• Title/Summary/Keyword: rDNA sequencing

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Screening of BCL-2 associated X protein gene polymorphism associated with scrotal hernia in domesticated swine using polymerase chain reaction-restriction fragment length polymorphism

  • Manalaysay, Jessica G.;Antonio, Nathaniel D.;Apilado, Ralph Lorenz R.;Bambico, Joseph F.;Mingala, Claro N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.262-266
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    • 2017
  • Objective: This study was conducted to screen scrotal hernia in domesticated swine from selected breeders in the Philippines. This defect is associated with a cytosine to thymine mutation in the BCL-2 associated X protein (BAX) gene of swine. Methods: Genetic screening was done by DNA extraction followed by amplification and digestion using polymerase chain reaction-restriction fragment length polymorphism, amplifying the 416 bp region of the BAX gene that was subjected to digestion using the Ear I enzyme. Sequencing was also conducted to validate the results. Results: Results revealed that out of 538 samples tested, 411 (76.4%) of the samples were found to be normal whereas the remaining were carriers of the mutation in which 80 (14.9%) were heterozygous mutants and 47 (8.7%) were homozygous mutants. Pietrain breed was found to have the highest incidence. Conclusion: Having a scrotal hernia eliminates the chances of using the boar as a breeder stock because the following generations arising from it would most likely exhibit herniation. It is therefore advised to establish a genetic screening method for Scrotal Hernia in the Philippines to eliminate the negative gene from the herd.

Heterotrophic Bacterial Growth on Hoses in a Neonatal Water Distribution System

  • Buffet-Bataillon, Sylvie;Bonnaure-Mallet, Martine;De La Pintiere, Armelle;Defawe, Guy;Gautier-Lerestif, Anne Lise;Fauveau, Severine;Minet, Jacques
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.779-781
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    • 2010
  • After preliminary tests indicated an increased number of heterotrophic bacteria, we investigated possible sources of contamination in a neonatal intensive care unit (NICU) water distribution system. Scanning electron microscopic examination of flexible metallic hoses associated with the system revealed the presence of a biofilm; partial 16S rDNA sequencing revealed that the biofilm contained Blastomonas natatoria. Purgation of the water system three times a day, reinforced faucet cleaning, decreasing the cold water temperature to $12^{\circ}C$, and six repeated chlorinations at concentrations as high as 2 mg/l were not sufficient to eradicate the bacterial contamination. Replacing all of the rubber-interior flexible metallic hoses with teflon-lined hoses, followed by heating the water to $70^{\circ}C$, successfully controlled the bacteria.

Two- Dimensional Electrophoresis Analysis of Proteins; Bacillus subtilis LTD and Its Antifungal Activity Deficient Mutant

  • Lee, Young-Keun;Dinh, Le Thi;Jang, Yu-Sin;Chung, Hye-Young;Chang, Hwa-Hyoung
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.487-493
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    • 2004
  • To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.

Metabolic Flux Shift of Weissella kimchii sk10 Grown Under Aerobic Conditions

  • Park, Sun-Mi;Kang, Hye-Sun;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.919-923
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    • 2004
  • The sk10 isolated from kimchi was identified as W. kimchii on the basis of l6s-rDNA sequencing. Studies were made to analyze the metabolic flux shift of the sk10 on glucose under aerobic growth conditions. The sk10 produced 38.2 mM acetate, 16.3 mM ethanol, and 33.2 mM lactate under aerobic conditions, but 2.4 mM acetate, 48.0 mM ethanol, and 44.1 mM lactate under anaerobic conditions. The NADH peroxidase (NADH-dependent hydrogen peroxidase) activity of sk10 grown under aerobic conditions was 11 times higher than that under anaerobic conditions. Under the low ratio of $NADH/NAD^+$, the metabolic flux toward lactate and ethanol was shifted to the flux through acetate kinase without NADH oxidation. The kinds of enzymes and metabolites of sk10 were close to those in the pathway of Leuconostoc sp., but the metabolites produced under aerobic growth conditions were different from those of Leuconostoc sp. The stoichiometric balance calculated using the concentrations of metabolites and substrate was about 97%, coincident with the theoretical values under both aerobic and anaerobic conditions. From these results, it was concluded that the metabolic flux of W. kimchii sk10 was partially shifted from lactate and ethanol to acetate under aerobic conditions only.

