scholarly journals Isolation and characterization of enterohaemorrhagic Escherichia coli O157[ratio ]H7 from cattle in Belgium and Poland

2002 ◽  
Vol 129 (1) ◽  
pp. 41-47 ◽  
Author(s):  
A. V. TUTENEL ◽  
D. PIERARD ◽  
J. URADZINSKI ◽  
E. JOZWIK ◽  
M. PASTUSZCZAK ◽  
...  
Keyword(s):  
Phage Type ◽  
Immunomagnetic Separation ◽  
E Coli ◽  
Slaughter Cattle ◽  
Isolation And Characterization ◽  
Enterohaemorrhagic Escherichia Coli ◽  
Faecal Samples ◽  
Peptone Water ◽  
Ferment Lactose

EHEC O157 were isolated from faeces of Belgian and Polish beef slaughter cattle. In Belgium, 1281 faecal samples were analysed by immunomagnetic separation [IMS] after enrichment in buffered peptone water from June 1998 till July 1999. Eighty-one samples (6.3%) were positive for E. coli O157. Phage type 8 was most frequently found. Bulls between 1 and 2 years old, slaughtered in September and October were most frequently found positive. Atypical biochemical features were observed in some isolates: 22 (27%) isolates were urease positive and 1 (1.2%) isolate was unable to ferment lactose. In Poland, 551 faecal samples, taken from January 1999 till December 1999, were examined using exactly the same techniques. Four faecal samples (0.7%) were positive for O157 EHEC, yielding seven phage type 8 isolates. All positive samples were from cattle younger than 2 years. Positive samples occurred in August, September and October.

Isolation and characterization of Escherichia coli O157:H7 and shiga toxin-converting bacteriophages from strains of human, bovine and porcine origin

2002 ◽  
Vol 2 (3) ◽  
pp. 29-38 ◽  
Author(s):  
E.E. Müller ◽  
M.B. Taylor ◽  
W.O.K. Grabow ◽  
M.M. Ehlers
Keyword(s):  
Escherichia Coli ◽  
Pcr Analysis ◽  
Sewage Sample ◽  
Natural Environments ◽  
Escherichia Coli O157 ◽  
Electron Microscopic ◽  
E Coli ◽  
Isolation And Characterization ◽  
Faecal Samples

Toxin-converting bacteriophages encoding the Stx2 gene were induced from strains of Escherichia coli O157:H7 isolated from sewage, bovine and porcine faeces. Toxin synthesis can be stimulated by the induction of integrated toxin-converting phages from the host E. coli O157:H7 organism by ultra-violet (UV) exposure. The UV-mediated DNA damage of E. coli O157:H7 triggers a bacterial SOS response resulting in phage release. Free ranging phages outside their E. coli O157:H7 hosts were detected but could not be isolated directly from environmental samples such as sewage and river water. E. coli O157:H7 colonies carrying the genes coding for Stx2 were isolated from 1 sewage sample (0.76% of positive samples), 8 cattle faecal samples (16.67% of positive samples) and 2 pig faecal samples (14.28% of positive samples). Characterization of E. coli O157:H7 was done by repetitive sequence analysis using ERIC-PCR to determine the relationships between the individual E. coli O157:H7 strains. The ERIC-PCR analysis revealed distinct patterns for all E. coli O157:H7 strains with some small differences between the strains. DNA sequencing of some of the E. coli O157:H7 positive isolates carrying the Stx2 genes were performed confirming the amplified DNA nucleotide sequences of Stx2. Electron microscopic analysis revealed, for the first time in South Africa, that Stx2-converting phages induced from E. coli O157:H7 have different morphologies to that of phage lambda which was previously described. The role of the induced integrated Stx2 phages in natural environments such as river and dam water remains unclear. With the induction of Stx2-converting phages from environmental E. coli O157:H7 isolates, it is now possible to determine the potential of these phages to convert non-pathogenic E. coli strains and other enterobacteriaciae into pathogenic strains.

scholarly journals Immunomagnetic separation as a sensitive method for isolatingEscherichia coliO157 from food samples

