Levan Production by Suhomyces kilbournensis Using Sugarcane Molasses as a Carbon Source in Submerged Fermentation.

The valorization of byproducts from the sugarcane industry represents a potential alternative method with a low energy cost for the production of metabolites that are of commercial and industrial interest. The production of exopolysaccharides (EPSs) was carried out using the yeast Suhomyces kilbournensis isolated from agro-industrial sugarcane, and the products and byproducts of this agro-industrial sugarcane were used as carbon sources for their recovery. The effect of pH, temperature, and carbon and nitrogen sources and their concentration in EPS production by submerged fermentation (SmF) was studied in 170 mL glass containers of uniform geometry at 30 °C with an initial pH of 6.5. The resulting EPSs were characterized with Fourier-transform infrared spectroscopy (FT-IR). The results showed that the highest EPS production yields were 4.26 and 44.33 g/L after 6 h of fermentation using sucrose and molasses as carbon sources, respectively. Finally, an FT-IR analysis of the EPSs produced by S. kilbournensis corresponded to levan, corroborating its origin. It is important to mention that this is the first work that reports the production of levan using this yeast. This is relevant because, currently, most studies are focused on the use of recombinant and genetically modified microorganisms; in this scenario, Suhomyces kilbournensis is a native yeast isolated from the sugar production process, giving it a great advantage in the incorporation of carbon sources into their metabolic processes in order to produce levan sucrose, which uses fructose to polymerize levan.

Soybean molasses can be used as a substitute for corn in grazing beef cattle supplements during the rainy season.

Soybean molasses (SBMO) is a byproduct derived from the production of soy protein concentrate, obtained through solubilization in water and alcohol. The utilization of SBMO as an animal feed ingredient shows promising potential, primarily due to its low cost and as a potential energy concentrate. This study aimed to assess the intake, digestibility, ruminal parameters (pH and ruminal ammonia - NH3), nitrogen retention (NR) and microbial protein synthesis in grazing beef cattle supplemented with SBMO as a substitute for corn during the rainy season. Five Nellore (10-month-old) bulls with an average initial weight of 246 ± 11.2 kg were utilized in a 5 × 5 Latin square design. The animals were housed in five paddocks, each consisting of 0.34 ha of Marandu grass (Urochloa brizantha). Five isonitrogenous protein-energy supplements (300 g crude protein [CP]/kg supplement) were formulated, with SBMO replacing corn at varying levels (0, 0.25, 0.50, 0.75, or 1.00 g-1 g). The supplements were provided daily at a quantity of 2.0 kg-1 animal. The inclusion of SBMO at any level of corn substitution did not significantly affect the intake of pasture dry matter or total dry matter (P > 0.10). Likewise, the intake of CP and, consequently, the ruminal concentration of NH3 did not differ among the SBMO levels. Increasing the inclusion of SBMO did not have a significant impact on NR (P > 0.10), indicating that animals receiving supplements containing 100% SBMO as a substitute for corn may perform similarly to animals receiving supplements with 100% corn (0% SBMO). Soybean molasses represents a viable alternative energy source for grazing beef cattle during the rainy season and can entirely replace corn without adversely affecting animal nutritional performance.

Flavours and flavourings in waterpipe products: a comparison between tobacco, herbal molasses and steam stones.

ObjectivesFlavoured products are especially appealing to youth and contribute to the onset of waterpipe smoking and continued use of waterpipe tobacco. The goal of database and chemical analysis was to provide a clear overview of commonly used flavours and flavourings in tobacco and related waterpipe products, that is, herbal molasses and steam stones. METHODS: In 2019, 249 waterpipe tobacco products were registered in the European Common Entry Gate by manufacturers to be marketed in The Netherlands. Flavour categories were assigned to the registered products based on their brand names and product descriptions. Nicotine and eleven 1111 flavourings were identified and quantified in waterpipe tobacco (n=8), herbal molasses (n=7) and steam stones (n=4) by extraction and gas chromatography-mass spectrometry (GC-MS) analysis. RESULTS: Flavour categories could be assigned to 237 of 249 registered waterpipe tobacco products. Eight flavour main categories and 48 unique subcategories were identified and presented in a flavour wheel. All registered waterpipe tobacco products were flavoured, and the majority (78%) was fruit flavoured. Herbal molasses contained similar median flavouring levels, and steam stones contained lower median levels compared with waterpipe tobacco. Flavourings in waterpipe products were almost exclusively fruity and sweet, often in combination with menthol/mint flavourings. CONCLUSIONS: This study is the first to present a waterpipe tobacco flavour wheel, providing a quick overview of waterpipe tobacco flavours and thereby aiding communication among experts around the globe. GC-MS analysis revealed that the most prevalent flavourings are present in similar levels in herbal and tobacco waterpipe products. Banning flavourings in all waterpipe products would be a good strategy to reduce waterpipe smoking among youth.

