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G . Z H A O A N D G . Z H A N G . 2005.Aims: The effects of protective agents, rehydration media and freezing temperature on the viabilities of Lactobacillus brevis and Oenococcus oeni H-2 when subjected to freeze-drying were investigated. Methods and Results: Several protectants and rehydration media were tested to improve the survival after freezedrying. The cells were also frozen at )65 and )20°C to check the effect of freezing temperature on the viability. Conclusions: The best protectant and rehydration medium to obtain the highest viability after freeze-drying varied with the species of bacteria. Yeast extract (4AE0%) and sodium glutamate (2AE5% ) gave maximum viability of L. brevis and O. oeni (67AE8% and 53AE6% respectively). The highest survival of L. brevis and O. oeni were obtained when rehydrated with 10% sucrose and MGY medium respectively. When the bacterial cells were frozen quickly ()65°C) than slowly ()20°C), L. brevis and O. oeni both showed increased viability after freeze-drying. Significance and Impact of the Study: The viabilities of L. brevis and O. oeni after freeze-drying were shown to be strain specific and dependent on protective agents, rehydration media and freezing temperature.

While green discoloration during garlic processing is of a major concern, this greening is desirable and required for the traditional homemade Chinese "Laba" garlic. To obtain insights into the mechanism of color formation, simulation of the greening of "Laba" garlic was carried out in the laboratory by soaking aged garlic in 5% (v/v, pH 2.33) acetic acid solution. After 2 days, the garlic cloves turned green. Up to 4 days, pigment(s) diffused from garlic cloves to the pickling solution. The solution exhibits two maximal absorbances at approximately 440 and approximately 590 nm, corresponding to yellow and blue species, respectively, the combination of which creates the green coloration. With increasing time from 4 to 25 days, the concentration of both yellow and blue species increases at nearly the same rate, while after 25 days, the concentration of the yellow species increases faster than that of the blue species. Interestingly, most thiosulfinates ( approximately 85%) in garlic cloves were converted within 4 days, suggesting that thiosulfinate conversion is proportional to the formation of the pigments. Consistent with this conclusion, alliinase and acetic acid were required for the color formation. UV-vis spectral measurements and pH results suggest that the color formation occurs by two kinds of processes: one enzymatic and the other nonenzymatic. Low pH (2.0-3.0) favors nonenzymatic reactions, while high pH (6.0 or above) is conducive to enzymatic reactions. Thus, the ideal pH for the entire process of garlic greening is between 4.0 and 5.0, which is a compromise of the optimal pH of both the enzymatic and nonenzymatic reactions.

Four adult Simmental male cattle (376 ± 9.0 kg initial BW), fitted with permanent rumen cannulas, were used in a 4 × 4 Latin square design to investigate the effects of dietary supplementing tannic acid (TA) on rumen fermentation, methane (CH ) production, rumen microbes, nutrient digestibility and plasma biochemical parameters. Four levels of TA, that is 0, 6.5, 13.0 or 26.0 g/kg dry matter (DM), were added to the basal ration (composed of corn silage and concentrate mixture) as experimental treatments respectively. Each experimental period consisted of a 12-day adaptation phase followed by a 3-day sampling phase. The results showed that supplementing TA at 26.0 g/kg DM decreased the relative abundance of protozoa, methanogens and Ruminococcus albus to the total ruminal bacterial 16S rDNA in beef cattle (p < 0.05). The results also showed that supplementing TA at 6.5, 13.0 or 26.0 g/kg DM decreased (p < 0.01) the CH production (l/kg DM intake) by 11.1%, 14.7% and 33.6% respectively. Supplementing TA at 13.0 or 26.0 g/kg DM decreased the ratio of acetate to propionate and ammonia nitrogen (NH -N) (p < 0.05) and tended to decrease the total volatile fatty acid (VFA) concentration of rumen fluid (p = 0.07). Supplementing TA at 26.0 g/kg DM decreased DM and organic matter (OM) digestibility (p < 0.05), supplementing TA at 6.5, 13.0 or 26.0 g/kg DM decreased (p < 0.01) crude protein (CP) digestibility by 5.0%, 8.6% and 15.7%, respectively, and supplementing TA at 6.5, 13.0 or 26.0 g/kg DM increased (p < 0.05) the plasma total antioxidant capability. It was concluded that supplementing TA in the ration of beef cattle decreased the CH production and digestibility of CP of beef cattle. Supplementing TA could be an effective option to mitigate CH emission form cattle, further research is necessary to study the effects of TA on the performance of cattle.

Modified guanidine-based polymers with chain extension were synthesized by condensation and cross-linking polymerizations in an attempt to increase molecular weight and charge density of the antimicrobial polymers. The antimicrobial activity and the corresponding mechanisms were investigated by several approaches. The results indicated that the antimicrobial activities of the modified guanidine-based polymer, based on the minimum inhibition concentration (MIC) against E.coli, varied with alkyl monomer ratios. UV absorption at 260 nm further quantified the amount of intracellular components leaked into bacteria suspension. The UV absorption measurements were also used to monitor inhibition processes dynamically. It was found that the modified guanidine-based polymer inhibited the growth of bacteria by causing membrane compromised and intracellular leaked. Dual fluorescent dyes were used to stain all bacteria including the dead ones, which enabled us to utilize CLSM to visualize the viability of bacteria in the presence of various modified guanidine-based polymers without causing any damage. The morphologies of bacteria untreated and treated with modified guanidine-based polymer were observed using an atomic force microscope (AFM), which further demonstrated the damage of E.coli membrane and the leakage of intracellular component induced by the modified guanidine-based polymers.