An in vitro gas production technique was used in this study to elucidate the effect of two strains of active live yeast on methane ($CH_4$) production in the large intestinal content of pigs to provide an insight to whether active live yeast could suppress $CH_4$ production in the hindgut of pigs. Treatments used in this study include blank (no substrate and no live yeast cells), control (no live yeast cells) and yeast (YST) supplementation groups (supplemented with live yeast cells, YST1 or YST2). The yeast cultures contained $1.8{\times}10^{10}$ cells per g, which were added at the rates of 0.2 mg and 0.4 mg per ml of the fermented inoculum. Large intestinal contents were collected from 2 Duroc${\times}$Landrace${\times}$Yorkshire pigs, mixed with a phosphate buffer (1:2), and incubated anaerobically at $39^{\circ}C$ for 24 h using 500 mg substrate (dry matter (DM) basis). Total gas and $CH_4$ production decreased (p<0.05) with supplementation of yeast. The methane production reduction potential (MRP) was calculated by assuming net methane concentration for the control as 100%. The MRP of yeast 2 was more than 25%. Compared with the control group, in vitro DM digestibility (IVDMD) and total volatile fatty acids (VFA) concentration increased (p<0.05) in 0.4 mg/ml YST1 and 0.2 mg/ml YST2 supplementation groups. Proportion of propionate, butyrate and valerate increased (p<0.05), but that of acetate decreased (p<0.05), which led to a decreased (p<0.05) acetate: propionate (A: P) ratio in the both YST2 treatments and the 0.4 mg/ml YST 1 supplementation groups. Hydrogen recovery decreased (p<0.05) with yeast supplementation. Quantity of methanogenic archaea per milliliter of inoculum decreased (p<0.05) with yeast supplementation after 24 h of incubation. Our results suggest that live yeast cells suppressed in vitro $CH_4$ production when inoculated into the large intestinal contents of pigs and shifted the fermentation pattern to favor propionate production together with an increased population of acetogenic bacteria, both of which serve as a competitive pathway for the available H2 resulting in the reduction of methanogenic archaea.
Pinus rigida ${\times}$ P. taeda seedlings in a nursery was inoculated with basidiospores of Pisolithus tinctorius (Pt) either collected from Suweon, Korea or introduced from U.S.A. to compare the effectiveness of the spores from two different origins as mycorrhizal inocula. Nursery beds were fumigated with methyl bromide and 1g of spores was used to inoculate $1m^2$ of soil surface just before seed sowing. Seedlings inoculated with American Pt (#250 strain from Georgia, U.S.A.) were 15% taller than Korean Pt at the end of the first growing season. The seedlings from fumigation treatment only (no inoculation involved) was slightly taller (statistically unsignificant) than those with Korean Pt, but slightly smaller than those with American Pt. In a subsequent year experiment, the seedlings inoculated with American and Korean Pt after soil fumigation were 66% and 60% taller, respectively, than seedlings infected by natural fungi without soil fumigation, suggesting the dual effects of Pt and fumigation on the seedling growth. Therefore potential of Pt spores for an effective inoculum exists and selection of Pt strains which have adapted to specific local environments is needed to develop better sources of mycorrhizal inocula.
Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.
The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.
Kim, Junhyung;Kim, Young-Eun;Park, Myeonghwa;Song, Young Eun;Seol, Eunhee;Kim, Jung Rae;Oh, You-Kwan
New & Renewable Energy
/
v.16
no.1
/
pp.58-67
/
2020
Microbial electrosynthesis has recently been considered a potentially sustainable biotechnology for converting carbon dioxide (CO2) into valuable biochemicals. In this study, bioelectrochemical acetate production from CO2 was studied in an H-type two-chambered reactor system with an anaerobic microbial consortium. Metal-rich mud flat was used as the inoculum and incubated electrochemically for 90 days under a cathode potential of -1.1 V (vs. Ag/AgCl). Four consecutive batch cultivations resulted in a high acetate concentration and productivity of 93 mmol/L and 7.35 mmol/L/day, respectively. The maximal coulombic efficiency (rate of recovered acetate from supplied electrons) was estimated to be 64%. Cyclic voltammetry showed a characteristic reduction peak at -0.2~-0.4 V, implying reductive acetate generation on the cathode electrode. Furthermore, several electroactive acetate-producing microorganisms were identified based on denaturing- gradient-gel-electrophoresis (DGGE) and 16S rRNA sequence analyses. These results suggest that the mud flat can be used effectively as a microbial source for bioelectrochemical CO2 conversion.