Fermenting molasses and a synthetic odour blend to attract blood-fed Anopheles coluzzii.

Collecting blood-fed mosquitoes to monitor pathogen presence or to gather information on the host blood meal is often challenging. Fermenting molasses can be used to produce carbon dioxide to attract host-seeking mosquitoes, however, earlier work indicated that it may also attract blood-fed mosquitoes in the field. In the current study, these field results were validated in an experimental setting using a large cage setup with Anopheles coluzzii (Diptera, Culicidae). Blood-fed mosquitoes were indeed attracted to fermenting molasses with the highest attraction at 72 hours post feeding, which was used for subsequent experiments. Next, it was tested if fermentation of molasses is required for attraction, and whether it acts as an oviposition attractant, increases egg laying, or increases mosquito survival. The compounds that could be responsible for attraction were identified by combined electrophysiology and chemical analyses and formulated into a synthetic blend. Fermenting molasses attracted blood-fed mosquitoes in the large cage study, while fermenting sugar and non-fermenting molasses did not. The fecundity of blood-fed mosquitoes increased after feeding on fermenting molasses, however, compounds emanating from molasses did not trigger oviposition. The synthetic blend attracted blood-fed mosquitoes and may be used to determine mosquito host selection and for xenomonitoring, as 'flying syringes' to detect non-vector borne pathogens.

Effectively Converting Cane Molasses into 2,3-Butanediol Using Clostridiumljungdahlii by an Integrated Fermentation and Membrane Separation Process.

Firstly, 2,3-butanediol (2,3-BDO) is a chemical platform used in several applications. However, the pathogenic nature of its producers and the expensive feedstocks used limit its scale production. In this study, cane molasses was used for 2,3-BDO production by a nonpathogenic Clostridium ljungdahlii. It was found that cane molasses alone, without the addition of other ingredients, was favorable for use as the culture medium for 2,3-BDO production. Compared with the control (i.e., the modified DSMZ 879 medium), the differential genes are mainly involved in the pathways of carbohydrate metabolism, membrane transport, and amino acid metabolism in the case of the cane molasses alone. However, when cane molasses alone was used, cell growth was significantly inhibited by KCl in cane molasses. Similarly, a high concentration of sugars (i.e., above 35 g/L) can inhibit cell growth and 2,3-BDO production. More seriously, 2,3-BDO production was inhibited by itself. As a result, cane molasses alone with an initial 35 g/L total sugars was suitable for 2,3-BDO production in batch culture. Finally, an integrated fermentation and membrane separation process was developed to maintain high 2,3-BDO productivity of 0.46 g·L-1·h-1. Meanwhile, the varied fouling mechanism indicated that the fermentation properties changed significantly, especially for the cell properties. Therefore, the integrated fermentation and membrane separation process was favorable for 2,3-BDO production by C. ljungdahlii using cane molasses.

Effect of sugarcane bagasse as industrial by-products treated with Lactobacillus casei TH14, cellulase and molasses on feed utilization, ruminal ecology and milk production of mid-lactating Holstein Friesian cows.

