scholarly journals Molecular characterization of enterohemorrhagic Escherichia coli isolated from diarrhea samples from human, livestock, and ground beef in North Jordan

2021 ◽  
pp. 2827-2832
Author(s):  
Yaser H. Tarazi ◽  
Saeb N. El-Sukhon ◽  
Zuhair Bani Ismail ◽  
Amani A. Almestarehieh
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogen ◽  
Ground Beef ◽  
Enterohemorrhagic Escherichia Coli ◽  
Diffusion Test ◽  
Disk Diffusion Test ◽  
Fecal Samples ◽  
E Coli ◽  
Beef Meat ◽  
Meat Samples

Background and Aim: Enterohemorrhagic Escherichia coli (EHEC) is an important foodborne pathogen with worldwide distribution. Data regarding its presence, distribution, virulence, and antimicrobial susceptibility among various animal species and humans in Jordan are lacking. Therefore, the objectives of this study were to isolate and characterize EHEC from human and animal diarrhea fecal samples and ground beef samples. Materials and Methods: A total of 100 and 270 diarrhea fecal samples from humans and animals, respectively, were collected. In addition, 40 ground beef meat samples were collected from retail markets. EHEC was positively identified by detecting Shiga toxins (stx1 and stx2) genes using multiplex polymerase chain reaction (PCR). Antimicrobial susceptibility patterns were determined using the disk diffusion test. Beta-lactamase production was detected using the double disk diffusion test and the extended-spectrum beta-lactamases (ESBLs) were identified by detection of blaTEM, blaSHV, and OXA-1 genes using multiplex PCR. Pulsed-field gel electrophoresis (PFGE) was used to investigate the relatedness of EHEC isolates from different sources. Results: Out of 410 samples, 194 E. coli isolates were positively identified, of which 57 isolates (29%) were classified as EHEC. Thirty-five (61%) of EHEC isolates were serotyped as O157 (19: O157:H7 and 16: O157:NM). The stx1 gene was detected only among the sheep and goats isolates at a rate of 7.6% and 5.2%, respectively, while the stx2 gene was detected in only one ground beef meat sample. EHEC isolates showed high resistance patterns against amoxicillin, gentamycin, cephalexin, and doxycycline. Twenty-four out of 32 EHEC isolates were determined as ESBL producers, among which 14 isolates expressed the blaSHV gene and 19 isolates expressed the blaTEM while four expressed both genes. PFGE analysis revealed two clusters with high similarity (92%) originated from ground beef meat and cattle fecal samples. No similarities were found between human and animal E. coli isolates. Conclusion: Results of this study indicate widespread ESBL EHEC among humans, animals, and ground beef meat samples. These results represent an important alarm that requires the implementation of appropriate preventative measures by both human and animal health sectors to prevent the transmission of this important foodborne pathogen.

Comparison of Rapid Enzyme-Linked Immunosorbent Assay and Immunomagnetic Separation Methods for Detection of Escherichia coli O157 in Fecal, Hide, Carcass, and Ground Beef Samples

2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Immunomagnetic Separation ◽  
Separation Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Escherichia Coli O157 ◽  
Perfect Agreement ◽  
Fecal Samples ◽  
E Coli ◽  
Beef Products

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.

scholarly journals Detex for Detection of Escherichia coli O157 in Raw Ground Beef and Raw Ground Poultry

2001 ◽  
Vol 84 (3) ◽  
pp. 752-760 ◽  
Author(s):  
Yvette M Henry ◽  
Nandini Natrajan ◽  
Wendy F Lauer
Keyword(s):  
Escherichia Coli ◽  
False Positive Rate ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Department Of Agriculture ◽  
E Coli ◽  
Metal Plating ◽  
Positive Rate ◽  
Meat Samples ◽  
Inspection Service

Abstract A method for detection of Escherichia coli O157 in beef and poultry is presented. The method is antibody-based and uses a patented antibody-specific metal-plating procedure for the detection of E. coli O157 in enriched meat samples. Both raw ground beef and raw ground poultry were tested as matrixes for the organism. The sensitivity and specificity of the assay were 98 and 90%, respectively. The accuracy of the assay was 96%. Overall, the method agreement between the E. coli O157 Detex assay and the U.S. Department of Agriculture/Food Safety Inspection Service method was 96%. Sample temperature upon loading of the apparatus was critical to the observed false-positive rate of the system.

