Examination of marine-based cultivation of three demosponges for acquiring bioactive marine natural products

Marine sponges are an extremely rich and important source of natural products. Mariculture is one solution to the so-called "supply problem" that often hampers further studies and development of novel compounds from sponges. We report the extended culture (767 days) at sea in depths of 10 and 20 m of three sponge species: Negombata magnifica, Amphimedon chloros and Theonella swinhoei that produce latrunculin-B, halitoxin and swinholide-A, respectively. Since sponge-associated microorganisms may be the true producers of many of the natural products found in sponges and also be linked to the health of the sponges, we examined the stability of the bacterial communities in cultured versus wild sponges. Growth rate of the sponges (ranging from 308 to 61 and -19 (%)(year(-1)) in N. magnifica, A. chloros and T. swinhoei, respectively) differed significantly between species but not between the two depths at which the species were cultivated. Survivorship varied from 96% to 57%. During culture all species maintained the content of the desired natural product. Denaturing gradient gel electrophoresis analysis of the sponge-associated bacterial consortia revealed that differences existed between cultured and wild sponges in T. swinhoei and A. chloros but the communities remained quite stable in N. magnifica. The cultivation technique for production of natural products was found to be most appropriate for N. magnifica, while for T. swinhoei and A. chloros it was less successful, because of poorer growth and survival rates and shifts in their bacterial consortia.     

A retrospective study of clinical signs and epidemiology of chronic valve disease in a group of 207 Dachshunds in Poland

Background

Chronic mitral valve disease is frequently seen in the Dachshund. Dachshunds (n=207) made up 11.73% of the dogs admitted to the Cardiology Service at the Small Animal Clinic, Warsaw University of Life Sciences, Poland (first visits only).

Results

Of these, 35 dogs had no clinically detectable heart disease while 172 had chronic valve disease with the mitral valve affected most often (130 dogs), both mitral and tricuspid valves infrequently (39 dogs) and rarely the tricuspid valve (3 dogs). Males were affected more frequently than females and the average age of dogs with chronic valve disease was 11.9 years for females and 11.3 years for males. A majority of the diseased Dachshunds were classified as ISACHC 2 (79), followed by ISACHC 1 (60). Most frequent clinical signs noted by owners included coughing, exercise intolerance, dyspnea and tachypnea. Heart murmurs were generally louder with increased disease severity; however there were 20 dogs in the ISACHC 1 group with no audible heart murmurs. The most frequent electrocardiographic abnormalities included an increased P wave and QRS complex duration, increased R wave amplitude and tachycardia. With increased disease severity, echocardiography revealed an increase in heart size. A higher ISACHC class was related to increased heart size (based on echocardiography) and increased percentage of patients exhibiting enlargement of both left atrium and left ventricle (based on radiography).

Conclusions

The Dachshund is often affected by chronic mitral valvular disease with a late onset of associated clinical signs and few cardiac complications.     

Inhibition of Armillaria and Heterobasidion growth by Penicillium adametzii isolated from Pinus sylvestris forest soil

Penicillium adametzii significantly inhibited the growth of Heterobasidium annosum sensu stricto on 2% potato dextrose agar after 10 days at 20–25°C. Chloroform extracts from P. adametzii culture filtrate and from culture grown on rice often decreased the dry weight of Armillaria colony and diameter of Heterobasidion colony in vitro. The effect depended on species and isolate of Armillaria, Heterobasidion and P. adametzii and concentration of the extract. Higher concentration of chloroform extract increased the dry weight of stems and roots of Pinus sylvestris plants inoculated with either of two isolates of Armillaria ostoyae and one isolate of Armillaria gallica. Lower concentrations of chloroform extract increased the dry weight of stems and roots of P. sylvestris inoculated with another isolate of A. gallica. Culture filtrate of P. adametzii decreased the length of necrosis on P. sylvestris stem inoculated with either of two of four isolates of H. annosum s.s., but if applied 1 month before inoculation, it increased the length of necrosis caused by another H. annosum s.s. isolate. Twenty fractions from the P. adametzii chloroform extract were separated by thin layer chromatography. Carboxylic acids, flavonoids and glycosides were detected in chloroform extract of P. adametzii by high performance liquid chromatography/mass spectrometer analysis. This is the first report of interactions between P. adametzii and either Armillaria or Heterobasidion with P. sylvestris.     