BACKGROUND: The study aimed to evaluate the effect of Lactobacillus casei TH14, cellulase, and molasses combination fermented sugarcane bagasse (SB) as an exclusive roughage source in the total mixed ration (TMR) for mid-lactation 75% crossbred Holstein cows on feed intake, digestibility, ruminal ecology, milk yield and milk composition. Four multiparous mid-lactation crossbred (75% Holstein Friesian and 25% Thai native breed) dairy cows of 439 ± 16 kg body weight, 215 ± 5 days in milk and average milk yield 10 ± 2 kg d-1 were assigned to a 4 × 4 Latin square design. The unfermented SB (SB-TMR), SB fermented with cellulase and molasses (CM-TMR), SB fermented with L. casei TH14 and molasses (LM-TMR), and SB fermented with L. casei TH14, cellulase and molasses (LCM-TMR) were used as dietary treatments. RESULTS: CM-TMR, LM-TMR and LCM-TMR significantly (P < 0.01) increased dry matter and fiber digestibility, gross energy and metabolizable energy intake (P < 0.05), blood glucose, total volatile fatty acids (P < 0.05), propionic acid and milk yield, but decreased ammonia, acetic acid, acetic:propionic ratio and methane production (P < 0.05) when compared with the SB-TMR. Compared with fermented SB treatments, LCM-TMR had lower (P < 0.05) ruminal ammonia and greater blood glucose (P < 0.01); LCM-TMR showed (P < 0.05) greater volatile fatty acids, propionic acid, milk yield and total solids, and lower acetic:propionic ratio (P < 0.01); methane, protozoa and somatic cell count were found to be lowest in LCM-TMR. CONCLUSION: Combination of L. casei TH14 and additives (LCM-TMR) effectively enhanced feed use, rumen ecology and milk production of Holstein Friesian cows. © 2021 Society of Chemical Industry.

Lactobacillus plantarum and molasses alter dynamic chemical composition, microbial community, and aerobic stability of mixed (amaranth and rice straw) silage.

BACKGROUND: The objective was to determine how molasses and Lactobacillus plantarum affect chemical composition, fermentation quality, aerobic stability, and the microbial community of an ensiled mixture of amaranth (Amaranthus hypochondriaus, AF) and rice straw. Treatments were control (C, no addition), L. plantarum (L; 2 × 105 cfu g-1 fresh weight), molasses (M; 40 g kg-1 fresh matter), and their combination (LM). All treatments were ensiled for 1, 3, 5, 7, and 30 days. RESULTS: All additives improved fermentation quality with greater lactic acid (LA), acetic acid, and lower pH than C silage over the ensiling period. The LM silage combination optimized fermentability, manifested as greater LA contents and a more rapid pH reduction during the first 7 days of ensiling than L or M silages. After 30 days of ensiling, inoculant L. plantarum increased Lactobacillus abundance and reduced bacterial diversity and Enterobacteriaceae abundance compared with silage treated with molasses. Molasses addition reduced the relative concentration of structural carbohydrates (neutral and acid detergent fiber, and hemicellulose) after 30 days of ensiling. Finally, there was spoilage after 2 days and 4 days of aerobic exposure in C and LM silages respectively, whereas L silage had not spoiled after 4 days. CONCLUSIONS: Although the combination of L. plantarum and molasses further optimized fermentation characteristics, L silage had better aerobic stability. © 2021 Society of Chemical Industry.

Liquid molasses interacts with buffers to affect ruminal fermentation, milk fatty acid profile, and milk fat synthesis in dairy cows fed high-concentrate diets.

We aimed to evaluate the effects of feeding sugarcane liquid molasses (LM) with or without a commercial buffer mix (BFM) on ruminal fermentation parameters, milk fatty acid (FA) profile, and milk yield and composition in dairy cows fed high-concentrate diets (35:65 forage-to-concentrate ratio). Eight multiparous Holstein cows (4 ruminally cannulated) averaging 165 ± 12 d in milk at the beginning of the study were randomly assigned to a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Cows were fed the following diets: (1) no LM or BFM supplementation (CTRL), (2) LM without BFM supplementation (MOL), (3) BFM without LM supplementation (BUF), and (4) LM plus BFM supplementation (COMBO). These 4 isonitrogenous and isoenergetic diets were formulated by replacing (dry matter basis) 5% ground corn with LM, whereas BFM replaced wheat bran at 0.8% of the diet. Significant LM × BFM interactions were observed for the duration of ruminal pH below 5.8, molar proportion of propionate, acetate-to-propionate ratio, milk proportions of trans-10 18:1 and total trans FA, and concentration and yield of milk fat. Feeding MOL and BUF alone were effective on reducing the time that ruminal pH remained below 5.8 compared with the CTRL treatment, and the COMBO diet decreased it further. A similar pattern was observed for the ruminal molar proportion of propionate. The milk proportions of trans-10 18:1 and total trans FA dropped significantly with BFM or LM supplementation versus cows fed CTRL, and the COMBO diet decreased these variables further. Note, however, that these changes elicited by the COMBO diet were not in the same magnitude as those caused by MOL or BUF fed alone. The ruminal molar proportion of acetate increased with the BUF diet and that of butyrate increased in cows fed MOL, but mean ruminal pH was not affected by treatments. Diets with LM resulted in increased concentrations of short- and medium-chain FA in milk fat. The yield of 3.5% fat-corrected milk increased significantly in cows fed MOL or BUF due to the improved concentration of milk fat. A trend and a significant increase for energy-corrected milk were observed with feeding MOL or BUF, respectively. Overall, inclusion of LM and BFM appears to reduce milk trans-10 18:1 FA and total trans FA by modulating ruminal pH and volatile FA profile in cows fed high-concentrate diets.

