Genetic diversity of Fagopyrum homotropicum, a wild species related to common buckwheat

Nineteen natural populations of Fagopyrum homotropicum, a self-fertilizing close relative of common buckwheat, from the Yunnan and Sichuan provinces of China were investigated for their chromosome number and allozyme variation at 16 loci of 11 enzymes. Three populations, 'Deqin', 'Zhongdian', and 'Xiancheng', were revealed to be allotetraploid. Judging from allozyme constitution of the tetraploid and their possible progenitors, diploid progenitors are probably the diploid population of F. homotropicum from Lijiang and a natural population of F. esculentum ssp. ancestralis. Diploid populations of F. homotropicum are fixed for a given allele at almost all isozyme loci. Allozyme variation has been maintained in natural populations mainly by fixing different alleles in different populations, as they are highly differentiated among the populations (G_st = 0.969). The position of populations in the phylogenetic tree constructed from genetic distance nearly corresponded with the geographical position of the populations.     

Effect of <Emphasis Type="Italic">Lactococcus lactis</Emphasis> K-C2 on the growth performance,amino acid content and gut microflora of amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis>

This study aimed to evaluate the effect of Lactococcus lactis K-C2 on the growth performance, microbial diversity, and release of free amino acids in the intestinal tract and the edible parts of young amberjack, Seriola dumerili. Fish were fed a diet with or without strain K-C2 (2?×?10¹⁰ cfu/g feed) for 25 days. The results indicated that the growth performance of fish in the treated group was significantly higher than those in the control group (p?<?0.05). The amount of five amino acids (aspartate, sarcosine, taurine, alanine, and arginine) in the gut content and 13 of 21 amino acids in the edible parts of fish in the treated group were significantly higher (p?<?0.05) than those in the control group. Sphingomonas, Propionibacterium, and Mycobacterium were observed in gut microflora of fish in both the control and treated groups. Staphylococcus and Kocuria were detected in one sample from the control and treated groups; Acinetobacter and Acidobacteria were found in one sample from the control group. L. lactis was only found in one sample in the treated group. In conclusion, the dietary administration of probiotic L. lactis stimulated growth, reduced feed consumption, and improved the nutritional value of cultured amberjack.     

Comparison of New Ultrasonic Digestion Approaches for Plant Matrices in the Analysis of Trace Metals by Inductively Coupled Plasma–Optical Emission Spectroscopy Analysis: Contrast with USEPA Method 3050B

An ultrasonic method using two approaches, A and B, along with a reference Environmental Protection Agency (EPA) Method 3050B [i.e., a mixture of 30 mL of nitric acid–hydrochloric acid–hydrogen peroxide–water (HNO₃-HCl-H₂O₂-H₂O)] were contrasted for leaching of a plant matrix. The trace metals were arsenic, cadmium, cobalt, chromium, copper, mercury, manganese, nickel, lead, selenium, and zinc (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Se and Zn) and quantified by ICP-OES followed by an investigation into residue formation and the impact of digestion time. Approach B was the most accurate and precise with percent recoveries ranging between 99 and 120%, whereas ultrasonic approach A and the USEPA method 3050B gave similar results with poor accuracies and precisions. In the optimization of the digestion time using approach B, the total metal recovery was fairly the same over a period of 120 min except for Cr and Cu, which showed slight variations.     

Isolation and characterization of macrophages from a mixed primary culture of bovine liver cells

Previously, we developed a simple and efficient method to isolate liver macrophages from a mixed primary culture of adult rat liver cells. To extend the applicability of this method, we isolated macrophages from mixed primary cultures of bovine liver cells. Macrophage cells proliferated on the cell sheet of mixed bovine liver cells after 8-16d of culture. These cells were detached by shaking of the culture flasks. Subsequent transfer and brief incubation in plastic dishes resulted in selective adhesion of macrophages. After rinses with PBS, attached macrophages were harvested. More than 10(6) cells could be harvested from the culture flask at intervals of 2-3d for more than three weeks. The isolated cells were strongly positive for bovine macrophage markers, such as CD68, CD172a and Iba-1. These cells exhibited functional properties of macrophages, including active phagocytosis of polystyrene microbeads, proliferative response to recombinant bovine granulocyte-macrophage colony-stimulating factor, upregulation of specific inflammatory cytokine genes upon stimulation with lipopolysaccharide, and formation of multinucleated giant cells. The shaking and attachment method provides a simple and efficient alternative to obtain bovine liver macrophages without requiring complex equipment or specialized technical skills.     