Biodegradation of Phenanthrene by Psychrotrophic Bacteria from Lake Baikal

  • AHN TAE-SEOK;LEE GEON-HYOUNG;SONG HONG-GYU
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1135-1139
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    • 2005
  • Psychrotrophic phenanthrene-degrading bacteria were identified in the sediment samples collected from Lake Baikal, Russia. Among 70 phenanthrene-degrading isolates, the seven that had the highest phenanthrene-degradation rates were identified by 16S rDNA sequencing. Isolate P25, identified as the Gram-positive rod-shaped organism Rhodococcus erythropolis, had the highest growth and degradation rate at $15^{\circ}C$. It could remove $26.0\%$ of 100 mg $1^{-1}$ phenanthrene in 20 days at $15^{\circ}C$, and degradation was less at $5^{\circ}C\;and\;25^{\circ}C$. The addition of surfactants to enhance degradation was tested. Brij 30 and Triton X-100 inhibited degradation at all surfactant concentrations tested, but Tween 80 stimulated phenanthrene degradation, especially at low concentrations. When $20{\times}$ CMC (critical micelle concentration) of Tween 80 was added, $38.0\%$ of 100 mg $1^{-1}$ phenanthrene was degraded in 12 days at $15^{\circ}C$. This psychrotrophic phenanthrene-degrading bacterium is a candidate for use in bioremediation of polycyclic hydrocarbon contamination in low temperature environments.

A New Record of Nannizziopsis mirabilis from Field Soil in Korea

  • Adhikari, Mahesh;Gurung, Sun Kumar;Kim, Sang Woo;Lee, Hyun Goo;Ju, Han Jun;Gwon, Byeong Heon;Kosol, San;Bazie, Setu;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.249-254
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    • 2018
  • Nannizziopsis mirabilis (KNU17-67) was isolated from a field soil sample collected from Jejudo in 2017. This fungal isolate was identified and described by its morphological characters and the internal transcribed spacer rDNA gene sequencing. Potato dextrose agar medium was used to study its macro and micromorphology. In addition, a molecular phylogenetic tree was constructed for its precise confirmation as N. mirabilis. This fungal isolate has not officially been reported previously in Korea.

Characterization of Lactic Acid Bacteria Isolated from Sauce-type Kimchi

  • Jung, Suk-Hee;Park, Joung-Whan;Cho, Il-Jae;Lee, Nam-Keun;Yeo, In-Cheol;Kim, Byung-Yong;Kim, Hye-Kyung;Hahm, Young-Tae
    • Preventive Nutrition and Food Science
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    • v.17 no.3
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    • pp.217-222
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    • 2012
  • This study was carried out to investigate the isolation and characterization of lactic acid bacteria (LAB) from naturally fermented sauce-type kimchi. Sauce-type kimchi was prepared with fresh, chopped ingredients (Korean cabbage, radish, garlic, ginger, green onion, and red pepper). The two isolated bacteria from sauce-type kimchi were identified as Pediococcus pentosaceus and Lactobacillus brevis by 16S rDNA sequencing and tentatively named Pediococcus sp. IJ-K1 and Lactobacillus sp. IJ-K2, respectively. Pediococcus sp. IJ-K1 was isolated from the early and middle fermentation stages of sauce-type kimchi whereas Lactobacillus sp. IJ-K2 was isolated from the late fermentation stage. The resistance of Pediococcus sp. IJ-K1 and Lactobacillus sp. IJ-K2 to artificial gastric and bile acids led to bacterial survival rates that were 100% and 84.21%, respectively.

