Luminometric measurement of population activity of genetically modified Pseudomonas fluorescens in the soil

Genetically modified cells of Pseudomonas fluorescens, chromosomally marked with genes for bioluminescence, were inoculated into sterile soil microcosms. During incubation for 90 days, viable cell concentration did not change significantly but light output, measured by luminometry, decreased, indicating reduced metabolic activity due to lack of substrates. Amendment with nutrients resulted in parallel increases in both luminescence and dehydrogenase activity. Luminometry therefore enables rapid monitoring of the activity of populations of luminescence-marked microbial inocula in the soil, with greater sensitivity and selectivity than traditional techniques.     

Coupling of Fermentation and Esterification: Microbial Esterification of Decanoic Acid with Ethanol Produced via Fermentation

Two different kinds of bioprocess, ethanol fermentation and subsequent microbial esterification, were coupled using Issatchenkia terricola IFO 0933 in an interface bioreactor. The strain produced ethyl decanoate (Et-DA) by esterification of exogenous decanoic acid (DA) with ethanol produced via fermentation. The efficiency of the new coupling system depended on the concentration of glucose in a carrier and DA in an organic phase (decane) in an agar plate interface bioreactor. Optimum glucose content and DA concentration were 4% and 29 mM, respectively.     

Long-Term m5C Methylome Dynamics Parallel Phenotypic Adaptation in the Cyanobacterium Trichodesmium

A major challenge in modern biology is understanding how the effects of short-term biological responses influence long-term evolutionary adaptation, defined as a genetically determined increase in fitness to novel environments. This is particularly important in globally important microbes experiencing rapid global change, due to their influence on food webs, biogeochemical cycles, and climate. Epigenetic modifications like methylation have been demonstrated to influence short-term plastic responses, which ultimately impact long-term adaptive responses to environmental change. However, there remains a paucity of empirical research examining long-term methylation dynamics during environmental adaptation in nonmodel, ecologically important microbes. Here, we show the first empirical evidence in a marine prokaryote for long-term m5C methylome modifications correlated with phenotypic adaptation to CO₂, using a 7-year evolution experiment (1,000+ generations) with the biogeochemically important marine cyanobacterium Trichodesmium. We identify m5C methylated sites that rapidly changed in response to high (750 µatm) CO₂ exposure and were maintained for at least 4.5 years of CO₂ selection. After 7 years of CO₂ selection, however, m5C methylation levels that initially responded to high-CO₂ returned to ancestral, ambient CO₂ levels. Concurrently, high-CO₂ adapted growth and N₂ fixation rates remained significantly higher than those of ambient CO₂ adapted cell lines irrespective of CO₂ concentration, a trend consistent with genetic assimilation theory. These data demonstrate the maintenance of CO₂-responsive m5C methylation for 4.5 years alongside phenotypic adaptation before returning to ancestral methylation levels. These observations in a globally distributed marine prokaryote provide critical evolutionary insights into biogeochemically important traits under global change.     

Potential applications of natural origin polymer-based systems in soft tissue regeneration

Despite the many advances in tissue engineering approaches, scientists still face significant challenges in trying to repair and replace soft tissues. Nature-inspired routes involving the creation of polymer-based systems of natural origins constitute an interesting alternative route to produce novel materials. The interest in these materials comes from the possibility of constructing multi-component systems that can be manipulated by composition allowing one to mimic the tissue environment required for the cellular regeneration of soft tissues. For this purpose, factors such as the design, choice, and compatibility of the polymers are considered to be key factors for successful strategies in soft tissue regeneration. More recently, polysaccharide-protein based systems have being increasingly studied and proposed for the treatment of soft tissues. The characteristics, properties, and compatibility of the resulting materials investigated in the last 10 years, as well as commercially available matrices or those currently under investigation are the subject matter of this review.     

GROWTH AND ORGANIC OSMOLYTES OF GEOGRAPHICALLY DIFFERENT ISOLATES OF MICROCOLEUS CHTHONOPLASTES (CYANOBACTERIA) FROM BENTHIC MICROBIAL MATS:RESPONSE TO SALINITY CHANGE1

The comparative growth and osmotic acclimation often culture strains of the marine benthic cyanobacterium Microcoleus chthonoplastes Thuret isolated from microbial mats in Germany, Spain, Egypt, the United States, Mexico, Chile, and Australia were investigated in salinities ranging from freshwater to twice seawater. All isolates showed a broad growth versus salinity response consistent with the dominance of this species in intertidal and hypersaline microbial communities. Growth optima, salinity preferences, and maximum growth rates differed for each isolate and could be related to the habitat from which they were isolated. This is most obvious when comparing strains from brackish habitats with those from a hypersaline lake. While the former isolates exhibited sharply pronounced growth optima under hyposaline conditions, cultures from the hypersaline environment grew best in salinity more than double seawater. The major low-molecular weight organic compounds present in all M. chthonoplastes strains were the carbohydrates glycosylglycerol and trehalose. This was proven by using ¹³C-nuclear magnetic resonance spectroscopy. Glycosylglycerol was synthesized and accumulated with increasing salinities, indicating its role as an osmolyte. In contrast, trehalose was present in relatively high concentrations under hyposaline conditions only. Differences in the patterns of growth versus salinity, as well as in those of osmotic acclimation among the M. chthonoplastes isolates, point to the development of different physiological ecotypes within the species.     

