scholarly journals Molecular surveillance of shiga toxigenic Escherichia coli in selected beef abattoirs in Osun State Nigeria

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Femi Ayoade ◽  
Judith Oguzie ◽  
Philomena Eromon ◽  
Omolola E. Omotosho ◽  
Tosin Ogunbiyi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Sequence Analysis ◽  
Block Design ◽  
Latex Agglutination ◽  
Pcr Analysis ◽  
Accession Number ◽  
E Coli ◽  
Representative Isolate ◽  
Randomized Block Design ◽  
Toxigenic Strains

AbstractShiga toxigenic strains of E. coli (STEC) known to be etiological agents for diarrhea were screened for their incidence/occurrence in selected abattoirs sources in Osogbo metropolis of Osun State, Nigeria using a randomized block design. Samples were plated directly on selective and differential media and E. coli isolates. Multiplex PCR analysis was used to screen for the presence of specific virulence factors. These were confirmed serologically as non-O157 STEC using latex agglutination serotyping kit. Sequence analysis of PCR products was performed on a representative isolate showing the highest combination of virulence genes using the 16S gene for identification purposes only. Results showed that the average cfu/cm2 was significantly lower in the samples collected at Sekona-2 slaughter slab compared with those collected at Al-maleek batch abattoir and Sekona-1 slaughter slab in ascending order at P = 0.03. Moreover, the average cfu/cm2E. coli in samples collected from butchering knife was significantly lower when compared with that of the workers’ hand (P = 0.047) and slaughtering floor (P = 0.047) but not with the slaughter table (P = 0.98) and effluent water from the abattoir house (P = 0.39). These data suggest that the abattoir type may not be as important in the prevalence and spread of STEC as the hygiene practices of the workers. Sequence analysis of a representative isolate showed 100% coverage and 96.46% percentage identity with Escherichia coli O113:H21 (GenBank Accession number: CP031892.1) strain from Canada. This sequence was subsequently submitted to GenBank with accession number MW463885. From evolutionary analyses, the strain from Nigeria, sequenced in this study, is evolutionarily distant when compared with the publicly available sequences from Nigeria. Although no case of E. coli O157 was found within the study area, percent occurrence of non-O157 STEC as high as 46.3% at some of the sampled sites is worrisome and requires regulatory interventions in ensuring hygienic practices at the abattoirs within the study area.

Molecular Surveillance of Shiga Toxigenic Escherichia coli in selected beef abattoirs in Osun State Nigeria

2021 ◽  
Author(s):  
Femi Ayoade ◽  
Judith Oguzie ◽  
Philomena Eromon ◽  
Omolola Omotosho ◽  
Tosin Ogunbiyi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Block Design ◽  
Latex Agglutination ◽  
Pcr Analysis ◽  
Accession Number ◽  
E Coli ◽  
Randomized Block Design ◽  
Pcr Products ◽  
Toxigenic Strains ◽  
Etiological Agents

Abstract Shiga toxigenic strains of E. coli (STEC) known to be etiological agents for diarrhea were screened for their incidence/ occurrence in selected abattoirs and retail meat sources in Osogbo metropolis of Osun State, Nigeria using a randomized block design. Samples were plated directly on selective and differential media and confirmed serologically using latex agglutination serotyping kit, then, multiplex PCR analysis was used to screen for the presence of specific virulence factors. The results showed a percent occurrence of STEC at the sampled sites ranging from 25.8–46.3%. None of the strains showed any visible agglutination with the O157 latex reagent. Sequence analysis of PCR products was performed on a representative isolate showing the highest combination of virulence genes. This sequence was subsequently submitted to GenBank with accession number MW463885. The sequence showed 100% coverage and 96.46% percentage identity with Escherichia coli O113:H21 (GenBank Accession number: CP031892.1) strain from Canada. From evolutionary analyses, the strain from Nigeria, sequenced in this study, is evolutionarily distant when compared with the publicly available sequences from Nigeria. Although no case of E. coli O157 was found within the study area, percent occurrence of non-O157 STEC as high as 46.3% at some of the sampled sites is worrisome and requires regulatory interventions in ensuring hygienic practices at the abattoirs within the study area.

