Human enteric Adenovirus 41 (HueAdV-41) is a major waterborne virus that causes human gastroenteritis and is classified as a viral group I double-strand DNA virus, Adenoviridae. HueAdV-41 has been detected with the polymerase chain reaction (PCR) in various samples such as ground water. However, the PCR-based diagnostic method has problems such as reaction time, sensitivity, and specificity. Thus, the loop-mediated isothermal amplification (LAMP) assay has emerged as an excellent method for field applications. In this study, we developed a LAMP system that can rapidly detect HueAdV-41 with high specificity and sensitivity. HueAdV-41 specific LAMP primer sets were tested through a specific, non-specific selection and sensitivity test for three prepared LAMP primer sets, of which only one primer set and optimum reaction temperature were selected. The developed LAMP primer set condition was confirmed as 63℃, and the sensitivity was 1 copy. In addition, to confirm the system, a LAMP positive reaction was developed with the restriction enzyme Taq I (T/GCC). The developed method in this study was more specific, rapid (typically within 2 - 3 hours), and highly sensitive than that of the conventional PCR method. To evaluate and verify the developed LAMP assay, an artificial infection test was done with five cDNAs from groundwater samples, and the results were compared to those of the conventional PCR method. We expect the developed LAMP primer set will be used to diagnose HueAdV-41 from various samples.
Effects of oral administration with fermented product from sewage in land-based seawater fish farm on haematological disturbance in the olive flounder, Paralichthys olivaceus was investigated. After 4 weeks of conditioning with a basal diet, fish were divided into 4 groups and provided experimental diet (0.1, 0.5, 1.0 and 2.0%) supplement of fermented sewage for 80 days. Proximal analysis was performed for the product of sewage which was fermented by lactic acid and yeast. RBC count, hemoglobin concentration and hematocrit value were increased according to the treated periods, however, no statistical difference was observed between control and treatment groups. There were no significant difference in serum organic, inorganic compounds and enzyme activities between control and treatment groups. This study hypothesized that the supplement of fermented product from sewage in land-based seawater fish farm might be an additive supplement for source of fish diet in view of haematological examination. Recycling of the sewage may be an economic artificial sources of diet for fish aquaculture practices.
Hyeonjin Park;Jin-Kyung Cha;So-Myeong Lee;Youngho Kwon;Jisu Choi;Ki-Won Oh;Jong-Hee Lee
KOREAN JOURNAL OF CROP SCIENCE
/
v.68
no.3
/
pp.188-196
/
2023
Wheat (Triticum aestivum L.) is an important crop in Korea, with a per capita consumption of 31.6 kg in 2019. In the southern region, wheat is grown after paddy rice, and it is harvested during the rainy season in mid-June. This timing, in combination with high humidity and untimely rainfall, activates the enzyme alpha-amylase, which breaks down starch in the wheat grains. As a result, sprouted grains have lower quality and value for flour. However, seeds that absorb water before sprouting are expected to maintain better quality. The aim of the study was to identify the critical period during wheat maturation when rainfall has the greatest impact on grain quality, to prevent price declines due to quality deterioration. Two wheat cultivars, Jokyoung and Hwanggeumal, were grown in a speed breeding room, and artificial rainfall was applied at different times after heading (30, 35, 40, 45, 50, and 55 days). The proportion of vitreous grains decreased from 40 to 55 days after heading (DAH). Both cultivars had chalky grain sections from 35 DAH, with Hwanggeumal having a higher proportion of vitreous grains. Starch degradation was observed using FE-SEM (Field Emission Scanning Electron Microscope) at 40 DAH for Jokyoung and 50 DAH for Hwanggeumal. Color measurements indicated increased L and E values from 40 DAH, with rain treatment at 55 DAH leading to a significant increase in L values for both cultivars. Ash content increased at 45 DAH, whereas SDSS decreased at 35 DAH. Overall, grain quality from 40 DAH until harvest was found to be affected to the greatest extent by direct exposure of the spikes to moisture. Red wheat showed better quality than white wheat. These findings have implications for the cultivation of high-quality wheat and can guide future research efforts in this area.
