Influence of cell density of Escherichia coli and the dinoflagellate Crypthecodinium cohnii on life history traits of the nematode Panagrolaimus sp. strain NFS 24-5, a potential larval food for marine aquaculture

Nematology ◽  
2014 ◽  
Vol 16 (4) ◽  
pp. 419-426 ◽  
Author(s):  
Farhana Ayub ◽  
Olaf Strauch ◽  
Laurent Seychelles ◽  
Ralf-Udo Ehlers
Keyword(s):  
Escherichia Coli ◽  
Life History ◽  
Cell Density ◽  
Mass Production ◽  
Life History Traits ◽  
Dry Mass ◽  
Marine Aquaculture ◽  
Crypthecodinium Cohnii ◽  
E Coli ◽  
A Cell

The nematode Panagrolaimus sp. NFS 24-5 has potential for use as living food for larval shrimps and fish in marine aquaculture. The nematodes are usually produced on bacterial or yeast cells. Nematodes cannot synthesise the long chain fatty acid docosahexaenoic acid (DHA) which is essential for feeding marine aquaculture organisms. The eukaryotic, heterotrophic dinoflagellate Crypthecodinium cohnii consists of approximately 20% DHA. To culture the nematodes and simultaneously enrich them with DHA, single adult male and female individuals were cultured in hanging drops with variable cell density of C. cohnii. Life history traits, such as net reproductive rate (), population doubling time (PDT) and intrinsic rate of natural increase (), were assessed and compared with data obtained from cultures on Escherichia coli. A maximum was recorded at a cell density of 4 × 106 C. cohnii cells ml−1, corresponding to 2478.82 μg dry mass ml−1. The same was achieved with 7× lower biomass of E. coli at a cell density of 3 × 109 cells ml−1, corresponding to 335.63 μg dry mass ml−1. The results exclude the use of the dinoflagellate culture from application in mass production of the nematode for aquaculture food and limit the use to post-harvest enrichment of the nematodes with essential fatty acids. At a density of 3 × 109 E. coli cells ml−1 the PDT was lowest and the was highest, indicating that this cell density might be closest to optimum conditions for nematode reproduction. Exceeding this cell density yielded fewer offspring within a longer time period. Implications for mass production in monoxenic liquid cultures are discussed.

Studies on the Growth of Escherichia coli O157:H7 Strains at 45.5°C

2000 ◽  
Vol 63 (9) ◽  
pp. 1173-1178 ◽  
Author(s):  
JASON FERENC ◽  
JASON OLIVER ◽  
RUTH WITKOWSKI ◽  
LYNNE McLANDSBOROUGH ◽  
ROBERT E. LEVIN
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Bile Salts ◽  
Present Report ◽  
Initial Density ◽  
Sensitive Strain ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Gas Formation ◽  
A Cell

The objectives of the present report were to examine the ability of 18 strains of Escherichia coli O157:H7 to grow in EC broth at 42.4, 43.5, 44.5, and 45.5°C, and to document the incidence of phenotypic variants present in low numbers that are capable of growth at 45.5°C in EC broth. Among the 18 strains of E. coli O157:H7 studied, only 3 were capable of producing turbid growth with gas formation in EC broth at 45.5°C with 1 × 102 initial CFU/ml. Higher initial densities of CFU resulted in turbid growth and gas formation in EC broth at 45.5°C with all strains. The presence of bile salts #3 in EC broth was found to be inhibitory at 45.5° C. All 18 strains were found to be capable of growth at 45.5°C in nonselective media. The ability of at least one sensitive strain to grow in EC broth at 45.5°C was found to be dependent on the initial number of CFU/ml. Prior growth of cells of a sensitive strain in EC broth at 45.5°C from a cell density of 2.0 × 107 to 8.0 × 107 CFU/ml followed by removal of cells and reinoculation at a cell density of 2.0 × 106 CFU/ml resulted in growth at 45.5°C that did not occur without such conditioning of the inhibitory medium. These results indicate that the ability of most strains of E. coli O157:H7 to grow in EC broth at 45.5°C is dependent on the initial density of CFU and that at low densities of CFU the ability to initiate growth is dependent on either low numbers of phenotypic variants tolerant to the presence of bile salts #3 in EC broth at 45.5°C or to conditioning of the medium with prior elevated numbers of cells.

