• Title/Summary/Keyword: Agriculture and Forestry

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Expression of Codon Optimized β2-Adrenergic Receptor in Sf9 Insect Cells for Multianalyte Detection of β-Agonist Residues in Pork

  • Liu, Yuan;Wang, Jian;Liu, Yang;Yang, Liting;Zhu, Xuran;Wang, Wei;Zhang, Jiaxiao;Wei, Dong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1470-1477
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    • 2019
  • ${\beta}_2$-adrenergic receptor (${\beta}_2-AR$) was expressed efficiently using Bac-to-Bac Baculovirus Expression System in Sf9 cells as a bio-recognition element for multianalyte screening of ${\beta}$-agonist residues in pork. Sf9 cells were selected as the expression system, and codon optimization of wild-type nucleic acid sequence and time-dependent screening of expression conditions were then carried out for enhancing expression level and biological activity. Under optimum conditions of multiplicity of infection (MOI) = 5 and 48 h post transfection, the protein yield was up to 1.23 mg/ml. After purification by chromatographic techniques, the purified recombinant protein was applied to develop a direct competitive enzyme-linked receptor assay (ELRA) and the efficiency and reliability of the assay was determined. The IC50 values of clenbuterol, salbutamol, and ractopamine were 28.36, 50.70, and $59.57{\mu}g/l$, and clenbuterol showed 47.61% and 55.94% cross-reactivities with ractopamine and salbutamol, respectively. The limit of detection (LOD) was $3.2{\mu}g/l$ and the relevant recoveries in pork samples were in the range of 73.0-91.2%, 69.4-84.6%, and 63.7-80.2%, respectively. The results showed that it had better performance compared with other present nonradioactive receptorbased assays, indicating that the genetically modified ${\beta}_2-AR$ would have great application potential in detection of ${\beta}$-agonist residues.

Mass spectrometry-based ginsenoside profiling: Recent applications, limitations, and perspectives

  • Hyun Woo Kim;Dae Hyun Kim;Byeol Ryu;You Jin Chung;Kyungha Lee;Young Chang Kim;Jung Woo Lee;Dong Hwi Kim;Woojong Jang;Woohyeon Cho;Hyeonah Shim;Sang Hyun Sung;Tae-Jin Yang;Kyo Bin Kang
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.149-162
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    • 2024
  • Ginseng, the roots of Panax species, is an important medicinal herb used as a tonic. As ginsenosides are key bioactive components of ginseng, holistic chemical profiling of them has provided many insights into understanding ginseng. Mass spectrometry has been a major methodology for profiling, which has been applied to realize numerous goals in ginseng research, such as the discrimination of different species, geographical origins, and ages, and the monitoring of processing and biotransformation. This review summarizes the various applications of ginsenoside profiling in ginseng research over the last three decades that have contributed to expanding our understanding of ginseng. However, we also note that most of the studies overlooked a crucial factor that influences the levels of ginsenosides: genetic variation. To highlight the effects of genetic variation on the chemical contents, we present our results of untargeted and targeted ginsenoside profiling of different genotypes cultivated under identical conditions, in addition to data regarding genome-level genetic diversity. Additionally, we analyze the other limitations of previous studies, such as imperfect variable control, deficient metadata, and lack of additional effort to validate causation. We conclude that the values of ginsenoside profiling studies can be enhanced by overcoming such limitations, as well as by integrating with other -omics techniques.

Differential Frost Tolerance and Enzymatic Activities in the Leaves and Immature Fruits of Loquat (Eriobotrya japonica Lindl.)