Nutrient and bulk density characteristics of soil profiles in six land use systems along topo-sequences in inland valley watersheds of Ashanti region,Ghana

The bulk density and nutrient distribution in soil profiles as affected by various land use systems were studied. Mean bulk densities of the various soil profiles, 0-20, 20-40, and 4060 cm in the selected five land use systems along the upland / lowland topo-sequences were, 1.26, 1.73, and 1.44 Mg m^-3 for the primary forest (PF) plot in the upland area, 1.13, 1.51, and 1.50 Mg m^-3 for the cacao farm (CP) plot, 1.16, 1.63, and 1.26 Mg m^-3 for the mixed cropping (MC) plot, 1.39, 1.61, and 1.50 Mg m-a for the fallow (Fallow) plot, all three land use systems being in the upland / fringe areas, and 1.46, 1.72, and 1.60 Mg m^-3 for the lowland traditional rice farming (TR) plot, respectively. The very high bulk density of the second layer, especially in the PF plot, may be related to the hardening of the iron / quartz stone layer due to prolonged dry conditions, which may affect plant growth and hydrological cycles in the benchmark inland valley watersheds. The TC contents of the topsoil samples were 43, 32, 24, 26, 18, and 15 g kgm^-1 for the PF, CP, Fallow, MC, TR, and sawah (Sawah) plots, respectively. The C / N ratios of the topsoil samples were 10, 10, 9.8, 9.1, 8.8, and 8.8 for the CP, Fallow, MC, Sawah, TR, and PF plots, respectively, displaying a higher natural fertility, especially for the N supply in the PF plot in the upland area. The available phosphorus levels were low even for the topsoil samples, 1.8 to 3.4 mg kg^-1 for the upland soils while the lowland TR and Sawah plots recorded values of 4.4 and 4.9 mg kg^-1, respectively. The contents of exchangeable K, 0.1-0.4 cmole kg^-1, of the topsoil samples were in the range of those of normal inland valley soils in West Africa. In contrast to the content of available P, although the MC plot showed a depletion, the PF, Fallow, and CP plots showed a replenishment of available K through biological nutrient cycling. The levels of exchangeable Ca and Mg as well as eCEC, were also relatively high compared to those of mean inland valley soils in West Africa. The major component of eCEC was Ex. Ca. The levels of Ex. Ca in the topsoil samples were 11.1, 9.1, 7.8, 4.7, 5.1, and 3.9 cmol_c kg^-1 for the PF, CP, Fallow, MC, TR, and Sawah plots, respectively. The distinctive feature of Ex. Mg was its large distribution in the upper soil profiles in the CP plot, indicating that in the cacao farms, Ex. Mg was enriched, probably due to the high level of Mg in the leaf litter. The lowland soils, normally, should have been enriched in exchangeable bases such as Ca, K, and Mg, leading to a higher eCEC value and higher general fertility through the process of geological fertilization, Le., nutrient flows from upland to lowland areas, as in the case of lowland sawah in monsoon Asia. At the benchmark sites, however, the effects of the geological fertilization process were not evident. This is one of the most important characteristics of West African inland valley watersheds.     

Protozoan predation of Escherichia coli in hydroponic media of leafy vegetables

Multiple outbreaks of food poisoning associated with fresh vegetable consumptions have occurred in many countries. Numerous reports have described human pathogenic bacteria, such as Escherichia coli O157:H7 and Salmonella spp., that can internalize into fresh vegetables via root or leaf surfaces. While attempting to obtain the threshold concentration of internalization of E. coli inoculated into hydroponic medium during vegetable cultivation, we observed a rapid decrease in E. coli numbers. In the present study, we determined that the rapid decline in E. coli was not due to a physiological change into a viable but non-culturable (VNC) state. The population crash was instead caused by true bacterial death, as the rapid descent was also confirmed by micro-colony fluorescence in situ hybridization, a culture-independent method that can detect VNC cells. We next monitored the number of E. coli inoculated into intact or filter-sterilized hydroponic medium after cultivation of various types of plants. We found that the number of E. coli in intact hydroponic medium decreased markedly, whereas the level in filter-sterilized hydroponic medium was completely unchanged. This result suggests that biotic factors were present that could be eliminated by filtering. Robust predation of E. coli by protozoa (ciliates and flagellates) was observed using fluorescently labeled bacteria incorporated into the hydroponic medium. Finally, morphological identification of flagellates by scanning electron microscopy revealed the presence of a species of Stramenopiles. These findings suggest the importance of protozoa as bacterial feeders in hydroponic systems and hence the use of these organisms as potential control agents of human pathogenic bacteria.     

Selective staining and visualization of hyphal sheath of a white-rot fungus <Emphasis Type="Italic">Phanerochaete crassa</Emphasis> WD1694 with phloxine B

The hyphal sheath is a morphological feature of many kinds of fungi. Although the fine structures of the sheath have been studied in detail by a number of electron microscopy techniques, the function and physiology of the hyphal sheath are not yet clarified. One reason for this is that the hyphal sheath is a colorless, mucilaginous, and delicate material so that it is not easily identified. We developed a simple method to visualize and identify the hyphal sheath of the white-rot fungus Phanerochaete crassa WD1694. The small mycelial pellets in shaken liquid cultures of P. crassa WD1694 were stained directly with phloxine B. Both the hyphae and the hyphal sheath that filled the gaps between each of the hyphae were visualized and observed by light microscopy. The stained hyphae were further studied by transmission electron microscopy, atomic force microscopy, and fl uorescence microscopy. Based on these observations, we confirmed that the staining of the hyphae was also due to the presence of the hyphal sheath that closely covered the fungal cell wall. These results clearly showed that the hyphal sheath was selectively stained with phloxine B and could be observed and identified by conventional light microscopy. Part of this report was presented at the 50th Lignin Symposium, Nagoya, October 2005