scholarly journals Primer reporte de Escherichia coli Productora de Toxina Shiga no O157 que codifica el gen de la enterohemolisina en carne cruda en Colombia

2019 ◽  
Vol 69 (1) ◽  
pp. 59-67
Author(s):  
Fanny Herrera Arias ◽  
Jesús Santos Buelga ◽  
Raquel Villamizar Gallardo

Dentro de los patotipos de Escherichia coli, el grupo STEC puede producir en el ser humano desde diarrea hemorrágica hasta insuficiencia renal aguda e incluso la muerte; el ganado bovino es el principal reservorio de este agente patógeno y por ende la ingestión de alimentos derivados de estos animales de abasto son una fuente muy importante de infección para el hombre. El objetivo principal de este estudio fue determinar la prevalencia de STEC en muestras de carne cruda comercializada en Pamplona-Colombia y en cepas obtenidas a partir de las muestras. Se analizaron cien muestras de carne cruda aplicando la técnica de Reacción en Cadena de la Polimerasa para la detección de los siguientes genes en muestras y en cepas STEC: stx1, stx2, eae y hlyA. Adicionalmente, se estableció el patrón de resistencia-susceptibilidad antibiótica de cepas STEC aisladas empleando métodos regulados. En el 39% de las muestras fue posible detectar el gen stx2; en el 38%, de ellas, se detectaron los genes stx1 y stx2. Además, se aislaron cepas STEC en el 13% de las muestras analizadas, 85% de ellas portaban el gen hlyA. No se detectó la presencia del gen eae o del serogrupo O157. Las cepas aisladas demostraron resistencia frente a algunos antibióticos de primera y segunda generación. En conclusión, se detectó la presencia de genes que codifican factores de virulencia en las muestras de carne analizadas que representan un riesgo potencial para la salud de los consumidores. Este es el primer reporte de STEC no O157 que codifica el gen de la enterohemolisina en alimentos en Colombia. Within the Escherichia coli patotypes, the STEC group can produce in humans from hemorrhagic diarrhea to acute renal failure and even death; cattle are the main reservoir of this pathogen and therefore the ingestion of food derived from these stock animals are a very important source of infection for man. The main objective of this study was to determine the prevalence of STEC in raw meat samples marketed in Pamplona-Colombia and in strains obtained from those samples. One hundred raw meat samples were analyzed using the Polymerase Chain Reaction technique for the detection of the following genes in samples and in STEC strains: stx1, stx2, eae and hlyA. In addition, STEC strains were isolated in 13% of the analyzed samples, 85% of them carried the hlyA gene. The presence of the eae gene or serogroup O157 was not detected. The isolated strains demonstrated resistance against some first and second generation antibiotics. In conclusion, the presence of genes encoding virulence factors in the meat samples analyzed, that represent a potential health risk factor to consumers, was confirmed. This is the first report of STEC non-O157 that encodes the enterohemolysin gene in foods in Colombia.

2017 ◽  
Vol 11 (07) ◽  
pp. 549-556 ◽  
Author(s):  
Hajer Kilani ◽  
Mohamed Salah Abbassi ◽  
Sana Ferjani ◽  
Rakia Ben Salem ◽  
Riadh Mansouri ◽  
...  

Introduction: The virulent Escherichia coli strains responsible for extraintestinal infections were mainly belonged to B2 and D phylogroups. However, no past studies have determinate via the presence of virulence genes the frequency of E. coli pathovars recovered from animals housed in farms in Tunisia. The aims of this study were to investigate 26 E. coli isolated from healthy and diarrheic animals and to determinate via the presence of virulence genes the frequency of pathovars. Methodology: Twenty-six E. coli isolates of phylogroups B2 (n = 14), B22 (n = 9), B23 (n = 5), and D2 (n = 12) were characterized. Genes encoding virulence factors (fimH,eaeA,aggC,papC, papG allele III, hlyA, east1, cnf1, exhA,stx1, stx2, iutA, fyuA, ibeA,and ipaH), and antibiotic resistance as well as class 1 and 2 integrons were searched by polymerase chain reaction (PCR). The genetic relationship of isolates was done by PFGE. Results: According to the occurrence of specific genes the 26 isolates were classified as:9 EAEC, 2 EHEC, 4 UPEC, 3 EPEC/EHEC and 1 NTEC. Therefore, 2 Ex-PEC and 5 APEC were presented amongst our strains. Some isolates (12) were clonal and the remaining was unrelated. Conclusions: Higher diversity of pathovars which carried diverse combinations of virulence genes in healthy isolates. In addition, it seems that the infections were caused by different mechanisms.


2015 ◽  
Vol 2 (2) ◽  
pp. 26-31 ◽  
Author(s):  
A. Paliy ◽  
A. Zavgorodniy ◽  
B. Stegniy ◽  
A. Gerilovych

Due to the absence of elaborated effi cient means for specifi c prevention of bovine tuberculosis, it is ex- tremely important to detect and eliminate the source of infection and to take veterinary and sanitary preven- tive measures. Here the critical role is attributed to disinfection, which breaks the epizootic chain due to the elimination of pathogenic microorganisms in the environment and involves the application of disinfectants of different chemical groups. Aim. To study the tuberculocidal properties of new disinfectants DZPT-2 and FAG against atypical mycobacteria Mycobacterium fortitum and a TB agent Mycobacterium bovis. Methods. The bacteriological and molecular-genetic methods were used. Results. It was determined that DZPT-2 prepara- tion has bactericidal effect on M. fortuitum when used in the concentration of 2.0 % of the active ingredient (AI) when exposed for 5–24 h, while disinfectant FAG has a bactericidal effect in the concentration of 2.0 % when exposed for 24 h. Disinfectant DZPT-2 in the concentration of 2.0 % of the AI, when exposed for 5–24 h, and FAG preparation in the concentration of 2.0 %, when exposed for 24 h, and with the norm of consump- tion rate of 1 cubic decimeter per 1 square meter disinfect the test-objects (batiste, wood, glazed tile, metal, glass), contaminated with the TB agent M. bovis. Conclusions. Disinfecting preparations of DZPT-2 in the concentration of 2.0 % of AI when exposed for 5 h and FAG in the concentration of 2.0 % when exposed for 24 h may be used in the complex of veterinary and sanitary measures to prevent and control TB of farm ani- mals. The possibility of using the polymerase chain reaction as an additional method of estimating tuberculo- cide activity of disinfectants was proven.


