scholarly journals Protocol for evaluating the abilities of diverse nitroaromatic prodrug metabolites to exit a model Gram negative bacterial vector

2020 ◽  
Author(s):  
JVE Chan-Hyams ◽  
David Ackerley
Keyword(s):  
Escherichia Coli ◽  
Flow Cytometry ◽  
Bystander Effect ◽  
Recipient Strain ◽  
Converting Enzyme ◽  
Microtitre Plate ◽  
Enzyme Prodrug Therapy ◽  
Gram Negative ◽  
E Coli ◽  
Bacterial Vector

© 2020 The Authors Bacterial-directed enzyme-prodrug therapy (BDEPT) uses tumour-tropic bacteria armed with a genetically-encoded prodrug-converting enzyme to sensitise tumours to a systemically-administered prodrug. A strong bystander effect (i.e., efficient bacteria-to-tumour transfer of activated prodrug metabolites) is critical to maximise tumour cell killing and avoid bacterial self-sterilisation. To investigate the bystander effect in bacteria we developed a sensitive screen that utilised two Escherichia coli strains grown in co-culture. The first of these was an activator strain that overexpressed the E. coli nitroreductase NfsA, and the second was a nitroreductase null recipient strain bearing an SOS-GFP DNA damage responsive gene construct. In this system, induction of GFP by genotoxic prodrug metabolites can only occur following their transfer from the activator to the recipient cells. This can be monitored both in fluorescence based microtitre plate assays and by flow-cytometry, enabling modelling of the abilities of diverse nitroaromatic prodrug metabolites to exit a Gram negative vector.

scholarly journals Protocol for evaluating the abilities of diverse nitroaromatic prodrug metabolites to exit a model Gram negative bacterial vector

2020 ◽  
Author(s):  
JVE Chan-Hyams ◽  
David Ackerley
Keyword(s):  
Escherichia Coli ◽  
Flow Cytometry ◽  
Bystander Effect ◽  
Recipient Strain ◽  
Converting Enzyme ◽  
Microtitre Plate ◽  
Enzyme Prodrug Therapy ◽  
Gram Negative ◽  
E Coli ◽  
Bacterial Vector

© 2020 The Authors Bacterial-directed enzyme-prodrug therapy (BDEPT) uses tumour-tropic bacteria armed with a genetically-encoded prodrug-converting enzyme to sensitise tumours to a systemically-administered prodrug. A strong bystander effect (i.e., efficient bacteria-to-tumour transfer of activated prodrug metabolites) is critical to maximise tumour cell killing and avoid bacterial self-sterilisation. To investigate the bystander effect in bacteria we developed a sensitive screen that utilised two Escherichia coli strains grown in co-culture. The first of these was an activator strain that overexpressed the E. coli nitroreductase NfsA, and the second was a nitroreductase null recipient strain bearing an SOS-GFP DNA damage responsive gene construct. In this system, induction of GFP by genotoxic prodrug metabolites can only occur following their transfer from the activator to the recipient cells. This can be monitored both in fluorescence based microtitre plate assays and by flow-cytometry, enabling modelling of the abilities of diverse nitroaromatic prodrug metabolites to exit a Gram negative vector.

Synthesis of Chalcones and Nucleosides Incorporating [1, 3, 4]Oxadiazolenone Core and Evaluation of their Antifungal and Antibacterial Activities

2020 ◽  
Vol 15 (6) ◽  
pp. 665-679
Author(s):  
Alok K. Srivastava ◽  
Lokesh K. Pandey
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Antibacterial Activity ◽  
Bacillus Subtilis ◽  
Antifungal Activity ◽  
Aspergillus Niger ◽  
Gram Negative ◽  
E Coli ◽  
Gram Positive Bacteria ◽  
Good Antibacterial Activity