Short communication: Characterization of molasses chemical composition.

Beet and cane molasses are produced worldwide as a by-product of sugar extraction and are widely used in animal nutrition. Due to their composition, they are fed to ruminants as an energy source. However, molasses has not been properly characterized in the literature; its description has been limited to the type (sugarcane or beet) or to the amount of dry matter (DM), total or water-soluble sugars, crude protein, and ash. Our objective was to better characterize the composition of cane and beet molasses, examine possible differences, and obtain a proper definition of such feeds. For this purpose, 16 cane and 16 beet molasses samples were sourced worldwide and analyzed for chemical composition. The chemical analysis used in this trial characterized 97.4 and 98.3% of the compounds in the DM of cane and beet molasses, respectively. Cane molasses contained less DM compared with beet molasses (76.8 ± 1.02 vs. 78.3 ± 1.61%) as well as crude protein content (6.7 ± 1.8 vs. 13.5 ± 1.4% of DM), with a minimum value of 2.2% of DM in cane molasses and a maximum of 15.6% of DM in beet molasses. The amount of sucrose differed between beet and cane molasses (60.9 ± 4.4 vs. 48.8 ± 6.4% of DM), but variability was high even within cane molasses (39.2-67.3% of DM) and beet molasses. Glucose and fructose were detected in cane molasses (5.3 ± 2.7 and 8.1 ± 2.8% of DM, respectively), showing high variability. Organic acid composition differed as well. Lactic acid was more concentrated in cane molasses than in beet molasses (6.1 ± 2.8 vs. 4.5 ± 1.8% of DM), varying from 1.6 to 12.8% of DM in cane molasses. Dietary cation-anion difference showed numerical differences among cane and beet molasses (7 ± 53 vs. 66 ± 45 mEq/100 g of DM, on average). It varied from -76 to +155 mEq/100 g of DM in the cane group and from +0 to +162 mEq/100 g of DM in the beet group. Data obtained in this study detailed differences in composition between sources of molasses and suggested that a more complete characterization could improve the use of molasses in ration formulation.

Influence of caramel and molasses addition on acrylamide and 5-hydroxylmethylfurfural formation and sensory characteristics of non-centrifugal cane sugar during manufacturing.

BACKGROUND: The aims of this study are to (i) evaluate the effects of color enhancers, caramel (C) and molasses (M), on acrylamide and 5-hydroxylmethylfurfural (HMF) formation in non-centrifugal cane sugar (NCS) and to (ii) perform nine-point hedonic scale and evaluation of sensory attributes, encompassing the appearance, flavor, texture and aftertaste, by 71 consumers on NCS, NCS_C, and NCS products made with a blend of molasses and sugar (NCS_MS) and steam processing (NCS_S). RESULTS: With the addition of molasses and caramel at the maximum allowable level of 5 g kg-1 in sugarcane juice, significantly greater acrylamide or HMF did not accumulate in NCS_C and NCS_M during the thermal manufacturing process, while color values of NCS_C significantly changed (P < 0.05). The increases in acrylamide and HMF contents were influenced by pH because they were produced by the Maillard reaction. Hedonic responses showed that NCS_MS was rated with the highest score for overall acceptance, whereas NCS_S, with the lowest content of acrylamide, exhibited the lowest score for every attribute. In addition, the appearance acceptance score of NCS_C was significantly higher than that of NCS (P < 0.05). Significant differences were also found between NCS and NCS_C in the frequency of 9 of 16 items with which consumers selected to characterize the appearance in a check-all-that-apply questionnaire (P < 0.05). CONCLUSIONS: The association between hedonic evaluations and sensory profiles in visual attributes of NCS_C indicated that caramel could be a promising addition in Maillard reaction-mitigated NCS products to improve consumer preferences through color strengthening without safety concerns. © 2020 Society of Chemical Industry.

