Optimization of single plate-serial dilution spotting (SP-SDS) with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples

Thomas, Pious; Sekhar, Aparna C.; Upreti, Reshmi; Mujawar, M.M.; Pasha, Sadiq S.

doi:10.1016/j.btre.2015.08.003

Cited by 192 publications

(142 citation statements)

References 19 publications

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“…70 Plates were incubated overnight at 30 C before scoring for cell growth. The resulting cell suspensions were successively diluted 1:10 seven times, and from each of these dilutions six 5 μl spots were plated on a YPD agar plate.…”

Section: Heat Shock Tolerancementioning

confidence: 99%

Matrices (re)loaded: Durability, viability, and fermentative capacity of yeast encapsulated in beads of different composition during long‐term fed‐batch culture

Gulli

Yunker

Rosenzweig

2019

Biotechnology Progress

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Encapsulated microbes have been used for decades to produce commodities ranging from methyl ketone to beer. Encapsulated cells undergo limited replication, which enables them to more efficiently convert substrate to product than planktonic cells and which contributes to their stress resistance. To determine how encapsulated yeast supports long-term, repeated fed-batch ethanologenic fermentation, and whether different matrices influence that process, fermentation and indicators of matrix durability and cell viability were monitored in high-dextrose, fed-batch culture over 7 weeks. At most timepoints, ethanol yield (g/g) in encapsulated cultures exceeded that in planktonic cultures. And frequently, ethanol yield differed among the four matrices tested: sodium alginate crosslinked with Ca 2+ and chitosan, sodium alginate crosslinked with Ca 2+ , Protanal alginate crosslinked with Ca 2+ and chitosan, Protanal alginate crosslinked with Ca 2+ , with the last of these consistently demonstrating the highest values. Young's modulus and viscosity were higher for matrices crosslinked with chitosan over the first week; thereafter values for both parameters declined and were indistinguishable among treatments. Encapsulated cells exhibited greater heat shock tolerance at 50 C than planktonic cells in either stationary or exponential phase, with similar thermotolerance observed across all four matrix types. Altogether, these data demonstrate the feasibility of re-using encapsulated yeast to convert dextrose to ethanol over at least 7 weeks. K E Y W O R D Salginate, chitosan, fermentation, immobilization, yeast encapsulation

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Section: Heat Shock Tolerancementioning

confidence: 99%

Matrices (re)loaded: Durability, viability, and fermentative capacity of yeast encapsulated in beads of different composition during long‐term fed‐batch culture

Gulli

Yunker

Rosenzweig

2019

Biotechnology Progress

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“…The determination of the dominance of bacteria referred to Thomas et al (2015), the sample diluted through serial dilution as in the procedure by Madigan et al (2011) by plating in TSA media. The bacterial density for the initial dilution was 30-300 colonies; it used to find the dominance group of bacteria.…”