scholarly journals Pilot-Scale Production of Fatty Acid Ethyl Esters by an Engineered Escherichia coli Strain Harboring the p(Microdiesel) Plasmid

2010 ◽  
Vol 76 (13) ◽  
pp. 4560-4565 ◽  
Author(s):  
Yasser Elbahloul ◽  
Alexander Steinbüchel
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Carbon Sources ◽  
Dry Mass ◽  
Pilot Scale ◽  
Coli Strain ◽  
Fatty Acid Ethyl Esters ◽  
Ethyl Esters ◽  
Scale Production ◽  
A Cell

ABSTRACT Fatty acid ethyl esters (FAEEs) were produced in this study by the use of an engineered Escherichia coli p(Microdiesel) strain. Four fed-batch pilot scale cultivations were carried out by first using glycerol as sole carbon source for biomass production before glucose and oleic acid were added as carbon sources. Cultivations yielded a cell density of up to 61 ± 3.1 g of cell dry mass (CDM) per liter and a maximal FAEE content of 25.4% ± 1.1% (wt/wt) of CDM.

scholarly journals Induction of Viable but Nonculturable Escherichia coli O157:H7 in the Phyllosphere of Lettuce: a Food Safety Risk Factor

2011 ◽  
Vol 77 (23) ◽  
pp. 8295-8302 ◽  
Author(s):  
Laura-Dorina Dinu ◽  
Susan Bach
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Low Temperature ◽  
Cell Density ◽  
Prolonged Storage ◽  
Potential Hazard ◽  
Vbnc State ◽  
Viable But Nonculturable ◽  
Content Type ◽  
E Coli

ABSTRACTEscherichia coliO157:H7 continues to be an important human pathogen and has been increasingly linked to food-borne illness associated with fresh produce, particularly leafy greens. The aim of this work was to investigate the fate ofE. coliO157:H7 on the phyllosphere of lettuce under low temperature and to evaluate the potential hazard of viable but nonculturable (VBNC) cells induced under such stressful conditions. First, we studied the survival of six bacterial strains following prolonged storage in water at low temperature (4°C) and selected two strains with different nonculturable responses for the construction ofE. coliO157:H7 Tn7gfptransformants in order to quantitatively assess the occurrence of human pathogens on the plant surface. Under a suboptimal growth temperature (16°C), bothE. coliO157:H7 strains maintained culturability on lettuce leaves, but under more stressful conditions (8°C), the bacterial populations evolved toward the VBNC state. The strain-dependent nonculturable response was more evident in the experiments with different inoculum doses (109and 106E. coliO157:H7 bacteria per g of leaf) when strain BRMSID 188 lost culturability after 15 days and strain ATCC 43895 lost culturability within 7 days, regardless of the inoculum dose. However, the number of cells entering the VBNC state in high-cell-density inoculum (approximately 55%) was lower than in low-cell-density inoculum (approximately 70%). We recorded the presence of verotoxin for 3 days in samples that contained a VBNC population of 4 to 5 log10cells but did not detect culturable cells. These findings indicate thatE. coliO157:H7 VBNC cells are induced on lettuce plants, and this may have implications regarding food safety.

scholarly journals Heterogeneous Absorption of Antimicrobial Peptide LL37 inEscherichia coliCells Enhances Population Survivability

2018 ◽  
Author(s):  
Mehdi Snoussi ◽  
John Paul Talledo ◽  
Nathan-Alexander Del Rosario ◽  
Bae-Yeun Ha ◽  
Andrej Košmrlj ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mathematical Model ◽  
Minimum Inhibitory Concentration ◽  
Cell Density ◽  
Inhibitory Concentration ◽  
Single Cell Level ◽  
Cell Level ◽  
E Coli ◽  
Rapid Absorption ◽  
Growing Population

