• Title/Summary/Keyword: Korean indigenous species

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Screening of Biodegradable Function of Indigenous Ligno-degrading Mushroom Using Dyes

  • Jang, Kab-Yeul;Cho, Soo-Muk;Seok, Soon-Ja;Kong, Won-Sik;Kim, Gyu-Hyun;Sung, Jae-Mo
    • Mycobiology
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    • v.37 no.1
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    • pp.53-61
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    • 2009
  • The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).

Characteristics and Frequencies of Alternative Initiation Codon(AIC) of mtDNA ND2 in Five Pig Breeds (돼지 5품종에 있어서 mtDNA ND2 유전자의 선택적 개시코돈의 특성과 빈도)

  • Han, S.H.;Cho, I.C;Choi, Y.L.;Lee, C.E.;Ko, M,S.;Kim, J.H.;Seo, B.Y.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.903-908
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    • 2004
  • Alternative initiation codon(AlC) has been reported in the mitochondrial genes in various mammalian species. We investigated the AlC of mitochondrial NADH dehydrogenase 2 gene(rntDNA ND2) in five pig breeds. Two kinds of initiation codons(ATA/ATf) showing different frequencies among tested pig breeds were used. While all Large White pigs had ATA as an initiator methionine codon, all Landrace pigs had ATf. The other breeds(Berkshire, Duroc and Hampshire) had both initiation codons with the ATA frequencies, 91.9, 21.3 and 60.00/0, respectively. In the previous reports, all Chinese indigenous pig breeds were identified to have unique initiation codon ATA. Although the effect of Ale on the translation of mtDNA ND2 has not been studied in this study, AlC patterns in mtDNA ND2 will contribute to the maternity test using molecular markers in pig breeding.

Clean-up of the Crude Oil Contaminated Marine Sediments Through Biocarrier-Mediated Bioaugmentation (생물담체 활용 생물접종에 의한 원유로 오염된 해양토양의 정화)

  • Ekpeghere, Kelvin I.;Bae, Hwan-Jin;Kwon, Sung-Hyun;Kim, Byung-Hyuk;Park, Duck-Ja;Kim, Hee-Shik;Koh, Sung-Cheol
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.354-361
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    • 2009
  • This study was carried out to develop an effective biocarrier-mediated bioaugmentation technology which will be useful for remediation of the crude oil-contaminated marine sediments. Enrichment of several microbial communities was made from several oil-polluted seashore sites and the two distinctively functional consortia have been successfully selected. These two consortia were grown together and used to manufacture the microbial agents for bioaugmentation of marine sediments polluted with crude oil. The most dominant species in the mixed culture was identified as Alcanivorax borkumensis based on pure culture and DGGE analysis. Bioaugmentation of oil-polluted marine sediments with the microbial agent MA-2 formulated using the mixed culture and biocarriers (activated carbon and minerals) was more effective, especially in combination with an oxygen producing (releasing) compound (ORC). Ninty percent of TPH was removed in the presence of ORC in 35 days while 74% in the absence of ORC. This indicated that the indigenous consortial degraders could be immobilized on the active carbon as a biocarrier to manufacture microbial agents and then effectively bioaugmented for remediation of the oil-polluted sediments.

Investigation of genetic variability in commercial and invaded natural populations of red swamp crayfish(Procambarus clarkii) from South Korea (미국가재(Procambarus clarkii) 수족관 개체군 및 국내 침입 자연개체군의 유전적 변이 연구)

