Two-stage continuous fermentation of Saccharomycopsis fibuligeria and Candida utilis

Biomass production and carbohydrate reduction were determined for a two-stage continuous fermentation process with a simulated potato processing waste feed. The amylolytic yeast Saccharomycopsis fibuligera was grown in the first stage and a mixed culture of S. fibuligera and Candida utilis was maintained in the second stage. All conditions for the first and second stages were fixed except the flow of medium to the second stage was varied. Maximum biomass production occurred at a second stage dilution rate, D(2), of 0.27 h (-1). Carbohydrate reduction was inversely proportional to D(2), between 0.10 and 0.35 h (-1).     

Historical range and movements of the Elephants in Babile Elephant Sanctuary,Ethiopia

A study on historical range and movement patterns of the African savannah elephants (Loxodonta africana africana) in Ethiopia was made from December 2004 to November 2008. Babile Elephant Sanctuary is one of the strongholds of elephants in the country, where it now supports approximately 27% of the elephant population in Ethiopia. Elephant movement routes and ranges were identified based on interviews of local people and by tracking the animals in the field. Elephants of the Sanctuary were categorized into two distinctive groups or clans based on their movement patterns and their associations to each other. Three major valleys, namely Gobele, Erer and Dakata, were crossed by elephants. Elephants in the population had an estimated annual range of 3014 km². Of this, 85.7% (2583 km²) was inside the Sanctuary and 14.3% (430 km²) outside but adjacent to the Sanctuary. Elephant use outside of the Sanctuary included three different areas: Upper Erer Valley (8 km²), Upper Gobele Valley (48 km²) and west of Gobele Valley (374 km²). The extent of range varied seasonally. During the wet season, elephant range was restricted to Erer Valley and its tributaries (1266 km² and a density of approximately 0.25 animal km^?2), or 42% smaller than the dry season range (1417 km², 0.23 elephant km^?2). Range use was smallest (332 km²) in October to November and from April to May. During the dry season, elephants were found in two distinctive groups, one using Gobele Valley and the other Erer Valley.     

Evaluation of the Performance Characteristics of Bilayer Tablets: Part I. Impact of Material Properties and Process Parameters on the Strength of Bilayer Tablets

Bilayer tableting technology has gained popularity in recent times, as bilayer tablets offer several advantages over conventional tablets. There is a dearth of knowledge on the impact of material properties and process conditions on the performance of bilayer tablets. This paper takes a statistical approach to develop a model that will determine the effect of the material properties and bilayer compression process parameters on the bonding strength and mode of breakage of bilayer tablets. Experiments were carried out at pilot scale to simulate the commercial manufacturing conditions. As part of this endeavor, a seven-factor half-fraction factorial (2⁷⁻¹) design was executed to study the effect of bilayer tablet compression process factors on the bonding strength of bilayer tablets. Factors studied in this work include: material properties (plastic and brittle), layer ratio, dwell time, layer sequence, first- and second-layer forces, and lubricant concentration. Bilayer tablets manufactured in this study were tested using the axial tester, as it considers both the interfacial and individual layer bonding strengths. Responses of the experiments were analyzed using PROC GLM of SAS (SAS Institute Inc, Cary, North Carolina). A model was fit using all the responses to determine the significant interactions (p < 0.05). The results of this study indicated that nature of materials played a critical role on the strength of bilayer compacts and also on mode of fracture. Bilayer tablets made with brittle materials in both the layers are strongest, and fracture occurred in the first layer indicating that interface is stronger than layers. Significant interactions were observed between the selected factors and these results will provide an insight into the interplay of material properties, process parameters, and lubricant concentration on the bonding strength and mode of breakage of bilayer tablets.

Electronic supplementary material

The online version of this article (doi:10.1208/s12249-012-9845-9) contains supplementary material, which is available to authorized users.KEY WORDS: axial tester, bilayer tablets, DOE, interfacial strength, process parameters     

Rapid Evolution of Recombinant Saccharomyces cerevisiae for Xylose Fermentation through Formation of Extra-chromosomal Circular DNA

Circular DNA elements are involved in genome plasticity, particularly of tandem repeats. However, amplifications of DNA segments in Saccharomyces cerevisiae reported so far involve pre-existing repetitive sequences such as ribosomal DNA, Ty elements and Long Terminal Repeats (LTRs). Here, we report the generation of an eccDNA, (extrachromosomal circular DNA element) in a region without any repetitive sequences during an adaptive evolution experiment. We performed whole genome sequence comparison between an efficient D-xylose fermenting yeast strain developed by metabolic and evolutionary engineering, and its parent industrial strain. We found that the heterologous gene XylA that had been inserted close to an ARS sequence in the parent strain has been amplified about 9 fold in both alleles of the chromosomal locus of the evolved strain compared to its parent. Analysis of the amplification process during the adaptive evolution revealed formation of a XylA-carrying eccDNA, pXI2-6, followed by chromosomal integration in tandem arrays over the course of the evolutionary adaptation. Formation of the eccDNA occurred in the absence of any repetitive DNA elements, probably using a micro-homology sequence of 8 nucleotides flanking the amplified sequence. We isolated the pXI2-6 eccDNA from an intermediate strain of the evolutionary adaptation process, sequenced it completely and showed that it confers high xylose fermentation capacity when it is transferred to a new strain. In this way, we have provided clear evidence that gene amplification can occur through generation of eccDNA without the presence of flanking repetitive sequences and can serve as a rapid means of adaptation to selection pressure.     

