The current research is designed to analyze sterilization effects on mulberries in terms of storage temperature and storage period after treating with tap water (TW), electrolyzed water (EW) and aqueous chlorine dioxide ($ClO_2$). The treated $ClO_2$ concentrations are 10, 50, 100 and 200 ppm. In each concentration, the mulberries are soaked for 30 seconds respectively. The sterilization effects are being compared at room temperature ($25^{\circ}C$) and at $4^{\circ}C$, respectively. And the enzyme activity related to quality is also being investigated and analyzed about for browning inhibition effects. Microbial sterilizing power increases more in treating plots with EW and $ClO_2$ than treating plot with TW. Futhermore sterilizing power of $ClO_2$ increased sharply on high concentration treatment plot as well. Sterilization effects of $ClO_2$ during storage time are better at cold temperature. Pictures taken from scanning electron microscope reveal that there are no microbes in sterilizing solutions treatment plots. From measurement of the enzyme activity, it is concluded that activities decrease more in sterilizing solutions treatment plots as comparing with TW treated plot during the time. The amount of total polyphenolics decrease with the time passing and EW and $ClO_2$ treatment shows less contents than TW treatment. Thus, EW and $ClO_2$ treatment of mulberris are considered as method to improve safety by reducing total plate count and to contribute to quality maintenance and to extend storage time.
Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.
Effects of heat and gamma irradiation on chemical, microbiological, and immunological changes of raw milk were compared. Free fatty acid content of milk showed increasing tendency according to the increase of heating temperature and irradiation dose, and showed similarity in UHT (ultra high temperature) and 5 kGy irradiation. Total bacterial counts and coliforms were not detected after treatment of LTLT (low temperature long time), HTST (high temperature short time), UHT, and irradiation from 1 to 10 kGy in the milk with initial microbial load at $10^3$ CFU/mL initially, but after 7 day storage, were not detected in UHT milk and that irradiated at 3 kGy or above. Heat treatment decreased (p<0.05) arginine, asparate, iso-leucine, lysine, and methionine content compared to raw milk while irradiation decreased (p<0.05) asparate, histidine, iso-luecine, leucine, and lysine content, which means irradiation could change primary structure of milk proteins. It was concluded that f kGy gamma irradiation treatment of raw milk could give a similar effect to UHT treatment in chemical and microbiological viewpoint, and may reduce allergenicity of raw milk.
Although bacterial outbreaks from ready-to-eat foods such as sprouts have increased, the information on microbial biocontrol by means of sanitizers is limited. Twenty sprouts of red cabbage, baby radish, alfalfa, and broccoli each were collected from the wholesale markets in Seoul. Ethanol and organic acids including acetic acid, citric acid, and lactic acid were used to control the amount of mesophilic bacteria and Bacillus cereus. Sanitizer mixtures of ethanol and organic acids showed a remarkable reduction of viable counts and, in particular, the sanitizer mixture comprised of 20% ethanol and 1% lactic acid seemed to be best by a reduction of 7~8 log CFU/g on the sprouts after a 10 minute exposure. At the same time, the sanitizer comprised of 20% ethanol and 1% lactic acid showed the same reduction for Escherichia coli, Salmonella typhimurium, B. cereus, Staphylococcus aureus, and Enterococcus faecalis, with more effect toward the Gram-negative bacteria. Sensory evaluation by texture, browning, off-flavour, and overall acceptability seemed better just after treatment when compared to one day after treatment with the sanitizer. Therefore, the sanitizers of ethanol-organic acid might be an effective means to control the bacterial contamination of sprouts in palce of sodium hypochloric acid.