  • Ji Hyoun Kang;Jeong Mi Hwang;Soon-Jik Kwon;Min Jeong Baek;Sun-Jae Park;Changseob Lim;Yeon Jae Bae
    • Korean Journal of Environmental Biology
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    • v.41 no.3
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    • pp.325-334
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    • 2023
  • The invasive red swamp crayfish, Procambarus clarkii, is native to south-central United States and northeastern Mexico. Recently, it has been being spreading in the wild in South Korea. However, its primary sources, introduction routes, establishment, and expansion in South Korea remain unclear. Here, we analyzed genetic diversity and population genetic structures of its domestic natural populations during early invasion, commercial stock from local aquaria (a suspected introduction source), and original United States population using mitochondrial COI gene sequences for 267 individuals and eight microsatellite markers for 158 individuals. Natural and commercial populations of P. clarkii showed reduced genetic diversity (e.g., haplotype diversity and allelic richness). The highest genetic diversity was observed in one original source population based on both genetic markers. Despite a large number of individuals in commercial aquaria, we detected remarkably low genetic diversity and only three haplotypes among 226 individuals, suggesting an inbred population likely originating from a small founder group. Additionally, the low genetic diversity in the natural population indicates a small effective population size during early establishment of P. clarkii in South Korea. Interestingly, genetic differentiation between natural populations and the United States population was lower than that between natural populations and aquarium populations. This suggests that various genetic types from the United States likely have entered different domestic aquariums, leading to distinct natural populations through separate pathways. Results of our study will provide an insight on the level of genetic divergence and population differentiation during the initial stage of invasion of non-indigenous species into new environments.

A study on Restoration Plan of Cultural Forest and Change of $Pinus$ $densiflora$ Forest in Inwangsan(Mt.), Seoul (인왕산 소나무림의 경관변화와 문화경관림 복원방안 연구)

  • Cho, Jun-Soo;Lee, Kyong-Jae;Han, Bong-Ho;Ki, Kyong-Seok
    • Korean Journal of Environment and Ecology
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    • v.26 no.2
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    • pp.219-232
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    • 2012
  • The purpose of this study was to find characteristics of $Pinus$ $densiflora$ forests in previous records and cultural landscape regions and to establish a management method for recovery of $P.$ $densiflora$ landscape. A total of 16 landscape paintings were analyzed to study $P.$ $densiflora$ forests in Inwangsan(Mt.) shown in figure data. As a result of analyzing figure data, $P.$ $densiflora$ $forests$ were found to be distributed mainly in the ridge line of east slope side of Inwangsan(Mt.) from landscape paintings of Jung Seon, Jang Si Heung and Gang Hee Un. In order to analyze the landscape of $P.$ $densiflora$ forests shown in photograph data on Inwangsan(Mt.), photographs of Inwangsan(Mt.) since late 1800s were searched and a total of 24 photographs were used for analysis. As a result of photographic analysis, $P.$ $densiflora$ forests were found to be distributed mainly in the low area and ridge line of western slope side of Inwangsan(Mt.). As a result of analyzing the distribution status of 10 cultural heritages of Inwangsan(Mt.), cultural heritages in the mountain were mainly established with rocks and $P.$ $densiflora$ forests as the background and are establishing their landscape not only as a form of cultural heritage but also including the natural landscape in the background. A method of restoring cultural landscape of P. densiflora forests was necessary. As a result of analyzing 26 rock beds of Inwangsan(Mt.), most of $P.$ $densiflora$ forests were distributed nearby rock beds since the past. Among such regions, restoration and management of introduced species in the accumulated soil regions to $P.$ $densiflora$ forests were deemed necessary. As a result of analyzing current vegetation of $P.$ $densiflora$ forest areas of the past, 51.87% of areas maintained $P.$ $densiflora$ forest and 25.80% were changed to artificial forests. Most of low areas of the mountain were turned into urban area (10.16%). Management type for restoration of $P.$ $densiflora$ landscape of Inwangsan(Mt.) was classified into preservation, $P.$ $densiflora$ restoration and maintenance. Preservation management was configured as $P.$ $densiflora$ distribution and rocky area distribution. The targets of $P.$ $densiflora$ restoration were indigenous broad-leaf trees, vegetation areas with introduced species and damaged regions within the mountain. The targets of maintenance were urban area in the low area of Inwangsan(Mt.) and military base at the top of the mountain.