Field Evidence Supporting Conventional Onion Curing Practices as a Strategy To Mitigate Escherichia coli Contamination from Irrigation Water

2018 ◽  
Vol 81 (3) ◽  
pp. 369-376 ◽  
Author(s):  
Daniel Wright ◽  
Erik Feibert ◽  
Stuart Reitz ◽  
Clint Shock ◽  
Joy Waite-Cusic
Keyword(s):  
Escherichia Coli ◽  
Irrigation Water ◽  
Block Design ◽  
Microbiological Quality ◽  
Poor Quality ◽  
Inoculum Level ◽  
E Coli ◽  
Bulb Onion ◽  
Field Evidence ◽  
Randomized Block Design

ABSTRACT The Produce Safety Rule of the U.S. Food Safety Modernization Act includes restrictions on the use of agricultural water of poor microbiological quality. Mitigation options for poor water quality include the application of an irrigation-to-harvest interval of <4 days; however, dry bulb onion production includes an extended irrigation-to-harvest interval (<30 days). This study evaluated conventional curing practices for mitigating Escherichia coli contamination in a field setting. Well water inoculated with rifampin-resistant E. coli (1, 2, or 3 log CFU/mL) was applied to onion fields (randomized block design; n = 5) via drip tape on the final day of irrigation. Onions remained undisturbed for 7 days and were then lifted to the surface to cure for an additional 21 days before harvest. Water, onions, and soil were tested for presence of rifampin-resistant E. coli. One day after irrigation, 13.3% of onions (20 of 150) receiving the poorest quality water (3 log CFU/mL) tested positive for E. coli; this prevalence was reduced to 4% (6 of 150 onions) after 7 days. Regardless of inoculum level, E. coli was not detected on any onions beyond 15 days postirrigation. These results support conventional dry bulb onion curing practices as an effective strategy to mitigate microbiological concerns associated with poor quality irrigation water.

scholarly journals Virulence Factors and Antibiotic Resistance of Escherichia coli O157:H7 Isolated from Ogun River, Abeokuta Metropolis, Nigeria

2019 ◽  
pp. 22-30
Author(s):  
Lateefat M. Ewuoso ◽  
Saka A. Balogun ◽  
Sarafadeen O. Kareem ◽  
Temilade F. Akinhanmi
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Shiga Toxin ◽  
Latex Agglutination ◽  
Diffusion Method ◽  
Pcr Analysis ◽  
E Coli ◽  
Multiple Drugs ◽  
Ogun River

Although Escherichia coli (E. coli)is a harmless gut microbe in man, some strains of this bacterium are pathogenic due to the acquisition of virulence factors. The aim of this study isto investigate E. coli O157:H7 strains isolated from Ogun River, Abeokuta metropolis, for virulence factors and antibiotic resistance. Water samples were collected bimonthly from six different locations over a period of six months. The samples were cultured on Sorbitol MacConkey Agar and E. coli O157:H7 isolates were confirmed through serological characterization using the latex agglutination test. The presence of virulence genes (stx1, stx2, eae, and hylA) in the isolates was analyzed through polymerase chain reaction (PCR). Further, antibiotic susceptibility of the isolates was tested using the disc diffusion method. The PCR analysis revealed that the five E. coli O157:H7 strains isolated possessed Shiga toxin 1 (stx1), Shiga toxin 2 (stx2), and Haemolysin (hlyA) genes. Additionally, the isolates were resistant to Augmentin, Ceftriaxone, Nitrofurantoin, Gentamycin, Amoxicillin, and Pefloxacin. This study shows that E. coli O157:H7 strains are present in Ogun River and that these strains possess multiple virulence factors and are resistant to multiple drugs.

