Substrate Supplementation Research Articles

Algae Biomass Based Media for Poly(3-hydroxybutyrate) (PHB) Production by Escherichia coli

In addition to biodiesel production from algae, the production of other valuable bioproducts facilitates the development of an algae-based biorefinery platform. The goal of this study was to utilize the aqueous fraction from a novel algal wet lipid extraction procedure as the medium for the production of a bio product, poly(3-hydroxybutyrate) (PHB), via the growth of recombinant Escherichia coli. PHB yield was measured at 34 % of the E. coli dry cell mass, and was increased to 51 % when the algae aqueous medium was supplemented with glucose. While the addition of inorganic nutrients to the aqueous phase did not increase PHB production or growth of E. coli, growth of E. coli was observed to increase with the supplementation of carbon substrate (glucose). The addition of carbon rich waste to the aqueous fraction of wastewater-derived algae may in the future provide a sustainable alternative. Future research will be directed at evaluating this concept to develop a sustainable process for the production of bioplastics through an algae-based biorefinery platform.

The effect of the timing of meal intake on energy metabolism during moderate exercise.

Although the intake of carbohydrates is important for the supplementation of energy substrate utilized during exercise, fat oxidation is possibly prevented by an elevation of insulin, and whether or not the timing of the intake of meals affects energy metabolism during exercise has not been clarified. The purpose of this study was to investigate the effect of the timing of the intake of meals taken at different times before exercise on the carbohydrate and fat metabolism during aerobic exercise. The subjects were eight young trained athletes who performed cycling exercise at the lactate threshold (LT) intensity for 60 min. They performed under five conditions consisting of a no-meal (water) trial, and four meal trials that had a normal meal at 1, 2, 3, and 4 h before the exercise. There were no significant changes for any trial in respiratory exchange ratio, or carbohydrate or fat oxidation rates during exercise. The serum insulin level before exercise in the meal trials was more elevated, the shorter the time to the start of the exercise from meal intake. A tendency for higher blood glucose was shown during exercise with a shorter interval time in the meal trials. No alterations were demonstrated for the serum free fatty acids in any of the groups. These results showed that the timing of the pre-exercise meal taken within a 4-h period before exercise did not affect the energy metabolism of the trained subjects during exercise at LT intensity.

Effect of Different Substrate Supplements on Oyster Mushroom (Pleurotus spp.) Production

st flush showing no significant differences with treatment T1 whereas T1 took maximum yield in 2 nd flush and 3 rd flush. P. djmor showed the highest percentage of dry matter (17.23%) and moisture content was found high in P. sajor-caju (87.37%). P. ostreatus and P. sajor-caju showed the maximum protein (27.23%) and fiber (26.28%) contents. The ash contents were found maximum P. sajor-caju (9.08%).The highest fat and carbohydrate contents were found in Pleurotus djmor (3.07%) and P. djmor (37.69) respectively.

Effects of a commercial blend of essential oils and monensin in a high-grain diet containing wheat distillers’ grains on in vitro fermentation

Li, Y. L., Li, C., Beauchemin, K. A. and Yang, W. Z. 2013. Effects of a commercial blend of essential oils and monensin in a high-grain diet containing wheat distillers’ grains on in vitro fermentation. Can. J. Anim. Sci. 93: 387–398. Our objective was to evaluate in vitro effects of a commercial blend of essential oils (BEO) versus monensin (MON) on fermentation of a high-grain diet containing wheat distillers’ dried grains with solubles (DDGS). Two experiments were conducted. The first experiment was designed as a short-term batch culture to determine the optimum dose of BEO (0, 45, 90 or 180 mg kg−1DM) on fermentation variables. The second experiment was a completely randomized block design with a 2×2 factorial arrangement of treatments using the rumen simulation technique. A high-grain dietary substrate containing wheat DDGS was supplemented with 0 or 28 mg MON kg−1DM combined with 0 and 90 mg BEO kg−1DM. Monensin did not affect the total volatile fatty acid (VFA) concentration or the molar proportions of individual VFA, but reduced CH4production (mL L−1gas; P=0.01) and disappearance of DM (P=0.04). Supplementation of BEO did not affect VFA concentration, but decreased (P=0.01) the molar proportion of acetate and reduced (P=0.02) the acetate to propionate concentration ratio and CH4production (mL g−1digested organic matter). The BEO increased (P=0.01) disappearance of neutral detergent fiber without affecting disappearance of other nutrients. These results indicate that supplementation of high-grain dietary substrate containing wheat DDGS with BEO improved fermentation pattern by increasing propionate concentration, reducing CH4production, and increasing fiber digestibility. This commercial BEO could be a potential substitute for MON for beef cattle fed high-grain diets containing DDGS.

