Characterization of an Insoluble and Soluble Form of Melanin Produced by Streptomyces cavourensis SV 21, a Sea Cucumber Associated Bacterium

Melanin is a widely distributed and striking dark-colored pigment produced by countless living organisms. Although a wide range of bioactivities have been recognized, there are still major constraints in using melanin for biotechnological applications such as its fragmentary known chemical structure and its insolubility in inorganic and organic solvents. In this study, a bacterial culture of Streptomyces cavourensis SV 21 produced two distinct forms of melanin: (1) a particulate, insoluble form as well as (2) a rarely observed water-soluble form. The here presented novel, acid-free purification protocol of purified particulate melanin (PPM) and purified dissolved melanin (PDM) represents the basis for an in-depth comparison of their physicochemical and biological properties, which were compared to the traditional acid-based precipitation of melanin (AM) and to a synthetic melanin standard (SM). Our data show that the differences in solubility between PDM and PPM in aqueous solutions may be a result of different adjoining cation species, since the soluble PDM polymer is largely composed of Mg²⁺ ions and the insoluble PPM is dominated by Ca²⁺ ions. Furthermore, AM shared most properties with SM, which is likely attributed to a similar, acid-based production protocol. The here presented gentler approach of purifying melanin facilitates a new perspective of an intact form of soluble and insoluble melanin that is less chemical altered and thus closer to its original biological form.     

Genetic Diversity Analysis of Indonesian Aromatic Rice Varieties (Oryza sativa L.) Using RAPD

Journal of Crop Science and Biotechnology - In this research, we investigated genetic diversity of 21 rice genotypes (Oryza sativa L.) assessed using 38 decamer RAPD primers. A total of 405 bands...     

Seasonal changes in the spatial structures of N2O, CO2, and CH4 fluxes from Acacia mangium plantation soils in Indonesia

We evaluated the spatial structures of nitrous oxide (N₂O), carbon dioxide (CO₂), and methane (CH₄) fluxes in an Acacia mangium plantation stand in Sumatra, Indonesia, in drier (August) and wetter (March) seasons. A 60 × 100-m plot was established in an A. mangium plantation that included different topographical elements of the upper plateau, lower plateau, upper slope and foot slope. The plot was divided into 10 × 10-m grids and gas fluxes and soil properties were measured at 77 grid points at 10-m intervals within the plot. Spatial structures of the gas fluxes and soil properties were identified using geostatistical analyses. Averaged N₂O and CO₂ fluxes in the wetter season (1.85 mg N m⁻² d⁻¹ and 4.29 g C m⁻² d⁻¹, respectively) were significantly higher than those in the drier season (0.55 mg N m⁻² d⁻¹ and 2.73 g C m⁻² d⁻¹, respectively) and averaged CH₄ uptake rates in the drier season (−0.62 mg C m⁻² d⁻¹) were higher than those in the wetter season (−0.24 mg C m⁻² d⁻¹). These values of N₂O fluxes in A. mangium soils were higher than those reported for natural forest soils in Sumatra, while CO₂ and CH₄ fluxes were in the range of fluxes reported for natural forest soils. Seasonal differences in these gas fluxes appears to be controlled by soil water content and substrate availability due to differing precipitation and mineralization of litter between seasons. N₂O fluxes had strong spatial dependence with a range of about 18 m in both the drier and wetter seasons. Topography was associated with the N₂O fluxes in the wetter season with higher and lower fluxes on the foot slope and on the upper plateau, respectively, via controlling the anaerobic-aerobic conditions in the soils. In the drier season, however, we could not find obvious topographic influences on the spatial patterns of N₂O fluxes and they may have depended on litter amount distribution. CO₂ fluxes had no spatial dependence in both seasons, but the topographic influence was significant in the drier season with lowest fluxes on the foot slope, while there was no significant difference between topographic positions in the wetter season. The distributions of litter amount and soil organic matter were possibly associated with CO₂ fluxes through their effects on microbial activities and fine root distribution in this A. mangium plantation.     

Thermal properties of partially hydrolyzed starch-glycerophosphatidylcholine complexes with various acyl chains

Complexes of starch and monoacyl-sn-glycerophosphatidylcholine (GPC) containing various acyl (myristoyl, palmitoyl, and stearoyl) chains were subjected to hydrolysis with glucoamylase (EC 3.2.1.3). The enzyme hydrolyzed approximately 40% of starch control and 20-28% of starch-GPC complexes. Among the GPCs examined, 1- and 2-monomyristoyl-sn-GPC showed the highest resistance to enzyme hydrolysis, and the hydrolysis rate of starch-GPCs was greater with longer chains. Enzymatic hydrolysis strongly affected the thermal properties of the starch. After enzymatic hydrolysis of starch-GPC complexes for 24 h, their thermograms had broader peaks with lower enthalpies than the corresponding starch without enzyme; however, the starch-GPC complexes showed little change. The surface of starch-GPC granules was less eroded. These results showed that the increasing amount of starch-GPC complexes could be more resistant to hydrolysis.     