Comparative Evaluation of a Phage Protein Ligand Assay with Real-Time PCR and a Reference Method for the Detection of Escherichia coli O157:H7 in Raw Ground Beef and Trimmings

2011 ◽  
Vol 74 (1) ◽  
pp. 6-12 ◽  
Author(s):  
F. SAVOYE ◽  
P. FENG ◽  
C. ROZAND ◽  
M. BOUVIER ◽  
A. GLEIZAL ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Ground Beef ◽  
Reference Method ◽  
Enterohemorrhagic Escherichia Coli ◽  
Detection Methods ◽  
Escherichia Coli O157 ◽  
Rt Pcr ◽  
Raw Meat ◽  
E Coli

Enterohemorrhagic Escherichia coli O157:H7 is an important pathogen associated with infections caused by consumption of undercooked raw meat. Sensitive and rapid detection methods for E. coli O157:H7 are essential for the meat industry to ensure a safe meat supply. This study was conducted to compare the sensitivity of the VIDAS ultraperformance E. coli test (ECPT UP) with a noncommercial real-time (RT) PCR method and the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) reference method for detecting E. coli O157:H7 in raw ground beef. Optimal enrichment times and the efficacy of testing different types of raw meat, either as individual samples (25 g) or as composites (375 g), were examined. For 25-g samples of each type of raw ground beef tested, 6 h of enrichment was sufficient for both the VIDAS ECPT UP and RT-PCR methods, but for 375-g samples, 24 h of enrichment was required. Both the VIDAS ECPT UP and RT-PCR methods produced results similar to those obtained with the USDA-FSIS reference method after 18 to 24 h of enrichment. The primer specificity of the RT-PCR assay and the highly specific phage ligand used in the VIDAS ECPT UP for target recognition enabled the detection of low levels of E. coli O157:H7 in 25 g of various types of raw ground beef. The tests also allowed the detection of E. coli O157:H7 in composite raw ground beef and trimmings in samples of up to 375 g.

scholarly journals Characterization and Antimicrobial-Resistance Profile of Escherichia coli O157 and O157: H7 Isolated from Modified Atmosphere Packaged Meat Samples

2017 ◽  
Vol 5 (10) ◽  
pp. 1142
Author(s):  
Özgür Çadırcı ◽  
Ali Gücükoğlu ◽  
Göknur Terzi Güzel ◽  
Tolga Uyanık ◽  
Abdulaziz Abdulahi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Modified Atmosphere Packaging ◽  
Meat Products ◽  
Ground Beef ◽  
Inhibitory Effect ◽  
Modified Atmosphere ◽  
O157 Strain ◽  
E Coli ◽  
O157 Isolate ◽  
Meat Samples

Shiga-like toxin producing Escherichia coli is still an important public issue which causes extremely dangerous health problems. This study was planned in order to examine the inhibitory effect of Modified Atmosphere Packaging application on E. coli O157 and O157: H7. The purposes of the present study were to detect E. coli O157 and O157: H7 strains from ground and cubed beef. A total of 100 MAP cattle meat products (50 minced meat, 50 meat cubes) were collected from the markets and butchers in Samsun province between May and October 2013. According to results, 1(1/50-2%) E. coli O157 and 1(1/50-2%) E. coli O157: H7 strains isolated from 50 ground beef samples, while 1 (1/50-2%) E. coli O157 strain was identified from 50 cubed beef samples. It was determined that E. coli O157 isolate obtained from the MAP ground beef carried stx1, stx2 genes; E. coli O157: H7 isolate carried stx1, stx2, eaeA and hylA genes while E. coli O157 isolate obtained from the MAP cubed meat only carried the stx2 gene. In antibiogram test, both E. coli O157 isolates were resistant to streptomycin and one E. coli O157: H7 isolate was resistant to streptomycin, cephalothin and tetracycline. As a consequence; in order to protect public health, products should be kept in proper hygienic and technical conditions during sale and storage and use of uncontrolled antibiotics should be avoided.