About the Treatment of Kasabach-Merritt Syndrome

To the Editor, Dr. Emre and colleagues briefly reported on a 24-year-old female with Kasabach-Merritt syndrome in the most recent issue of this journal [1]. Three units of fresh frozen plasma were administrated within 3 days to the patient for the correction of severe hypofibrinogenemia despite a very high D-dimer level (5000 ng/mL), both of which reflected consumption coagulopathy, as stated by the authors. Although post-transfusion laboratory findings were not reported and consumption coagulopathy did not seem to be aggravated in the authors’ patient, I would like to draw attention to the complication of substrate supplementation in consumption coagulopathy cases without the taking of necessary precautions. On such an occasion, I urge our approach of mega-dose methylprednisolone treatment (MDMP; daily, 30 mg/kg for 3 days, then 20 mg/kg for 4 days, and subsequently 10, 5, 2, and 1 mg/kg with each dose administered for 1 week, around 6 AM as a single dose). This was originally given intravenously, but the oral route is now recommended. Although both routes seem to be equally effective, oral administration is more practical, is cheaper, and does not require patient admission [2,3,4,5].

Supplementation at casing to improve yield and quality of white button mushroom

Supplementation of substrate at casing to increase the yield and quality of mushroom [Agaricus bisporus (Lange) Sing] is an important practice in commercial production of white button mushroom. This project was done to study the effects of supplementing the compost at casing with ground corn and soybean seed applied at: 0 gas control, 17, 34 and51 gper17 kgcompost on production and harvest quality of A. bisporus. There were significant differences between supplemented and non-supplemented substrates. The34 gsoybean and51 gcorn treatments had the highest yield. There were significant differences in quality indices of mushroom due to the type and amount of the supplement. The51 gsoybean supplement produced higher protein compared with other substrates. The highest vitamin C, total phenol, total soluble solids and antioxidant capacity obtained of 34 and51 gsoybeans or 34 and51 gcorn, respectively. Increased production and quality of A. bisporus may be achieved by addition of suitable amounts of supplements.

Eastern Redcedar (Juniperus virginiana) as a Substrate Component Effects Growth of Three Tree Species

Sustainable and local alternative substrates are being explored for nursery crop production due to concern over pine bark (PB) supplies and costs. This study evaluated a tree species which is weedy in the Great Plains region of the United States, eastern redcedar, processed through a hammer mill equipped with a 19 mm (3/4 in) screen size to create six substrates consisting of 0, 5, 10, 20, 40, and 80% eastern redcedar chips (ERC) and 20% sand; the remaining volume was composed of PB. Each of these substrates were then used to grow baldcypress (Taxodium distichum), Chinese pistache (Pistacia chinensis), and silver maple (Acer saccharinum) under two fertilizer rates: either a 4.5 kg·m−3 (7.5 lbs·yd−3) low fertilizer rate or a 8.9 kg·m−3 (15 lbs·yd−3) high fertilizer rate. Substrates composed of 40 and 80% ERC had reduced container capacity, resulting in less growth of all three species. Plants responded similarly to both fertilizer rates suggesting that the limiting factor to plant growth is substrate physical properties. Plants grown in 5–20% ERC were of comparable size and quality to those grown in the control substrate. Therefore, ERC can be recommended as a PB substrate supplement, but not as a full replacement at this time.

The Effect of Supplementation of Micro Nutrient on Nutrient Rice Bran Which Fermented by Bacillus amyloliquefaciens

An experiment was conducted to improve the nutrient content of rice bran which supplemented with Zn, Sulfur and urea through fermentation by using cellulolytic bacteria (Bacillus amyloliquefaciens) a s inoculums. The objective of this experiment was to study the effect of supplementation of catalytic substrate consisting of Zn, sulfur and urea on the substrata characteristics and its microbial population.The experiment was determination of the optimum conditions (dosage of Zn, Sulfur and urea) for rice bran fermentation based on nutrient quality and quantity of these fermented products. The study was conducted in experimental methods, using the completely randomize design in factorial with 3 treatments were : 1) A factor (Dose of urea: A1 = 1.0%, A2 = 1.5%, A3 = 2.0%), 2) B factor (Dose sulfur: B1 = 0.2%, B2 = 0.4%, B3 = 0.8%) and 3) C factor (Dose Zn: C1 = 0.0025%, C2 = 0.005%, C3 = 0.0075%). Results of study showed that optimum conditions of the fermentation of rice brand was at 2% urea, 0.0025% Zn and 0.2% sulfur. The protein level increased 100% and the mineral content 60%, accompanied by 8.2% of increase in the digestibility. The level of phytic acid decreased 97%. This conditions can increase 36% of metabolizable energy, 15% nitrogen retention, 98% crude fiber digestible.