The Influence of Biochar Enriched with Magnesium and Sulfur on the Amount of Perennial Ryegrass Biomass and Selected Chemical Properties and Biological of Sandy Soil

Biochar used in this study was prepared from wheat straw and enriched with 10% solutions of chemically pure salts: ammonium sulfate [(NH₄)₂SO₄] and magnesium sulfate (MgSO₄ 7H₂O). The aim of this study was to determine the growth response of Perennial ryegrass L. as well as identify changes in chemical and biological properties of sandy soil after the application of biochar. A significant increase in the amount of Perennial ryegrass L. biomass was determined after amending the soil with unenriched biochar and biochar enriched with (NH₄)₂SO₄. The highest immobilization of available forms of Mg was observed for biochar enriched with (NH₄)₂SO₄. The highest dehydrogenase activity was determined in the soil with biochar enriched with MgSO₄ 7H₂O.     

Point mutations in BHV-1 Us3 gene abolish its ability to induce cytoskeletal changes in various cell types

The Us3 gene is conserved among alphaherpesviruses and codes for a protein kinase, a multifunctional protein involved in many phases of virus infection, like nuclear egress, modulation of apoptosis and modification of the cellular cytoskeleton. Bovine herpesvirus (BHV-1), a member of the Alphaherpesvirinae, contains an open reading frame homologous to Us3 of other herpesviruses, which has been identified as a serine/threonine kinase (Takashima, Y., Tamura, H., Xuan, X., Otsuka, H., 1999. Identification of the Us3 gene product of BHV-1 as a protein kinase and characterization of BHV-1 mutants of the Us3 gene. Virus Res. 59, 23–34). To study the activity of BHV-1 Us3, we have cloned its sequence under control of the human cytomegalovirus (HCMV) promoter/enhancer and introduced it into a recombinant baculovirus (Bac Us3). Confocal microscopy analysis showed profound cytoskeletal modifications in various BHV-1-permissive and non-permissive cells transduced with BacUs3. We observed that Us3 expression changed cellular shape and induced formation of long microtubule-containing cell projections, a phenomenon which had also been observed in cells expressing pseudorabies virus Us3. The intracellular localization of Us3 was mostly nuclear but when the protein accumulated it could be detected in the cytoplasm, cell membranes and projections. Mutated forms of BHV-1 Us3 with point mutations near or within the kinase catalytic domain did not affect cell morphology indicating that kinase activity of BHV-1 Us3 is required for its cytoskeleton remodelling function.     

Influence of bitches sex pheromones on the heart rate and other chosen parameters of blood flow in stud dogs (Canis familiaris)

The aim of this study was to verify the usefulness of the measurement of chosen cardiovascular activity parameters for examination of sex pheromones effects in male dogs. We evaluated the influence of the bitches' sexual pheromones (BSP) on heart rate (HR) with the use of a Holter monitor, and chosen parameters of blood flow in vessels of penis, testes and kidney, with color-coded and pulsed Doppler ultrasonography. We found that the BSP increased HR in all examined males even without showing any other signs of arousal. There were no changes in the RI (resistant index) and PI (pulsative index) in any trials during the pheromone presentation. The increase in blood flow intensity was noted in penile vessels but not in the testes and kidney. We concluded that measurement of flow intensity in the penis as well as the changes in heart rate as the male's reaction to the BSP can be useful in research concerning sex pheromones in dogs.     

Within-Field Variability of Winter Wheat Yield and Grain Quality versus Soil Properties

Within-field variability in wheat grain yield and its quality always exists in production fields and depends, among other factors, mainly on various soil properties related to nutrients and water availability. The aim of the research was to examine the relationships between selected soil properties such as texture; pH; content of the available phosphorus (P), potassium (K), magnesium (Mg), and organic carbon; and winter wheat grain yield and quality under rainfed conditions. Six crop fields with winter wheat (Triticum aestivum, L.) in three sites located in different regions of Poland were examined during two seasons. The grain yield was mainly determined by the soil texture, and the majority of the chemical soil properties did not have a significant effect on grain yield. The grain quality traits were determined by the examined soil properties to a smaller degree than grain yield. The relationships were not consistent across sites and years.     