Genetic mapping of the stem rust (Puccinia graminis f. sp. tritici Eriks. & E. Henn) resistance gene Sr13 in wheat (Triticum aestivum L.)

Puccinia graminis f. sp. tritici, the causative agent of stem rust in wheat, is known for its high virulence variability and ability to evolve new virulence to resistance genes. Thus, pyramiding of several resistance genes in a single line is the best strategy for a sustainable control of wheat stem rust. Sr13 is one of the few resistance genes that are effective against wide ranging P. graminis f. sp. tritici races, including the pestilent race Ug99. Its effectiveness to Ug99 makes it a valuable source for resistance to stem rust. Molecular markers play a pivotal role in the genetic characterization of the new sources of resistance as well as in stacking two or more resistance genes in a single line. Therefore, the aim of this study was to develop molecular markers for Sr13 facilitating efficient pyramiding of Sr genes. Based on the 158 F₂ individuals derived from a cross of Khapstein/9*LMPG × Morocco and SSR analyses, the Sr13 locus was mapped on chromosome 6A of wheat, and a genetic map comprising about 90 cM was constructed with the closest marker barc37 being located 4.0 cM distally of Sr13. Of the nine mapped markers, barc37 amplified an allele specific for the presence of Sr13 as shown by testing different cultivars and breeding lines. These newly developed markers will increase the efficiency of incorporating Sr13 into cultivars that are widely adopted, but susceptible to hazardous Ug99 and/or assist for the development of new elite lines that are resistant to Ug99.     

Otolith age determination for adult tilapia, Oreochromis niloticus L. from Lake Awassa (Ethiopian Rift Valley) by interpreting biannuli and differentiating biannual recruitment

Age of mature Oreochromis niloticus in Lake Awassa, Ethiopia, was estimated by analysing optical macrozones (translucent and opaque) in sagittal otoliths from fish sampled over a 12-month period. Seasonal record on the type of macrozone at the edge of otoliths suggested that two translucent macrozones associated with biannuli were formed each year; one during January and February and another during June and July. Formation of translucent macrozones coincided with minimum water temperature, spawning associated loss in condition and presumably with reduction in the quantity and quality of the food consumed by the fish. Relative marginal increment analysis showed that biannulus formation may be completed in March and in August each year. A concurrent study has confirmed a biannual recruitment in O. niloticus in Lake Awassa, from which mid-February and mid-August were taken as median hatch-dates. A procedure to assign otolith age is described which considers median hatch-dates and uses the number of biannuli in otoliths without discriminating between fish from the two recruitment cohorts. We also show a procedure to discriminate between the two cohorts by using the number of biannuli, conditions on the edge of the otolith and time of capture to assign age for the two recruitment cohorts separately.     

The seasonal abundance of the phototrophic ciliate Mesodinium rubrum in Southampton Water, England

The abundance of the marine phototrophic planktonic ciliateMesodinium rubrum was monitored throughout an annual cycle attwo stations in the Southampton Water estuary. Seasonal changesin the concentration of nitrate, ammonia and phosphate weremonitored both at the inner estuary station (NW Netley) andouter estuary station (Calshot). Nutrient levels in the winterwere similar at both stations, and were diminished during sequentialdiatom blooms dominated initially by Sketetonema costatum andthen by Rhizosolenia dclicatula. Nitrate was reduced to a seasonalminimum in the outer estuary following these spring diatom blooms,but in the inner estuary was sustained >500 µg I^–1until the onset of the M.rubrum bloom. During the developmentof a visible red tide of M.rubrum in June/July at NW Netley,nutrient concentrations were considerably reduced. Cell numbersof M.rubrum varied between 2 and 3 cells ml^– during winterto >400 cells ml^–1 during the bloom at NW Netley, whereasat Calshot cell abundance did not increase above 25 cells ml^–1at any time of the year. At NW Netley, dense accumulations ofthe ciliate occurred over restricted depth intervals duringthe bloom and possible factors influencing the observed verticaldistribution of cells are considered. ¹Present address: Biology Department, Faculty of Science, AddisAbaba University, PO Box 1176, Addis Ababa, Ethiopia     

Evaluation of growth and biomass production of Arthrospira (Spirulina) fusiformis in laboratory cultures using waters from the Ethiopian soda lakes Chitu and Shala

The cost of nutrient media is the major challenge for biomass production of Spirulina. Although much effort has been made to use enriched seawater for the cultivation of this microalga, little attention has been given to the potential of water of soda lakes. In this study, growth (μ, day^?1) and biomass production (B) of Arthrospira fusiformis cultivated using waters of the soda lakes Chitu and Shala with or without supplementation were evaluated. Comparable μ and B values were achieved in both Lake Chitu water-based media (CBM) and Lake Shala water-based media (SBM), with slightly higher values in the latter. Both CBM and SBM supplemented with the standard Spirulina medium (SM) by 25 % and 50 % supported considerably higher μ and B. The pH and salinity of the cultures showed significant variations (P?μ and B. The observed higher μ and B were probably associated with the reduction in pH and salinity of the supplemented media due to addition of bicarbonate–carbonates and dilution, and provision of the limiting nutrient nitrogen. The higher μ and B in SBM may have resulted from some of their aggregate chemical parameters, which were closer to those in the SM, and abundant PO₄-P. This seems to suggest that Lake Shala water is more conducive to Arthrospira. We contend that 25 % and 50 % supplemented Lake Shala water can be preferably used to produce Arthrospira biomass, thereby reducing the cost of nutrients by 75 % and 50 %, respectively.