Effect of culture conditions on growth and production of docosahexaenoic acid (DHA) usingThraustochytrium aureum ATCC 34304

Environmental and medium factors were investigated as basic data for optimizing DHA production when usingThraustochytrium aureum. To study the effect of environmental conditions, the rotation speed and culture temperature were, changed. Plus the trend of the growth characteristics, lipid content in the biomass, and DHA content in lipids were evaluated according to various initial glucose concentrations. The biomass, lipid, and DHA analyses showed that the physiological characteristics ofT. aureum were closely related with the environmental and medium conditions, as in the case of other marine microorganisms. For example, a low rotation speed of 50 rpm lowered the cell growth rate as well as the DHA content in the lipids. A low temperature had a negative effect on the cell growth, yet a positive effect on the lipid content in the biomass. Different initial glucose concentrations had no effect on the lipid content in the biomass or DHA content in the lipids, yet did affect the cell growth. Accordingly, these results show that environmental and medium factors must be synthetically considered in order to optimize DHA production when usingT. aureum.     

Bacillus licheniformis protease-catalyzed peptide synthesis via the kinetically controlled approach using the carbamoylmethyl ester as an acyl donor in anhydrous acetonitrile

Summary The couplings ofN-protected amino acid esters with amino acid amides proved to be carried out in anhydrous acetonitrile in the presence ofBacillus licheniformis protease (subtilisin Carlsberg) immobilized on Celite. The maximal peptide yields were obtained with the immobilized enzyme prepared through lyophilization from a pH 10.7 buffer solution. A series of dipeptide syntheses and several segment condensations were achieved generally in high yields by the combined use of the immobilized enzyme prepared from this pH and the carbamoylmethyl ester as the acyl donor.     

Competitive ability of Daphnia under dominance of non-toxic filamentous cyanobacteria

It is generally assumed that Daphnia is more susceptible to the inhibitory effects of filamentous cyanobacteria than small cladocerans since daphnids have a larger gape size and filtrate the filaments, whereas small cladocerans do not. This study addresses the question whether food limitation has the potential to modify this scenario of cladoceran response to dominance of non-toxic filamentous cyanobacteria. Daphnia galeatawas grown under limited (0.1 mg C l^–1) and unlimited concentrations (1.0 mg C l^–1) of high-quality food algae both in the absence/presence of non-toxic filamentous Aphanizomenon flexuosum. As the effects of these cyanobacteria on D. galeatawere positive under food limiting conditions and negative at the high food density, it was concluded that D. galeatawas mainly affected by nutritional quality due to its ability to ingest the filaments, while mechanical interference with food collection was not important. In competition experiments between D. galeataand Bosmina longirostris, D. galeatawas the dominant species at regular additions of food (1.0 mg C l^–1) in the absence of Aphanizomenon. In the presence of these cyanobacteria, D. galeatawas inhibited during the first days of the experimental period. However, the negative effect at the initially high food density was outweighed by nutrition at food limiting conditions and the outcome in competitive dominance was not changed. The results demonstrate that the ability of D. galeata to ingest large-sized non-toxic cyanobacteria can be considered as advantageous under food limiting conditions.     

Influence of Environmental Factors on the Chemotaxis of Bradyrhizobium japonicum

Dependence of motility and chemotaxis was studied in two strains of Bradyrhizobium japonicum upon several environmental factors. In both strains, chemotaxis was found to increase with an increasing concentration of the attractant (glucose) to 5.5 × 10^–2 M. Both motility and chemotaxis reached their maximum in the two- to three-day cultures at neutral pH. The maximum motility of these bacteria occurred at 40°C. The maximum values of chemotaxis in these microorganisms were, however, observed at 20–25°C. Chemotaxis in acidic or alkaline media and at low temperatures was found to be markedly weaker. Nonoptimal values of these parameters in soil may be a limiting factor for the interaction of the given bacteria with soybean roots.     

Effects of activated charcoal, explant size, explant position and sucrose concentration on plant and shoot regeneration of Lilium longiflorum via young stem culture

An efficient system for the in vitro plant and shootregeneration of Lilium longiflorum was developed andaccomplished using transverse thin cell layers (tTCL) of young stems.tTCLs were cut transversely along young stems from which the shoot-tipshad been removed. Sections were measured accurately using a graded gridand were cut in 4 mm × 4 mm × 1 mm cubes, eliminatingepidermal tissue, and were cultured on one-half MS medium containing 8 gl^–1 agar, different sucrose concentrations (10, 20, 30 or 40g l^–1), and with or without 1 mg l^–1 activatedcharcoal (AC). Plants formed on the surface of tTCLs within 60 days onone-half MS medium containing 8 g l^–1 agar and 20 gl^–1 sucrose. Sections of 1 mm taken just below the apicalarea developed buds within 15 days, whereas the sections closer to thebase required about 45 days. Shoot regeneration was enhanced whensucrose concentration was used at 30 or 40 g l^–1 after 60days of culture. No root formation occurred. Both shooting and rootingoccurred when sucrose was used at 20 g l^–1. The plantletswere transferred to soil and grew well under greenhouseconditions.     