Dietary supplementation of beta-glucan-rich molasses yeast powder on antibody response to swine fever virus and hematology of starter-grower pigs.

This research investigated the impact of dietary beta-glucan-rich molasses yeast powder (MYP) supplementation on the antibody response to swine fever virus (Titer) and hematology of starter-grower pig. Sixteen cross pigs (30 kg body weight) were equally split into four groups; each group with four replicates and fed four dietary treatments that consisted of basal diets (control) and the basal diets added with 2.5, 5.0, and 7.5% MYP. Feed and water were consumed ad libitum for 44 days. Feed intake (FI), MYP intake (MYPI), beta-glucan intake (BGI), and Mannan-oligosaccharide intake (MOSI) were recorded daily. Titer was evaluated after 15 (Titer15) and 30 (Titer30) days after vaccination, while hematology was analyzed at the end of the experiment. The results indicated that it was unchangeable for ADFI (P > 0.05). No impacts were observed on hematological variables and Titer15 in MYP fed pigs (P > 0.05). However, supplementation with 7.5% MYP increased platelet count (PC) and Titer30 (P < 0.01), but decreased hematocrit (Hct) (P < 0.05). Titer 30 and titer 15 were linked to MYPI, BGI, and MOSI (P < 0.05). Based on the study, feeding starter-grower pigs diets supplemented with 7.5% MYP might enhance the antibody response to swine fever virus 30 days after vaccination, and it has a potential role in the application in prevention of swine fever virus disease.

Development of food-grade expression system for d-allulose 3-epimerase preparation with tandem isoenzyme genes in Corynebacterium glutamicum and its application in conversion of cane molasses to D-allulose.

D-Allulose 3-epimerase (DAE) has been applied to produce D-allulose, a low-calorie and functional sweetener. In this study, a new DAE from Paenibacillus senegalensis was characterized in Escherichia coli. Furthermore, we presented a tandem isoenzyme gene expression strategy to express multiple DAEs in one cell and construct food-grade expression systems based on Corynebacterium glutamicum. Seventeen expression cassettes based on three DAE genes from different organisms were constructed. Among all recombinant strains, DAE16 harboring three DAE genes in an expression vector exhibited the highest enzyme activity with 22.7 U/mg. Whole-cell transformation of DAE16 produced 225 g/L D-allulose with a volumetric productivity of 353 g·g -1 ·hr -1 . The catalytic efficiency of strain C-DAE9 integrating total 11 DAE genes in chromosome was 16.4-fold higher than strains carrying one DAE. Fed-batch culture of C-DAE9 gave enzyme activity of 44,700 U/L. We also expressed a thermostable invertase in C. glutamicum and obtained enzyme activity of 29 U/mg. Immobilized cells expressing DAE or invertase exhibited 80% of retained activity after 30 cycles of catalytic reactions. Those immobilized cells were coupled to produce 61.2 g/L D-allulose from cane molasses in a two-step reaction process. This study provided an efficient approach for enzyme preparation and allowed access to produce D-allulose from other abundant and low-cost feedstock enriched with sucrose.

Selection of thermotolerant Saccharomyces cerevisiae for high temperature ethanol production from molasses and increasing ethanol production by strain improvement.