2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.


2008 ◽  
Vol 21 (10) ◽  
pp. 1325-1336 ◽  
Author(s):  
Jorrit-Jan Krijger ◽  
Ralf Horbach ◽  
Michael Behr ◽  
Patrick Schweizer ◽  
Holger B. Deising ◽  
...  

The hemibiotroph Colletotrichum graminicola is the causal agent of stem rot and leaf anthracnose on Zea mays. Following penetration of epidermal cells, the fungus enters a short biotrophic phase, followed by a destructive necrotrophic phase of pathogenesis. During both phases, secreted fungal proteins are supposed to determine progress and success of the infection. To identify genes encoding such proteins, we constructed a yeast signal sequence trap (YSST) cDNA-library from RNA extracted from mycelium grown in vitro on corn cell walls and leaf extract. Of the 103 identified unigenes, 50 showed significant similarities to genes with a reported function, 25 sequences were similar to genes without a known function, and 28 sequences showed no similarity to entries in the databases. Macroarray hybridization and quantitative reverse-transcriptase polymerase chain reaction confirmed that most genes identified by the YSST screen are expressed in planta. Other than some genes that were constantly expressed, a larger set showed peaks of transcript abundances at specific phases of pathogenesis. Another set exhibited biphasic expression with peaks at the biotrophic and necrotrophic phase. Transcript analyses of in vitro-grown cultures revealed that several of the genes identified by the YSST screen were induced by the addition of corn leaf components, indicating that host-derived factors may have mimicked the host milieu.


1999 ◽  
Vol 62 (10) ◽  
pp. 1115-1122 ◽  
Author(s):  
A. E. HEUVELINK ◽  
J. T. M. ZWARTKRUIS-NAHUIS ◽  
R. R. BEUMER ◽  
D E. de BOER

In 1996 and 1997, 2,941 fresh and processed meat products obtained from supermarkets and butcher shops in The Netherlands were examined for the presence of verocytotoxin-producing Escherichia coli of serogroup O157 (O157 VTEC). Additionally, the fate of O157 VTEC in raw meat products stored at low temperatures and the effect of different additives were evaluated. O157 VTEC strains were isolated from 6 (1.1%) of 571 samples of raw minced beef, 2 (0.5%) of 402 samples of raw minced mixed beef and pork, 1 (1.3%) of 76 samples of raw minced pork, 1 (0.3%) of 393 samples of other raw pork products, and 1 (0.3%) of 328 samples of cooked or fermented ready-to-eat meats. Other raw beef products (n = 223) and meat samples originating from poultry (n = 819), sheep or lamb (n = 46), or wild animals (n = 83) were all found to be negative for O157 VTEC. For the survival experiments we used tartaar (minced beef with a fat content of less than 10%) and filet americain (tartaar mixed with a mayonnaise-based sauce [80 to 20%]). The O157 VTEC strain tested was able to survive in tartaar and filet americain stored at −20, 0, 5, or 7°C for 3 days. At both 7 and at 15°C, O157 VTEC counts in tartaar and filet americain remained virtually unchanged throughout a storage period of 5 days. Addition of acetic acid (to pH 4.0), sodium lactate (1 and 2% [wt/wt]), or components of the lactoperoxidase–thiocyanate–hydrogen peroxide system to filet americain did not result in a reduction of viable O157 VTEC cells during storage at 7 or 15°C. It was concluded that raw meat contaminated with O157 VTEC will remain a hazard even if the meat is held at low or freezing temperatures.


2005 ◽  
Vol 37 (4) ◽  
pp. 265-269 ◽  
Author(s):  
Xi-Qiang Zhu ◽  
Su-Xia Li ◽  
Hua-Jun He ◽  
Qin-Sheng Yuan

Abstract The EC-SOD cDNA was cloned by polymerase chain reaction (PCR) and inserted into the Escherichia coli expression plasmid pET-28a(+) and transformed into E. coli BL21(DE3). The corresponding protein that was overexpressed as a recombinant His6-tagged EC-SOD was present in the form of inactive inclusion bodies. This structure was first solubilized under denaturant conditions (8.0 M urea). Then, after a capture step using immobilized metal affinity chromatography (IMAC), a gradual refolding of the protein was performed on-column using a linear urea gradient from 8.0 M to 1.5 M in the presence of glutathione (GSH) and oxidized glutathione (GSSG). The mass ratio of GSH to GSSG was 4:1. The purified enzyme was active, showing that at least part of the protein was properly refolded. The protein was made concentrated by ultrafiltration, and then isolated using Sephacryl S-200 HR. There were two protein peaks in the A280 profile. Based on the results of electrophoresis, we concluded that the two fractions were formed by protein subunits of the same mass, and in the fraction where the molecular weight was higher, the dimer was formed through the disulfide bond between subunits. Activities were detected in the two fractions, but the activity of the dimer was much higher than that of the single monomer. The special activities of the two fractions were found to be 3475 U/mg protein and 510 U/mg protein, respectively.


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