Culture of unicellular protists in marine broth v3

protocols.io ◽  
2016 ◽  
Author(s):  
Maria Rubio-Brotons
Keyword(s):  
Marine Broth

scholarly journals ISOLASI BAKTERI YANG BERSIMBION DENGAN ASCIDIAN Herdmania momus YANG MEMILIKI AKTIVITAS ANTIBAKTERI

2020 ◽  
Vol 8 (2) ◽  
pp. 20
Author(s):  
Silvia E Koyongian ◽  
Deiske A Sumilat ◽  
Rosita A J Lintang ◽  
Stenly Wullur ◽  
Sandra O Tilaar ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Marine Invertebrates ◽  
Nutrient Agar ◽  
Symbiotic Bacteria ◽  
Bacterial Isolates ◽  
Marine Broth ◽  
Antibacterial Screening ◽  
Coral Reef Ecosystems ◽  
And Staphylococcus Aureus

 Ascidian is marine invertebrates in coral reef ecosystems that produce many bioactive compounds for pharmacology. The presence of symbiotic bacteria with marine organisms is protected the host biota by producing secondary metabolites. The purpose of this study is to obtain symbiotic bacterial isolates with Herdmania momus ascidian, then to observe the antibacterial activity of these bacterial isolates against Escherichia coli, and Staphylococcus aureus. Isolation and culture of the symbiotic bacteria were made on Nutrient Agar and Zobell Marine Broth media. The antibacterial screening showed that the Herdmania momus symbiotic bacteria were able to inhibit the growth of Staphylococcus aureus and Escherichia coli.Keywords: ascidians, Herdmania momus, bacteria, isolation, antibacterialAbstak          Ascidian adalah avetebrata laut di ekosistem terumbu karang yang banyak menghasilkan senyawa bioaktif untuk bidang farmakologi. Keberadaan bakteri yang bersimbion dengan organisme laut pada umumnya untuk melindungi biota yang ditumpanginya dan dirinya dengan cara menghasilkan senyawa metabolit sekunder. Tujuan dari penelitian ini yaitu untuk mendapatkan isolat bakteri yang bersimbion dengan ascidian Herdmania momus, kemudian mengamati aktivitas antibakteri dari isolat bakteri tersebut terhadap Escherichia coli, dan Staphylococcus aureus. Isolasi dan kultur bakteri yang bersimbion dengan ascidian dibuat pada media Nutrient Agar dan Zobell Marine Broth. Skrining aktivitas antibakteri menunjukkan isolat bakteri yang bersimbion dengan ascidian Herdmania momus mampu menghambat pertumbuhan organisme uji Staphylococcus aureus dan Escherichia coli.Kata kunci: ascidian, Herdmania momus, bakteri, isolasi, antibakteri

Uptake and Conversion of Selenium by a Marine Bacterium

1983 ◽  
Vol 40 (S2) ◽  
pp. s215-s220 ◽  
Author(s):  
A. Foda ◽  
J. H. Vandermeulen ◽  
J. J. Wrench
Keyword(s):  
Amino Acid ◽  
Marine Bacterium ◽  
Lipid Fraction ◽  
Water Soluble ◽  
Gas Liquid Chromatography ◽  
Bacterial Cells ◽  
Marine Broth ◽  
Simultaneous Measurements ◽  
Uptake Studies ◽  
Bacterial Lipid

Bio-conversion of Se was examined by incubating Pseudomonas marina in seawater containing either selenite (Na2SeIVO3) or selenate (Na2SeVIO4). At the concentrations of selenite and selenate used (10−4–10−7 mol/L), the growth of P. marina was not inhibited. Under these conditions, selenite was taken up by P. marina, but selenate was not found to enter the cells. Pseudomonas marina incorporated selenite from filtered seawater into sub-cellular fractions, primarily protein (30–50%) and amino acids (44–70%). When incubated in marine broth, P. marina incorporated 75selenite primarily into protein (up to 75%), with a lesser amount into the amino acid fraction (approximately 25%). Insignificant amounts were associated with the bacterial lipid fraction. SeIV was found in the protein and amino acid fractions within 10 min of incubation in medium containing selenite. In uptake studies the level of SeIV in the incubating medium decreased markedly, corresponding presumably to SeIV entering the bacterial cells from the medium. However, simultaneous measurements of total Se in the medium (gas–liquid chromatography following photooxidation) revealed an increasing amount of non-SeIV species of Se in the medium throughout the same period. These results were interpreted as due to the bioconversion of selenite by P. marina into water-soluble non-SeIV metabolite(s) and their subsequent release back into the medium. Up to 35% of the total Se found in the medium after 24 h can be accounted for by this conversion of SeIV into another oxidation state. Pseudomonas marina is also capable of reducing SeIV to elemental Se; this pathway becomes increasingly evident at higher concentrations of Na2SeO3.