Effects of wheat dried distillers' grains with solubles and cinnamaldehyde on in vitro fermentation and protein degradation using the Rusitec technique

This study was conducted to evaluate the effect of wheat dried distillers' grains with solubles (DDGS) and cinnamaldehyde (CIN) on in vitro fermentation and microbial profiles using the rumen simulation technique. The control substrate (10% barley silage, 85% barley grain and 5% supplement, on dry matter basis) and the wheat DDGS substrate (30% wheat DDGS replaced an equal portion of barley grain) were combined with 0 and 300 mg CIN/l of culture fluid. The inclusion of DDGS increased (p < 0.05) the concentration of volatile fatty acids (VFA) and the molar proportion of acetate and propionate. Dry matter disappearance (p = 0.03) and production of bacterial protein (p < 0.01) were greater, whereas the disappearances of crude protein (CP) and neutral detergent fibre were less (p < 0.01) for the DDGS than for the control substrate. With addition of CIN, concentration of total VFA decreased and fermentation pattern changed to greater acetate and less propionate proportions (p < 0.01). The CIN reduced (p < 0.01) methane production and CP degradability. The copy numbers of Fibrobacter, Prevotella and Archaea were not affected by DDGS but were reduced (p < 0.05) by CIN. The results indicate that replacing barley grain by DDGS increased nutrient fermentability and potentially increase protein flows to the intestine. Supplementation of high-grain substrates with CIN reduced methane production and potentially increased the true protein reaching the small intestine; however, overall reduction of feed fermentation may lower the feeding value of a high-grain diet.

Effects of Supplemental Levels of Spent Mushroom (Flammulina velutipes) Substrates on Chemical Composition and Quality of Whole Crop Sorghum Silage

This study was carried out to determine the supplemental level of spent mushroom (Flammulina velutipes) substrates as an energy source in manufacturing of high moisture sorghum whole crop silage. Whole crop sorghum was harvested at heading stage and ensiled with spent mushroom substrates of 20% (S-20), 40% (S-40) and 60% (S-60) as fresh matter basis. Each silage was manufactured in plastic buckets included vinyl bag by three replications and stored for 0, 3, 6 and 9 weeks, respectively. Fermentation characteristics and quality of sorghum silages manufactured by supplemental level of spent mushroom substrates were as follows. Moisture contents of whole crop sorghum and spent mushroom substrates were 83.85% and 54.3%, respectively, and that of silages was 78% for S-20, 71% for S-40 and 68% for S-60. Ether extracts content of silages was significantly (P<0.05) increased during the fermentation periods. The pH in silages fermented for 3 weeks and above ranged from 4.24 to 4.42, and the decrease of pH by fermentation was relatively greater in S-40 compare to the other treatments. The lactic acid content of silage inclined that the S-40 was higher compared to the other treatments and decreased with elongation of fermentation period of silage. The contents of acetic acid and propionic acid of silages were not influenced by treatments and fermentation period. Flieg's score for estimation of silage quality ranged from 60 to 83, and was relatively high quality in the S-40 fermented for 9 week, and was relatively low quality in the S-60 fermented for 9 week. From above results, we suggest that 40% supplementation of spent Flammulina velutipes mushroom substrates as an energy source is resonable level in manufacturing of high moisture sorghum whole crop silage.

Oral l-serine supplementation reduces production of neurotoxic deoxysphingolipids in mice and humans with hereditary sensory autonomic neuropathy type 1

