Effect of microbial inoculants on the quality and aerobic stability of bermudagrass round-bale haylage.

The objective of this study was to compare the efficacy of using 4 commercially available microbial inoculants to improve the fermentation and aerobic stability of bermudagrass haylage. We hypothesized that the microbial inoculants would increase the fermentation and aerobic stability of the haylages. Bermudagrass (4-wk regrowth) was harvested and treated with (1) deionized water (control); (2) Buchneri 500 (B500; Lallemand Animal Nutrition, Milwaukee, WI) containing 1×10(5) of Pediococcus pentosaceus and 4×10(5) of Lactobacillus buchneri 40788; (3) Biotal Plus II (BPII; Lallemand Animal Nutrition) containing 1.2×10(5) of P. pentosaceus and Propionibacteria freudenreichii; (4) Silage Inoculant II (SI; AgriKing Inc., Fulton, IL) containing 1×10(5) of Lactobacillus plantarum and P. pentosaceus; and (5) Silo King (SK; AgriKing Inc.), containing 1×10(5) of L. plantarum, Enterococcus faecium, and P. pentosaceus, respectively. Forty round bales (8 per treatment; 441±26kg; 1.2×1.2 m diameter) were made and each was wrapped with 7 layers of plastic. Twenty bales were stored for 112 d and the remaining 20 were stored for 30 d and sampled by coring after intermediary storage periods of 0, 3, 7, and 30 d. The pH of control and inoculated haylages sampled on d 3 did not differ. However, B500 and BPII had lower pH (5.77±0.04 vs. 6.16±0.04; 5.06±0.13 vs. 5.52±0.13) than other treatments by d 7 and 30, respectively. At final bale opening on d 112, all treatments had lower pH than the control haylage (4.77±0.07 vs. 5.37±0.07). The B500, BPII, and SI haylages had greater lactic acid and lactic-to-acetic acid ratios than SK and control haylages. No differences were detected in neutral detergent fiber digestibility, dry matter losses, dry matter, lactic and acetic acid concentrations, and yeast and coliform counts. The SK haylage had lower clostridia counts compared with the control (1.19±0.23 vs. 1.99±0.23 cfu/g). Treatments B500, BPII, SI, and SK tended to reduce mold counts and they improved aerobic stability by 236, 197, 188, and 95%, respectively, compared with the control (276±22 vs. 99±22h).

Interactions between oil substrates and glucose on pure cultures of ruminal lipase-producing bacteria.

The hydrolysis of free fatty acids from lipids is a prerequisite for biohydrogenation, a process that effectively saturates free fatty acids. Anaerovibrio lipolyticus 5s and Butyrivibrio fibrisolvens have long been thought to be the major contributors to ruminal lipolysis; however, Propionibacterium avidum and acnes recently have been identified as contributing lipase activity in the rumen. In order to further characterize the lipase activity of these bacterial populations, each was grown with three different lipid substrates, olive oil, corn oil, and flaxseed oil (3 %). Because different finishing rations contain varying levels of glycogen (a source of free glucose) this study also documented the effects of glucose on lipolysis. P. avidum and A. lipolyticus 5s demonstrated the most rapid rates (P < 0.05) of lipolysis for cultures grown with olive oil and flaxseed oil, respectively. A. lipolyticus, B. fibrisolvens, and P. avidum more effectively hydrolyzed flaxseed oil than olive oil or corn oil, especially in the presence of 0.02 % glucose. Conversely, P. acnes hydrolyzed corn oil more readily than olive oil or flaxseed oil and glucose had no effect on lipolytic rate. Thus, these bacterial species demonstrated different specificities for oil substrates and different sensitivities to glucose.

An economical biorefinery process for propionic acid production from glycerol and potato juice using high cell density fermentation.

An economically sustainable process was developed for propionic acid production by fermentation of glycerol using Propionibacterium acidipropionici and potato juice, a by-product of starch processing, as a nitrogen/vitamin source. The fermentation was done as high-cell-density sequential batches with cell recycle. Propionic acid production and glycerol consumption rates were dependent on initial biomass concentration, and reached a maximum of 1.42 and 2.30 g L(-1) h(-1), respectively, from 50 g L(-1) glycerol at initial cell density of 23.7 gCDW L(-1). Halving the concentration of nitrogen/vitamin source resulted in reduction of acetic and succinic acids yields by ~39% each. At glycerol concentrations of 85 and 120 g L(-1), respectively, 43.8 and 50.8 g L(-1) propionic acid were obtained at a rate of 0.88 and 0.29 g L(-1) h(-1) and yield of 84 and 78 mol%. Succinic acid was 13 g% of propionic acid and could represent a potential co-product covering the cost of nitrogen/vitamin source.

Formulation and evaluation of herbal anti-acne moisturizer.