scholarly journals Global Transcriptomic Responses of Roseithermus sacchariphilus Strain RA in Media Supplemented with Beechwood Xylan

2020 ◽  
Vol 8 (7) ◽  
pp. 976
Author(s):  
Kok Jun Liew ◽  
Neil C. Bruce ◽  
Rajesh Kumar Sani ◽  
Chun Shiong Chong ◽  
Amira Suriaty Yaakop ◽  
...  
Keyword(s):  
New Genus ◽  
Enzymatic Activities ◽  
Glycosyl Hydrolases ◽  
Marine Broth ◽  
Beechwood Xylan ◽  
Transcriptomic Responses ◽  
Membrane Components ◽  
Upregulated Genes ◽  
Current Report

The majority of the members in order Rhodothermales are underexplored prokaryotic extremophiles. Roseithermus, a new genus within Rhodothermales, was first described in 2019. Roseithermus sacchariphilus is the only species in this genus. The current report aims to evaluate the transcriptomic responses of R. sacchariphilus strain RA when cultivated on beechwood xylan. Strain RA doubled its growth in Marine Broth (MB) containing xylan compared to Marine Broth (MB) alone. Strain RA harbors 54 potential glycosyl hydrolases (GHs) that are affiliated with 30 families, including cellulases (families GH 3, 5, 9, and 44) and hemicellulases (GH 2, 10, 16, 29, 31,43, 51, 53, 67, 78, 92, 106, 113, 130, and 154). The majority of these GHs were upregulated when the cells were grown in MB containing xylan medium and enzymatic activities for xylanase, endoglucanase, β-xylosidase, and β-glucosidase were elevated. Interestingly, with the introduction of xylan, five out of six cellulolytic genes were upregulated. Furthermore, approximately 1122 genes equivalent to one-third of the total genes for strain RA were upregulated. These upregulated genes were mostly involved in transportation, chemotaxis, and membrane components synthesis.

scholarly journals Influence of Iron on Production of the Antibacterial Compound Tropodithietic Acid and Its Noninhibitory Analog in Phaeobacter inhibens

2015 ◽  
Vol 82 (2) ◽  
pp. 502-509 ◽  
Author(s):  
Paul W. D'Alvise ◽  
Christopher B. W. Phippen ◽  
Kristian F. Nielsen ◽  
Lone Gram
Keyword(s):  
Mass Spectrometry ◽  
Antibacterial Activity ◽  
High Resolution Mass Spectrometry ◽  
Biologically Active ◽  
Brown Pigment ◽  
Marine Broth ◽  
Content Type ◽  
Tropodithietic Acid ◽  
Antibacterial Compound ◽  
Phaeobacter Inhibens

ABSTRACTTropodithietic acid (TDA) is an antibacterial compound produced by somePhaeobacterandRuegeriaspp. of theRoseobacterclade. TDA production is studied in marine broth or agar since antibacterial activity in other media is not observed. The purpose of this study was to determine how TDA production is influenced by substrate components. High concentrations of ferric citrate, as present in marine broth, or other iron sources were required for production of antibacterially active TDA. However, when supernatants of noninhibitory, low-iron cultures ofPhaeobacter inhibenswere acidified, antibacterial activity was detected in a bioassay. The absence of TDA in nonacidified cultures and the presence of TDA in acidified cultures were verified by liquid chromatography–high-resolution mass spectrometry. A noninhibitory TDA analog (pre-TDA) was produced byP. inhibens,Ruegeria mobilisF1926, andPhaeobactersp. strain 27-4 under low-iron concentrations and was instantaneously converted to TDA when pH was lowered. Production of TDA in the presence of Fe3+coincides with formation of a dark brown substance, which could be precipitated by acid addition. From this brown pigment TDA could be liberated slowly with aqueous ammonia, and both direct-infusion mass spectrometry and elemental analysis indicated a [FeIII(TDA)2]xcomplex. The pigment could also be produced by precipitation of pure TDA with FeCl3. Our results raise questions about how biologically active TDA is produced in natural marine settings where iron is typically limited and whether the affinity of TDA to iron points to a physiological or ecological function of TDA other than as an antibacterial compound.