Milk production responses and rumen fermentation of dairy cows supplemented with summer brassicas

Forage brassicas, such as summer turnip (ST; Brassica rapa) and forage rape (FR; Brassica napus), are used as supplementary crops during summer. However, studies with lactating dairy cows fed these forages are limited and report inconsistent productive responses. The aim of this study was to determine dry matter intake, rumen fermentation and milk production responses of dairy cows in mid-lactation supplemented with and without summer (‘ST’ or ‘FR’) brassicas. Twelve multiparous lactating dairy cows were randomly allocated to three dietary treatments in a replicated 3 × 3 Latin square design balanced for residual effects over three 21-day periods. The control diet consisted of 16.2 kg DM of grass silage, 2.25 kg DM of commercial concentrate and 2.25 kg DM solvent-extracted soybean meal. For the other two dietary treatments, 25% of the amounts of silage and concentrates were replaced with FR or ST. The inclusion of forage brassicas had no effects on milk production (24.2 kg cow/day average) and composition (average milk fat and protein 43.2 and 33.6 g/l, respectively). Dry matter intake was 0.98 kg and 1.12 kg lower for cows supplemented with FR and ST, respectively, resulting in a greater feed conversion efficiency (1.35 kg milk/kg DM for ST and FR v. 1.27 kg milk/kg DM for the control diet). Intraruminal pH was lower for cows supplemented with ST compared to the control diet; however, it did not decrease below pH 5.8 at any time of the day. After feeding, the concentrations of total short-chain fatty acids (SCFAs) in rumen contents increased with ST supplementation compared to the control diet. Inclusion of FR in the diet increased the molar proportion of acetate (68.5 mmol/100 mmol) in total SCFA at the expense of propionate, measured 6 h after feeding of the forage. The molar proportion of butyric acid was greater with ST and FR supplementation (13.1 and 12 mmol/100 mmol, respectively) than in control cows. The estimated microbial nitrogen (N) flow was 89.1 g/day greater when supplementing FR compared to the control diet. Based on the haematological measures, the inclusion of summer brassica forages did not affect the health status of the animals. These results indicate that mid-lactation dairy cows fed brassicas are able to maintain production despite the reduced intake, probably due to improved rumen fermentation and therefore nutrient utilization.     

Effects of sodium selenite and coated sodium selenite on lactation performance,total tract nutrient digestion and rumen fermentation in Holstein dairy cows

Se can enhance lactation performance by improving nutrient utilization and antioxidant status. However, sodium selenite (SS) can be reduced to non-absorbable elemental Se in the rumen, thereby reducing the intestinal availability of Se. The study investigated the impacts of SS and coated SS (CSS) supplementation on lactation performance, nutrient digestibility, ruminal fermentation and microbiota in dairy cows. Sixty multiparous Holstein dairy cows were blocked by parity, daily milk yield and days in milk and randomly assigned to five treatments: control, SS addition (0.3 mg Se/kg DM as SS addition) or CSS addition (0.1, 0.2 and 0.3 mg Se/kg DM as CSS addition for low CSS (LCSS), medium CSS (MCSS) and high CSS (HCSS), respectively). Experiment period was 110 days with 20 days of adaptation and 90 days of sample collection. Dry matter intake was higher for MCSS and HCSS compared with control. Yields of milk, milk fat and milk protein and feed efficiency were higher for MCSS and HCSS than for control, SS and LCSS. Digestibility of DM and organic matter was highest for CSS addition, followed by SS addition and then control. Digestibility of CP was higher for MCSS and HCSS than for control, SS and LCSS. Higher digestibility of ether extract, NDF and ADF was observed for SS or CSS addition. Ruminal pH decreased with dietary Se addition. Acetate to propionate ratio and ammonia N were lower, and total volatile fatty acids (VFAs) concentration was greater for SS, MCSS and HCSS than control. Ruminal H ion concentration was highest for MCSS and HCSS and lowest for control. Activities of cellobiase, carboxymethyl-cellulase, xylanase and protease and copies of total bacteria, fungi, Ruminococcus flavefaciens, Fibrobacter succinogenes and Ruminococcus amylophilus increased with SS or CSS addition. Activity of α-amylase, copies of protozoa, Ruminococcus albus and Butyrivibrio fibrisolvens and serum glucose, total protein, albumin and glutathione peroxidase were higher for SS, MCSS and HCSS than for control and LCSS. Dietary SS or CSS supplementation elevated blood Se concentration and total antioxidant capacity activity. The data implied that milk yield was elevated due to the increase in total tract nutrient digestibility, total VFA concentration and microorganism population with 0.2 or 0.3 mg Se/kg DM from CSS supplementation in dairy cows. Compared with SS, HCSS addition was more efficient in promoting lactation performance of dairy cows.