Background: [1, 3, 4]oxadiazolenone core containing chalcones and nucleosides were synthesized by Claisen-Schmidt condensation of a variety of benzaldehyde derivatives, obtained from oxidation of substituted 5-(3/6 substituted-4-Methylphenyl)-1, 3, 4-oxadiazole-2(3H)-one and various substituted acetophenone. The resultant chalcones were coupled with penta-O-acetylglucopyranose followed by deacetylation to get [1, 3, 4] oxadiazolenone core containing chalcones and nucleosides. Various analytical techniques viz IR, NMR, LC-MS and elemental analysis were used to confirm the structure of the synthesised compounds.The compounds were targeted against Bacillus subtilis, Staphylococcus aureus and Escherichia coli for antibacterial activity and Aspergillus flavus, Aspergillus niger and Fusarium oxysporum for antifungal activity. Methods: A mixture of Acid hydrazides (3.0 mmol) and N, Nʹ- carbonyl diimidazole (3.3 mmol) in 15 mL of dioxane was refluxed to afford substituted [1, 3, 4]-oxadiazole-2(3H)-one. The resulted [1, 3, 4]- oxadiazole-2(3H)-one (1.42 mmol) was oxidized with Chromyl chloride (1.5 mL) in 20 mL of carbon tetra chloride and condensed with acetophenones (1.42 mmol) to get chalcones 4. The equimolar ratio of obtained chalcones 4 and β -D-1,2,3,4,6- penta-O-acetylglucopyranose in presence of iodine was refluxed to get nucleosides 5. The [1, 3, 4] oxadiazolenone core containing chalcones 4 and nucleosides 5 were tested to determined minimum inhibitory concentration (MIC) value with the experimental procedure of Benson using disc-diffusion method. All compounds were tested at concentration of 5 mg/mL, 2.5 mg/mL, 1.25 mg/mL, 0.62 mg/mL, 0.31 mg/mL and 0.15 mg/mL for antifungal activity against three strains of pathogenic fungi Aspergillus flavus (A. flavus), Aspergillus niger (A. niger) and Fusarium oxysporum (F. oxysporum) and for antibacterial activity against Gram-negative bacterium: Escherichia coli (E. coli), and two Gram-positive bacteria: Staphylococcus aureus (S. aureus) and Bacillus subtilis(B. subtilis). Result: The chalcones 4 and nucleosides 5 were screened for antibacterial activity against E. coli, S. aureus and B. subtilis whereas antifungal activity against A. flavus, A. niger and F. oxysporum. Compounds 4a-t showed good antibacterial activity whereas compounds 5a-t containing glucose moiety showed better activity against fungi. The glucose moiety of compounds 5 helps to enter into the cell wall of fungi and control the cell growth. Conclusion: Chalcones 4 and nucleosides 5 incorporating [1, 3, 4] oxadiazolenone core were synthesized and characterized by various spectral techniques and elemental analysis. These compounds were evaluated for their antifungal activity against three fungi; viz. A. flavus, A. niger and F. oxysporum. In addition to this, synthesized compounds were evaluated for their antibacterial activity against gram negative bacteria E. Coli and gram positive bacteria S. aureus, B. subtilis. Compounds 4a-t showed good antibacterial activity whereas 5a-t showed better activity against fungi.

scholarly journals Proteomic response of Escherichia coli to a membrane lytic and iron chelating truncated Amaranthus tricolor defensin

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Tessa B. Moyer ◽  
Ashleigh L. Purvis ◽  
Andrew J. Wommack ◽  
Leslie M. Hicks
Keyword(s):  
Escherichia Coli ◽  
Antibacterial Activity ◽  
Antimicrobial Peptides ◽  
Mechanism Of Action ◽  
Gram Negative Bacteria ◽  
Amaranthus Tricolor ◽  
Core Motif ◽  
Gram Negative ◽  
E Coli ◽  
Membrane Lysis

Abstract Background Plant defensins are a broadly distributed family of antimicrobial peptides which have been primarily studied for agriculturally relevant antifungal activity. Recent studies have probed defensins against Gram-negative bacteria revealing evidence for multiple mechanisms of action including membrane lysis and ribosomal inhibition. Herein, a truncated synthetic analog containing the γ-core motif of Amaranthus tricolor DEF2 (Atr-DEF2) reveals Gram-negative antibacterial activity and its mechanism of action is probed via proteomics, outer membrane permeability studies, and iron reduction/chelation assays. Results Atr-DEF2(G39-C54) demonstrated activity against two Gram-negative human bacterial pathogens, Escherichia coli and Klebsiella pneumoniae. Quantitative proteomics revealed changes in the E. coli proteome in response to treatment of sub-lethal concentrations of the truncated defensin, including bacterial outer membrane (OM) and iron acquisition/processing related proteins. Modification of OM charge is a common response of Gram-negative bacteria to membrane lytic antimicrobial peptides (AMPs) to reduce electrostatic interactions, and this mechanism of action was confirmed for Atr-DEF2(G39-C54) via an N-phenylnaphthalen-1-amine uptake assay. Additionally, in vitro assays confirmed the capacity of Atr-DEF2(G39-C54) to reduce Fe3+ and chelate Fe2+ at cell culture relevant concentrations, thus limiting the availability of essential enzymatic cofactors. Conclusions This study highlights the utility of plant defensin γ-core motif synthetic analogs for characterization of novel defensin activity. Proteomic changes in E. coli after treatment with Atr-DEF2(G39-C54) supported the hypothesis that membrane lysis is an important component of γ-core motif mediated antibacterial activity but also emphasized that other properties, such as metal sequestration, may contribute to a multifaceted mechanism of action.