A thermotolerant ethanol fermenting yeast strain is a key requirement for effective ethanol production at high temperature. This work aimed to select a thermotolerant yeast producing a high ethanol concentration from molasses and increasing its ethanol production by mutagenesis. Saccharomyces cerevisiae DMKU 3-S087 was selected from 168 ethanol producing strains because it produced the highest ethanol concentration from molasses at 40 °C. Optimization of molasses broth composition was performed by the response surface method using Box-Behnken design. In molasses broth containing optimal total fermentable sugars (TFS) of 200 g/L and optimal (NH4)2SO4 of 1 g/L, with an initial pH of 5.5 by shaking flask cultivation at 40 °C ethanol, productivity and yield were 58.4 ± 0.24 g/L, 1.39 g/L/h and 0.29 g/g, respectively. Batch fermentation in a 5 L stirred-tank fermenter with 3 L optimized molasses broth adjusted to an initial pH of 5.5 and fermentation controlled at 40 °C and 300 rpm agitation resulted in 72.4 g/L ethanol, 1.21 g/L/h productivity and 0.36 g/g yield at 60 h. Strain DMKU 3-S087 improvement was performed by mutagenesis using ultraviolet radiation and ethyl methane sulfonate (EMS). Six EMS mutants produced higher ethanol (65.2 ± 0.48-73.0 ± 0.54 g/L) in molasses broth containing 200 g/L TFS and 1 g/L (NH4)2SO4 by shake flask fermentation at 37 °C than the wild type (59.8 ± 0.25 g/L). Among these mutants, only mutant S087E100-265 produced higher ethanol (62.5 ± 0.26 g/L) than the wild type (59.5 ± 0.02 g/L) at 40 °C. In addition, mutant S087E100-265 showed better tolerance to high sugar concentration, furfural, hydroxymethylfurfural and acetic acid than the wild type.

Profiling Metabolites and Biological Activities of Sugarcane (Saccharum officinarum Linn.) Juice and its Product Molasses via a Multiplex Metabolomics Approach.

Sugarcane (Saccharum officinarum L.) is an important perennial grass in the Poaceae family cultivated worldwide due to its economical and medicinal value. In this study, a combined approach using mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy was employed for the large-scale metabolite profiling of sugarcane juice and its by-product molasses. The polyphenols were analysed via UPLC-UV-ESI-MS, whereas the primary metabolites such as sugars and organic and amino acids were profiled using NMR spectroscopy and gas chromatography/mass spectrometry (GC/MS). UPLC/MS was more effective than NMR spectroscopy or GC/MS for determining differences among the metabolite compositions of the products. Under the optimized conditions, UPLC/MS led to the identification of 42 metabolites, including nine flavonoids, nine fatty acids, and two sterols. C/O Flavone glycosides were the main subclass detected, with tricin-7-O-deoxyhexosyl glucuronide being detected in sugarcane and molasses for the first time. Based on GC/MS analysis, disaccharides were the predominant species in the sugarcane juice and molasses, with sucrose accounting for 66% and 59%, respectively, by mass of all identified metabolites. The phenolic profiles of sugarcane and molasses were further investigated in relation to their in vitro antioxidant activities using free radical scavenging assays such as 2,2-Diphenyl-1-picrylhydrazyl free radical-scavenging ability (DPPH), Trolox equivalent antioxidant capacity (TEAC) and ferric reducing antioxidant power (FRAP). In view of its higher total phenolic content (TPC) (196 ± 2.1 mg GAE/100 g extract) compared to that of sugarcane juice (93 ± 2.9 mg GAE/100 g extract), molasses exhibited a substantially higher antioxidant effect. Interestingly, both extracts were also found to inhibit α-glucosidase and α-amylase enzymes, suggesting a possible antihyperglycaemic effect. These findings suggest molasses may be a new source of natural antioxidants for functional foods.

Stability of pyrrolizidine alkaloids from Senecio vernalis in grass silage under different ensilage conditions.

BACKGROUND: This study evaluated the degradation of pyrrolizidine alkaloids (PAs) from eastern groundsel (Senecio vernalis) in grass silage prepared with different inoculants. Silages were produced from ryegrass with 230 g kg-1 dry matter (DM) content and mixed with eastern groundsel (9:1; w/w fresh matter basis) containing 5.5 g kg-1 DM PA. Treatments were: CON (untreated control), LP (3.0 × 105 cfu g-1 Lactobacillus plantarum DSMZ 8862/8866) or LBLC (7.3 × 104 cfu g-1 Lactobacillus buchneri LN40177 / Lactobacillus casei LC32909), and each of the treatments in combination with 30 g kg-1 molasses. Silages were prepared in glass jars and opened after 3, 10, and 90 days. Fermentation characteristics were determined and the PAs analyzed. RESULTS: Although the levels of fermentation acids differed between treatments, results indicated good quality of all silages during 90 days. Significant time (P < 0.001) and treatment (P < 0.001) effects were observed for PAs. Concentrations of senecionine and seneciphylline decreased with molasses, declined over time, and were negatively correlated with lactic, propionic, and butyric acid, or with lactic and butyric acid in case of seneciphylline. In all silages, seneciphylline and senecionine N-oxides were undetectable after 3 days, whereas senkirkine, the most abundant PA, remained stable. CONCLUSIONS: Silage prepared from grass contaminated with eastern groundsel still contained high PA levels, and was hence a potential health hazard. Molasses supplementation reduced concentrations of senecionine and seneciphylline, while the bacterial inoculants had no effect. Other potentially toxic PA metabolites were not analyzed in the present study and further research is needed. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Bioprocessing of agro-industrial residues into lactic acid and probiotic enriched livestock feed.

