• Title/Summary/Keyword: 16S-rRNA

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Nicotinic acid changes rumen fermentation and apparent nutrient digestibility by regulating rumen microbiota in Xiangzhong black cattle

  • Zhuqing Yang;Linbin Bao;Wanming Song;Xianghui Zhao;Huan Liang;Mingjin Yu;Mingren Qu
    • Animal Bioscience
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    • v.37 no.2
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    • pp.240-252
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    • 2024
  • Objective: The aim of this study was to investigate the impact of dietary nicotinic acid (NA) on apparent nutrient digestibility, rumen fermentation, and rumen microbiota in uncastrated Xiangzhong black cattle. Methods: Twenty-one uncastrated Xiangzhong black cattle (385.08±15.20 kg) aged 1.5 years were randomly assigned to the control group (CL, 0 mg/kg NA in concentrate diet), NA1 group (800 mg/kg NA in concentrate diet) and NA2 group (1,200 mg/kg NA in concentrate diet). All animals were fed a 60% concentrate diet and 40% dried rice straw for a 120-day feeding experiment. Results: Supplemental NA not only enhanced the apparent nutrient digestibility of acid detergent fiber (p<0.01), but also elevated the rumen acetate and total volatile fatty acid concentrations (p<0.05). 16S rRNA gene sequencing analysis of rumen microbiota revealed that dietary NA changed the diversity of rumen microbiota (p<0.05) and the abundance of bacterial taxa in the rumen. The relative abundances of eight Erysipelotrichales taxa, five Ruminococcaceae taxa, and five Sphaerochaetales taxa were decreased by dietary NA (p<0.05). However, the relative abundances of two taxa belonging to Roseburia faecis were increased by supplemental 800 mg/kg NA, and the abundances of seven Prevotella taxa, three Paraprevotellaceae taxa, three Bifidobacteriaceae taxa, and two operational taxonomic units annotated to Fibrobacter succinogenes were increased by 1,200 mg/kg NA in diets. Furthermore, the correlation analysis found significant correlations between the concentrations of volatile fatty acids in the rumen and the abundances of bacterial taxa, especially Prevotella. Conclusion: The results from this study suggest that dietary NA plays an important role in regulating apparent digestibility of acid detergent fiber, acetate, total volatile fatty acid concentrations, and the composition of rumen microbiota.

Effects of supplemental bacteriophage on the gut microbiota and nutrient digestibility of ileal-cannulated pigs

  • Hyunwoong Jo;Geongoo Han;Eun Bae Kim;Changsu Kong;Beob Gyun Kim
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.340-352
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    • 2024
  • This study measured the potential changes of the microbiota in the gastrointestinal tract and energy and nutrient digestibility by supplemental bacteriophages in pigs. Twelve castrated male pigs (initial mean body weight = 29.5 ± 2.3 kg) were surgically cannulated using T-cannula. The animals were housed individually in pens equipped with a feeder and a nipple waterer. The pigs were allotted to 1 of 3 experimental diets in a quadruplicated 3 × 2 Latin square design with 3 experimental diets, 2 periods, and 12 pigs resulting in 8 replicates per diet. The 3 diets were a control mainly based on corn and soybean meal with no antibiotics or bacteriophages, a diet containing 0.1% antibiotics, and a diet containing 0.2% bacteriophages. On day 5 of the experimental period, feces were collected and on days 6 and 7, ileal digesta were collected. Genomic DNA for bacteria were extracted from the ileal digesta and feces and the V4 region of the 16S rRNA gene was amplified. The ileal and fecal digestibility of energy, dry matter, organic matter, crude protein, and fiber was unaffected by dietary antibiotics or bacteriophages. At the phylum level, the supplemental antibiotic or bacteriophage tended to result in a higher proportion of Firmicutes (p = 0.059) and a lower proportion of Bacteroidetes (p = 0.099) in the ileal digesta samples compared with the control group with no difference between the antibiotic and bacteriophage groups. At the genus level, the supplemental antibiotic or bacteriophage tended to result in a higher proportion of Lactobacillus (p = 0.062) and a lower proportion of Bacteroides (p = 0.074) and Streptococcus (p = 0.088) in the ileal digesta compared with the control group with no difference between the antibiotic and bacteriophage groups. In the feces, supplemental antibiotics or bacteriophages reduced the proportion of Bifidobacterium compared with the control group (p = 0.029) with no difference between the antibiotic and bacteriophage groups. Overall, supplemental antibiotics and bacteriophages showed positive effect on the microbiota of in the ileal digesta without largely affecting energy or nutrient digestibility, with no differences between the antibiotic and bacteriophage groups in growing pigs.