1994 ◽  
Vol 113 (1) ◽  
pp. 31-39 ◽  
Author(s):  
D. J. Wright ◽  
P. A. Chapman ◽  
C. A. Siddons
Keyword(s):  
Magnetic Beads ◽  
Phage Type ◽  
Dairy Herd ◽  
Immunomagnetic Separation ◽  
Epidemiological Studies ◽  
Food Samples ◽  
Tween 20 ◽  
Specific Method ◽  
E Coli ◽  
Peptone Water

SUMMARYMinced beef samples inoculated withEscherichia coliO157 were cultured in buffered peptone water supplemented with vancomycin, cefsulodin and cefixime (BPW-VCC) and subcultured to cefixime tellurite sorbitol MacConkey (CT-SMAC) agar both directly and after immunomagnetic separation (IMS) of the organism with magnetic beads coated with an antibody againstE. coliO157 (Dynabeads anti-E. coli0157, Dynal, Oslo).E. coliO157 was recovered from initial inocula of 200 organisms/g by direct subculture and 2 organisms/g by IMS. Twelve strains ofE. coli0157 of different combinations of phage type, H antigen and toxin genotype were all recovered from initial inocula of two organisms/g by IMS. Nonspecific binding of other organisms to the magnetic beads could be reduced by washing of the beads in PBS with Tween-20 0·002–0.005%E. coliO157 was not bound by magnetic coated with an unrelated antibody.During investigation of a dairy herd that was possibly linked to a small outbreak of infection withE. coliO157, the organism was isolated from 2 of 279 forestream milk samples from individual cattle; both isolates were made only by the IMS technique. IMS is rapid, technically simple, and a specific method for isolation ofE. coliO157 and will be useful in epidemiological studies.

scholarly journals Infection with verocytotoxin-producing Escherichia coli O157 during a visit to an inner city open farm

2000 ◽  
Vol 125 (3) ◽  
pp. 531-536 ◽  
Author(s):  
P. A. CHAPMAN ◽  
J. CORNELL ◽  
C. GREEN
Keyword(s):  
Escherichia Coli ◽  
Inner City ◽  
Magnetic Beads ◽  
Profile Analysis ◽  
Enrichment Culture ◽  
Phage Type ◽  
Immunomagnetic Separation ◽  
E Coli ◽  
Zoonotic Infections ◽  
Faecal Samples

Two cases of Escherichia coli O157 infection occurred in children after visiting an inner city open farm. Subsequently faecal samples collected from animal pens and samples of composted mixed animal manure and vegetable waste were examined for E. coli O157 by enrichment culture, immunomagnetic separation and culture of magnetic beads to cefixime tellurite sorbitol MacConkey agar. Strains of E. coli O157 were characterized by hybridization with DNA probes for VT1, VT2 and eaeA, plasmid profile analysis, phage typing and pulsed field gel electrophoresis (PFGE). Verocytotoxin-producing E. coli O157 strains were isolated from faecal samples from a cow, a horse, 3 breeds of pigs, 2 breeds of sheep and 2 breeds of goats and from 2 samples of compost which had been processed for 3 months. All strains were phage type 21, hybridized with probes for VT2 and eaeA but not with one for VT1, harboured 92 and 2 kb plasmids and gave indistinguishable banding patterns with PFGE. Although only two culture-confirmed cases of infection had been identified, the farm had over 100 000 visitors per year and so it was closed as a precaution both to allow a thorough investigation and to prevent further cases. The investigation identified many factors which may have contributed to transmission of E. coli O157 infection. Most of these were readily resolved by appropriate corrective measures and as there were no further cases associated with the farm during the ensuing 4 weeks it then re-opened. These cases highlight the risk, especially to young children, of acquiring zoonotic infections during visits to open farms and emphasize the need for adequate guidance and supervision before and during such visits.

scholarly journals Isolation and characterization of escherichia coli in ready-to-eat foods vended in Islamic University, Kushtia