BACKGROUND: Growing challenges of resource depletion, food security and environmental protection are putting stress on the development of biorefinery processes for bioprocessing of residues from food and agro-industry into value-added products. In this study, the simultaneous production of lactic acid (LA) and livestock feed on a combined substrate based on molasses and potato stillage by Lactobacillus paracasei NRRL B-4564 immobilized onto sunflower seed hull (SSH), brewer's spent grain (BSG) and sugar beet pulp (SBP) was studied. RESULTS: The highest total LA concentration of 399 g L-1 with overall productivity of 1.27 g L-1  h-1 was achieved in repeated batch fermentation by SBP-immobilized biocatalyst, followed by BSG- and SSH-immobilized cells. Fermentation improved the content of proteins and ash, and decreased the content of fibers in all three support materials. In addition, the fermentation had favorable effect on in vitro dry matter digestibility and energy values of SSH and BSG. According to assessment of probiotic potential, L. paracasei demonstrated a favorable probiotic profile, exhibiting high resistance to simulated ruminant digestive tract and significant antioxidant and antimicrobial activity. CONCLUSIONS: The proposed strategy enables valorization of agro-industrial residues as value-added ruminant feed and simultaneous LA production. Following principles of circular economy, the developed process combines different raw materials and integrates them into a biorefinery process, improving the overall profitability and productivity. © 2019 Society of Chemical Industry.

Chemical composition, in vitro gas production, methane production and fatty acid profile of canola silage (Brassica napus) with four levels of molasses.

The objective of this study was to investigate the effect of four levels of molasses on chemical composition, in vitro digestibility, methane production and fatty acid profile of canola silages. A canola (Brassica napus var. Monty) crop was established in a small-scale agricultural farm and harvested 148 days after sowing. Four levels of molasses were tested with respect to the fresh weight (1.5 kg); these were 1% (CS-1), 2% (CS-2), 3% (CS-3) and 4% (CS-4) molasses, and 0% molasses (CS-0) was included as a control. A total of 45 microsilages were prepared using PVC pipes (4 in. of diameter × 20 cm of length), and the forage was compressed using a manual press. The effects of control and treatments were tested using the general linear model Y = µ + Ti + Eij. The linolenic acid (C18:3n3), palmitic acid (C16:0) and linoleic acid methyl ester (C18:2n6c) accounted for 30%, 21% and 10.5% of total fatty acids, respectively; the fermentation parameters and in vitro methane production were not affected (P > 0.05) by treatments; in vitro digestibility decreased significantly (P < 0.05) as the level of molasses increased. It was concluded that CS-4 improved the DM content by 9% and showed high content of linolenic acid methyl ester. The gross energy of canola silages could favour the oleic acid methyl ester.

Effects of monensin on growth performance of beef heifers consuming warm-season perennial grass and supplemented with sugarcane molasses.