Ji, Seung-Cheol;Takaoka, Osamu;Takii, Kenji;Jeong, Gwan-Sik;Han, Seock-Jung
Journal of Aquaculture
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v.21
no.4
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pp.272-277
/
2008
We investigated the growth and survival in seedling production, and growth performance was compared with the different rearing tanks and protein source of formulated feed for juvenile Pacific bluefin tuna Thunnus orientalis (PBT). The survival rate at the end of nursery culture at 30 days after hatching was $0.69{\pm}0.40%$, and total length and mean body weight were $49.83{\pm}2.52\;mm$ and $1.03{\pm}0.09\;g$, respectively. Growth performance has no significant difference in fish reared by different tanks forms for 10 days. In order to develop an artificial diet, we evaluated the dietary utility of enzyme treated fish meal (Bio-CP, BIO) for juvenile PBT. Only diet BIO sustained similar growth and higher feed efficiency, and final carcass lipid content as compared to those of Sand lance (SL) These results revealed that BIO-CP a suitable dietary protein source, could sustain growth of PBT.
For selecting Bacillus strains producing high-quality Cheonggukjang, 8 strains were isolated from the different Cheonggukjang samples. Seven of them exhibited the highest 16S rRNA gene sequence similarity value of over 99.9% to Bacillus subtilis subsp. subtilis and one of them showed the similarity to B. licheniformis. All the strains showed positive activities for amylase, cellulase, protease and lipase, and 6 strains are positive for fibrinolytic activity. To confirm the safety of the strains isolated from the samples of Cheonggukjang which are manufactured by traditional method, strains were analyzed for the presence of seven toxin genes of Bacillus cereus and results were found negative. And 7 strains did not produce at all or merely produce both histamine and tyramine, the representative biogenic amines. Biogenic amine degradation analysis by HPLC revealed that, most of them exhibited tyramine degradation activity. For Cheonggukjang fermented by artificial inoculation of selected strains, fermentation property, sensory test, volatile basic nitrogen production and metabolic profiles by $^1H-NMR$ were tested. Seven strains were confirmed to make high-quality Cheonggukjang.
For the quality enhancement of harvested-year leaf tobacco to the quality of 2-year naturally aged leaf tobacco, cellulose and nicotine degradative bacteria were isolated and identified. Effects of artificial fermentation treated cellulase and nicotine degradative bacteria on the quality of leaf tobacco were investigated from the chemical and sensory points of view. 1, Changes in chemical composition of leaf tobacco resulted from the addition of cellulase extracted from Cellulomonas sp. [3ml(${\mu}{\textrm}{m}$ D-glucose/ml. mil-1) of enzymes solution 11009 of leaf tobacco] and nicotine degradative bacteria, Pseudomonas sp. 2ml(IX109 cells$\div$ 100g of leaf tobacco), and subsequently fermented at 40${\mu}{\textrm}{m}$$^{\circ}C$, 65% R. H. for 40 days are as follows : 1) Content of crude fiber decreased 12% It took 9 min, 53 sec. to reach full combustion in control group but took only 7 min. 47 sec. in the treated group, taking almost equal time to 2-year naturally aged leaf tobacco(7 min. 35sec.). 2) Light intensity of control group was 60.96% with bright lemon color but that of treated leaf tobacco accounted for 47.69 with orange to dark brown color series, which was almost equal to the value, 45.69, of 2-year naturally aged leaf tobacco. 3) Linoleic acid, serving mild taste among organic acids, amounted to 1.llmg/g in control group but increased to 1.35m9/9 in the treated leaf tobacco, identical to the content(1.35mg/g) of 2-year naturally aged leaf tobacco. 4) Content of solanone, on of the typical leaf tobacco flavor compounds, accounted for 2.95% in control group but increased to 2.87% in treated group. 5) Methyl furan, useful flavor compound in smoke composition, accounted for 17.6$\mu\textrm{g}$/cig. in control group but increased to 25.9$\mu\textrm{g}$/cig. in treated group. However, acroleine decreased from 69.3$\mu\textrm{g}$/cig. in control group to 58.6$\mu\textrm{g}$/cig. in treated group 2. In sonsory test, mild taste evaluation of control group scored 5.47 and treated group 7.93 which was evaluted almost equal to the value(8.00) of 2-year naturally aged leaf tobacco. Aroma evaluation of control group scored 5.60, treated group 8.20, and 2-year naturally aged leaf tobacco 8.33. In addition, total harmony taste of control group showed 5.67, treated group 8.07 (p<0.01), and 2-year naturally aged leaf tobacco 8.00. From these results, it can be said that quality of treated leaf tobacco is not inferior to that 2-year naturally aged leaf tobacco.