scholarly journals Tateyamaria pelophila sp. nov., a facultatively anaerobic alphaproteobacterium isolated from tidal-flat sediment, and emended descriptions of the genus Tateyamaria and of Tateyamaria omphalii

2010 ◽  
Vol 60 (8) ◽  
pp. 1770-1777 ◽  
Author(s):  
Henrik Sass ◽  
Beate Köpke ◽  
Heike Rütters ◽  
Theresa Feuerlein ◽  
Stefan Dröge ◽  
...  
Keyword(s):  
Tidal Flat ◽  
Novel Species ◽  
Major Fatty Acid ◽  
Physiological Data ◽  
Most Probable Number ◽  
Marine Broth ◽  
Number Series ◽  
Probable Number ◽  
Carbon Compounds ◽  
Constant Growth

A Gram-negative motile rod, strain SAM4T, was isolated from the highest positive dilution of a most probable number series inoculated with tidal-flat sediments from the German North Sea coast. The isolate grew at 4–35 °C and showed constant growth yields throughout almost the whole temperature range. Growth was observed between pH 6 and 9 and at salinities of 0.3–10.2 %. Strain SAM4T required Na+ for growth, contained bacteriochlorophyll a and was catalase- and oxidase-positive. It was nutritionally versatile growing on a variety of carbon compounds including carbohydrates, amino acids and organic acids like lactate or succinate. It grew anaerobically on complex media such as marine broth, indicating fermentation, and by reducing trimethylammonium oxide. The dominant phospholipids were phosphatidylethanolamine and phosphatidylglycerol, whereas only traces of phosphatidylcholine and an unidentified lipid were found. The major fatty acid was n-C18 : 1 ω7c. The DNA G+C content was 56.4 mol%. The isolate was identified as a member of the Roseobacter clade within the class Alphaproteobacteria. However, based on phylogenetic, phenotypic and physiological data, it clearly differs from its closest relative Tateyamaria omphalii. Therefore, a novel species is proposed: Tateyamaria pelophila sp. nov., with strain SAM4T (=DSM 17270T=LMG 23018T) as the type strain. Emended descriptions of the genus Tateyamaria and of Tateyamaria omphalii are also presented.

scholarly journals Deteksi bakteri pembentuk amina biogenik pada ikan Scombridae secara multiplex PCR

2020 ◽  
Vol 23 (2) ◽  
pp. 359-371
Author(s):  
Rizsa Mustika Pertiwi ◽  
Mala Nurilmala ◽  
Asadatun Abdullah ◽  
Nurjanah ◽  
Roza Yusfiandayani ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Histidine Decarboxylase ◽  
Lysine Decarboxylase ◽  
Tyrosine Decarboxylase ◽  
Marine Broth