Hereditary sensory and autonomic neuropathy type 1 (HSAN1) causes sensory loss that predominantly affects the lower limbs, often preceded by hyperpathia and spontaneous shooting or lancinating pain. It is caused by several missense mutations in the genes encoding 2 of the 3 subunits of the enzyme serine palmitoyltransferase (SPT). The mutant forms of the enzyme show a shift from their canonical substrate L-serine to the alternative substrate L-alanine. This shift leads to increased formation of neurotoxic deoxysphingolipids (dSLs). Our initial analysis showed that in HEK cells transfected with SPTLC1 mutants, dSL generation was modulated in vitro in the presence of various amino acids. We therefore examined whether in vivo specific amino acid substrate supplementation influenced dSL levels and disease severity in HSAN1. In mice bearing a transgene expressing the C133W SPTLC1 mutant linked to HSAN1, a 10% L-serine–enriched diet reduced dSL levels. L-serine supplementation also improved measures of motor and sensory performance as well as measures of male fertility. In contrast, a 10% L-alanine–enriched diet increased dSL levels and led to severe peripheral neuropathy. In a pilot study with 14 HSAN1 patients, L-serine supplementation similarly reduced dSL levels. These observations support the hypothesis that an altered substrate selectivity of the mutant SPT is key to the pathophysiology of HSAN1 and raise the prospect of l-serine supplementation as a first treatment option for this disorder.

Effects of carbon sources and lipids on freeze-drying survival of Lactobacillus bulgaricus in growth media

The aim of this study was to investigate the effects of various substrate supplements in the growth media on the synthesis of cyclopropane fatty acid (cycC19:0) and freeze-drying survival rate of Lactobacillus bulgaricus L2 and their relatedness. Survival rate during freeze-drying was enhanced markedly (P < 0.05) when cells were grown in the presence of Tween 20 and sucrose as compared with the control. However, the survival rate of L. bulgaricus was lower when cells were grown in the presence of sugars (mannitol, maltose, dextrin and trehalose) and Tween (Tween 40 and Tween 60) relative to the control. Compared with the control, cells with a higher freeze-drying survival rate exhibited a higher cycC19:0 content of change. There was no linear correlation between these two parameters.

Metabolic engineering for the production of prenylated polyphenols in transgenic legume plants using bacterial and plant prenyltransferases

Prenylated polyphenols are secondary metabolites beneficial for human health because of their various biological activities. Metabolic engineering was performed using Streptomyces and Sophora flavescens prenyltransferase genes to produce prenylated polyphenols in transgenic legume plants. Three Streptomyces genes, NphB, SCO7190, and NovQ, whose gene products have broad substrate specificity, were overexpressed in a model legume, Lotus japonicus, in the cytosol, plastids or mitochondria with modification to induce the protein localization. Two plant genes, N8DT and G6DT, from Sophora flavescens whose gene products show narrow substrate specificity were also overexpressed in Lotus japonicus. Prenylated polyphenols were undetectable in these plants; however, supplementation of a flavonoid substrate resulted in the production of prenylated polyphenols such as 7- O-geranylgenistein, 6-dimethylallylnaringenin, 6-dimethylallylgenistein, 8-dimethylallynaringenin, and 6-dimethylallylgenistein in transgenic plants. Although transformants with the native NovQ did not produce prenylated polyphenols, modification of its codon usage led to the production of 6-dimethylallylnaringenin and 6-dimethylallylgenistein in transformants following naringenin supplementation. Prenylated polyphenols were not produced in mitochondrial-targeted transformants even under substrate feeding. SCO7190 was also expressed in soybean, and dimethylallylapigenin and dimethylallyldaidzein were produced by supplementing naringenin. This study demonstrated the potential for the production of novel prenylated polyphenols in transgenic plants. In particular, the enzymatic properties of prenyltransferases seemed to be altered in transgenic plants in a host species-dependent manner.

Biodegradation of BTE in soil by indigenous microbial populations with and without biogenic substrates

Degradation of benzene, toluene, and ethylbenzene (BTE) by microbial populations indigenous to the soil and populations proliferated from the indigenous using biogenic substrates were compared. The reaction system consisted of aerobic microcosms representing an unsaturated soil. Microcosms supplemented with glucose and citrate, when compared to the unsupplemented microcosms, showed increases in bacterial counts, but the overall degradation rates for B, T, or E were reduced in spite of shorter lag times. Both biogenic substrate supplements were non-beneficial for BTE degradation due largely to the preferential and healthy growth of the indigenous populations on the biogenic substrates, and thus the urgency of developing a favorable amount of BTE degraders was reduced.

Ethanol production from high dry matter corncob using fed-batch simultaneous saccharification and fermentation after combined pretreatment

To obtain high concentration of ethanol from cellulose, corncob was pretreated with acid and alkali to remove non-cellulose components, and then subjected to simultaneous saccharification and fermentation (SSF). An ethanol concentration as high as 69.2g/L was achieved with 19% dry matter (DM) using batch SSF, resulting in an 81.2% overall ethanol yield. A fed-batch process using a high solid concentration was also investigated. Fresh substrate was pretreated with dilute sulfuric acid–sodium hydroxide, and then added at different amounts during the first 24h, to yield a final dry matter content of 25% (w/v). SSF conditions with cellulose loading of 22.8 FPU/g glucan, dry yeast (Saccharomyces cerevisiae) loading of 5g/L and substrate supplementation every 4h yielded the highest ethanol concentration of 84.7g/L after 96h. This corresponded to a 79% overall ethanol yield.

