Exposing Artemia Salina to Chattonella Subsalsa: A General Toxicity Test

The raphidophyte Chattonella subsalsa has been reported to cause harmful algal blooms in every major ocean. In South Carolina, C. subsalsa blooms have been observed in brackish stormwater detention ponds as well as estuarine waters neighboring urbanized areas. Blooms frequently cause fish kills although the fish kill mechanism of C. subsalsa is currently unknown. In many harmful species, the lethality of algal cells is thought to correspond with algal growth phase. Algal growth is known to progress through five distinct phases; lag, early exponential, late exponential, stationary, and decline. In nature, harmful algal blooms commonly occur in the late exponential or stationary growth phases; however, in vitro studies of Chattonella have identified the early exponential phase as most lethal. The strain of C. subsalsa used for this study was found to progress through the five growth phases in a period of twenty days. To examine the lethality of C. subsalsa at various growth phases, the zooplankton species Artemia salina was exposed to C. subsalsa culture at two-day intervals for twenty days. Deaths fluctuated among the growth phases of C. subsalsa with the highest mortalities observed in the late exponential and stationary growth phases. The late exponential and stationary growth phases were found to have significantly greater percent mortalities than the early exponential, lag phase, and control groups (Kruskal-Wallis rank sum test, p=0.05)

Enhanced catharanthine and vindoline production in suspension cultures of Catharanthus roseus by ultraviolet-B light

Suspension cultures of Catharanthus roseus were used to evaluate ultraviolet-B (UV-B) treatment as an abiotic elicitor of secondary metabolites. A dispersed cell suspension culture from C. roseus leaves in late exponential phase and stationary phase were irradiated with UV-B for 5 min. The stationary phase cultures were more responsive to UV-B irradiation than late exponential phase cultures. Catharanthine and vindoline increased 3-fold and 12-fold, respectively, on treatment with a 5-min UV-B irradiation

ISOLATION AND ANTIBACTERIAL ACTIVITY TEST OF ASSOCIATED BACTERIA FROM MARINE SPONGE Tetilla sp.

The marine sponge of Tetilla sp. collected near Gunung Kidul coast was studied for isolation of associated bacterial and its antibacterial activity. Sixteen bacterial strains were isolated from sponge Tetilla sp., four isolates among it showed ability to produce antibacterial metabolite. Cell extract of isolates were tested for antibacterial activity against Escherichia coli and Staphylococcus aureus using disc diffusion agar method. Cells extract of four selected isolate from late exponential phase and stationary phase broth culture showed antibacterial activity against Escherichia coli and Staphylococcus aureus. Thin layer chromatography analysis detected alkaloid and terpenoid in cell extract from stationary phase of isolates culture and none of it was detected in cell extract from late exponential phase of isolates culture

Increased L-ornithine production by an arg mutant of Acinetobacter lwoffi

The metabolic production of L-ornithine by an arg mutant of Acinetobacter Iwoffi using n-hexadecane as sole carbon source was studied. Time course experiments under optimised conditions showed that L-ornithine production was growth related, with maximum concentrations (10.5gl-1) accumulating in the late exponential phase of growth

Improved mass cultivation of the marine diatom Chaetoceros calcitrans for shellfish hatcheries : a thesis presented in partial fulfilment of the requirements for the degree of Master of Philosophy in Biotechnology at Massey University, Palmerston North, New Zealand

A medium for the optimal growth of Chaetoceros calcitrans in batch and continuous culture systems was developed. A method was developed for continuous culture of C. calcitrans that was free from detrimental infection by bacteria. The concentration of tested nutrients in the developed medium were sodium nitrate, 160 mg/L; sodium dihydrogen orthophosphate, 40 mg/L; and the molar Si:N ratio was 0.25 (99.9 mg/L sodium metasilicate). Isolated bacterial strains were shown to be detrimental to the growth of C. calcitrans in batch and continuous culture. Electrolytically treated water was suitable for the growth of C. calcitrans, but a subsequent flourish of bacterial growth at the late exponential phase reduced the quality of the algal cells and made the culture unsuitable for feeding to shellfish larvae. Heat treated water (95°C for ten and a half minutes) gave stable growth for the continuous culture of C. calcitrans in 38 L plastic bioreactor bags for at least 38 days. The superficial gas velocity in the culture bags was 0.09 L/min. Higher superficial gas velocities (e.g. 0.40 L/min were detrimental to C. calcitrans

Enhancement of Recombinant Protein Production in Transgenic Nicotiana benthamiana Plant Cell Suspension Cultures with Co-Cultivation of Agrobacterium Containing Silencing Suppressors.