scholarly journals A High-Throughput Approach To Identify Compounds That Impair Envelope Integrity in Escherichia coli

2016 ◽  
Vol 60 (10) ◽  
pp. 5995-6002 ◽  
Author(s):  
Kristin R. Baker ◽  
Bimal Jana ◽  
Henrik Franzyk ◽  
Luca Guardabassi
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
High Throughput Screening ◽  
Gram Negative Bacteria ◽  
Chromogenic Substrate ◽  
Intrinsic Resistance ◽  
Gram Negative ◽  
Content Type ◽  
E Coli ◽  
Screening Platform

ABSTRACTThe envelope of Gram-negative bacteria constitutes an impenetrable barrier to numerous classes of antimicrobials. This intrinsic resistance, coupled with acquired multidrug resistance, has drastically limited the treatment options against Gram-negative pathogens. The aim of the present study was to develop and validate an assay for identifying compounds that increase envelope permeability, thereby conferring antimicrobial susceptibility by weakening of the cell envelope barrier in Gram-negative bacteria. A high-throughput whole-cell screening platform was developed to measureEscherichia colienvelope permeability to a β-galactosidase chromogenic substrate. The signal produced by cytoplasmic β-galactosidase-dependent cleavage of the chromogenic substrate was used to determine the degree of envelope permeabilization. The assay was optimized by using known envelope-permeabilizing compounds andE. coligene deletion mutants with impaired envelope integrity. As a proof of concept, a compound library comprising 36 peptides and 45 peptidomimetics was screened, leading to identification of two peptides that substantially increased envelope permeability. Compound 79 reduced significantly (from 8- to 125-fold) the MICs of erythromycin, fusidic acid, novobiocin and rifampin and displayed synergy (fractional inhibitory concentration index, <0.2) with these antibiotics by checkerboard assays in two genetically distinctE. colistrains, including the high-risk multidrug-resistant, CTX-M-15-producing sequence type 131 clone. Notably, in the presence of 0.25 μM of this peptide, both strains were susceptible to rifampin according to the resistance breakpoints (R> 0.5 μg/ml) for Gram-positive bacterial pathogens. The high-throughput screening platform developed in this study can be applied to accelerate the discovery of antimicrobial helper drug candidates and targets that enhance the delivery of existing antibiotics by impairing envelope integrity in Gram-negative bacteria.

scholarly journals Multiplex Polymerase Chain Reaction Assay for Identification of Enterotoxigenic Escherichia Coli Strains

2001 ◽  
Vol 13 (4) ◽  
pp. 308-311 ◽  
Author(s):  
Jacek Osek
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Direct Determination ◽  
Enteric Bacteria ◽  
Enterotoxigenic Escherichia Coli ◽  
Chain Reaction ◽  
Gram Negative ◽  
E Coli ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.

scholarly journals GREEN SYNTHESIS OF MAGNETIC IRON NANOPARTICLES USING MEDICINAL PLANT TRIDAX PROCUMBENS LEAF EXTRACTS AND ITS APPLICATION AS AN ANTIMICROBIAL AGENT AGAINST E. COLI

2020 ◽  
pp. 34-39
Author(s):  
YOJANA Y. PATIL ◽  
VAISHNVI B. SUTAR ◽  
ARPITA P. TIWARI
Keyword(s):  
Escherichia Coli ◽  
Magnetic Nanoparticles ◽  
Iron Nanoparticles ◽  
Most Probable Number ◽  
Leaf Extracts ◽  
Probable Number ◽  
Gram Negative ◽  
E Coli ◽  
Tridax Procumbens ◽  
Escherichia Coli Bacteria