The moisture content present in human skin makes it look young and the use of moisturizer results in fastening the moisture with a surface film of oil. Acne vulgaris is one of the most commonly seen diseases among the youth. The present study is focused on the use of herbs as moisturizer for acne treatment. The anti-acne moisturizer was formulated from herbal crude extracts and investigated the physico-chemical parameters as well as antibacterial activity of the formulation. The study revealed that ethanol extract of Andrographis paniculata, Glycyrrhiza glabra, Ocimum sanctum, Azadiracta indica and Green tea possessed the potential for inhibiting acne. It was observed that the optimal formula of anti-acne moisturizer was satisfactorily effective to control acne inducing bacteria i.e., Staphylococcus epidermis and Propionibacterium. The physico-chemical parameters of the formulation were also optimal with no signs of irritation.

Production of cis-9,trans-11-conjugated linoleic acid in camelina meal and okara by an oat-assisted microbial process.

A method to obtain cis-9,trans-11-conjugated linoleic acid (c9,t11-CLA) into camelina meal and okara, the byproducts of plant oil processing, is described. The triacylglycerols in these materials were hydrolyzed with the aid of lipolytically active oat flour for 3 weeks at a water activity of 0.70. The resulting free linoleic acid was then isomerized predominantly to c9,t11-CLA by resting cells of Propionibacterium freudenreichii ssp. shermanii in 5% aqueous camelina meal and okara slurries. In camelina meal slurries, c9,t11-CLA content after 21 h of fermentation was 0.83 mg/mL and 96 mg/g of total lipids. In okara slurries, the content of c9,t11-CLA was 1.1 mg/mL and 78 mg/g of total lipids. Doubling the hydrolysis time in okara increased the subsequent content of c9,t11-CLA to 1.4 mg/mL, corresponding to 110 mg/g of total lipids. After isomerization, CLA was concentrated into a particulate material of the slurries by acidification. The results suggest that the method is applicable to a wide spectrum of lipid-containing plant materials to further increase their nutritional value.

Effects of copper supplement on growth and viability of strains used as starters and adjunct cultures for Emmental cheese manufacture.

AIMS: To determine the effects of supplemented copper (Cu2+) on growth and viability of strains used as starters and adjunct cultures for Emmental cheese manufacture. METHODS AND RESULTS: Thirteen strains belonging to Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus rhamnosus, Streptococcus thermophilus or Propionibacterium freudenreichii species were exposed to various copper concentrations in the proper growth medium at relevant growth temperatures, and the effects of supplemented copper on bacterial growth and cell viability were determined by optical density and pH measurements, also by platings. Among the species considered, L. delbrueckii was the most copper resistant and S. thermophilus the most sensitive to copper. Anaerobic conditions increased this sensitivity significantly. There was also a considerable amount of variation in copper resistance at strain level. CONCLUSIONS: Copper resistance is both a species- and strain-dependent property and may reflect variability in copper-binding capacities by cell wall components among species and strains. In addition, the chemical state of copper may be involved. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed that copper resistance is a highly variable property among starter and adjunct strains, and this variability should be considered when strains are selected for Emmental cheese manufacture.

Production of Propionibacterium shermanii biomass and vitamin B12 on spent media.

AIMS: The propionibacteria are commercially important due to their use in the cheese industry, and there is a growing interest for their probiotic effects. Stimulatory effects of lactic acid bacteria (LAB) on propionic acid bacteria have been observed. This study was designed to examine the possibility of using spent media previously used to grow LAB for the production of biomass and metabolites of Propionibacterium freudenreichii subsp. shermanii. METHODS AND RESULTS: Seventeen MRS and vegetable juice media were prefermented by various LAB and evaluated for their ability to subsequently support the growth of Propionibacterium, using automated spectrophotometry (AS). Growth of Propionibacterium in spent media was strongly affected by the LAB strain used to produce the spent medium. The native MRS medium (not prefermented) yielded the highest optical density values followed by prefermented media by Lactobacillus acidophilus, Bifidobacterium longum and Lactococcus lactis. Prefermented cabbage juice enabled good growth of Propionibacterium. For the production of organic acids and vitamin B12, cells of Propionibacterium were concentrated and immobilized in alginate beads in the aim of accelerating the bioconversions. More propionic acid was obtained in spent media than in native MRS. The concentration of vitamin B12 was higher in media fermented with free cells than those with immobilized cultures; with the free cells, its concentration varied from 900 to 1800 ng ml(-1) of media. CONCLUSIONS: It was demonstrated that spent media could be recycled for the production of Propionibacterium and metabolites, depending on the LAB strain that was previously grown. Media remediation is needed to improve the production of vitamin B12, especially with immobilized cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents an option for recycling of spent media generated by producers of LAB or producers of fermented vegetables. The propionic fermentation may result in three commercial products: biomass, vitamin B12 or organic acids, which may be used as starters, supplements or food preservatives. It is an attractive process from economical and environmental standpoints.