Amina biogenik merupakan komponen basa nitrogen yang terbentuk oleh dekarboksilasi asam amino. Amina biogenik yang sering terdeteksi pada produk perikanan di antaranya histamin, tiramin dan kadaverin. Gen pengkode dekarboksilasi asam amino tersebut yaitu histidine decarboxylase (hdc), tyrosine decarboxylase (tdc) serta lysine decarboxylase (ldc). Penelitian ini bertujuan untuk mendapatkan isolat DNA bakteri beserta konsentrasi dan kemurniannya, menyeleksi media pertumbuhan bakteri, mendeteksi gen hdc, tdc dan ldc menggunakan multiplex PCR serta identifikasi bakteri.. Sampel yang digunakan yaitu ikan tuna beku, tongkol beku, cakalang beku, tuna loin, pindang tongkol potong dan pindang tongkol bumbu kuning serta kontrol positif menggunakan ikan tuna yang disimpan pada suhu ruang selama 3 hari. Penelitian terdiri dari tahap pra-pengayaan bakteri pada media marine broth dan lactose broth, isolasi DNA sampel tanpa dan hasil pra-pengayaan, uji kemurnian dan konsentrasi isolat serta amplifikasi gen target hdc, tdc, ldc menggunakan multiplex PCR. Hasil yang diperoleh yaitu kualitas isolat DNA bakteri pada umumnya sudah murni, dengan konsentrasi isolat 15,75-157,84 ng/µL. Teknik multiplex PCR berhasil mendeteksi gen hdc, tdc dan ldc pada ikan scombridae. Kondisi optimum amplifikasi PCR yaitu annealing pada suhu 50°C selama 90 detik. Gen yang terdeteksi pada sampel tanpa pra-pengayaan yaitu tdc, sedangkan ldc dan hdc ditemukan pada sampel hasil pra-pengayaan media lactose broth dan marine broth. Bakteri pembentuk amina biogenik teridentifikasi sebagai Enterococcus fecalis, M. morganii, E. aerogenes, Citrobacter sp., Hafnia paralvei dan Obesumbacterium proteus. Amina biogenik merupakan komponen basa nitrogen yang terbentuk oleh dekarboksilasi asam amino. Amina biogenik yang sering terdeteksi pada produk perikanan di antaranya histamin, tiramin dan kadaverin. Gen pengkode dekarboksilasi asam amino tersebut yaitu histidine decarboxylase (hdc), tyrosine decarboxylase (tdc) serta lysine decarboxylase (ldc). Penelitian ini bertujuan untuk mendapatkan isolat DNA bakteri beserta konsentrasi dan kemurniannya, menyeleksi media pertumbuhan bakteri, mendeteksi gen hdc, tdc dan ldc menggunakan multiplex PCR serta identifikasi bakteri.. Sampel yang digunakan yaitu ikan tuna beku, tongkol beku, cakalang beku, tuna loin, pindang tongkol potong dan pindang tongkol bumbu kuning serta kontrol positif menggunakan ikan tuna yang disimpan pada suhu ruang selama 3 hari. Penelitian terdiri dari tahap pra-pengayaan bakteri pada media marine broth dan lactose broth, isolasi DNA sampel tanpa dan hasil pra-pengayaan, uji kemurnian dan konsentrasi isolat serta amplifikasi gen target hdc, tdc, ldc menggunakan multiplex PCR. Hasil yang diperoleh yaitu kualitas isolat DNA bakteri pada umumnya sudah murni, dengan konsentrasi isolat 15,75-157,84 ng/µL. Teknik multiplex PCR berhasil mendeteksi gen hdc, tdc dan ldc pada ikan scombridae. Kondisi optimum amplifikasi PCR yaitu annealing pada suhu 50°C selama 90 detik. Gen yang terdeteksi pada sampel tanpa pra-pengayaan yaitu tdc, sedangkan ldc dan hdc ditemukan pada sampel hasil pra-pengayaan media lactose broth dan marine broth. Bakteri pembentuk amina biogenik teridentifikasi sebagai Enterococcus fecalis, M. morganii, E. aerogenes, Citrobacter sp., Hafnia paralvei dan Obesumbacterium proteus.

scholarly journals Preliminary Detection of Antibacterial Activity of Fishpond Water Bacteria against Aquaculture Pathogenic Bacteria

Biota ◽  
2020 ◽  
Vol 13 (2) ◽  
pp. 91-102
Author(s):  
Stella Magdalena ◽  
Sindy Sindy ◽  
Yogiara Yogiara
Keyword(s):  
Pathogenic Bacteria ◽  
Bacterial Resistance ◽  
16S Rrna Gene Sequencing ◽  
Luria Broth ◽  
Rrna Gene ◽  
Clear Zone ◽  
Marine Broth ◽  
Antibacterial Compounds ◽  
Rrna Gene Sequencing ◽  
Potential Use

Aquaculture is currently experiencing massive loss due to the outbreak of pathogenic bacteria. One of the outbreak causes is the development of pathogenic bacterial resistance to the antibacterial. The problem can be solved using microorganisms that can produce new antibacterial compounds. The purpose of this research was to obtain bacteria from fishpond water that could produce antibacterial compounds. About two out of 81 isolates could produce antibacterial compounds. Those two isolates were obtained from saltwater fishponds in North Jakarta (TS2) and Harapan Island (PHY). All fishpond water was grown in marine broth or Luria broth. Extraction of antibacterial compounds was performed using four types of solvents: chloroform, dichloromethane, ethyl acetate, and methanol. Each of the solvents showed a different result. The extraction can only be successfully performed using chloroform and dichloromethane. Extraction using dichloromethane showed a larger inhibitory clear zone than chloroform. Based on 16S rRNA gene sequencing, PHY isolate was identified as Bacillus sp. and TS2 as Acinetobacter sp. In conclusion, isolate TS2 and PHY, which produced antibacterial compounds, showed potential use as aquaculture probiotics.