Optimizing In Vitro Propagation of Sophora koreensis Nakai using Statistical Analysis (다양한 통계분석 기법을 이용한 개느삼(Sophora koreensis Nakai)의 기내 증식 최적 조건 구명)

  • Jeong, Ukhan;Lee, Hwa;Park, Sanghee;Cheong, Eun Ju
    • Journal of Korean Society of Forest Science
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    • v.110 no.1
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    • pp.53-63
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    • 2021
  • Sophora koreensis Nakai is an indigenous plant in Koreawith a restricted natural range, part of which is in Gangwon province. The species is known to contain phytochemicals that have beneficial effects on human health, and it is economically important in bioindustry. Because of the limited number of plants in a small range of habitats, the mass-propagation method should be developed for use and conservation. In vitro tissue culture is a reliable method in terms of mass propagation from selected clones of the species. We investigated the optimal conditions of the medium in this process, especially focusing on the concentrations of plant growth regulators(PGRs) in the culture of stem-containing axillary buds. Three statistical methods, i.e., ANOVA, response surface method(RSM), and fuzzy clustering were used to analyze the plant growth, number of shoots induced, and shoot length with various combinations of PGRs. Results from the RSM differed from those of the other two methods; thus, the method was not suitable. ANOVA and fuzzy clustering showed similar results. However, more accurate results were obtained using fuzzy clustering because it provided a probability for each treatment. On the basis of the fuzzy clustering analysis, stem tissue produced the greatest number of shoots(11.03 per explant; 63.33%) on a medium supplemented with 5-��M 6-benzylaminopurine and 2.5-��M thidiazuron(TDZ). Proliferation of shoots(2.18 ± 0.21 cm, 63.33%) was attained on a medium supplemented with 2.5-��M BA, 2.5-��M TDZ, and 2.5-��M gibberellic acid.

An Investigation of Local Naming Issue of Tamarix aphylla (에셀나무(Tamarix aphylla)의 명칭문제에 대한 고찰)

  • Kim, Young-Sook
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.37 no.1
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    • pp.56-67
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    • 2019
  • In order to investigate the issue with the proper name of eshel(Tamarix aphylla) mentioned in the Bible, analysis of morphological taxonomy features of plants, studies on the symbolism of the Tamarix genus, analysis of examples in Korean classics and Chinese classics, and studies on the problems found in translations of Korean, Chinese and Japanese Bibles. The results are as follows. According to plant taxonomy, similar species of the Tamarix genus are differentiated by the leaf and flower, and because the size is very small about 2-4mm, it is difficult to differentiate by the naked eye. However, T. aphylla found in the plains of Israel and T. chinensis of China and Korea have distinctive differences in terms of the shape of the branch that droops and its blooming period. The Tamarix genus is a very precious tree that was planted in royal courtyards of ancient Mesopotamia and the Han(漢) Dynasty of China, and in ancient Egypt, it was said to be a tree that gave life to the dead. In the Bible, it was used as a sign of the covenant that God was with Abraham, and it also symbolized the prophet Samuel and the court of Samuel. When examining the example in Korean classics, the Tamarix genus was used as a common term in the Joseon Dynasty and it was often used as the medical term '$Ch{\bar{e}}ngli{\check{u}}$(檉柳)'. Meanwhile, the term 'wiseonglyu(渭城柳)' was used as a literary term. Upon researching the period and name of literature related to $Ch{\bar{e}}ngli{\check{u}}$(檉柳) among Chinese medicinal herb books, a total of 16 terms were used and among these terms, the term Chuísīliǔ(垂絲柳) used in the Chinese Bible cannot be found. There was no word called 'wiseonglyu(渭城柳)' that originated from the poem by Wang Wei(699-759) of Tang(唐) Dynasty and in fact, the word 'halyu(河柳)' that was related to Zhou(周) China. But when investigating the academic terms of China currently used, the words Chuísīliǔ(垂絲柳) and $Ch{\bar{e}}ngli{\check{u}}$(檉柳) are used equally, and therefore, it appears that the translation of eshel in the Chinese Bible as either Chuísīliǔ (垂絲柳) or $Ch{\bar{e}}ngli{\check{u}}$(檉柳) both appear to be of no issue. There were errors translating tamarix into 'やなぎ(willow)' in the Meiji Testaments(舊新約全書 1887), and translated correctly 'ぎょりゅう(檉柳)' since the Colloquial Japanese Bible(口語譯 聖書 1955). However, there are claims that 'gyoryu(ぎょりゅう 檉柳)' is not an indigenous species but an exotics species in the Edo Period, so it is necessary to reconsider the terminology. As apparent in the Korean classics examples analysis, there is high possibility that Korea's T. chinensis were grown in the Korean Peninsula for medicinal and gardening purposes. Therefore, the use of the medicinal term $Ch{\bar{e}}ngli{\check{u}}$(檉柳) or literary term 'wiseonglyu' in the Korean Bible may not be a big issue. However, the term 'wiseonglyu' is used very rarely even in China and as this may be connected to the admiration of China and Chinese things by literary persons of the Joseon Dynasty, so the use of this term should be reviewed carefully. Therefore, rather than using terms that may be of issue in the Bible, it is more feasible to transliterate the Hebrew word and call it eshel.