AbstractAntimicrobial peptides (AMPs) are broad spectrum antibiotics that selectively target bacteria. Here we investigate the activity of human AMP LL37 againstEscherichia coliby integrating quantitative, population and single-cell level experiments with theoretical modeling. Our data indicate an unexpected, rapid absorption and retention of a large number of LL37 byE. colicells upon the inhibition of their growth, which increases the chance of survival for the rest of population. Cultures with high-enough cell density exhibit two distinct subpopulations: a non-growing population that absorb peptides and a growing population that survive owing to the sequestration of the AMPs by others. A mathematical model based on this binary picture reproduces the rather surprising behaviors ofE. colicultures in the presence of LL37, including the increase of the minimum inhibitory concentration with cell density (even in dilute cultures) and the extensive lag in growth introduced by sub-lethal dosages of LL37.

scholarly journals Development of a semi-defined medium for high cell density cultivation of Escherichia coli in shake flasks: Part 1

2016 ◽  
Author(s):  
Wenfa Ng ◽  
Yen-Peng Ting
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Yeast Extract ◽  
High Cell Density ◽  
Defined Medium ◽  
High Cell ◽  
High Cell Density Cultivation ◽  
E Coli ◽  
Carbon And Nitrogen ◽  
Shake Flasks

Sufficient quantities of cells of consistent characteristics are needed for studying biological processes (at the population level) in many areas of applied microbiology. However, generating the requisite biomass by cell culture is usually the rate-limiting step of a project given the relatively low biomass yield of many commercial culture media in shake flasks. This work reports the formulation of a semi-defined medium that enabled aerobic high cell density cultivation of Escherichia coli DH5α (ATCC 53868) in shake flasks. The formulated medium (FM) comprises: a buffer system (K2HPO4: 12.54 g/L and KH2PO4: 2.31 g/L); vitamins and trace elements (yeast extract: 12.0 g/L); salts (NaCl: 5.0 g/L and MgSO4: 0.24 g/L); and carbon and nitrogen sources (D-Glucose: 6.0 g/L and NH4Cl: 1.5 g/L). Notable characteristics of this medium are: high buffer capacity (89 mM phosphate), 1:1 molar ratio between D-Glucose and NH4Cl, and yeast extract providing trace elements and a secondary source of carbon and nitrogen. Preliminary data revealed that an OD600nm of 9 was attained after 24 hours of cultivation at 37 oC, with glucose and NH4Cl as the main nutrients. At 48 hours, the OD600nm reached a maximum value of 11 with yeast extract providing the necessary nutrients for cell growth and biomass formation. The broth’s pH varied between 5.5 and 7.8 during cultivation. For comparison, the maximum OD600nm of E. coli grown in three commonly used complex media: Nutrient Broth, LB Lennox, and Tryptic Soy Broth (TSB) were 1.4, 3.2 and 9.2, respectively, under identical culture conditions. Finally, FM maintained the viability of a larger population of cells for three days - compared to a population collapse observed in TSB after one day. Collectively, the present findings suggested that the formulated medium might find use as a high cell density aerobic growth medium for E. coli in shake flasks. Part 2 of this work describes improvements in medium performance - specifically, higher cell yield as well as a shorter diauxic lag phase and total culture period – achieved through a small reduction in D-Glucose and NH4Cl concentrations in the medium composition. An abstract preprint of Part 2 is available at https://peerj.com/preprints/117/

scholarly journals Development of a semi-defined medium for high cell density cultivation of Escherichia coli in shake flasks: Part 1

2017 ◽  
Author(s):  
Wenfa Ng
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Yeast Extract ◽  
High Cell Density ◽  
Defined Medium ◽  
High Cell ◽  
High Cell Density Cultivation ◽  
E Coli ◽  
Carbon And Nitrogen ◽  
Shake Flasks