Section: The Dominance Of Bacteriamentioning

confidence: 99%

Characterization of pathogenic bacteria in eel Anguilla bicolor bicolor

Wahjuningrum¹,

Hidayat²,

Budiardi³

2018

Jurnal Akuakultur Indonesia

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This research aimed to characterize bacteria caused disease in eel Anguilla bicolor bicolor. The research was conducted in two steps. The first step included the isolation and identification of bacteria from the disease infected glass eel (average body length: 5.0 ± 0.5 cm, average weight: 0.5 ± 0.1 g). The observation were colony and cell morphology, physiology, and biochemical characterization of bacteria, hemolysis test, and bacteria identification performed by KIT API 20 E, KIT API 20 Strep, and KIT API 20 Listeria. The second step was Koch's postulate, tested on healthy elver with an average length of 15.00 ± 0.65 cm and weight of 3.00 ± 0.75 g. The results showed three dominant species of bacteria suspected as a causative agent in eel, namely: Aeromonas hydrophila, Streptococcus agalactiae, and Listeria grayi. Koch's postulates test proved that the Aeromonas hydrophila and Streptococcus agalactiae were virulent to Anguilla bicolor bicolor. Thus, A. hydrophila and S. agalactiae were disease-causing agent bacteria in eel.Keywords: Anguilla bicolor bicolor, bacteria, A. hydrophila, S. agalactiae. ABSTRAKPenelitian ini bertujuan untuk mengkarakterisasi bakteri penyebab penyakit pada ikan sidat Anguilla bicolor bicolor. Penelitian ini dilakukan dalam dua tahap. Tahap pertama meliputi isolasi dan identifikasi bakteri dari ikan sidat kondisi sakit pada stadia glass eel. Ukuran panjang ikan sidat rata-rata 5 ± 0,5 cm dan bobot ratarata 0,5 ± 0,08 g, pengamatan bentuk morfologi koloni dan morfologi sel, karakterisasi fisiologi, dan biokimia bakteri, serta uji hemolisis, dan identifikasi jenis bakteri dengan KIT API 20 E, KIT API 20 Strep, dan KIT API 20 Listeria. Tahap kedua yaitu uji postulat Koch pada ikan sidat kondisi sehat stadia elver yang berukuran panjang rata-rata 15 ± 0,65 cm dan bobot rata-rata 3 ± 0,75 g. Hasil penelitian diperoleh tiga jenis bakteri dominan yaitu Aeromonas hydrophila, Streptococcus agalactiae, dan Listeria grayi. Uji postulat Koch membuktikan bahwa bakteri A. hydrophila dan S. agalactiae bersifat virulen pada ikan sidat Anguilla bicolor bicolor. Dengan demikian maka bakteri A. hydrophila dan S. agalactiae sebagai bakteri penyebab penyakit pada ikan sidat.Kata kunci: Anguilla bicolor bicolor, bakteri, A. hydrophila, S. agalactiae

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“…The phenotype of the ost2 knockout expressing DAD1 or OST2 was analyzed in the presence or absence of 1 µg/mL tunicamycin (Sigma). Cell viability was analyzed by single plate-serial dilution spotting (SP-SDS) (Thomas et al, 2015) after tunicamycin treatment using a starting OD 600 = 0.6 and 5 µl of dilutions (10 1 –10 6 ) on the plates, which were incubated at 28 °C for 48 h.…”

Section: Methodsmentioning

confidence: 99%

Stress responses of the oil-producing green microalgaBotryococcus brauniiRace B

Cornejo-Corona

Thapa

Browne

et al. 2016

PeerJ

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Plants react to biotic and abiotic stresses with a variety of responses including the production of reactive oxygen species (ROS), which may result in programmed cell death (PCD). The mechanisms underlying ROS production and PCD have not been well studied in microalgae. Here, we analyzed ROS accumulation, biomass accumulation, and hydrocarbon production in the colony-forming green microalga Botryococcus braunii in response to several stress inducers such as NaCl, NaHCO3, salicylic acid (SA), methyl jasmonate, and acetic acid. We also identified and cloned a single cDNA for the B. braunii ortholog of the Arabidopsis gene defender against cell death 1 (DAD1), a gene that is directly involved in PCD regulation. The function of B. braunii DAD1 was assessed by a complementation assay of the yeast knockout line of the DAD1 ortholog, oligosaccharyl transferase 2. Additionally, we found that DAD1 transcription was induced in response to SA at short times. These results suggest that B. braunii responds to stresses by mechanisms similar to those in land plants and other organisms.

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Optimization of single plate-serial dilution spotting (SP-SDS) with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples

Abstract: Graphical abstract

Cited by 192 publications

References 19 publications

Matrices (re)loaded: Durability, viability, and fermentative capacity of yeast encapsulated in beads of different composition during long‐term fed‐batch culture

Matrices (re)loaded: Durability, viability, and fermentative capacity of yeast encapsulated in beads of different composition during long‐term fed‐batch culture

Characterization of pathogenic bacteria in eel Anguilla bicolor bicolor

Stress responses of the oil-producing green microalgaBotryococcus brauniiRace B

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