The objective of this study was to evaluate the effects of monensin on performance of beef heifers fed warm-season forages. Brangus heifers (n = 24) were stratified by BW and BCS, and randomly assigned into 1 of 12 bahiagrass pastures (1.2 ha and 2 heifers/pasture). Heifers were offered 14 kg of sugarcane molasses and 3.5 kg of cottonseed meal weekly from day 0 to 84. Treatments were randomly assigned to pastures (6 pastures/treatment) and consisted of heifers supplemented with or without 200 mg/day of monensin. On d 85, heifers were allocated to individual drylot pens, provided free choice access to bermudagrass hay, and received their respective treatment for 10 d of adaptation and 11 d of data collection. Monensin did not impact (P ≥ 0.13) heifer BW, BCS, overall ADG, bahiagrass IVDOM, CP, herbage mass, and allowance. Supplement disappearance after 10 and 34 h of supplementation was greater for control vs. monensin heifers (P = 0.04) and tended to be greater for control vs. monensin heifers 24 h post-supplementation (P = 0.07). Plasma concentrations of glucose, IGF-1, and BUN (P ≥ 0.24) did not differ between treatments. From d 85 to 106, forage and total DM intake, in vivo DM digestibility, and heifer growth performance did not differ between treatments (P ≥ 0.12). Therefore, adding monensin to sugarcane molasses-based supplements decreased supplement consumption rate, but did not impact growth and blood parameters of heifers grazing warm-season grasses with limited nutritive value.

Engineered E. coli W enables efficient 2,3-butanediol production from glucose and sugar beet molasses using defined minimal medium as economic basis.

BACKGROUND: Efficient microbial production of chemicals is often hindered by the cytotoxicity of the products or by the pathogenicity of the host strains. Hence 2,3-butanediol, an important drop-in chemical, is an interesting alternative target molecule for microbial synthesis since it is non-cytotoxic. Metabolic engineering of non-pathogenic and industrially relevant microorganisms, such as Escherichia coli, have already yielded in promising 2,3-butanediol titers showing the potential of microbial synthesis of 2,3-butanediol. However, current microbial 2,3-butanediol production processes often rely on yeast extract as expensive additive, rendering these processes infeasible for industrial production. RESULTS: The aim of this study was to develop an efficient 2,3-butanediol production process with E. coli operating on the premise of using cost-effective medium without complex supplements, considering second generation feedstocks. Different gene donors and promoter fine-tuning allowed for construction of a potent E. coli strain for the production of 2,3-butanediol as important drop-in chemical. Pulsed fed-batch cultivations of E. coli W using microaerobic conditions showed high diol productivity of 4.5 g l-1 h-1. Optimizing oxygen supply and elimination of acetoin and by-product formation improved the 2,3-butanediol titer to 68 g l-1, 76% of the theoretical maximum yield, however, at the expense of productivity. Sugar beet molasses was tested as a potential substrate for industrial production of chemicals. Pulsed fed-batch cultivations produced 56 g l-1 2,3-butanediol, underlining the great potential of E. coli W as production organism for high value-added chemicals. CONCLUSION: A potent 2,3-butanediol producing E. coli strain was generated by considering promoter fine-tuning to balance cell fitness and production capacity. For the first time, 2,3-butanediol production was achieved with promising titer, rate and yield and no acetoin formation from glucose in pulsed fed-batch cultivations using chemically defined medium without complex hydrolysates. Furthermore, versatility of E. coli W as production host was demonstrated by efficiently converting sucrose from sugar beet molasses into 2,3-butanediol.

Graded replacement of maize grain with molassed sugar beet pulp modulated ruminal microbial community and fermentation profile in vitro.

BACKGROUND: Molassed sugar beet pulp (Bp) is a viable alternative to grains in cattle nutrition for reducing human edible energy input. Yet little is known about the effects of high inclusion rates of Bp on rumen microbiota. This study used an in vitro approach and the quantitative polymerase chain reaction technique to establish the effects of a graded replacement of maize grain (MG) by Bp on the ruminal microbial community, fermentation profile and nutrient degradation. RESULTS: Six different amounts of Bp (0-400 g kg-1 ), which replaced MG in the diet, were tested using the in vitro semi-continuous rumen simulation technique. The increased inclusion of Bp resulted in greater dietary content and degradation of neutral detergent fibre (P < 0.01). Further, Bp feeding enhanced (P < 0.01) the abundance of genus Prevotella and shifted (P < 0.01) the short-chain fatty acid patterns in favour of acetate and propionate and at the expense of butyrate. A total replacement of MG with Bp resulted in an increased daily methane production (P < 0.01). CONCLUSION: Results suggest positive effects of the replacement of MG by Bp especially in terms of stimulating ruminal acetate and propionate fermentation. However, high replacement rates of Bp resulted in lowered utilization of ammonia and higher ruminal methane production. © 2017 Society of Chemical Industry.