The study was carried out to investigate the variations in the fibroin hydrolysis of different cocoon shell layers, and the relationship between the hydrolysis and silk physical property. The obtained results are as follows: 1. The fibroin hydrolyzing ratio was highest at the inner layer of cocoon shell and. next in order. at the middle layer and at the outer layer. 2. The fibroin hydrolyzing ratio of abnormal cocoons was higher than normal cocoon and it was different among them, tile highest was double coroon, thin shell cocoon and perforated cocoon in order. 3. The sericin content and fibroin hydrolyzing ratio of Jam 111 and Jam 112 was higher than that of Jam 111 $\times$ Jam 112. 4. The fibroin hydroiyzing ratio of the cocoons fed with the artificial diets was increased at the inner layer. The sericin content of those lessened at the inner layer, however, it slightly increased at the most inner layer more than at the inner layer. 5. The breaking strength of the degummed silk fiber of different cocoon layers was reduced at the inner layer. The breaking Strength of abnormal silk fiber was less than that of normal silk fiber. 6. A negative correlation (r= -0.8) )was approved between fibroin hydrolyzing ratio and breaking strength of silk fiber, and the regression line was Y=-0.29x+5.07.
Paenibacillus polymyxa is a plant growth-promoting rhizobacterium (PGPR) which can be used for biological control of plant diseases. Several bacterial strains were isolated from rotten roots of Korean ginseng (Panax ginseng C. A. Meyer) that were in storage. These strains were identified as P. polymyxa, based on a RAPD analysis using a P. polymyxa-specific primer, cultural and physiological characteristics, an analysis utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME), and the 16S rDNA sequence analysis. These strains were found to cause the rot in stored ginseng roots. Twenty-six P. polymyxa strains, including twenty GBR strains, were phylogenetically classified into two groups according to the ERIC and BOX-PCR analyses and 16S rDNA sequencing, and the resulting groupings systematized to the degrees of virulence of each strain in causing root rot. In particular, highly virulent GBR strains clustered together, and this group may be considered as subspecies or biovar. The virulence of the strains seemed to be related to their starch hydrolysis enzyme activity, but not their cellulase or hemicellulase activity, since strains with reduced or no starch-hydrolytic activity showed little or no virulence. Artificial inoculation of the highly virulent strain GBR-1 onto the root surfaces of Korean ginseng resulted in small brown lesions which were sunken and confined to the outer portion of the root. Ginseng root discs inoculated in vitro or two-year-old roots grown in soil drenched with the inoculum developed significant rot only when the inoculum density was $10^{6}-10^{7}$ or more colony-forming units (CFU) per ml. These results suggest that P. polymyxa might induce ginseng root rot if their population levels are high. Based on these results, it is recommended that the concentration of P. polymyxa should be monitored, when it is used as a biocontrol agent of ginseng, especially in the treatment of stored roots.
Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.
Purpose: This study aimed to evaluate the biocompatibility and the mechanical properties of ultraviolet (UV) cross-linked and biphasic calcium phosphate (BCP)-added collagen membranes and to compare the clinical results of ridge preservation to those obtained using chemically cross-linked collagen membranes. Methods: The study comprised an in vitro test and a clinical trial for membrane evaluation. BCP-added collagen membranes with UV cross-linking were prepared. In the in vitro test, scanning electron microscopy, a collagenase assay, and a tensile strength test were performed. The clinical trial involved 14 patients undergoing a ridge preservation procedure. All participants were randomly divided into the test group, which received UV cross-linked membranes (n=7), and the control group, which received chemically cross-linked membranes (n=7). BCP bone substitutes were used for both the test group and the control group. Cone-beam computed tomography (CBCT) scans were performed and alginate impressions were taken 1 week and 3 months after surgery. The casts were scanned via an optical scanner to measure the volumetric changes. The results were analyzed using the nonparametric Mann-Whitney U test. Results: The fastest degradation rate was found in the collagen membranes without the addition of BCP. The highest enzyme resistance and the highest tensile strength were found when the collagen-to-BCP ratio was 1:1. There was no significant difference in dimensional changes in the 3-dimensional modeling or CBCT scans between the test and control groups in the clinical trial (P>0.05). Conclusions: The addition of BCP and UV cross-linking improved the biocompatibility and the mechanical strength of the membranes. Within the limits of the clinical trial, the sites grafted using BCP in combination with UV cross-linked and BCP-added collagen membranes (test group) did not show any statistically significant difference in terms of dimensional change compared with the control group.
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