  • Zheng, Guohua;Niu, Xianqian;Zhang, Jinbiao;Wu, Hanwen;Lin, Xiuxiang;Pan, Dongming
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.309-316
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    • 2015
  • In this experiment, five commercial cultivars and one wild species of loquat were used to investigate frost tolerance and enzymatic activities in leaves and young fruits under cold stress at $-3^{\circ}C$. The frost injury, malondialdehyde (MDA) content, and oxygen-scavenging enzyme activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were studied. This results showed that the wild species 'Wild Oak-leaf' loquat was the most frost tolerant among accessions tested, followed by the cultivar 'Golden Block'. Other cultivars, 'Wu Gong Bai', 'Taicheng 4', 'Xiangzhong 11' and 'Zaozhong 6', were relatively weak in frost tolerance. The enzymatic activities of SOD, POD and CAT increased initially and then decreased as the exposure time increased. However, the enzymatic peak occurred later in the frost-tolerant accession than in the frost-sensitive accession. The correlation coefficients of MDA contents between leaves and immature fruits were from 0.93 to 0.99 in the five commercial loquat cultivars. For the 'Wild Oak-leaf' loquat, the correlation coefficients of MDA and POD were 0.98 and 0.95, respectively, but the coefficients for SOD, CAT and APX were relatively low. In general, there were good correlations between loquat leaves and immature fruits in MDA content and enzyme activities. These results indicate that analysis of these physiological and biochemical activities in loquat leaves could potentially be used to predict the cold tolerance in loquat at immature fruit stage and to accelerate breeding programs for cold tolerance in loquat.

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping

  • Ali, Habib;Muhammad, Abrar;Hou, Youming
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.796-808
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    • 2018
  • The intracellular bacterium Wolbachia pipientis is widespread in arthropods. Recently, possibilities of novel Wolbachia-mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting Wolbachia surface protein (wsp) and multilocus sequence type (MLST) genotypic systems, we determined the Wolbachia infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of B. longissima were infected with the same Wolbachia strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles (ftsZ-234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of B. longissima and Wolbachia. The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the wsp and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the Wolbachia endosymbiont in B. longissima, which demonstrated that Wolbachia is ubiquitous across all developmental stages and distributed in the entire body of B. longissima. Understanding the Wolbachia infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.

Phage types of Salmonella enteritidis isolated from animal and human in Korea (한국의 동물과 사람에서 분리한 Salmonella enteritidis의 phage types)

  • Woo, Yong-ku;Park, Mi-sun;Woo, Seung-ryong;Kim, Bong-hwan;Kim, Jae-hak
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.515-524
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    • 2000
  • The present study was conducted to investigate the phage types(PT) of Salmonella enteritidis(SE) isolated from animals and human patients in Korea during the period 1993-1999. SE were isolated from chickens including the layer and broiler, duck, rat, swine and human patients in nationwide scale. A total of 231 non-human and 14 human SE strains were phage typed and 80.4%(197/245) of SE isolates tested could discriminate as a ten SEPTs including the RDNC by the 10 standard phages, which were brought from the Central Public Health Laboratory in England. In analysing of phage types both animal and human SE isolates, SEPT 4(55.4%) was the most prevalent in domestic animals including the layer(39.3%) and broiler(64.5%), duck(55.5%), and swine(75.0%) since 1993. About 57% of the S enteritidis from human was SEPT 4. Along with SEPT 4, SEPT 1, SEPT 6a, SEPT 7 and SEPT 7a var, SEPT 9b, SEPT 15, SEPT 21b and SEPT 22 were also found from animal and human SE isolates. SEPT 4(40.0%) was isolated from frozen chicken meats imported from China along with SEPT 7 and 7a var strains in this study. In conclusion, SEPT 4 was the most predominant SEPT in our country regardless of animal or human hosts including the Che-Ju island and almost exclusively found in domestic poultry(layer and broiler). During the review period, SEPT 4 has already been spreading continually among the animal hosts since 1993.

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Effect of Nonstarch Polysaccharide-Rich By-Product Diets on Nitrogen Excretion and Nitrogen Losses from Slurry of Growing-Finishing Pigs