Plant growth-promoting effects of rhizobacterial inoculation obtained in pot experiments cannot always be dependably reproduced in fields. In this study, we investigated the effect of inoculation with Azospirillum brasilense and Methylobacterium oryzae, which have displayed growth promoting effects in several pot experiments, on growth and fruit yield of red pepper under field condition in a plastic-film house. Four rows spaced 90 cm apart were prepared after application of compost ($10Mg\;ha^{-1}$), and red pepper seedlings (Capsicum annum L., Nocgwang) were transplanted in each row with 40-cm space. Experimental treatments were consisted of A. brasilense CW903 inoculation, M. oryzae CBMB20 inoculation, and uninoculated control. Twelve plots, 10 plants per plot, were allotted to the three treatments with four replicates in a completely randomized design. At the time of transplanting, 50 mL of each inoculum ($1{\times}10^8cells\;mL^{-1}$) was introduced into root zone soil of each plant, and re-inoculated at 7 and 14 days after transplant. Plant growth and fruit yield were measured during the experiment. Both A. brasilense CW903 and M. oryzae CBMB20 could not promote growth of red pepper plants. All growth parameters measured were not significantly different among treatments. There were large variations in fruit yield recorded on plot basis, and no statistically significant differences were found among treatments. The failure to demonstrate the expected plant growth promoting effect of the inoculants is possibly due to various environmental factors, including weather and soil characteristics, reducing the possibility to express the potential of the inoculated bacterial strains.
The morphology of filamentous fungi closely correlates with the productivity in submerged culture. Using itaconic acid (IA) production by Aspergillus terreus as a research model, the quantitative relationship between the growth form of A. terreus and IA production was investigated. IA fermentation was scaled up from shake flasks to a 7 L stirred tank bioreactor based on the quantitative relationship. Our results demonstrated the following: (1) Three morphologies of A. terreus were formed by changing the inoculum level and shape of the flask. (2) Investigation of the effects of the three morphologies on broth rheology and IA production revealed the higher yield of IA on dry cell weight (DCW, IA/DCW) and yield of glucose on DCW (consumed glucose/DCW) were achieved during clump growth of A. terreus. (3) By varying the $KH_2PO_4$ concentration and culture temperature, the relationships between clump diameter and IA production were established, demonstrating that the yield of IA on DCW ($R^2$ = 0.9809) and yield of glucose on DCW ($R^2$ = 0.9421) were closely correlated with clump diameter. The optimum clump diameter range for higher IA production was 0.40-0.50 mm. (4) When the clump diameter was controlled at 0.45 mm by manipulating the mechanical stress in a 7 L fermentor, the yield of IA on DCW and yield of glucose on DCW were increased by 25.1% and 16.3%, respectively. The results presented in this study provide a potential approach for further enhancement of metabolite production by filamentous fungi.
Bacterial populations from the rhizosphere were obtained and the efficacy of the bacterial wilt suppression, root colonizing ability and resistance to three kinds of chemical pesticides were assayed. According to these results, SKU48-2 was selected as a potential biological agent to control the bacterial wilt caused by Ralstonia solanacearum. SKU48-2 strain at $10^8CFU/ml$ inoculum was able to suppress the bacterial wilt up to 60% in greenhouse trials. Also, the resistance of SKU48-2 to chemical pesticides make possible to use in combination with chemical pesticides for the control of bacterial wilt. Three different powder formulations of SKU48-2 were developed. The shelf-life of powder formulations was effective up to 6 months of storage. Unformulated bacterial suspension could not be stored for 2 weeks, at which time cell viability was completely lost. According to 16S rDNA sequence data, the SKU48-2 stain was identified as Bacillus subtilis.
Kim Chang Kyu;Ra Dong Soo;Min Hong Sik;Lee Young Hee;Lee Eun Jong
Korean Journal Plant Pathology
/
v.1
no.3
/
pp.169-172
/
1985
Natural sclerotia of Rhizoctonia solani causing rice sheath blight were inoculated at 10 day intervals from June 15 to July 15 in paddy field, Icheon, Korea. Percentage of infected stems, top lesion height and percentage of. lesion height vs. plant height were higher in the early inoculated plots than in the late inoculated ones. However, rio significant differences among inoculation dates of sclerotia were found on the basis of degree of damage at maturing stage and rice yield. These results suggest that the time of initial symptom appearance under the same inoculum potential may not affect the damage of rice plants by the fungus.
Purpose: Essential oils are secondary metabolites of herbs and have antibacterial activities against foodborne pathogens. However, their applications for food protection are limited due to the hydrophobic and volatile natures of essential oils. Methods: In this study, essential oil nanoemulsions of rosemary and lavender were formulated with non-ionic surfactant Tween 80 and water using ultrasonic emulsification, and their antibacterial effects were determined. Results: The antibacterial activities of nanoemulsions were evaluated against 12 strains of 10 bacterial species, and significant antibacterial effects were observed against four Gram-positive and four Gram-negative bacteria but not against Streptococcus mutans and Shigella sonnei. In the disc diffusion test, the diameter of the inhibition zone proportionally increased with the concentration of nanoemulsions. Using cell turbidity measurement, minimum bactericidal concentration (MBC) of the nanoemulsions, which is the lowest concentration reducing viability of the initial bacterial inoculum by ${\geq}99.9%$, was significantly higher than the minimum inhibitory concentration (MIC) of the nanoemulsions. The largest bactericidal effects of lavender and rosemary essential oil nanoemulsions were observed against S. enterica and S. aureus, respectively. Conclusion: Nanoemulsion technique could improve antibacterial activity of essential oil nanoemulsions by increasing the solubility and stability of essential oils. Our findings shed light on the potential use of essential oil nanoemulsions as an alternative to chemical sanitizers in food protection.
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