Variability of fat content and fatty acids profiles in seeds of a Polish white lupin (<Emphasis Type="Italic">Lupinus albus</Emphasis> L.) collection

The paper assess the variability of fat content and fatty acids profiles in seeds of a white lupin (Lupinus albus L.) domestic collection. The initial material comprised 371 accessions originated from 30 countries of Europe, Asia, Africa, North- and South America and Australia. According to data given by accession donors the material is divided into four classes of origin: wild lines, landraces, lines created by man and cultivars. Variability of fat content and fatty acids composition were estimated in seeds of each accession. The average fat content for analyzed collection is 9.81%. The broadest range of fat content was noticed for landraces and cultivars as compared to narrowest represented by lines created by man. Fat content ranged from 6.9% (induced mutant Wt 95497) to 14.1% (Polish cultivar Wt 95420 and the landrace Wt 95212 from Jordan). From a dietetic point of view, oil quality is more important than oil quantity in lupin seeds. On average the fatty acid (FA) in examined accessions ranked in following order of abundance: oleic acid (C_18:1) > linoleic acid (C_18:2) > linolenic acid (C_18:3) > palmitic acid (C_16:0) > eicosenoic acid (C_20:1) > stearic acid (C_18:0) ≈ erucic acid (C_22:1). In respect to unsaturated fatty acid (UFA), monounsaturated oleic acid in each of estimated classes of accessions was predominant and most abundant (55.7%) in broad range of minimum–maximum values from 41.2 to 66.2%. The second examined monounsaturated fatty acid was erucic acid (1.74%) found in seeds of almost all studied accessions. An exception were four accessions defined similarly to rapeseeds as “zero erucic” forms. In seeds of few accessions a content of erucic acid exceeded 3%. Among polyunsaturated fatty acids linoleic FA (ω?6) dominated followed by linolenic FA (ω?3). Both FA were in the range 13.7–33.2% and 5.6–12.8% with mean values on the level 19.6 and 10.1%, respectively. As a consequence, the examined white lupin seeds showed a very favourable ω?3/ω?6 FA ratio (0.51), ranging from 0.21 to 0.87, much higher than that of most vegetable oils. Fat content was positively correlated with stearic and oleic fatty acids and negatively with palmitic, linoleic, linolenic and erucic acid.     

Restoration of Vegetation in Relation to Soil Properties of Spoil Heap Heavily Contaminated with Heavy Metals

The main objectives of our study were to evaluate soil contamination on a zinc-lead spoil heap in the Upper Silesian Industrial Region in southern Poland using pollution indices, and to investigate the relation between soil properties and the natural succession of vegetation. Organic carbon and nitrogen, pH, soil texture, base cations, and heavy metal content were analyzed in soil samples at depths of 0–15 cm below the organic horizon over a regular grid of 14 sampling plots. The contents of Zn, Pb, and Cd exceeded by several times the acceptable thresholds. Measurements of soil enzyme activity were used to evaluate the progress of vegetation development in relation to soil chemical properties. The results indicate that heavy metals had a significant impact on soil enzyme activity and the development of vegetation cover. High contents of Pb and Cd reduced enzyme activity, while this activity increased with increasing amounts of soil organic matter. Further, the accumulative capacities of heavy metals in needles of Scots pine (Pinus sylvestris L.) and aboveground biomass of bladder campion (Silene vulgaris (Moench) Garcke) were examined. A high accumulation of Zn, Pb, and Cd in the aboveground tissues of S. vulgaris indicated an unusual tolerance of this species to heavy metals and the possibility of using this species in phytoremediation of post-industrial sites.     

Genome analysis of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">lachrymans</Emphasis> strain 814/98 indicates diversity within the pathovar

Although many Pseudomonas syringae strains have already been determined, only a few genomes of strains belonging to pathovar lachrymans have been sequenced so far. In this study we report the genome sequence of P. syringae pv. lachrymans strain 814/98, which is highly virulent to cucumber. The genome size was estimated to be 6.58 Mb, with 57.97% GC content. In total, 6024 genes encoding proteins and 92 genes encoding RNAs were identified in this genome. Comparisons with the available sequenced genomes of pathovar lachrymans as well as with other P. syringae pathovars were conducted, revealing the presence of three unique plasmids and 24 type III effector proteins (TTEs) in strain 814/98. The phylogenetic analyses of MLST loci and TTEs clearly showed the existence of two distinct clusters of strains within pathovar lachrymans, which were grouped into either phylogroup 1 or 3, supporting non-monophyly within this pathovar.