The Growth and EPA Synthesis of Shewanella oneidensis MR-1 and Expectation of EPA Biosynthetic Pathway

  • Jeong, Young-Su;Song, Sang-Kyu;Lee, Su-Jin;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.127-133
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    • 2006
  • Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles of S. oneidensis MR-1 under a variety of temperature conditions. The fastest growth of S. oneidensis MR-1 was observed at $30^{\circ}C$, with a specific growth rate and doubling time of $0.6885h^{-1}\;and\;1.007 h$. The maximum cell mass of this microorganism was elicited at a temperature of $4^{\circ}C$. The eicosapentaenoic acid (EPA) synthesis of S. oneidensis MR-1 was evaluated under these different culture temperatures. S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of $30^{\circ}C$, but was shown to synthesize EPA at temperatures below $30^{\circ}C$. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs between S. oneidensis MR-1 and Shewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to both Moritella sp., which is known to synthesize DHA via a PKS-like pathway, and S. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module of S. putrefaciens SCRC-2738 and the entire genome of S. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway in S. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding that S. oneidensis MR-1 was able to synthesize EPA without the expression of $dihomo-{\gamma}-linoleic$ acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

Isolation of Agarivorans sp. KC-1 and Characterization of Its Thermotolerant β-Agarase (한천분해세균 Agarivorans sp. KC-1의 분리 및 내열성 β-아가라제의 특성 규명)

  • Min, Kyung-Cheol;Lee, Chang-Eun;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1056-1061
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    • 2018
  • This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

An Analysis of the Antibiotic Resistance Genes of Multi-Drug Resistant (MDR) Acinetobacter baumannii (다제내성 Acinetobacter baumannii 의 항생제 내성 유전자 분석)

  • Lim, Jina;Lee, Gyusang;Choi, Yeonim;Kim, Jongbae
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.217-224
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    • 2016
  • Acinetobacter baumannii (A. baumannii) is prevalent in hospital environments and is an important opportunistic pathogen of nosocomial infection. It is known that this pathogen cause herd infection in hospitals, and the mortality rate is remarkably higher for patients infected with this pathogen and already have other underlying diseases. Herein, we investigated the antibiotic resistance rate and the type of resistance genes in 85 isolates of multi-drug resistant A. baumannii from the samples commissioned to laboratory medicine in two university hospitals-in hospital A and hospital B-located in Cheonan and Chungcheong provinces, respectively, in Korea. As a result, $bla_{OXA-23-like}$ and $bla_{OXA-51-like}$ were detected in 82 stains (96.5%). These 82 strains of $bla_{OXA-23-like}$ producing A. baumannii were confirmed with the ISAba1 gene found at the top of the $bla_{OXA-23-like}$ genes by PCR, inducing the resistance against carbapenemase. The armA, AME gene that induces the resistance against aminoglycoside was detected in 34 strains out of 38 strains from Hospital A (89.5%), and in 40 strains out of 47 strains from Hospital B (85.1%), while AMEs were found in 33 strains out of 38 strains from Hospital A (70.2%) and in 44 strains out of 47 strains in Hospital B (93.6%). Therefore, it was found that most multi-drug resistant A. baumannii from the Cheonan area expressed both acethyltransferase and adenyltransferase. This study investigated the multi-drug resistant A. baumannii isolated from Cheonan and Chungcheong provinces in Korea, and it is thought that the results of the study can be utilized as the basic information to cure multi-drug resistant A. baumannii infections and to prevent the spread of drug resistance.

Mitigation Effect of Drought Stress by Plant Growth-promoting Bacterium Bacillus sp. SB19 on Kale Seedlings in Greenhouse (식물생장촉진 Bacillus sp. SB19 균주의 케일 처리에 대한 가뭄 스트레스 완화 효과)