We have previously demonstrated that the inducible plant viral vector (CMViva) in transgenic plant cell cultures can significantly improve the productivity of extracellular functional recombinant human alpha-1-antiryspin (rAAT) compared with either a common plant constitutive promoter (Cauliflower mosaic virus (CaMV) 35S) or a chemically inducible promoter (estrogen receptor-based XVE) system. For a transgenic plant host system, however, viral or transgene-induced post-transcriptional gene silencing (PTGS) has been identified as a host response mechanism that may dramatically reduce the expression of a foreign gene. Previous studies have suggested that viral gene silencing suppressors encoded by a virus can block or interfere with the pathways of transgene-induced PTGS in plant cells. In this study, the capability of nine different viral gene silencing suppressors were evaluated for improving the production of rAAT protein in transgenic plant cell cultures (CMViva, XVE or 35S system) using an Agrobacterium-mediated transient expression co-cultivation process in which transgenic plant cells and recombinant Agrobacterium carrying the viral gene silencing suppressor were grown together in suspension cultures. Through the co-cultivation process, the impacts of gene silencing suppressors on the rAAT production were elucidated, and promising gene silencing suppressors were identified. Furthermore, the combinations of gene silencing suppressors were optimized using design of experiments methodology. The results have shown that in transgenic CMViva cell cultures, the functional rAAT as a percentage of total soluble protein is increased 5.7 fold with the expression of P19, and 17.2 fold with the co-expression of CP, P19 and P24

Clustering and dynamics of cytochrome bd-I complexes in the Escherichia coli plasma membrane in vivo

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Evidence of an association between poly(3-hydroxybutyrate) accumulation and phosphotransbutyrylase expression in Bacillus megaterium

Molecular analysis of a genomic region of Bacillus megaterium, a polyhydroxybutyrate (PHB)- producing microorganism, revealed the presence of a gene coding for the enzyme phosphotransbutyrylase (Ptb). Enzyme activity was measured throughout the different growth phases of B. megaterium and was found to correlate with PHB accumulation during the late-exponential growth phase. Ptb expression was repressed by glucose and activated by the branched amino acids isoleucine and valine. Overexpression of ActBm, a σ 54 regulator from B. megaterium whose gene is located upstream from ptb, caused an increase in Ptb activity and PHB accumulation in B. megaterium

Discovery Of Ethanol-Responsive Small Rnas In Zymomonas Mobilis

Zymomonas mobilis is a bacterium that can produce ethanol by fermentation. Due to its unique metabolism and efficient ethanol production, Z. mobilis has attracted special interest for biofuel energy applications; an important area of study is the regulation of those specific metabolic pathways. Small RNAs (sRNAs) have been studied as molecules that function as transcriptional regulators in response to cellular stresses. While sRNAs have been discovered in various organisms by computational prediction and experimental approaches, their discovery in Z. mobilis has not yet been reported. In this study, we have applied transcriptome analysis and computational predictions to facilitate identification and validation of 15 novel sRNAs in Z. mobilis. We furthermore characterized their expression in the context of high and low levels of intracellular ethanol. Here, we report that 3 of the sRNAs (Zms2, Zms4, and Zms6) are differentially expressed under aerobic and anaerobic conditions, when low and high ethanol productions are observed, respectively. Importantly, when we tested the effect of ethanol stress on the expression of sRNAs in Z. mobilis, Zms2, Zms6, and Zms18 showed differential expression under 5% ethanol stress conditions. These data suggest that in this organism regulatory RNAs can be associated with metabolic functions involved in ethanol stress responses.NSF CBET-1254754Welch Foundation F-1756Cellular and Molecular BiologyChemical Engineerin