Improvement of yield of Pleurotus eryngii var. eryngii by substrate supplementation and use of a casing overlay

Improved yield and biological efficiency (BE) of Pleurotus eryngii var. eryngii were achieved by supplementation of substrate with a commercial delayed-release nutrient and use of a casing overlay. Yield increases of 14% were achieved from cased substrates that were supplemented at time of casing with delayed-release nutrient (Remo’s). Use of a casing layer enhanced yield by 141% over non-cased substrates. When casing and substrate supplementation were combined, yield increased 179% over non-cased/non-supplemented substrates. Mushrooms harvested from cased substrates were darker in color and solids contents were lower compared to non-cased substrates. An additional break of mushrooms was harvested from non-cased “spent” substrate by fragmenting and re-supplementing the substrate prior to the application of a casing overlay. Three production methods were compared for their effect on mushroom yield: “standard”, “casing” and “casing after first break”. Casing of the substrate before first break (“casing” production method) resulted in the highest yield and biological efficiency.

Production of plant-specific tyramine derivatives by dual expression of tyramine N-hydroxycinnamoyltransferase and 4-coumarate:coenzyme A ligase in Escherichia coli

Plant-specific bioactive compounds including feruloyltyramine (FT), 4-coumaroyltyramine (CT), and caffeoyltyramine (CaT) were simultaneously produced in Escherichia coli by heterologous expression of two biosynthetic genes encoding 4-coumarate:coenzyme A ligase and tyramine N-hydroxycinnamoyltransferase (THT) cloned from Arabidopsis thaliana and pepper, respectively. Simultaneous supplementation of substrates to the recombinant E. coli resulted in the secretion of multiple tyramine derivatives into the medium at high yield: CT (189 mg l(-1)), FT (135 mg l(-1)), CaT (40 mg l(-1)). In addition, the recombinant E. coli also produced, albeit at low concentration, a range of dopamine derivatives such as feruloyldopamine due to THT's ability to accept dopamine as a substrate.

GROWING MEDIA FOR THE NURSERY INDUSTRY: USE OF AMENDMENTS IN TRADITIONAL BARK-BASED MEDIA

GROWING MEDIA FOR THE NURSERY INDUSTRY: USE OF AMENDMENTS IN TRADITIONAL BARK-BASED MEDIA

Treatment of textile wastewater with an anaerobic fluidized bed reactor

The discharge of textile wastewater containing dyes in the environment is worrying for both toxicological and esthetical reasons as dyes impede light penetration, damage the quality of the receiving streams and are toxic to food chain organisms. A fluidized bed reactor (FBR) with activated carbon as support material has been used to investigate the removal efficiency of chemical oxygen demand (COD), biochemical oxygen demand (BOD), and color of textile wastewater. The FBR was fed continuously without supplemented (Phase 1 and 2) and supplemented with glucose (Phase 3–5) at hydraulic retention times (HRT) of 4 h, and increased to 8 and 12 h with flow rates of 36, 18,12 L/d, respectively. The HRT was operated for a duration of 14 days. The effect of operational conditions such as organic loading rate (OLR), (HRT), and influence of glucose concentration as substrate additives was investigated to achieve the maximum removal efficiency in the reactor. Results indicated that anaerobic treatment of textile wastewater was possible with the supplementation of substrate additives as external carbon sources such as glucose (about 0.6 g/l). A further increase in the external carbon source added to textile wastewater did not improve the color removal efficiency of the anaerobic FBR reactor. The study implied that 98% soluble COD, 95% BOD 5, and 65% color reduction were possible by an anaerobic FBR.

The Hereditary Inclusion Body Myopathy Enigma and its Future Therapy

Hereditary inclusion body myopathy (HIBM) is a genetic muscle disease due to mutations in the gene encoding the enzyme complex UDP-N-acetylglucosamine 2 epimerase-N-acetylmannosamine kinase (GNE), which catalyzes the rate-limiting step in sialic acid production. The review describes some of the disease features that may be relevant for further understanding of the metabolic impairment of HIBM and its future therapy. It also addresses the biochemical basis behind the substrate supplementation therapy designed for this condition.