  • Kim, Dayeon;Lee, Sang-Yeob;Kim, Jung-Jun;Han, Ji-Hee
    • Korean Journal of Organic Agriculture
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    • v.24 no.4
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    • pp.833-847
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    • 2016
  • Drought stress is a major agricultural limitation to crop productivity worldwide, especially by which leafy vegetables, plant leaves eaten as vegetable, could be more lethal. The study was carried out to know the effect of drought tolerance plant growth promoting bacteria (PGPB) on water stress of kale seedlings. A total of 146 morphologically distinct bacterial colonies were isolated from bulk soil and rhizosphere soil of leafy vegetables and screened for plant growth promoting microbioassay in greenhouse. Out of them the isolate SB19 significantly promoted the growth of kale seedlings in increasement of about 42% of plant height (14.1 cm), 148% of leaf area ($19.0cm^2$) and 138% of shoot fresh weight (1662.5 mg) attained by the bacterially treated plants compared to distilled water treated control (9.9 cm, $7.7cm^2$, 698.8 mg). Shoot water content of SB19 treated kale seedlings (1393.8 mg) was also increased about 152% compared with control (552.5 mg). The SB19 isolated from bulk soil of kale plant in Iksan, Korea, was identified as species of Bacillus based on 16S rRNA gene sequencing analysis. We evaluated the effect of drought tolerance by the Bacillus sp. SB19 on kale seedlings at 7th and 14th days following the onset of the water stress and watering was only at 7th day in the middle of test. In the survey of 7th and 14th day, there were mitigation effect of drought stress in kale seedlings treated with $10^6$ and $10^7cell\;mL^{-1}$ of SB19 compared to distilled water treated control. Especially, there were more effective mitigation of drought damage in kale seedlings treated with $10^7cell\;mL^{-1}$ than $10^6cell\;mL^{-1}$. Further, although drought injury of bacterially treated kale seedlings were not improved at 14th day compared with 7th day, drought injury of $10^7cell\;mL^{-1}$ of SB19 treated kale seedlings were not happen rapidly but developed over a longer period of time than $10^6cell\;mL^{-1}$ of SB19 or control. The diffidence of results might be caused by the concentration of bacterial suspension. This study suggests that beneficial plant-microbe interaction could be a important role of enhancement of water availability and also provide a good method for improving quality of leafy vegetables under water stress conditions.

Isolation and biochemical characterization of acid tolerance xylanase producing Bacteria, Bacillus sp. GJY from city park soil (도심공원으로부터 산내성 xylanase를 생산하는 박테리아 분리 및 효소학적 특성)

  • Jang, Min-Young;Park, Hwa Rang;Lee, Chong Gyu;Choo, Gab-Chul;Cho, Hyun Seo;Park, Sam-Bong;Oh, Ki-Cheol;Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.60 no.1
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    • pp.79-86
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    • 2017
  • Microbes in forest are very important due to not only to enhance soil fertility but also maintain a healthy ecosystem by supplying the energy available to living organisms by producing various kinds of enzymes related to degradation of lignocellulosic biomass. In order to isolate a lignocellulosic biomass degrading bacterial strain from the Jurassic park located in Gyeongnam National University of Science and Technology, We used the Luria-Bertani-Carboxymethyl cellulose (CMC) agar trypan blue method containing 0.4 % carboxymethyl cellulose and 0.01 % trypan blue. As a result, we isolated a bacterial strain showing both activity on the CMC and xylan. To identify the isolated strain, 16S rRNA sequencing and API kit analysis were used. The isolated strain turned out to belong to Bacillus species and then named Bacillus sp. GJY. In the CMC zymogram analysis, it showed that one active band of about 28kDa in size is present. Xylan zymogram analysis also showed to have one active band of about 25kDa in size. The optimal growth temperature of Bacillus sp. GJY was $37^{\circ}C$. The maximal activities of CMCase and xylanase were 12 hour after incubation. The optimal pH and temperature for CMCase were 5.0 and $40^{\circ}C$, respectively, whereas the optimal pH and temperature for xylanase was 4.0 and $40^{\circ}C$. Both activities for CMCase and xylanase showed to be thermally stable at 40and $50^{\circ}C$, while both activities rapidly decreased at over $60^{\circ}C$.

Phylogentic Position, Pigment Content and Optimal Growth Condition of the Unicellular Hydrogen-Producing Cyanobacterial Strains from Korean Coasts (한국 연안산 단세포성 수소생산 남세균 종주들의 분류계통, 색소함량 및 최적성장 환경)