Sufficient quantities of cells of consistent characteristics are needed for studying biologicalprocesses (at the population level ) in many areas of applied microbiology. However, generating the requisite biomass by cell culture is usually the rate-limiting step of a project given the relatively low biomass yield of many commercial culture media in shake flask culture systems. This work reports the formulation of a semi-defined medium that enabled aerobic high cell density cultivation of Escherichia coli DH5α (ATCC 53868) in shake flasks. The formulated medium (FM) comprises: a buffer system (K2HPO4 : 12.54 g/L and KH2 PO4 : 2.31 g/L); vitamins and trace elements (yeast extract: 12.0 g/L); salts (NaCl: 5.0 g/L and MgSO4 : 0.24 g/L); and carbon and nitrogen sources (D-Glucose: 6.0 g/L and NH4Cl: 1.5 g/L). Notable characteristics of this medium are: high buffer capacity (89 mM phosphate), 1:1 molar ratio between D-Glucose and NH4Cl, and yeast extract providing trace elements and a secondary source of carbon and nitrogen. Preliminary data revealed an OD 600nm of 9 after 24 hours of cultivation at 37 oC, presumably with glucose and NH4Cl as the main nutrients. At 48 hours, an OD 600nm of 11 was attained with yeast extract providing the necessary nutrients for cell growth and biomass formation. The broth’s pH varied between 5.5 and 7.8 during cultivation. On the other hand, the maximum OD 600nm of E. coli grown in three commonly used complex media: Nutrient Broth, LB Lennox, and Tryptic Soy Broth (TSB) were 1.4, 3.2 and 9.2, respectively, under identical culture conditions. Finally, FM maintained the viability of a larger population of cells for three days, compared to a population collapse in TSB broth after one day. Collectively, the results suggested that the formulated medium might find use as a high cell density aerobic growth medium for E. coli in shake flasks. Part 2 of this work describes improvements in medium performance ; specifically, higher cell yield as well as a shorter diauxic lag phase and total culture period achieved through a small reduction in D-Glucose and NH4Cl concentrations in the medium composition. An abstract preprint of Part 2 is available at https://peerj.com/preprints/117/

scholarly journals Development of a semi-defined medium for high cell density cultivation of Escherichia coli in shake flask culture system

2015 ◽  
Author(s):  
Wenfa Ng ◽  
Yen-Peng Ting
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Yeast Extract ◽  
High Cell Density ◽  
Growth Media ◽  
Buffer System ◽  
High Cell ◽  
High Cell Density Cultivation ◽  
E Coli ◽  
Carbon And Nitrogen

Microbes in environmental studies should be cultured in growth media with characteristics as close to their original habitat as possible, and which also allows a high cell density to be attained for providing enough cells in subsequent experiments. This in-progress report describes the formulation of a medium with an environmentally-relevant composition, and which also affords aerobic high cell density cultivation of Escherichia coli DH5α in shake flasks. The formulated medium comprises four components: a buffer system (K2HPO4: 12.54 g/L and KH2PO4: 2.31 g/L), vitamins (yeast extract: 12.0 g/L), salts (NaCl: 5.0 g/L and MgSO4: 0.24 g/L), and carbon and nitrogen sources (D-Glucose: 6.0 g/L and NH4Cl: 1.5 g/L). Notable characteristics of this medium were: high capacity phosphate buffer system (89 mM phosphate); 1:1 molar ratio between D-Glucose and NH4Cl; and yeast extract providing trace elements and a secondary carbon and nitrogen source. Growth experiments revealed that an OD600nm of 9 was attained after 24 hours of cultivation at 37 oC. This phase of growth was largely fuelled by glucose and NH4Cl. After 48 hours, the OD600nm reached 11, which was fuelled by the mixture of carbohydrates, lipids and proteins in yeast extract. Broth’s pH varied between 5.5 and 7.8 during cultivation, which was in the range conducive for growth of E. coli. In comparison, the OD600nm of E. coli reached 1.4, 3.2, and 9.2 for three commonly used complex media; Nutrient Broth, LB Lennox, and Tryptic Soy Broth, respectively, over 48 hours under identical culture conditions. In addition, the formulated medium was able to maintain a large viable cell population for a longer period of time (three days) relative to Tryptic Soy Broth. Thus, preliminary data suggested that the formulated medium holds potential for use as a high cell density aerobic growth medium for Gram-negative bacteria.

scholarly journals Characterization of the Roles of Hemolysin and Other Toxins in Enteropathy Caused by Alpha-Hemolytic Escherichia coli Linked to Human Diarrhea