Objective: The present study was aimed at the biological synthesis of magnetic iron nanoparticles by using the plant extract of Tridax procumbens and also to study their antimicrobial property against gram-negative bacteria (Escherichia coli). Methods: The synthesis of magnetic iron nanoparticles was carried out by the co-precipitation method using biological methods like plant extract as reducing agent and capping agents are biocompatible and non-hazardous. These nanoparticles were characterized by UV-Visible spectroscopy, XRD (X-Ray Diffraction), and SEM (Scanning Electron Microscope). As well as antibacterial activity of the nanoparticles was carried out by agar well diffusion method and Most Probable Number (MPN) method against gram-negative E. coli (Escherichia coli) bacteria. Results: The average crystallite size of Magnetic Nanoparticles (MNPs) was found to be 72 nm by X-ray diffraction. The optical absorption band at wavelengths of 240 nm and 402 nm was obtained from the UV Visible spectrum. Spherical shape morphology was observed in SEM studies. The antibacterial assay clearly expressed that E. coli showed a maximum zone of inhibition (15±0.15 mm) at 2 mg/ml and 1 mg/ml concentration was found for Magnetic Nanoparticles. In the Most Probable Number (MPN) test it is seen that the bacterial count is reduced after adding synthesized NPs into the water sample. Conclusion: The results of the present study conclude that the Magnetic Nanoparticles synthesized using Tridax procumbens leaf extracts is found to be stable and show good antibacterial activity against gram-negative (Escherichia coli) bacteria.

scholarly journals Evaluating the abilities of diverse nitroaromatic prodrug metabolites to exit a model Gram negative vector for bacterial-directed enzyme-prodrug therapy

2020 ◽  
Author(s):  
JVE Chan-Hyams ◽  
JN Copp ◽  
JB Smaill ◽  
AV Patterson ◽  
David Ackerley
Keyword(s):  
Human Cell ◽  
Bacterial Cell ◽  
Bystander Effect ◽  
Cell Transfer ◽  
Bacterial Strains ◽  
Enzyme Prodrug Therapy ◽  
E Coli ◽  
Human Cell Cultures ◽  
Cell To Cell Transfer ◽  
Nitro Reduction

© 2018 Elsevier Inc. Gene-directed enzyme-prodrug therapy (GDEPT) employs tumour-tropic vectors including viruses and bacteria to deliver a genetically-encoded prodrug-converting enzyme to the tumour environment, thereby sensitising the tumour to the prodrug. Nitroreductases, able to activate a range of promising nitroaromatic prodrugs to genotoxic metabolites, are of great interest for GDEPT. The bystander effect (cell-to-cell transfer of activated prodrug metabolites) has been quantified for some nitroaromatic prodrugs in mixed multilayer human cell cultures, however while these provide a good model for viral DEPT (VDEPT) they do not inform on the ability of these prodrug metabolites to exit bacterial vectors (relevant to bacterial-DEPT (BDEPT)). To investigate this we grew two Escherichia coli strains in co-culture; an activator strain expressing the nitroreductase E. coli NfsA and a recipient strain containing an SOS-GFP DNA damage responsive gene construct. In this system, induction of GFP by reduced prodrug metabolites can only occur following their transfer from the activator to the recipient cells. We used this to investigate five clinically relevant prodrugs: metronidazole, CB1954, nitro-CBI-DEI, and two dinitrobenzamide mustard prodrug analogues, PR-104A and SN27686. Consistent with the bystander efficiencies previously measured in human cell multilayers, reduced metronidazole exhibited little bacterial cell-to-cell transfer, whereas nitro-CBI-DEI was passed very efficiently from activator to recipient cells post-reduction. However, in contrast with observations in human cell multilayers, the nitrogen mustard prodrug metabolites were not effectively passed between the two bacterial strains, whereas reduced CB1954 was transferred efficiently. Using nitroreductase enzymes that exhibit different biases for the 2- versus 4-nitro substituents of CB1954, we further showed that the 2-nitro reduction products exhibit substantially higher levels of bacterial cell-to-cell transfer than the 4-nitro reduction products, consistent with their relative bystander efficiencies in human cell culture. Overall, our data suggest that prodrugs may differ in their suitability for VDEPT versus BDEPT applications and emphasise the importance of evaluating an enzyme-prodrug partnership in an appropriate context for the intended vector.

scholarly journals TOTAL Escherichia coli PADA SOSIS IKAN YANG DI-COATING DENGAN MIOFIBRIL ASAP CAIR SELAMA PENYIMPANAN