Escherichia coli O157:H7 Prevalence in Fecal Samples of Cattle from a Southeastern Beef Cow-Calf Herd

2003 ◽  
Vol 66 (10) ◽  
pp. 1778-1782 ◽  
Author(s):  
D. G. RILEY ◽  
J. T. GRAY ◽  
G. H. LONERAGAN ◽  
K. S. BARLING ◽  
C. C. CHASE
Keyword(s):  
Escherichia Coli ◽  
Control Point ◽  
Foodborne Pathogen ◽  
Beef Cows ◽  
Escherichia Coli O157 ◽  
Fecal Samples ◽  
E Coli ◽  
Critical Control Point ◽  
Cow Calf ◽  
Culture Positive

The proportion of fecal samples culture-positive for Escherichia coli O157:H7 was determined for samples collected from 296 beef cows on pasture in a single Florida herd in October, November, and December 2001. The overall proportion of samples that cultured positive was 0.03. The proportion of cows that were culture-positive on at least one occasion was 0.091. No effect of pregnancy status or nutritional regimen on the proportion of culture-positive samples for E. coli O157:H7 was detected. We detected a breed effect on the shedding of E. coli O157, with Romosinuano cows having a lower (P < 0.01) proportion of samples culture-positive than Angus or Brahman cows. This difference might have resulted from the presence of confounding variables; however, it also might represent evidence of breed-to-breed genetic variation in E. coli O157 shedding. Further research is warranted to evaluate breed as a possible risk factor for shedding of this important foodborne pathogen. Further substantiated findings could indicate that breed is a cow-calf–level critical control point of E. coli O157:H7.

scholarly journals Prevalence and Loads of Fecal Pollution Indicators and the Antibiotic Resistance Phenotypes of Escherichia coli in Raw Minced Beef in Lebanon

Foods ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1543 ◽  
Author(s):  
Issmat I. Kassem ◽  
Nivin A Nasser ◽  
Joanna Salibi
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Food Safety ◽  
Food Systems ◽  
Meat Products ◽  
Fecal Coliforms ◽  
E Coli ◽  
Beef Meat ◽  
Minced Beef ◽  
Meat Samples

Meat is an important source of high biological value proteins as well as many vitamins and minerals. In Lebanon, beef meats, including raw minced beef, are among the most consumed of the meat products. However, minced beef meat can also be an important source of foodborne illnesses. This is of a major concern, because food safety in Lebanon suffers from well-documented challenges. Consequently, the prevalence and loads of fecal coliforms and Escherichia coli were quantified to assess the microbiological acceptability of minced beef meat in Lebanon. Additionally, antibiotic resistance phenotypes of the E. coli were determined in response to concerns about the emergence of resistance in food matrices in Lebanon. A total of 50 meat samples and 120 E. coli isolates were analyzed. Results showed that 98% and 76% of meat samples harbored fecal coliforms and E. coli above the microbial acceptance level, respectively. All E. coli were resistant to at least one antibiotic, while 35% of the isolates were multidrug-resistant (MDR). The results suggest that Lebanon needs to (1) update food safety systems to track and reduce the levels of potential contamination in important foods and (2) implement programs to control the proliferation of antimicrobial resistance in food systems.

Molecular detection of multidrug and colistin-resistant Escherichia coli isolated from house flies in various environmental settings

2019 ◽  
Vol 14 (10) ◽  
pp. 847-858 ◽  
Author(s):  
Abdus Sobur ◽  
Zobayda F Haque ◽  
Abdullah AM Sabuj ◽  
Samina Ievy ◽  
AMM Taufiqur Rahman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multidrug Resistant ◽  
Diffusion Test ◽  
Isolation And Identification ◽  
Antibiotic Resistant ◽  
Disk Diffusion Test ◽  
House Flies ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Antimicrobial Resistance Genes

Aim: To detect antibiotic-resistant pathogenic Escherichia coli in house flies captured from various environmental settings. Materials & methods: Isolation and identification of E. coli was performed based on culture and PCR. Antimicrobial susceptibility testing was conducted using disk diffusion test. Detection of virulence and antimicrobial resistance genes was carried out using PCR methods. Results: The prevalence of E. coli in flies was 61% with highest in dairy farms (70.67%) followed by food centers (65.33%), dustbins (64%) and area close to hospital (44%). The prevalence rates of tetA, tetB, ereA, CITM, SHV, mcr-3 and catA1 resistance determinants were 88.75, 48.52, 41.67, 37.17, 27.77, 18.37 and 14.81%, respectively. Conclusion: Multidrug-resistant pathogenic E. coli are widely distributed in house flies, which requires a one-health approach to delineate their control.