Kim, E.T.;Min, K.S.;Kim, C.H.;Moon, Y.H.;Kim, S.C.;Lee, Sung-Sill
Asian-Australasian Journal of Animal Sciences
/
v.26
no.4
/
pp.517-522
/
2013
The effect on methanogens attached to the surface of rumen ciliate protozoa by the addition of plant extracts (pine needles and ginkgo leaves) was studied with particular reference to their effectiveness for decreasing methane emission. The plant extracts (pine needles and ginkgo leaves) were added to an in vitro fermentation incubated with rumen fluid. The microbial population including bacteria, ciliated-associated methanogen, four different groups of methanogens and Fibrobacter succinogenes were quantified by using the real-time PCR. Gas profiles including methane, carbon dioxide and hydrogen, and runinal fermentation characteristics were observed in vitro. The methane emission from samples with an addition of individual juices from pine needles, ginkgo leaves and 70% ethanol extract from ginko leaves was significantly lower (p<0.05, 27.1, 28.1 and 28.1 vs 34.0 ml/g DM) than that of the control, respectively. Total VFAs in samples with an addition of any of the plant extracts were significantly lower than that of the control (p<0.05) as well. The order Methanococcales and the order Methanosarcinales were not detected by using PCR in any incubated mixtures. The ciliate-associated methanogens population decreased from 25% to 49% in the plant extacts as compared to control. We speculate that the supplementation of juice from pine needles and ginkgo leaves extract (70% ethanol extract) decreased the protozoa population resulting in a reduction of methane emission in the rumen and thus inhibiting methanogenesis. The order Methanobacteriales community was affected by addition of all plant extracts and decreased to less than the control, while the order Methanomicrobiales population showed an increase to more than that of the control. The F. succinogenes, the major fibrolytic microorganism, population in all added plant extracts was increased to greater than that of the control. In conclusion, pine needles and ginkgo leaves extracts appear to have properties that decrease methanogenesis by inhibiting protozoa species and may have a potential for use as additives for ruminants.
A cultivation-based approach was employed to compare the culturable bacterial diversity associated with two phylogenetically closely related marine sponges, Spirastrella abata and Spirastrella panis, which have geologically overlapping distribution patterns. The bacteria associated with sponge were cultivated using MA medium supplemented with 3% sponge extracts. Community structures of the culturable bacteria of the two sponge species were analyzed with PCR-RFLP (restriction fragment length polymorphism) based on 16S rDNA sequences. The RFLP fingerprinting of 16S rDNA digested with HaeIII and MspI, revealed 24 independent RFLP types, in which 1-5 representative strains from each type were partially sequenced. The sequence analysis showed >98.4% similarity to known bacterial species in public databases. Overall, the microbial populations of two sponges investigated were found to be the members of the classes; Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria. The Alphaproteobacteria were predominant in the bacterial communities of the two sponges. Gammaproteobacteria represented 38.5% of bacterial community in S. abata. Whereas only 1.6% of this class was present in S. panis. Bacillus species were dominat in S. panis. Bacillus species were found to be 44.3% of bacterial species in S. panis, while they were only 9.7% in S. abata. It is interesting to note that Planococcus maritimus (8.1%, phylum Firmicutes) and Psychrobacter nivimaris (28.9%, phylum Gammaproteobacteria) were found only in S. abata. This result revealed that profiles of bacterial communities from the sponges with a close phylogenetic relationship were highly species-specific.
Staphylococcus aureus is a major human pathogen that produces a wide array of toxins, leading to a number of adverse symptoms. We examined 275 strains of Staphylococcus aureus isolated from various foods between 2006 and 2008 for antimicrobial susceptibility. At least 259 (94.2%) of the tested strains showed antibiotic resistant properties, and 106 (40.7%) of them showed multiple antibiotic resistance. Eleven of the tested strains were resistant to oxacillin and mec A-positive. Moreover, oxacillin-resistant strains were significantly more likely to be multi-drug resistant (p < 0.01). Of the 275 isolates tested, 24.4% were noted as being positive for slime production and 30.5% were positive for biofilm assay. Antibiotic resistance was not associated with a significantly higher prevalence of biofilm formation. Twenty strains were classified using the DiversiLab system. Most of the strains could be classified into 2 clusters and 4 unique types. All 10 mec A-positive strains (cluster I) were grouped together into the same sub-cluster. Cluster II (6 strains) was not found to be resistant to oxacillin in this study. Although the prevalence of methicillin-resistant S. aureus in food is currently low, the risk of its transmission through the food chain cannot be disregarded.