Development of transferred xenogeneic vole embryos in mouse uteri

An experimental model to study interspecific pregnancy using voles, Microtus arvalis, and green fluorescent protein gene‐induced transgenic mice is presented. Xenogeneic blastocysts from the vole were transferred into the uteri of pseudopregnant mice along with allogeneic blastocysts from green fluorescent protein gene‐induced transgenic mice. The uteri containing xeno‐allo combined transfers were examined from day 6 to 13 of gestation. Although the vole embryos implanted, the uteri containing vole embryos were smaller compared with those having allogeneic mouse embryos. On day 8, the uteri containing vole embryos hemorrhaged internally and no vole embryo was found in the pregnant uterus after day 11. Allogeneic mouse embryos developed normally despite the presence and abortion of the vole embryos. In uteri implanted with vole embryos, decidua were formed and numerous blood vessels were distributed around the embryo. Maternal blood cells infiltrated into the celomic cavity of the vole embryo through the discontinuous region of trophoblast. Periodic acid‐Schiff‐positive granulated metrial gland cells were remarkably increased in the decidual sites. These findings suggest that a disorder of embryo–maternal interaction might induce the appearance of numerous granulated metrial gland cells and rejection of the embryos.     

Variability in in vitro macrophage activation by commercially diverse bulk echinacea plant material is predominantly due to bacterial lipoproteins and lipopolysaccharides

We previously reported that the majority of in vitro monocyte/macrophage activation exhibited by extracts of Echinacea and other botanicals depends upon bacterial lipopolysaccharides and Braun-type bacterial lipoproteins. We determined the contribution made by these bacterial components to the overall immune-enhancing activity detected in E. purpurea and E. angustifolia bulk root and aerial material obtained from six major growers/suppliers in North America. Substantial variation in activity (up to 200-fold) was observed in extracts of these materials when tested in two monocyte/macrophage cell lines. The majority of activity was negated by treatment with agents that target bacterial lipoproteins (lipoprotein lipase) and lipopolysaccharides (polymyxin B). Experiments comparing the activity of freeze-dried, freshly harvested Echinacea plants to those harvested and dried using various commercially relevant conditions suggest that postharvesting procedures do not substantially contribute to the variation observed in the commercial material.     

Effects of Fiber Additives on the Desiccation Crack Behavior of the Compacted Akaboku Soil as A Material for Landfill Cover Barrier

In the daily and final landfill cover barrier system, the hydraulic properties of compacted soil liners and the strength of soil can be adversely affected by desiccation cracking, resulting in the loss of effectiveness and integrity of the containment system as a barrier. Recently, there is an interest of using fiber additive to overcome the desiccation cracking problem. In this study, the desiccation crack test was conducted to investigate the effect of fiber additive on suppressing desiccation cracks in compacted Akaboku soils. Polypropylene (C₃H₆) fiber was used as an additive material for soil sample. The percentages of fiber used were varied as 0.0%, 0.2%, 0.4%, 0.6%, 0.8%, 1.0% and 1.2% (by dry weight of samples). The soil specimens were compacted under the conditions of maximum dry density and optimum water content. The surficial cracking area was measured to determine the crack intensity factor (CIF) of the soil samples. The desiccation crack test results indicated that the percentage of volume change of the compacted soil specimen decreased with addition of fiber. The change in the soil surface area decreased with increasing in the fiber content (FC), and consequently, the volumetric shrinkage strain decreased. The CIF for the soil without fiber (FC?=?0.0%) were significantly higher than the soil with fiber additive. The CIF of soil at FC?=?0.0% decreased from 2.75% to 0.6% for the soil at FC?=?0.2%. It was also found that the maximum crack depth reaches almost 50% of the thickness of the soil without fiber additive. This study suggests the potential application of the fiber additives to soils as an available method to suppress desiccation cracks encountered in landfill cover barriers.     

Evaluation of Alternative Protein Sources to Replace Fish Meal in Practical Diets for Juvenile Tilapia,Oreochromis spp

Two feeding experiments were conducted to evaluate if methionine is limiting in practical grow‐out diets for tilapia, Oreochromis spp. Four diets containing 32% protein and 5% lipid were designed to compare the use of diets high in dehulled solvent‐extracted soybean meal (DSESM) and expeller pressed soybean meal (EPSM) compared with a diet containing 6% fish meal (FM). Tilapia (4.78 ± 0.07 g, mean ± SD) were randomly stocked into twelve 600‐L flow‐through tanks at 20 fish per tank. After 6 wk, there were no notable trends or statistically significant differences (P > 0.05) in final mean weight, survival rate, and feed conversion ratio (FCR) among the treatments. Because results of this study indicated that DSESM could totally replace FM in practical diets for juvenile tilapia, a second batch of diets were formulated using other protein sources. Typical levels of cottonseed meal (CSM), DSESM, and meat and bone meal (MBM) were used to evaluate whether methionine could be limiting. Two basal diet formulations were tested either without or with methionine supplement (0.06/100 g diet). The first diet contained 15% CSM, 27% DSESM, and 10% MBM and the second diet contained 15% CSM and 37% DSESM. These diets contained 28% protein and 5% lipid. Tilapia (3.90 ± 0.05 g) were randomly stocked into twelve 60‐L glass aquaria of a recirculation system at 18 fish per aquarium for 5 wk and then moved to the 600‐L flow‐through tanks for five more weeks. After 10 wk, there were no statistically significant differences (P > 0.05) in final mean weight, survival rate, and FCR among the four treatments. Results of the present study indicated that DSESM and EPSM could totally replace FM’s inclusion rate in commercial diets for juvenile tilapia. Furthermore, methionine did not appear to be limiting in practical diets using typical levels of CSM, DSESM, and MBM as primary protein sources.