  • PARK, JONG-WOO;KIM, JU HEE;CHO, AE-RA;JUNG, YUN-DUK;KIM, PYOUNG JOONG;KIM, HYUNG-SEOP;YIH, WONHO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.3
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    • pp.131-140
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    • 2015
  • To set up unicellular cyanobacterial strains with photo-biological $H_2$ production potential, live samples were repeatedly collected from 68 stations in the coastal zone of Korea for the four years since 2005. Among 77 cyanobacterial strains established six (KNU strains, CB-MAL002, 026, 031, 054, 055 and 058) were finally chosen as the excellent strains for $H_2$ production with $H_2$ accumulation over 0.15 mL $H_2\;mL^{-1}$ under general basic $H_2$ production conditions as well as positive $H_2$ production for more than 60 hr. To explore optimum procedures for higher $H_2$ production efficiency of the six cyanobacterial strains, the inter-strain differences in the growth rate under the gradients of water temperature and salinity were investigated. The maximum daily growth rates of the six strains ranged from 1.78 to 2.08, and all of them exhibited $N_2-fixation$ ability. Based on the similarity of the 16S rRNA sequences, all the test strains were quite close to Cyanothece sp. ATCC51142 (99%). The six strains, however, were grouped into separate clades from strain ATCC51142 in the molecular phylogeny diagram. Chlorophyll- a content was 3.4~7.8% of the total dried weight, and the phycoerythrin and phycocyanin contents were half of those in the Atlantic strain, Synechococcus sp. Miami BG03511. The growth of the six strains was significantly suppressed at temperatures above the optimal range, $30{\sim}35^{\circ}C$, to be nearly stopped at $40^{\circ}C$. The growth was not inhibited by high salinities of 30 psu salinity in all the strains while strain CB055 maintained its high growth rate at low salinities down to 15 psu. The euryhaline strains like CB055 might support massive biotechnological cultivation systems using natural basal seawater in temperate latitudes. base seawater. The biological and ecophysiological characteristics of the test strains may contribute to designing the optimal procedures for photo-biological $H_2$ production by unicellular cyanobacteria.

Microbial Community Composition Associated with Anaerobic Oxidation of Methane in Gas Hydrate-Bearing Sediments in the Ulleung Basin, East Sea (동해 울릉분지 가스 하이드레이트 매장 지역의 메탄산화 미생물 군집 조성 및 분포)

  • Cho, Hyeyoun;Kim, Sung-Han;Shin, Kyung-Hoon;Bahk, Jang-Jun;Hyun, Jung-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.1
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    • pp.53-62
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    • 2015
  • To elucidate the microbial consortia responsible for the anaerobic methane oxidation in the methane hydrate bearing sediments, we compared the geochemical constituents of the sediment, the rate of sulfate reduction, and microbial biomass and diversity using an analysis of functional genes associated with the anaerobic methane oxidation and sulfate reduction between chimney site (UBGH2-3) on the continental slope and non-chimney site (UBGH2-10) on the basin of the Ulleung Basin. From the vertical profiles of geochemical constituents, sulfate and methane transition zone (SMTZ) was clearly defined between 0.5 and 1.5 mbsf (meters below seafloor) in the UBGH2-3, and between 6 and 7 mbsf at the UBGH2-10. At the UBGH2-3, the sulfate reduction rate (SRR) in the SMTZ exhibited was appeared to be $1.82nmol\;cm^{-3}d^{-1}$ at the depth of 1.15 mbsf. The SRR in the UBHG2-10 showed a highest value ($4.29nmol\;cm^{-3}d^{-1}$) at the SMTZ. The 16S rRNA gene copy numbers of total Prokaryotes, mcrA, (methyl coenzyme M reductase subunit A), and dsrA (dissimilatory sulfite reductase subunit A) showed the peaks in the SMTZ at both sites, but the maximum mcrA gene copy number of the UBGH2-10 appeared below the SMTZ (9.8 mbsf). ANME-1 was a predominant ANME (Anaerobic MEthanotroph) group in both SMTZs of the UBGH2-3 and -10. However, The sequences of ANME-2 were detected only at 2.2 mbsf of the UBGH2-3 where high methane flux was observed because of massive amount of gas hydrate at shallow depth. And Desulfosarcina-Desulfococcus (DSS) that is associated with ANME-2 was detected in 2.2 mbsf of the UBHG2-3. Overall results demonstrate that ANME-1 and ANME-2 are considered as significant archaeal groups related to methane cycle in the subsurface sediment of the East Sea, and ANME-2/DSS consortia might be more responsible for methane oxidation in the methane seeping region than in non-seeping region.

Biodegradation of Phenol by Comamonas testosteroni DWB-1-8 Isolated from the Activated Sludge of Textile Wastewater (섬유 폐수 활성 슬러지에서 분리한 Comamonas testosteroni의 생물학적 페놀 분해)

  • Kwon, Hae Jun;Choi, Doo Ho;Kim, Mi Gyeong;Kim, Dong-Hyun;Kim, Young Guk;Yoon, Hyeokjun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.30 no.2
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    • pp.156-161
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    • 2020
  • Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K2HPO4, 0.6% NaH2PO4, 0.1% NH4NO3, 0.015% MgSO4·7H2O, 0.001% FeSO4·7H2O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.