scholarly journals Isolasi Bakteri Endofit Dari Sea Grass Yang Tumbuh Di Kawasan Pantai Pulau Lombok Dan Potensinya Sebagai Sumber Antimokroba Terhadap Bakteri Patogen

2016 ◽  
Vol 16 (2) ◽  
Author(s):  
Lalu Zulkifli, Dwi Soelistya Diah Jekti, Nur Lestari dan Dewa Ayu Citra Rasmi
Keyword(s):  
Escherichia Coli ◽  
Antibacterial Activity ◽  
Endophytic Bacteria ◽  
Klebsiella Pneumonia ◽  
Diffusion Test ◽  
Biochemical Tests ◽  
Disk Diffusion Test ◽  
E Coli ◽  
Activity Test ◽  
Sea Grass

ABSTRAK Telah dilakukan isolasi bakteri endofit dari seagrass jenis Enhalus acoroides (Linneaus f.) Royle, yang tumbuh di perairan pantai Kuta Lombok Tengah. Media yang digunakan adalah NA, BHI, TSA dan Mac Conkey’ Agar, dengan menggunakan bagian akar batang, dan daun sebagai sumber isolat. Dilakukan subkultur berulang hingga diperoleh isolat endofit yang membentuk koloni tunggal.Terhadap koloni tunggal ini dilakukan identifikasi morfologi dan uji biokimia. Selanjutnya dilakukan uji aktivitas antibakteri dengan metode Difusi Agar (Disk diffusion test) dengan teknik sumuran menggunakan lima bakteri uji klinis yaitu : Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Shygella dysentriae, dan Staphylococcus. aureus. Kategori sensitivitas bakteri uji klinis terhadap ekstrak bakteri endofit mengacu kepada Mukherjee (1989). Identifikasi morfologi, uji biokimia dan uji aktifitas antibakteri dilakukan terhadap 11 isolat dari 28 isolat awal yang diperoleh diperoleh, Hasil menunjukkan bahwa semua isolat bakteri endofit tergolong dalam bentuk batang, 8 diantaranya termasuk Gram Positif. Hasil uji antibakteri menggunakan metode Difusi agar dengan teknik sumuran menunjukkan bahwa hampir semua isolat memiliki aktivitas antibakteri terhadap bakteri uji dengan daya hambat yang bervariasi. Diperoleh satu isolat yang palingkuat efeknya dan paling luas spektrumnya yaitu isolate 1 yang mampu menghambat pertumbuhan 4 jenis bakteri uji (E. coli, P. aeruginosa, K. pneumonia, S. dysentriae, dan S. aureus) hingga sampai pada kategori sensitif (dimeter zona hambat lebih dari 12 mm). Dapat disimpulkan bahwa isolat bakteri endofit yang berasal dari seagrass sukup potensial untuk dikembangkan sebagai salah satu alternatif sumber antibakteri untuk mengatasi penyakit yang disebabkan oleh bakteri pathogen. Kata Kunci: Antimikroba, bakteri endofit, lamun (sea grass), aktivitas antibakteri.ABSTRACTHave done the isolation of endophytic bacteria on seagrass kind Enhalus acoroides (Linnaeus f.) Royle, who grew up in the coastal waters of Kuta Lombok. The medium used is NA, BHI, TSA and Mac Conkey 'Agar, using the roots stems and leaves as a source of isolates. Subcultures performed repeatedly to obtain isolates endophytic form colonies tunggal.Terhadap single colony is to identify morphological and biochemical tests. Furthermore, the antibacterial activity test by the method Diffusion Agar (Disk diffusion test) with wells technique uses five clinical trials that bacteria: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Shygella dysentriae, and Staphylococcus. aureus. Categories sensitivity of bacteria clinical trials of endophytic bacteria extract refers to Mukherjee (1989). Identification of morphological, biochemical tests and antibacterial activity test conducted on 11 isolates of 28 isolates obtained initial obtained results showed that all isolates of endophytic bacteria belong in the form of rods, 8 of which include Gram Positive. Antibacterial test results using the diffusion method in order to pitting technique showed that almost all isolates possess antibacterial activity against bacteria inhibition test with varied. Retrieved palingkuat one isolate its effects and the most extensive spectrum that isolate one capable of inhibiting the growth of four kinds of test bacteria (E. coli, P. aeruginosa, K. pneumoniae, S. dysentriae, and S. aureus) to arrive at a sensitive category (dimeter inhibitory zone more than 12 mm). It can be concluded that endophytic bacteria isolates originating from sukup seagrass potential to be developed as an alternative source of antibacterial to treat diseases caused by pathogenic bacteria.Keywords: Antimicrobial, endophytic bacteria, seagrass (sea grass), antibacterial activity.