Nectar Features in Esterhazya macrodonta, a Hummingbird-Pollinated Scrophulariaceae in Southeastern Brazil

Esterhazya macrodonta is scarcely visited by hummingbirds, a fact that may be related to its low flower production and nectar features. Received 14 September 2000/ Accepted in revised form 31 January 2001     

Modulation of human humoral immune response through orally administered bovine colostrum

Eighteen healthy volunteers were randomized into two treatment groups and consumed liquid prepackaged bovine colostrum whey and placebo for 7 days. On days 1, 3 and 5, an attenuated Salmonella typhi Ty21a oral vaccine was given to all subjects to mimic an enteropathogenic infection. The circulating antibody secreting cells and the expression of phagocytosis receptors of the subjects before and after oral immunization were measured with the ELISPOT assay and flow cytometry. All subjects responded well to the vaccine. No significant differences were observed in ELISPOT values for IgA, IgG, IgM, Fcgamma and CR receptor expression on neutrophils and monocytes between the two groups. There was a trend towards greater increase in specific IgA among the subjects receiving their vaccine with bovine colostrum. These results suggest that bovine colostrum may possess some potential to enhance human special immune responses.     

Antagonistic effect of enterocin CCM 4231 from Enterococcus faecium on “bryndza”, a traditional Slovak dairy product from sheep milk

“Bryndza” is a traditional Slovak dairy product (type of soft cheese) made from sheep cheese which was ripened for 14 days. Because its manufacture, transporting and/or storing represent conditions which facilitate contamination, the effect of enterocin CCM4231 in “bryndza” was investigated with the aim to reduce the contaminant agents. “Bryndza” was divided into equal portions (50 g). The experimental sample (ES) as well as the control sample one (C1) were inoculated with Listeria innocua Li1 strain. The other control samples C2 and C3 were without Li1 strain. C3 control was selected as a reference control. ES and C2 portions were treated with purified enterocin CCM4231 in a concentration of 6400 AU/ml. Before the experimental inoculation, “bryndza” was checked for the presence of contaminant agents. The experiment lasted 1 week and the samples were stored in the refrigerator at 4 °C. Sampling was performed on day 1, on day 4 and on day 7. The control samples C2 and C3 were checked only on day 1 and then after 1 week. The following contaminant agents were detected in “bryndza” before its experimental inoculation with L. innocua Li1 strain: Escherichia coli in the amount 10³ cfu/ml/g, Staphylococcus aureus (10² cfu/ml/g) and enterococci (10⁴ cfu/ml/g). In the control sample C2, the number of E. coli was reduced to 10² cfu/ml/g. Enterococci and staphylococci were totally eliminated there. Concerning C3 control, natural decrease of bacteria was found and/or their unchanged counts. The value of pH (5) was stable during the whole experiment. In the experimental sample inoculated with Li1 strain, its counts were decreased immediately after enterocin CCM4231 addition approximately by one order of magnitude. This inhibitory effect was also detectable on day 4 by the difference of one order of magnitude between ES and C1. On day 7, 10³ cfu/ml/g of Li1 strain were detected in both samples (ES, C1). The difference by one order of magnitude indicated, an inhibitory effect of enterocin CCM4231 in “bryndza”. However, bacteriocin activity was not determined by laboratory analyses.     

土壤可溶性无机磷对微生物生物量磷测定的干扰

对于可溶性无机磷含量高于50mg/kg土的土壤,薰蒸处理前必须去除这部分磷,否则将严重干扰微生物量磷的测定,讨论了去除土壤中可溶性无机磷的方法。     

Growth inhibition by glucose oxidase system of enterotoxic Escherichia coli and Salmonella derby: in vitro studies

The antibacterial effect of different glucose oxidase (GOX)/glucose combinations was studied on two food-poisoning organisms, enterotoxic Escherichia coli PM 015 and Salmonella derby BP 177. Growth of E. coli in nutrient broth (NB) was clearly inhibited by 4.0 mg/ml glucose after 24 h when combined with 2.0 U/ml GOX and after 48 h when combined with 0.5 or 1.0 U/ml GOX. Salmonella derby was more resistant than E. coli, but showed clear growth inhibition only after 48 h when inoculated in tubes where 2 mg glucose/ml and 2 U GOX/ml (or 4 mg glucose/ml and 1 U GOX/ml) were combined. In order to understand if the enzyme effect on microbial growth can be attributed to hydrogen peroxide or to pH decrease as a result of the production of gluconic acid, catalase (CAT) was added to GOX/glucose system. Since CAT decomposes H₂O₂ to H₂O and O₂, the antibacterial effect was ascribed to a pH decrease as a result of gluconic acid in the system.