1970 ◽  
Vol 18 ◽  
pp. 99-103 ◽  
Author(s):  
S Biswas ◽  
MAK Parvez ◽  
M Shafiquzzaman ◽  
S Nahar ◽  
MN Rahman
Keyword(s):  
Escherichia Coli ◽  
Pattern Analysis ◽  
University Campus ◽  
Rflp Pattern ◽  
E Coli ◽  
Fish Meat ◽  
Isolation And Characterization ◽  
Food Borne ◽  
Identification And Characterization

Context: Escherichia coli is shed in the feces of warm blooded animals and humans and thus potential for public health. Detection and characterization of E. coli in the ready-to-eat (RTE) foods concerns due to their presence indicates fecal contamination of the food.   Objective: To identify, characterize and RFLP pattern analysis of E. coli isolated from RTE foods vended in Islamic University campus, Kushtia.   Materials and Methods: Fifty samples from four types of consumed foods in six student halls of residence, some temporary restaurants of Islamic University, Kushtia were assessed for bacterial contamination by standard methods. Identification and characterization of E. coli isolates were performed using IMViC tests. Genomic DNA was used to perform RFLP pattern analysis.   Results: Thirty seven out of 50 (74%) examined samples of RTE foods had E. coli contamination. The highest number of E. coli was isolated from vegetable oriented RTE foods (90.90%) and fish, meat and cereals samples were also significantly E. coli positive. RFLP profiling of two E. coli isolates were observed.   Conclusion: The results of this study provide evidence that some RTE foods had unsatisfactory levels of contamination with E. coli. Thus street vended RTE food could be important potential vehicles for food-borne diseases. Molecular characterization may be exploited to identify food borne pathogen among different species.  Keywords: Ready-to-eat foods; Escherichia coli; RFLP pattern DOI: http://dx.doi.org/10.3329/jbs.v18i0.8783 JBS 2010; 18(0): 99-103

Isolation and characterization of lytic bacteriophages against enterohaemorrhagic Escherichia coli

2011 ◽  
Vol 110 (5) ◽  
pp. 1323-1331 ◽  
Author(s):  
S. Viazis ◽  
M. Akhtar ◽  
J. Feirtag ◽  
A.D. Brabban ◽  
F. Diez-Gonzalez
Keyword(s):  
Escherichia Coli ◽  
Isolation And Characterization ◽  
Enterohaemorrhagic Escherichia Coli

scholarly journals Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species

2023 ◽  
Vol 83 ◽  
Author(s):  
M. Mushtaq ◽  
S. M. Bukhari ◽  
S. Ahmad ◽  
A. Khattak ◽  
M. B. Chattha ◽  
...  
Keyword(s):  
Bacterial Species ◽  
Gut Contents ◽  
Phasianus Colchicus ◽  
Gut Content ◽  
E Coli ◽  
Colony Forming Unit ◽  
Isolation And Characterization ◽  
Staphylococcus Spp ◽  
Campylobacter Spp

Abstract There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Isolation and Characterization of Levoglucosan Metabolizing Bacteria

2021 ◽  
Author(s):  
Ajay S. Arya ◽  
Minh T. H. Hang ◽  
Mark A. Eiteman
Keyword(s):  
Recombinant Expression ◽  
Bacterial Species ◽  
Soluble Carbohydrate ◽  
Rrna Gene ◽  
Gene Products ◽  
16S Rrna Gene Sequences ◽  
E Coli ◽  
Isolation And Characterization ◽  
Charred Wood