Candida biofilms are self-organized microbial communities growing on the surfaces of host tissues and medical devices. These biofilms have been displaying increasing resistance against conventional antifungal agents. The roots of Scutellaria baicalensis have been widely used for medicinal purpose throughout East Asia. The aim of the present study was to evaluate the effect of S. baicalensis aqueous extract upon the preformed biofilms of 10 clinical C. albicans isolates, and assess the mechanism of the antibiofilm activity. Its effect on preformed biofilm was judged using an XTT reduction assay and the metabolic activity of all tested strains were reduced ($57.7{\pm}17.3$%) at MIC values. The S. baicalenis extract inhibited (1, 3)-${\beta}$-D-glucan synthase activity. The effect of S. baicalensis on the morphology of C. albicans was related to the changes in growth caused by inhibiting glucan synthesis; most cells were round and swollen, and cell walls were densely stained or ruptured. The anticandidal activity was fungicidal, and the extract also arrested C. albicans cells at $G_0/G_1$. The data suggest that S. baicalensis has multiple fatal effects on target fungi, which ultimately result in cell wall disruption and killing by inhibiting (1, 3)-${\beta}$-D-glucan synthesis. Therefore, S. baicalensis holds great promise for use in treating and eliminating biofilm-associated Candida infections.
Lichens are symbiotic organisms composed of fungi, algae, or cyanobacteria which are able to survive in extreme environmental conditions ranging from deserts to polar areas. Some lichen symbionts produce a wide range of secondary metabolites that have many biological activities such as antibacterial, antifungal, antiviral, antitumor, antioxidant and anti-inflammatory etc. Among the symbionts of lichens, of the bacterial communities of lichen symbionts little is known. In this study, we isolated 4 microbial species from the Arctic lichen Stereocaulon spp. and evaluated their antioxidant properties using 1,1-diphenyl-2-picryl-hydrazyl assay as well as 2,2'-azino-bis(3-ethyl benzothiazoline-6-sulphonic acid) assay. Total phenolic contents and total flavonoid contents were also measured. A potent radical scavenging activity was detected in a number of the lichen extracts. Among the 4 species tested in this study, the ethyl acetate extract of Bosea vestrisii 36546(T) exhibited the strongest free radical scavenging activity, with an inhibition rate of 86.8% in DPPH and 75.2% in ABTS assays. Overall, these results suggest that lichen-bacteria could be a potential source of natural antioxidants.
In this study, we attempted to find new and efficient microbial enzymes for producing rare sugars. A ribose-5-phosphate isomerase B (OsRpiB) was cloned, overexpressed, and preliminarily purified successfully from a newly screened Ochrobactrum sp. CSL1, which could catalyze the isomerization reaction of rare sugars. A study of its substrate specificity showed that the cloned isomerase (OsRpiB) could effectively catalyze the conversion of $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose, which was unconventional for RpiB. The optimal reaction conditions ($50^{\circ}C$, pH 8.0, and 1 mM $Ca^{2+}$) were obtained to maximize the potential of OsRpiB in preparing $\text\tiny{L}$-rhamnulose. The catalytic properties of OsRpiB, including $K_m$, $k_{cat}$, and catalytic efficiency ($k_{cat}/K_m$), were determined as 43.47 mM, $129.4sec^{-1}$, and 2.98 mM/sec. The highest conversion rate of $\text\tiny{L}$-rhamnose under the optimized conditions by OsRpiB could reach 26% after 4.5 h. To the best of our knowledge, this is the first successful attempt of the novel biotransformation of $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose by OsRpiB biocatalysis.
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