Characteristics of Bacterial Community for Biological Activated Carbon(BAC) by Culturable and Unculturable Methods. (배양적 및 비배양적 방법에 의한 생물활성탄 부착세균 군집 특성)

  • Park, Hong-Ki;Jung, Eun-Young;Jung, Mi-Eun;Jung, Jong-Moon;Ji, Ki-Won;Yu, Pyung-Jong
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1284-1289
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    • 2007
  • The Biological Activated Carbon (BAC) process in the water treatments represents a kind of biofiltration process which capabilities of bacteria to remove organic matters are maximized. It enables to eliminate organic matters and effectively reduce microbial regrowth potentials. As attached bacteria employ natural organic matter as a substrate, they are significantly dependent on indigenous microorganisms. In this study, characteristics of bacterial community by culturable and unculturable Methods have been conducted in a pilot plant using SAC in water treatment process at the downstream of the Nakdong River. Based on the results, HPC and bacterial- production for coal-based activated carbon material were $1.20{\sim}56.2{\times}l0^7$ cfu/g and $1.2{\sim}3.7\;mgC/m^{3}h$, respectively, in the SAC process. The highest level of attached bacteria biomass and organic carbon removal efficiency was found in the coal-based activated carbon. The genera Pseudomonas, Flavobacterium, Alcaligenes, Acilzetobacter, and Spingomonas were identified for each activated carbon material. Pseudomonas vesicularis was the dominant species in the coconut- and coal-based materials, where as Pseudomonas cepacia was the dominant species in the wood-based material. The Scanning Electron Microscope (SEM) observation of the activated carbon surface also found the widespread distribution of rod form and coccus. The community of attached bacteria was investigated by performing Fluorescent in situ hybridization (FISH) analysis. a group was dominant in coal, wood and coccunt-based materials, ${\alpha},\;{\beta}\;and\;{\gamma}$ group ranged from 27.0 ${\sim}$ 43.0%, 7.1 ${\sim}$ 22.0%, 11.3 ${\sim}$ 28.6%, respectively. These results suggest that a group bacterial community appears to be regulated removal efficiency of organic material in water treatment process.

Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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Current Status of Cattle Genome Sequencing and Analysis using Next Generation Sequencing (차세대유전체해독 기법을 이용한 소 유전체 해독 연구현황)

  • Choi, Jung-Woo;Chai, Han-Ha;Yu, Dayeong;Lee, Kyung-Tai;Cho, Yong-Min;Lim, Dajeong
    • Journal of Life Science
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    • v.25 no.3
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    • pp.349-356
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    • 2015
  • Thanks to recent advances in next-generation sequencing (NGS) technology, diverse livestock species have been dissected at the genome-wide sequence level. As for cattle, there are currently four Korean indigenous breeds registered with the Domestic Animal Diversity Information System of the Food and Agricultural Organization of the United Nations: Hanwoo, Chikso, Heugu, and Jeju Heugu. These native genetic resources were recently whole-genome resequenced using various NGS technologies, providing enormous single nucleotide polymorphism information across the genomes. The NGS application further provided biological such that Korean native cattle are genetically distant from some cattle breeds of European origins. In addition, the NGS technology was successfully applied to detect structural variations, particularly copy number variations that were usually difficult to identify at the genome-wide level with reasonable accuracy. Despite the success, those recent studies also showed an inherent limitation in sequencing only a representative individual of each breed. To elucidate the biological implications of the sequenced data, further confirmatory studies should be followed by sequencing or validating the population of each breed. Because NGS sequencing prices have consistently dropped, various population genomic theories can now be applied to the sequencing data obtained from the population of each breed of interest. There are still few such population studies available for the Korean native cattle breeds, but this situation will soon be improved with the recent initiative for NGS sequencing of diverse native livestock resources, including the Korean native cattle breeds.