Incidence of Enterohemorrhagic Escherichia coli, Escherichia coli O157, Salmonella, and Listeria monocytogenes in Retail Fresh Ground Beef, Sprouts, and Mushrooms

2006 ◽  
Vol 69 (2) ◽  
pp. 441-443 ◽  
Author(s):  
M. SAMADPOUR ◽  
M. W. BARBOUR ◽  
T. NGUYEN ◽  
T.-M. CAO ◽  
F. BUCK ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Ground Beef ◽  
Food Samples ◽  
Enterohemorrhagic Escherichia Coli ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Pcr Assays ◽  
Retail Food

The objective of this study was to determine the prevalence of enterohemorrhagic Escherichia coli (EHEC), E. coli O157, Salmonella, and Listeria monocytogenes in retail food samples from Seattle, Wash. A total of 2,050 samples of ground beef (1,750 samples), mushrooms (100 samples), and sprouts (200 samples) were collected over a 12-month period and analyzed for the presence of these pathogens. PCR assays, followed by culture confirmation were used to determine the presence or absence of each organism. Of the 1,750 ground beef samples analyzed, 61 (3.5%) were positive for EHEC, and 20 (1.1%) of these were positive for E. coli O157. Salmonella was present in 67 (3.8%) of the 1,750 ground beef samples. Of 512 ground beef samples analyzed, 18 (3.5%) were positive for L. monocytogenes. EHEC was found in 12 (6.0%) of the 200 sprout samples, and 3 (1.5%) of these yielded E. coli O157. Of the 200 total sprout samples, 14 (7.0%) were positive for Salmonella and none were positive for L. monocytogenes. Among the 100 mushroom samples, 4 (4.0%) were positive for EHEC but none of these 4 samples were positive for E. coli O157. Salmonella was detected in 5 (5.0%) of the mushroom samples, and L. monocytogenes was found in 1 (1.0%) of the samples.

Antibacterial Effect of Lactoferricin B on Escherichia coli O157:H7 in Ground Beef

1999 ◽  
Vol 62 (7) ◽  
pp. 747-750 ◽  
Author(s):  
KUMAR S. VENKITANARAYANAN ◽  
TONG ZHAO ◽  
MICHAEL P. DOYLE
Keyword(s):  
Escherichia Coli ◽  
Antibacterial Effect ◽  
Ground Beef ◽  
Enterohemorrhagic Escherichia Coli ◽  
Plate Count ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Significant Difference ◽  
Sufficient Magnitude ◽  
Peptone Medium

The antibacterial activity of lactoferricin B on enterohemorrhagic Escherichia coli O157:H7 in 1% peptone medium and ground beef was studied at 4 and 10°C. In 1% peptone medium, 50 and 100 μg of lactoferricin B per ml reduced E. coli O157:H7 populations by approximately 0.7 and 2.0 log CFU/ml, respectively. Studies comparing the antibacterial effect of lactoferricin B on E. coli O157:H7 in 1% peptone at pH 5.5 and 7.2 did not reveal any significant difference (P > 0.5) at the two pH values. Lactoferricin B (100 μg/g) reduced E. coli O157:H7 population in ground beef by about 0.8 log CFU/g (P < 0.05). No significant difference (P > 0.5) was observed in the total plate count between treatment and control ground beef samples stored at 4 and 10°C. The antibacterial effect of lactoferricin B on E. coli O157:H7 observed in this study is not of sufficient magnitude to merit its use in ground beef for controlling the pathogen.

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