scholarly journals Ecology, Inhibitory Activity, and Morphogenesis of a Marine Antagonistic Bacterium Belonging to the Roseobacter Clade

2005 ◽  
Vol 71 (11) ◽  
pp. 7263-7270 ◽  
Author(s):  
Jesper Bartholin Bruhn ◽  
Kristian Fog Nielsen ◽  
Mette Hjelm ◽  
Michael Hansen ◽  
José Bresciani ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
High Resolution Mass Spectrometry ◽  
Single Cells ◽  
Homoserine Lactone ◽  
Growth Conditions ◽  
Buffer System ◽  
Larval Rearing ◽  
Marine Broth ◽  
Total Peak Area ◽  
Antibacterial Compounds

ABSTRACT Roseobacter strain 27-4 has been isolated from a turbot larval rearing unit and is capable of reducing mortality in turbot egg yolk sac larvae. Here, we demonstrate that the supernatant of Roseobacter 27-4 is lethal to the larval pathogens Vibrio anguillarum and Vibrio splendidus in a buffer system and inhibited their growth in marine broth. Liquid chromatography (LC) with both UV spectral detection and high-resolution mass spectrometry (HR-MS) identified the known antibacterial compound thiotropocin or its closely related precursor tropodithietic acid in the bioactive fractions. Antibacterial activity correlated with the appearance of a brownish pigment and was only formed in marine broth under static growth conditions. A thick biofilm of multicellular star-shaped aggregated cells formed at the air-liquid interface under static growth conditions. Here, the bioactive compound was the base peak in the LC-UV chromatograms of the extracts where it constituted 15% of the total peak area. Aerated conditions results in 10-fold-higher cell yield, however, cultures were nonpigmented, did not produce antibacterial activity, and grew as single cells. Production of antibacterial compounds may be quorum regulated, and we identified the acylated homoserine lactone (3-hydroxy-decanoyl homoserine lactone) from cultures of Roseobacter 27-4 using LC-HR-MS. The signal molecule was primarily detected in stagnant cultures. Roseobacter 27-4 grew between 10 and 30°C but died rapidly at 37°C. Also, the antibacterial compounds was sensitive to heat and was inactivated at 37°C in less than 2 days and at 25°C in 8 days. Using Roseobacter 27-4 as a probiotic culture will require that is be established in stagnant or adhered conditions and, due to the temperature sensitivity of the active compound, constant production must be ensured.

scholarly journals Consideration of Influential Factors on Bioleaching of Gold Ore Using Iodide-Oxidizing Bacteria

Minerals ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 274 ◽  
Author(s):  
San Yee Khaing ◽  
Yuichi Sugai ◽  
Kyuro Sasaki ◽  
Myo Min Tun
Keyword(s):  
Potassium Iodide ◽  
Bacterial Cell ◽  
Cell Number ◽  
Culture Solution ◽  
Gold Dissolution ◽  
Marine Broth ◽  
Gold Leaching ◽  
Temperature Range ◽  
Gold Ore ◽  
Bacterial Cell Number

Iodide-oxidizing bacteria (IOB) oxidize iodide into iodine and triiodide which can be utilized for gold dissolution. IOB can be therefore useful for gold leaching. This study examined the impact of incubation conditions such as concentration of the nutrient and iodide, initial bacterial cell number, incubation temperature, and shaking condition on the performance of the gold dissolution through the experiments incubating IOB in the culture medium containing the marine broth, potassium iodide and gold ore. The minimum necessary concentration of marine broth and potassium iodide for the complete gold dissolution were determined to be 18.7 g/L and 10.9 g/L respectively. The initial bacterial cell number had no effect on gold dissolution when it was 1 × 104 cells/mL or higher. Gold leaching with IOB should be operated under a temperature range of 30–35 °C, which was the optimal temperature range for IOB. The bacterial growth rate under shaking conditions was three times faster than that under static conditions. Shaking incubation effectively shortened the contact time compared to the static incubation. According to the pH and redox potential of the culture solution, the stable gold complex in the culture solution of this study could be designated as gold (I) diiodide.

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