The in vivo assessment of safety and gastrointestinal survival of an orally administered novel probiotic, Propionibacterium jensenii 702, in a male Wistar rat model.

This study aimed to evaluate in vivo gastrointestinal survival and safety of orally administered probiotic bacterium, Propionibacterium jensenii 702, using a male Wistar rat model. A high dose of 10(10) cfu/rat/day of P. jensenii 702 was fed to each rat for 81 days. The repeated dose toxicity and translocation of P. jensenii 702 into rat tissues were evaluated, along with the rat faecal beta-glucuronidase activities and dairy propionibacteria counts. Results showed that P. jensenii 702 had no adverse effect on general health status, body weight gain, visceral organs and faecal beta-glucuronidase activities. No viable cells of P. jensenii 702 were recovered from blood and tissue samples (mesenteric lymph nodes, liver and spleen) of rats, and no treatment-associated illness or death was observed. Faecal dairy propionibacteria counts reached 10(8) cfu/g after 36 days treatment and remained between 10(8)-10(9) cfu/g till the end of 81 days treatment. The results indicate that P. jensenii 702 was able to survive the in vivo gastrointestinal tract transit of rats, with no adverse affects on the animals. However, further human clinical trials are required before strain P. jensenii 702 could be incorporated into food for human consumption as probiotics.

Use of lactic and propionic acid bacteria in the production of fermented parsley juice.

Taking into consideration the proliferation rate of Propionibacterium sp. cells, pH changes and sensory properties of the fermented product obtained, selection of propionic acid bacteria (PAB) was made in order to determine their usefulness for the production of fermented parsley juice. The analysis included 12 strains, belonging to the following species: P. thoenii (4 strains), P. jensenii (6 strains), P. freudenreichii (1 strain) and P. acidipropionici (1 strain). The experiments show that many strains of propionic acid rods develop well in parsley juice, allowing to achieve the desired taste qualities of the product. The Propionibacterium strains selected by elimination were used as components of vaccine containing: Lactobacillus plantarum, Lactococcus lactis ssp. lactis, Saccharomyces cerevisiae.

Nutritional requirements of anaerobic coryneforms.

The nutritional requirements of three species of anaerobic coryneforms and their serotypes (Propionibacterium acnes types I and II, P. avidum types I and II, and P. granulosum) were determined. Strains of P. avidum would consistently grow to a transmittance of 1 to 3% at 560 nm in a basal salts medium supplemented with glucose, pantothenate, biotin, thiamine, and 12 amino acids (alanine, arginine, cysteine, glutamine, glycine, histidine, isoleucine, methionine, phenylalanine, serine, tyrosine, and tryptophan). Strains of P. acnes and P. granulosum, however, failed to grow in this medium unless six additional amino acids were present (asparagine, leucine, lysine, proline, threonine, and valine). All three species grew equally well whether the 18 amino acids were supplied in the form of a casein hydrolysate supplemented with tryptophan or were added separately. Nicotinamide enhanced growth of P. acnes but had no effect on growth of P. avidum and P. granulosum. Other nutrients which were not absolute requirements, but which significantly improved growth of these species, included the purines guanine and/or adenine, Tween 80, which served as a source of oleic acid, sodium L-lactate, alpha-ketoglutarate, and pyruvate. Strains (86) comprising all five groups grew well in the defined medium, except four strains of P. acnes type II (29 tested), which failed to grow unless heme and vitamin K were added to the medium. One strain of P. granulosum (22 tested) failed to grow in any defined medium, suggesting an additional growth factor requirement.

Phosphorus pentoxide as a drying agent for bacterial culture extracts analyzed by gas-liquid chromatography.

The procedure for gas chromatographic analysis of metabolic products of microbial fermentation includes solvent extraction of the aqueous growth media, drying of the extract, and direct chromatographic analysis of the solvent. In this study, 2 drying agents, magnesium sulfate and phosphorus pentoxide, were compared. Both were effective in removing water; however, phosphorus pentoxide removed water more completely and at a faster rate than magnesium sulfate. When a thermal conductivity detector is used, it is important to completely remove water from the solvent to prevent interference with volatile acids and alcohols. When water is present, short-chain alcohols (C2-C5) are eluted together with the water, causing peak overlap and shoulder separations. Phosphorus pentoxide quickly and effectively removed water so that a baseline was established following the solvent front on the chromatogram. The use of phosphorus pentoxide is particularly advantageous for identification or fermentation studies on Clostridium and Propionibacterium when rapid identification is desired or when large numbers of cultures are to be tested.