Effects of the concentration and nature of total dissolved solids in drinking water on feed intake, nutrient digestion, energy balance, methane emission, ruminal fermentation, and blood constituents in different breeds of young goats and hair sheep.

Understanding how different livestock species and breeds respond to consumption of brackish water could improve usage of this resource. Therefore, Angora, Boer, and Spanish goat doelings and Dorper, Katahdin, and St. Croix ewe lambs (6 animals per animal type [AT]; initial age = 296 ± 2.1 days) consuming water with varying concentrations of minerals of a natural brackish water source (BR) and sodium chloride (NaCl; SL) were used to determine effects on water and feed intake, nutrient digestion, heat energy, methane emission, ruminal fluid conditions, and blood constituent concentrations. There were 6 simultaneous 6 (water treatments [WT]) × 6 (AT) Latin squares with 3-wk periods. The WT were fresh (FR), BR alone (100-BR), a similar total dissolved solids (TDS) concentration as 100-BR via NaCl addition to FR (100-SL), BR with concentrations of all minerals increased by approximately 50% (150-BR), a similar TDS level as 150-BR by NaCl addition to FR (150-SL), and a similar 150 TDS level achieved by addition of a 1:1 mixture of BR minerals and NaCl to 100-BR (150-BR/SL). Concentrations (mg/kg) in BR were 4928 TDS, 85.9 bicarbonate, 224.9 calcium, 1175 chloride, 60.5 magnesium, 4.59 potassium, 1387 sodium, 1962 sulfate, and 8.3 boron, and TDS in other WT were 209, 5684, 7508, 8309, and 7319 mg/kg for FR, 100-SL, 150-BR, 150-SL, and 150-BR/SL, respectively. There were very few significant effects of WT or AT × WT interactions, although AT had numerous effects. Water intake was affected by AT (P = 0.02) and WT (P = 0.04), with greater water intake for 150-SL than for FR, 100-BR, 100-SL, and 150-BR. Dry matter intake among AT was lowest (P < 0.05) for Angora. Digestion of organic matter and neutral detergent fiber and heat energy differed among AT (P < 0.05), but nitrogen digestion and ruminal methane emission were similar among AT. Blood aldosterone concentration was higher (P < 0.05) for FR than for other WT. In conclusion, all AT seemed resilient to these WT regardless of mineral source and concentrations, with TDS less than 8300 mg/kg, which did not influence nutrient utilization, ruminal fermentation, energy balance, or blood constituent levels.

The genus Geobacillus and their biotechnological potential.

The genus Geobacillus comprises a group of Gram-positive thermophilic bacteria, including obligate aerobes, denitrifiers, and facultative anaerobes that can grow over a range of 45-75°C. Originally classified as group five Bacillus spp., strains of Bacillus stearothermophilus came to prominence as contaminants of canned food and soon became the organism of choice for comparative studies of metabolism and enzymology between mesophiles and thermophiles. More recently, their catabolic versatility, particularly in the degradation of hemicellulose and starch, and rapid growth rates have raised their profile as organisms with potential for second-generation (lignocellulosic) biorefineries for biofuel or chemical production. The continued development of genetic tools to facilitate both fundamental investigation and metabolic engineering is now helping to realize this potential, for both metabolite production and optimized catabolism. In addition, this catabolic versatility provides a range of useful thermostable enzymes for industrial application. A number of genome-sequencing projects have been completed or are underway allowing comparative studies. These reveal a significant amount of genome rearrangement within the genus, the presence of large genomic islands encompassing all the hemicellulose utilization genes and a genomic island incorporating a set of long chain alkane monooxygenase genes. With G+C contents of 45-55%, thermostability appears to derive in part from the ability to synthesize protamine and spermine, which can condense DNA and raise its Tm.

Modular system for assessment of glycosyl hydrolase secretion in Geobacillus thermoglucosidasius.

The facultatively anaerobic, thermophilic bacterium Geobacillus thermoglucosidasius is being developed as an industrial micro-organism for cellulosic bioethanol production. Process improvement would be gained by enhanced secretion of glycosyl hydrolases. Here we report the construction of a modular system for combining promoters, signal peptide encoding regions and glycosyl hydrolase genes to facilitate selection of the optimal combination in G. thermoglucosidasius. Initially, a minimal three-part E. coli-Geobacillus sp. shuttle vector pUCG3.8 was constructed using Gibson isothermal DNA assembly. The three PCR amplicons contained the pMB1 E. coli origin of replication and multiple cloning site (MCS) of pUC18, the Geobacillus sp. origin of replication pBST1 and the thermostable kanamycin nucleotidyltransferase gene (knt), respectively. G. thermoglucosidasius could be transformed with pUCG3.8 at an increased efficiency [2.8×10(5) c.f.u. (µg DNA)(-1)] compared to a previously reported shuttle vector, pUCG18. A modular cassette for the inducible expression and secretion of proteins in G. thermoglucosidasius, designed to allow the simple interchange of parts, was demonstrated using the endoglucanase Cel5A from Thermotoga maritima as a secretion target. Expression of cel5A was placed under the control of a cellobiose-inducible promoter (Pßglu) together with a signal peptide encoding sequence from a G. thermoglucosidasius C56-YS93 endo-ß-1,4-xylanase. The interchange of parts was demonstrated by exchanging the cel5A gene with the 3' region of a gene with homology to celA from Caldicellulosiruptor saccharolyticus and substituting Pßglu for the synthetic, constitutive promoter PUp2n38, which increased Cel5A activity five-fold. Cel5A and CelA activities were detected in culture supernatants indicating successful expression and secretion. N-terminal protein sequencing of Cel5A carrying a C-terminal FLAG epitope confirmed processing of the signal peptide sequence.