1998 ◽  
Vol 66 (5) ◽  
pp. 2040-2051 ◽  
Author(s):  
Simon J. Elliott ◽  
S. Srinivas ◽  
M. John Albert ◽  
Khorshed Alam ◽  
Roy M. Robins-Browne ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Virulence Factor ◽  
Adult Rabbit ◽  
Cell Hyperplasia ◽  
E Coli ◽  
Alpha Hemolysin ◽  
A Cell ◽  
Cytotoxic Necrotizing Factor 1

ABSTRACT Escherichia coli strains producing alpha-hemolysin have been associated with diarrhea in several studies, but it has not been clearly demonstrated that these strains are enteropathogens or that alpha-hemolysin is an enteric virulence factor. Such strains are generally regarded as avirulent commensals. We examined a collection of diarrhea-associated hemolytic E. coli (DHEC) strains for virulence factors. No strain produced classic enterotoxins, but they all produced an alpha-hemolysin that was indistinguishable from that of uropathogenic E. coli strains. DHEC strains also produced other toxins including cytotoxic necrotizing factor 1 (CNF1) and novel toxins, including a cell-detaching cytotoxin and a toxin that causes HeLa cell elongation. DHEC strains were enteropathogenic in the RITARD (reversible intestinal tie adult rabbit diarrhea) model of diarrhea, causing characteristic enteropathies, including inflammation, necrosis, and colonic cell hyperplasia in both small and large intestines. Alpha-hemolysin appeared to be a major virulence factor in this model since it conferred virulence to nonpathogenic E. colistrains. Other virulence factors also appear to be contributing to virulence. These findings support the epidemiologic link to diarrhea and suggest that further research into the role of DHEC and alpha-hemolysin in enteric disease is warranted.

Determining Mechanical Properties of Escherichia Coli by Nanoindentation

2012 ◽  
Vol 1424 ◽  
Author(s):  
C. A. Wright ◽  
C.J. Sullivan ◽  
B. Crawford ◽  
L.D. Britt ◽  
M.A. Mamun ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mechanical Properties ◽  
Cell Wall ◽  
Contact Area ◽  
Solute Concentration ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
A Cell ◽  
Afm Cantilever

ABSTRACTEscherichia coli, like other gram-negative bacteria, is protected from the surrounding harsh environment by a cell wall consisting of the peptidoglycan and outer membrane. Whereas the cytoplasmic membrane is the selective barrier, the cell wall provides mechanical strength for the cell. As bacteria navigate various environments, osmotic pressure can change dramatically due to changes in local solute concentration. The peptidoglycan together with the cellular proteins mitigates the osmotic stress that would otherwise cause lysis. The mechanical properties of E. coli cells and its individual layers have been largely indeterminable until the recent development of probe-based measurement tools. Since their invention, scientists have reported significant data measuring elasticity, modulus, and stiffness using atomic force microscopy (AFM). Fundamentally, in order to determine these mechanical properties through probe-based techniques, the contact area and load should be well defined. The load can be precisely calculated through the AFM cantilever spring constant. However, the silicon tip contact area can only be estimated, potentially leading to compounding uncertainties. Therefore, we developed a methodology to determine nanomechanical properties of E. coli using a nanoindenter.

scholarly journals Transfer characteristics of two resistance determinants in a wild strain of Klebsiella aerogenes (V9A)

1970 ◽  
Vol 16 (2) ◽  
pp. 235-240 ◽  
Author(s):  
E. C. R. Reeve
Keyword(s):  
Escherichia Coli ◽  
Cell Attachment ◽  
Wild Strain ◽  
Klebsiella Aerogenes ◽  
Sex Factors ◽  
Chromosomal Gene ◽  
E Coli ◽  
Resistance Determinants ◽  
A Cell ◽  
Plasmid F

SUMMARYKlebsiella aerogenes strain V9A carries determinants AK and TK, giving resistance to ampicillin and tetracycline, and a plasmid FKlac, but no active sex factor. F− and I-type sex factors were able to transfer TK from V9A to Escherichia coli K12 and between strains of K12, and TK behaved as a separate plasmid with its own replicon. AK could not be transferred, except possibly by a sex factor carrying its own A determinant, but the evidence for such transfer was inconclusive. It is suggested that AK is either a chromosomal gene or is in a plasmid with a cell attachment in Klebsiella not represented in E. coli K12. AK produces a β-lactamase.

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