  • Canh, T.T.;Verstegen, M.W.A.;Mui, N.B.;Aarnink, A.J.A.;Schrama, J.W.;Van't Klooster, C.E.;Duong, N.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.573-578
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    • 1999
  • An experiment was conducted to investigate the effect of diet for growing-finishing pigs with high level of non-starch polysaccharides (NSP) from by-products on nitrogen excretion and nitrogen losses from slurry during storage. Sixteen commercial crossbred barrows of about 68 kg BW were randomly allotted to one of four diets. The control diet was formulated using tapioca and rice as basal energy sources. In the other diets, tapioca was replaced by either coconut expellar, rice bran or beer by-product. The diets differed mainly in the amount and compostition of NSP. After a 12-day adaptation period, urine and faeces were collected separately in metabolism cages for 9 days. Urine and faeces from the first four days were used to analyse the nitrogen partitioning. Urine and faeces from the last 5 days were mixed as slurry. The slurry was sampled at the end of the collection period and again after 30 days storage, to analyse for nitrogen to calculate the losses. Increasing dietary NSP reduced urinary nitrogen and nitrogen losses from the slurry during storage. The pigs fed the diet based on beer by-product excreted the most nitrogen via faeces and the least nitrogen via urine. Nitrogen losses from slurry of pigs fed the beer by-product were from 34 to 65% lower than from the other three diets. It is concluded that including NSP-rich by-products in the diet of growing-finishing pigs reduces urinary nitrogen excretion and nitrogen losses from slurry during storage.

Collective Forest Management System in Japan: a Case Study in Osawa Property Ward Forest

  • De Zoysa, Mangala Premakumara;Inoue, Makoto;Yamashita, Utako;Hironori, Okuda
    • Journal of Forest and Environmental Science
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    • v.29 no.1
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    • pp.58-70
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    • 2013
  • Iriai an Indigenous forest management system in Japan from the viewpoint of "common pool resources" was a success resilient institution and resulted with sustainable production system and environmental conservation. This study was conducted in Osawa of the Nagano prefecture through group discussions, field observations and an in-depth field survey. Osawa Property Ward Forest is managed under the concept very much similarly to traditional "Iriai". This study firstly examined the changes of collective forest management system in terms of awareness and interest in forest management; forest management activities; role of forest; and collection of forest products. Then it analyzed the current threats for collective forest management have been identified as: land abandonment due to loss of benefits and lack of active community participation; deterioration of forest environment particularly the micro-climate and aesthetic values; conflict with local government authorities restraining the use of money in property ward forest and conflict with outsiders on damping of the garbage. Community cantered forestry management rules; livelihood contribution; protection of environment; local initiatives for protection and economic activities are the prevailing opportunities for collective forest management. The main requirements for revitalization of collective forest management are explained as local reciprocity; imposition of community based forest rules; encouraging local innovations; and building partnerships with stakeholders. Collective forest management system addresses the limitations of conventional forestry models, which had invalidated traditional 'iriai' institutions, and key to restoring sustainable use of forest and environmental resources. Cross-institutional collaborations together with responsibilities of local communities would ensure the revitalization of forest resources.

Characterization of a Recombinant Thermostable Xylanase from Hot Spring Thermophilic Geobacillus sp. TC-W7

  • Liu, Bin;Zhang, Ningning;Zhao, Chao;Lin, Baixue;Xie, Lianhui;Huang, Yifan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1388-1394
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    • 2012
  • A xylanase-producing thermophilic strain, Geobacillus sp. TC-W7, was isolated from a hot spring in Yongtai (Fuzhou, China). Subsequently, the xylanase gene that encoded 407 amino acids was cloned and expressed. The recombinant xylanase was purified by GST affinity chromatography and exhibited maximum activity at $75^{\circ}C$ and a pH of 8.2. The enzyme was active up to $95^{\circ}C$ and showed activity over a wide pH range of 5.2 to 10.2. Additionally, the recombinant xylanase showed high thermostability and pH stability. More than 85% of the enzyme's activity was retained after incubation at $70^{\circ}C$ for 90 min at a pH of 8.2. The activity of the recombinant xylanase was enhanced by treatment with 10 mM enzyme inhibitors (DDT, Tween-20, 2-Me, or TritonX-100) and was inhibited by EDTA or PMSF. Its functionality was stable in the presence of $Li^+$, $Na^+$, and $K^+$, but inhibited by $Hg^{2+}$, $Ni^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Pb^{2+}$, $Fe^{3+}$, and $Al^{3+}$. The functionality of the crude xylanase had similar properties to the recombinant xylanase except for when it was treated with $Al^{2+}$ or $Fe^{2+}$. The enzyme might be a promising candidate for various industrial applications such as the biofuel, food, and paper and pulp industries.