and other putative lipases are closely related. Biochemical characterization demonstrated that the maximum activity of the recombinant Lip1C protein occurs at pH 8.0 and 30°C using 4-nitrophenyl butyrate as substrate. The putative lipase had an apparent K _m value of 0.88 mmol/L, a k _cat value of 212/min, and a k _cat/K _m value of 241 L/mmol/min. Lip1C exhibited habitat-specific characteristics with 5 mmol/L AlCl₃, CuCl₂, and LiCl. The characterization of the biochemical properties of Lip1C enhances our understanding of this novel moonlighting enzyme isolated from a soil metagenome.     

Genes of intestinal <Emphasis Type="Italic">Escherichia coli</Emphasis> and their relation to the inflammatory activity in patients with ulcerative colitis and Crohn’s disease

Escherichia coli gene fimA was the most frequent gene that occurred in the intestine of all investigated groups. All subjects with fimA gene had significantly higher values of tumor necrosis factor alpha (TNF-α) and CRP than those with other E. coli genes. There was also a tendency to increased serum interleukin (IL)-6 levels in patients carrying the fimA gene; however, no relation was observed to serum IL-8 and IL-10. Patients with Crohn’s disease had significantly higher IL-6 than those with ulcerative colitis (UC) and controls. The highest levels of TNF-α were detected in the UC group. There were no significant differences in serum IL-8 and IL-10 between all three groups. The presence of E. coli gene fimA in the large bowel of patients with IBD is related to the immunological activity of the disease which may be important from the aspect of therapeutical strategy.     

Diverse stages of sex-chromosome differentiation in tinamid birds: evidence from crossover analysis in <Emphasis Type="Italic">Eudromia elegans</Emphasis> and <Emphasis Type="Italic">Crypturellus tataupa</Emphasis>

All extant birds share the same sex-chromosome system: ZZ males and ZW females with striking differences in the stages of sex-chromosome differentiation between the primitive palaeognathus ratites and the large majority of avian species grouped within neognaths. Evolutionarily close to ratites is the neotropical order Tinamiformes that has been scarcely explored regarding their ZW pair morphology and constitution. Tinamous, when compared to ratites, constitute a large group among Palaeognathae, therefore, exploring the extent of homology between the Z and W chromosomes in this group might reveal key features on the evolution of the avian sex chromosomes. We mapped MLH1 foci that are crossover markers on pachytene bivalents to determine the size and localization of the homologous region shared by the Z and W chromosomes in two tinamous: Eudromia elegans and Crypturellus tataupa. We found that the homologous (pseudoautosomal) region differ significantly in size between these two species. They both have a single recombination event on the long arm of the acrocentric Z and W chromosomes. However, in E. elegans the pseudoautosomal region occupies one-fourth of the W chromosome, while in C. tataupa it is restricted to the tip of the long arm of the W. The W chromosomes in these two species differ in their heterochromatin content: in E. elegans it shows a terminal euchromatic segment and in C. tataupa is completely heterochromatic. These results show that tinamous have ZW pairs with more diversified stages of differentiation compared to ratites. Finally, the idea that the avian proto-sex chromosomes started to diverge from the end of the long arm towards the centromere of an acrocentric pair is discussed.     

Direct and indirect mortality in Florida during the 2004 hurricane season

Previous studies have shown that natural disasters, and hurricanes in particular, have led to more deaths than those usually documented in short post-storm surveys. Such indirect deaths, thought to be related to dietary, stress or pre-existing medical conditions, can exceed the number of direct deaths and may persist for weeks or even months beyond the event itself. In the present study, cumulative sum of deviations plots are used to quantify the number of direct and indirect deaths resulting from Hurricanes Charley, Frances, Ivan and Jeanne that made landfall in Florida in 2004. Results suggest that there was an elevated mortality for up to 2 months following each storm, resulting in a total of 624 direct and indirect deaths attributable to the storm. Trauma-related deaths that can be associated directly with the storm account for only ∼4% of the total storm-related mortality, while indirect mortality accounts for most storm-related deaths. Specifically, a large percentage of the elevated mortality was associated with heart (34%) and cancer-related deaths (19%), while diabetes (5%) and accident-related deaths (9%) account for a smaller but still significant percentage of the elevated mortality. The results further suggest that the elevated mortality was the result of additional deaths that would not have otherwise occurred within that 5 month period, and not simply a clustering of deaths that were inevitable between 1 August and 31 December 2004. The elevated mortality identified in this study is significantly greater than the official count of 31 direct and 113 indirect deaths resulting from the four hurricanes combined. This suggests a need for improved mortality counts and surveillance in order to better evaluate and identify effective prevention policies, and to identify preventable deaths.     

A new approach using the Pierce two-node model for different body parts

This paper presents a new approach, in applying the Pierce two-node model, to predict local skin temperatures of individual body parts with good accuracy. In this study, local skin temperature measurements at 24 sites on the bodies of 11 human subjects were carried out in a controlled environment under three different indoor conditions (i.e. neutral, warm and cold). The neutral condition measurements were used to adjust the local skin set-points in the model for each body part. Additional modifications to the calculation algorithm were introduced corresponding to different body parts. The local core set-points were then calculated, using a line search method, as the input values that allow the model to predict the skin temperatures with maximum deviation of ±0.1°C for the neutral condition. The model predictability was verified for the other two indoor conditions, and the results show that the modified model predicts local skin temperatures with average deviation of ±0.3°C.     

Carbon dioxide balneotherapy and cardiovascular disease

Carbon dioxide (CO(2)) balneotherapy is a kind of remedy with a wide spectrum of applications which have been used since the Middle Ages. However, its potential use as an adjuvant therapeutic option in patients with cardiovascular disease is not yet fully clarified. We performed a thorough review of MEDLINE Database, EMBASE, ISI WEB of Knowledge, COCHRANE database and sites funded by balneotherapy centers across Europe in order to recognize relevant studies and aggregate evidence supporting the use of CO(2) baths in various cardiovascular diseases. The three main effects of CO(2) hydrotherapy during whole body or partial immersion, including decline in core temperature, an increase in cutaneous blood flow, and an elevation of the score on thermal sensation, are analyzed on a pathophysiology basis. Additionally, the indications and contra-indications of the method are presented in an evidence-based way, while the need for new methodologically sufficient studies examining the use of CO(2) baths in other cardiovascular substrates is discussed.     

Statistical optimization of medium components for avilamycin production by <Emphasis Type="Italic">Streptomyces viridochromogenes</Emphasis> Tü57-1 using response surface methodology

A fermentation medium for avilamycin production by Streptomyces viridochromogenes Tü57-1 has been optimized. Important components and their concentrations were investigated using fractional factorial design and Box–Behnken Design. The results showed that soybean flour, soluble starch, MgSO₄·7H₂O and CaCl₂·2H₂O are important for avilamycin production. A polynomial model related to medium components and avilamycin yield had been established. A high coefficient of determination (R ² = 0.92) was obtained that indicated good agreement between the experimental and predicted values of avilamycin yield. Student’s T-test of each coefficient showed that all the linear and quadratic terms had significant effect (P > |T| < 0.05) on avilamycin yield. The significance of tested components was related to MgSO₄·7H₂O (0.37 g/L), CaCl₂·2H₂O (0.39 g/L), soybean flour (21.97 g/L) and soluble starch (37.22 g/L). The yield of avilamycin reached 88.33 ± 0.94 mg/L (p < 0.05) that was 2.8-fold the initial yield.