2016 ◽  
Vol 4 (1) ◽  
Author(s):  
Jupni Keno ◽  
Henny Adeleida Dien ◽  
Agnes Triasih Agustin
Keyword(s):  
Escherichia Coli ◽  
Nutritional Value ◽  
Room Temperature ◽  
Total Coliform ◽  
Fish Protein ◽  
Gram Negative ◽  
E Coli ◽  
Liquid Smoke ◽  
Refrigerator Temperature ◽  
Descriptive Method

Fish sausages are prepared foods that have a high nutritional value, but that is the weakness of this commodity is rapidly decaying nature. Bacterial pathogens that must be avoided include Escherichia coli. These bacteria are gram-negative, rod-shaped and motile spores are not. The purpose of this study is to calculate the total coliforms and E. coli in fish sausage coating of fish protein myofibrils Black Marlin (Makaira indica) during storage at room temperature (28–29°C), and refrigerator temperature (10–13°C). The method used is descriptive method, which is a study conducted to analyze an individual, the state, or the symptoms of a particular group. The results showed that the total coliform in fish sausage in coating with liquid smoke is stored at room temperature, the lowest value is 7 MPN/g, the highest of 120 MPN/g, while the lowest value refrigerator temperature is 7 MPN/g, the highest 93 MPN/g. Total coliform in fish sausage in smokeless liquid coating stored at room temperature with the lowest value is 7 MPN/g, the highest 210 MPN/g, while the lowest value refrigerator temperature is 7 MPN/g, and the highest is 120 MPN/g. Total coliform in fish sausages are not in the coating deposited at room temperature with the lowest value is 7 MPN/g, the highest of 240 MPN/g, at refrigerator temperature the lowest value is 7 MPN/g, and the highest is 150 MPN/g. Total E. coli showed that the fish sausage in coating with liquid smoke is stored at room temperature, the lowest value is 1 MPN/g, and the highest is 4 MPN/g, while the lowest value refrigerator temperature is <3 MPN/g, and The highest is 3 MPN/g. Total E. coli in fish sausage in smokeless coating liquid stored at room temperature, the lowest value is 2 MPN/g, and the highest is 4 MPN/g, while the temperature of the refrigerator lowest value is 1 MPN/g, and a high of 3 MPN/g. Total E. coli in sausages are not in the coating deposited at room temperature, the lowest value is 2 MPN/g, and the highest is 5 MPN/g, and the refrigerator temperature is the lowest rating 2 MPN/g, the highest is 4 MPN/g during storage .Keywords: fish sausage, coating, myofibril, Eschericia coli.  Sosis ikan merupakan makanan siap saji yang mempunyai nilai gizi tinggi, namun yang menjadi kelemahan dari komoniti ini adalah sifatnya yang cepat membusuk. Bakteri patogen yang harus dihindari antara lain Escherichia coli.  Bakteri ini bersifat gram negatif, berbentuk batang tidak spora dan bersifat motil. Tujuan penelitian ini yaitu untuk menghitung total koliform dan E. coli pada sosis ikan yang dicoating dari miofibril protein ikan Black Marlin (Makaira indica) selama penyimpanan suhu ruang (28–29°C), dan suhu kulkas (10–13°C). Metode penelitian yang digunakan adalah metode deskriptif, yaitu suatu penelitian yang dilakukan untuk menganalisa suatu individu, keadaan, gejala atau kelompok tertentu. Hasil penelitian menunjukkan bahwa total koliform pada sosis ikan yang dicoating dengan asap cair disimpan pada suhu ruang, nilai terendah yaitu 7 MPN/g, tertinggi 120 MPN/g, sedangkan pada suhu kulkas nilai yang terendah yaitu 7 MPN/g, tertinggi 93 MPN/g. Total koliform pada sosis ikan yang dicoating tanpa asap cair disimpan pada suhu ruang dengan nilai terendah yaitu 7 MPN/g, tertinggi 210 MPN/g, sedangkan pada suhu kulkas nilai yang terendah yaitu 7 MPN/g, dan tertinggi 120 MPN/g. Total koliform pada sosis ikan tidak dicoating disimpan pada suhu ruang dengan nilai terendah yaitu 7 MPN/g, tertinggi 240 MPN/g , pada suhu kulkas nilai terendah yaitu 7 MPN/g , dan tertinggi 150 MPN/g. Total E. coli menunjukkan bahwa pada sosis ikan yang dicoating dengan asap cair disimpan pada suhu ruang, yaitu nilai terendah 3 MPN/g, dan tertinggi 4 MPN/g, sedangkan pada suhu kulkas nilai terendah yaitu <3 MPN/g, dan tertinggi 3 MPN/g. Total E. coli pada sosis ikan yang dicoating tanpa asap cair disimpan pada suhu ruang, yaitu nilai terendah 3 MPN/g, dan tertinggi 4 MPN/g, sedangkan pada suhu kulkas nilai terendah yaitu <3 MPN/g , dan tertinggi 3 MPN/g. Total E. coli pada sosis tidak dicoatingdisimpan pada suhu ruang, yaitu nilai terendah 4 MPN/g, dan tertinggi 7 MPN/g, dan pada suhu kulkas yaitu nilai terendah 3 MPN/g, tertinggi 4 MPN/g selama penyimpanan.Kata kunci: sosis ikan, coating, myofibril, Eschericia coli.