Effects of Forest Management Practices and Environment on Occurrence of Armillaria Species

  • Kim, Mee-Sook;Klopfenstein, Ned B.;Mcdonald, Geral I.
    • Journal of Korean Society of Forest Science
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    • v.99 no.2
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    • pp.251-257
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    • 2010
  • Influences of environment (indicated by plant associations) and forest management practices on the distribution of Armillaria spp. and genets (vegetative clones) were investigated. A total of 142 isolates of Armillaria was collected from various host trees on pristine and managed sites (thinned and/or fertilized) growing in relatively wet and dry environments in eastern Washington, U.S.A. The incidence of Armillaria spp. was significantly higher in the relatively wetter sites than the relatively drier sites, as indicated by plant associations. However, no differences in Armillaria occurrence were found among different forest management practices (control vs. thinned vs. thinned and fertilized) within both wetter and drier sites. Incidence of Armillaria was significantly different among conifer and shrub species. The highest proportion with Armillaria was found on grand fir (Abies grandis). Based on pairing tests and rDNA sequencing, the 142 isolates were comprised in a total of 20 genets representing three Armillaria species. More diverse Armillaria spp. were found in both relatively wetter and relatively drier sites within the undisturbed control plots, compared to plots disturbed by forest management practices. The results from this study provide baseline information toward understanding how environment and forest management practices influence incidence and diversity of Armillaria species and genets.

Flora of drift plastics: a new red algal genus, Tsunamia transpacifica(Stylonematophyceae) from Japanese tsunami debris in the northeast Pacific Ocean

  • West, John A.;Hansen, Gayle I.;Hanyuda, Takeaki;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.31 no.4
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    • pp.289-301
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    • 2016
  • Floating debris provides substrates for dispersal of organisms by ocean currents, including algae that thrive on plastics. The 2011 earthquake and tsunami in Tohuku, Japan resulted in large amounts of debris carried by the North Pacific Current to North America from 2012 to 2016. In 2015-2016, the plastics in the debris bore a complex biota including pink algal crusts. One sample (JAW4874) was isolated into culture and a three-gene phylogeny (psbA, rbcL, and SSU) indicated it was an unknown member of the red algal class Stylonematophyceae. It is a small pulvinate crust of radiating, branched, uniseriate filaments with cells containing a single centrally suspended nucleus and a single purple to pink, multi-lobed, parietal plastid lacking a pyrenoid. Cells can be released as spores that attach and germinate to form straight filaments by transverse apical cell divisions, and subsequent longitudinal and oblique intercalary divisions produce masses of lateral branches. This alga is named Tsunamia transpacifica gen. nov. et sp. nov. Sequencing of additional samples of red algal crusts on plastics revealed another undescribed Stylonematophycean species, suggesting that these algae may be frequent on drift oceanic plastics.

Probiotic Properties of Lactobacillus plantarum NK181 Isolated from Jeotgal, a Korean Fermented Food

  • Lee, Na-Kyoung;Kim, Hyoun-Wook;Chang, Hyo-Ihl;Yun, Cheol-Won;Kim, Seung-Wook;Kang, Chang-Won;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.227-231
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    • 2006
  • Strain NK181 was isolated for probiotic use from jeotkal and based on results of API 50 CHL kit and 16S rDNA sequencing was tentatively named Lactobacillus plantarum NK181. L. plantarum NK181 was highly resistant to artificial gastric juice (pH 2.5) and bile acid and demonstrated strong adherence to Caco-2 cells. In test using API ZYM kit, eight enzymes were produced. Supernatant of L. plantarum NK181 exhibited about 30% 1,1-diphenyl-2-picyryl hedrazyl (DPPH) radical-scavenging activity and reduced cholesterol by 70%. These results demonstrate potential use of L. plantarum NK181 as health-promoting probiotic.