Bacteria were isolated from wastewater and soil containing charred wood remnants based on their ability to use levoglucosan as a sole carbon source and on their levoglucosan dehydrogenase (LGDH) activity. On the basis of their 16S rRNA gene sequences, these bacteria represented diverse genera of Microbacterium, Paenibacillus , Shinella , and Klebsiella . Genomic sequencing of the isolates verified that two isolates represented novel species, Paenibacillus athensensis MEC069 T and Shinella sumterensis MEC087 T , while the remaining isolates were closely related to either Microbacterium lacusdiani or Klebsiella pneumoniae . The genetic sequence of LGDH, lgdA , was found in the genomes of these four isolates as well as Pseudarthrobacter phenanthrenivorans Sphe3. The identity of the P. phenanthrenivorans LGDH was experimentally verified following recombinant expression in E. coli . Comparison of the putative genes surrounding lgdA in the isolate genomes indicated that several other gene products facilitate the bacterial catabolism of levoglucosan, including a putative sugar isomerase and several transport proteins. Importance Levoglucosan is the most prevalent soluble carbohydrate remaining after high temperature pyrolysis of lignocellulosic biomass, but it is not fermented by typical production microbes such as Escherichia coli and Saccharomyces cerevisiae . A few fungi metabolize levoglucosan via the enzyme levoglucosan kinase, while several bacteria metabolize levoglucosan via levoglucosan dehydrogenase. This study describes the isolation and characterization of four bacterial species which degrade levoglucosan. Each isolate is shown to contain several genes within an operon involved in levoglucosan degradation, furthering our understanding of bacteria which metabolize levoglucosan.

Paucimyces polynucleatus gen. nov, sp. nov., a novel polycentric genus of anaerobic gut fungi from the faeces of a wild blackbuck antelope

2021 ◽  
Vol 71 (6) ◽  
Author(s):  
Radwa A. Hanafy ◽  
Noha H. Youssef ◽  
Mostafa S. Elshahed
Keyword(s):  
Ribosomal Subunit ◽  
New Genus And Species ◽  
Isolation And Characterization ◽  
Faecal Samples ◽  
Wide Range ◽  
Culture Independent ◽  
Antilope Cervicapra ◽  
Spherical Vesicles ◽  
Sequence Similarities

The anaerobic gut fungi (AGF; phylum Neocallimastigomycota) reside in the alimentary tracts of herbivores. Multiple novel, yet-uncultured AGF taxa have recently been identified in culture-independent diversity surveys. Here, we report on the isolation and characterization of the first representative of the RH5 lineage from faecal samples of a wild blackbuck (Indian Antelope, Antilope cervicapra) from Sutton County, Texas, USA. The isolates displayed medium sized (2–4 mm) compact circular colonies on agar roll tubes and thin loose biofilm-like growth in liquid medium. Microscopic examination revealed monoflagellated zoospores and polycentric thalli with highly branched nucleated filamentous rhizomycelium, a growth pattern encountered in a minority of described AGF genera so far. The obtained isolates are characterized by formation of spherical vesicles at the hyphal tips from which multiple sporangia formed either directly on the spherical vesicles or at the end of sporangiophores. Phylogenetic analysis using the D1/D2 regions of the large ribosomal subunit (D1/D2 LSU) and the ribosomal internal transcribed spacer 1 (ITS1) revealed sequence similarities of 93.5 and 81.3%, respectively, to the closest cultured relatives (Orpinomyces joyonii strain D3A (D1/D2 LSU) and Joblinomyces apicalis strain GFH681 (ITS1). Substrate utilization experiments using the type strain (BB-3T) demonstrated growth capabilities on a wide range of mono-, oligo- and polysaccharides, including glucose, xylose, mannose, fructose, cellobiose, sucrose, maltose, trehalose, lactose, cellulose, xylan, starch and raffinose. We propose accommodating these novel isolates in a new genus and species, for which the name Paucimyces polynucleatus gen. nov., sp. nov. is proposed.

Cloning and expression of DiT33 from Dirofilaria immitis: a specific and early marker of heartworm infection

Parasitology ◽  
1996 ◽  
Vol 112 (3) ◽  
pp. 331-338 ◽  
Author(s):  
X. Q. Hong ◽  
J. Santiago Mejia ◽  
S. Kumar ◽  
F. B. Perler ◽  
C. K. S. Carlow
Keyword(s):  
Dirofilaria Immitis ◽  
Cloning And Expression ◽  
E Coli ◽  
Spliced Leader ◽  
Isolation And Characterization ◽  
The Family ◽  
Heartworm Infection ◽  
High Level ◽  
Control And Management

SUMMARYDirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6·5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvDSB) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described.‡ This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26·4 kDa-protein with a high level of similarity (87–89%) to other filarial members of the family. DJT33 was over-expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection.

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