Simplified scheme for identification of prompt lactose-fermenting members of the Enterobacteriaceae

1976 ◽  
Vol 4 (6) ◽  
pp. 511-514
Author(s):  
M J Hicks ◽  
K J Ryan
Keyword(s):  
Escherichia Coli ◽  
Ornithine Decarboxylase ◽  
Clinical Microbiology ◽  
Clinical Microbiology Laboratory ◽  
Microbiology Laboratory ◽  
Biochemical Tests ◽  
Gram Negative ◽  
E Coli ◽  
Colonial Morphology ◽  
Level Identification

A brief, simplified scheme involving the spot indole test and colonial morphology was evaluated for genus level identification of prompt lactose-fermenting (PLF) members of the Enterobacteriaceae. One hundred and ninety-four consecutive, clinically important PLF gram-negative rods isolated in a clinical microbiology laboratory were identified by this simplified scheme, as well as by standard biochemical tests, and the API 20E (Analytab Products, Inc., Plainview, N.Y.) system. In the simplified scheme a flat, spot indole-positive colony was identified as Escherichia coli. Spot indole-negative organisms forming nucoid colonies were identified as Klebsiella sp. or Enterobacter sp. on the basis of semisolid motility and ornithine decarboxylase tests. Approximately 94% of the study isolates followed reactions typical for E. coli, Klebsiella sp., and Enterobacter sp. as defined by this simplified scheme. When compared with the standard and Analytab Products Inc. identifications, the overall accuracy was 97.4%. The accuracy of identification of E. coli, Klebsiella sp., and Enterobacter sp. was 98.1%, 95.6%, and 87.5%, respectively. This simplified scheme is recommended for identification of selected PLF isolates in the clinical microbiology laboratory.

Methodology for Enteropathogenic Escherichia coli

1975 ◽  
Vol 58 (2) ◽  
pp. 283-292
Author(s):  
Ira J Mehlman ◽  
Nicholas T Simon ◽  
Arvey C Sanders ◽  
Morris Fishbein ◽  
Joseph C Olson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Elevated Temperature ◽  
Growth Temperature ◽  
Maximal Growth ◽  
Enteropathogenic Escherichia Coli ◽  
Pathogenic Potential ◽  
Gram Negative ◽  
Tentative Identification ◽  
E Coli ◽  
Capsular Antigens

Abstract Pathogenic biotypes of Escherichia coli grow poorly at temperatures greatly different from that of the host. Percentages quantitatively recovered at 42.0, 44.0, 44.5, and 45.5°C in lauryl tryptose broth were 100, 76, 76, and 42, respectively. Corresponding values for 175 strains of varied origin were 98, 89, 82, and 65%. Maximal growth temperature is dependent upon medium. Lauryl tryptose and elevated coliform broths were equivalent in the recovery of small inocula (100 cells/ml) at 41.5–44.5°. Mac-Conkey, enteric enrichment, and Gram-negative broths were inhibitory at corresponding values. Growth at elevated temperature in nutrient broth is enhanced by carbohydrate. Standard lactose enrichment media fail to recover slow lactose fermenters. An acidified glutamic acid medium was unsuitable for recovery of E. coli. The data suggest modification of standard temperatures for the recovery of pathogenic biotypes. Previously recommended analytical methods have been simplified and supplemented. The enhancement of motility in indole-nitrite broth at 35°C is recommended. A 4-tube semiquantitative test is offered for tentative identification of somatic and capsular antigens. Inclusion of Alkalescens-Dispar strains is warranted by their pathogenic behavior. Examination in Shigella and Alkalescens-Dispar sera is required to cover the dysentery-like biotypes. Pathogenic potential cannot be inferred from serotype.

Sign in / Sign up

Export Citation Format

Share Document