scholarly journals Antibiogram of Diarrhoeagenic Escherichia coli Patients under Five Years Attending Selected Hospitals in Kaduna State, Nigeria

2019 ◽  
pp. 1-11
Author(s):  
R. O. Chioma ◽  
I. Mzungu ◽  
J. B. Orpin
Keyword(s):  
Escherichia Coli ◽  
Nutritional Status ◽  
Good Health ◽  
Latex Agglutination ◽  
Public Health Issue ◽  
Chi Square ◽  
Children Under Five ◽  
E Coli ◽  
Under Five ◽  
Good Nutrition

Escherichia coli infections and poor nutritional status have implications on the growth and development of children under five years, physically, mentally and health wise with consequences such as diarrhoea, stunting, wasting, underweight and often times leading to death, depending on their severity. This study evaluated the antibiogram of Escherichia coli O157 and Verocytotoxigenic Escherichia coli (VTEC)and the nutritional status of diarrhoeic children under five years in Kaduna State, Nigeria, using Conventional isolation methods, latex agglutination tests, VTEC-ELISA tests, Chi-square (SPSS Version 19) and WHO Antro (Version 3.2.2). Purposive sampling was used to select 350 children presenting with diarrhoea in six government hospitals within the three senatorial zones of Kaduna State. The results obtained revealed that 76(21.7%) of the 350 stool samples were positive for E. coli and 28(36.8%) were positive for E. coli O157:H7serotype and 1(1.3%) verocytotoxigenic E. coli (VTEC) serotype. High susceptibility to ciprofloxacin, chloramphenicol and high resistance to sulphamethoxazole, cefotaxime, amoxicillin, gentamicin and tetracycline by the isolates were observed. The study concluded that antibiotics have not been very effective in the treatment of E. coli-related diarrhoea, with VTEC now emerging in this part of the world, making it a serious public health issue. The study therefore recommends the implementation of programmes geared towards good hygiene, good nutrition and good health.

scholarly journals Bacteriophage Lambda: a Paradigm Revisited

2010 ◽  
Vol 84 (13) ◽  
pp. 6876-6879 ◽  
Author(s):  
Paul C. M. Fogg ◽  
Heather E. Allison ◽  
Jon R. Saunders ◽  
Alan J. McCarthy
Keyword(s):  
Escherichia Coli ◽  
Sequence Analysis ◽  
High Frequency ◽  
Southern Hybridization ◽  
Integration Site ◽  
Bacteriophage Lambda ◽  
Rare Event ◽  
Low Frequency ◽  
E Coli ◽  
Very Low Frequency

ABSTRACT Bacteriophage lambda has an archetypal immunity system, which prevents the superinfection of its Escherichia coli lysogens. It is now known that superinfection can occur with toxigenic lambda-like phages at a high frequency, and here we demonstrate that the superinfection of a lambda lysogen can lead to the acquisition of additional lambda genomes, which was confirmed by Southern hybridization and quantitative PCR. As many as eight integration events were observed but at a very low frequency (6.4 × 10−4) and always as multiple insertions at the established primary integration site in E. coli. Sequence analysis of the complete immunity region demonstrated that these multiply infected lysogens were not immunity mutants. In conclusion, although lambda superinfection immunity can be confounded, it is a rare event.

scholarly journals Impact of Ceftiofur Injection on Gut Microbiota and Escherichia coli Resistance in Pigs

2015 ◽  
Vol 59 (9) ◽  
pp. 5171-5180 ◽  
Author(s):  
M. A. Fleury ◽  
G. Mourand ◽  
E. Jouy ◽  
F. Touzain ◽  
L. Le Devendec ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gut Microbiota ◽  
Treated Group ◽  
Conjugative Plasmid ◽  
Health Concern ◽  
Pcr Analysis ◽  
Contamination Level ◽  
Content Type ◽  
E Coli ◽  
The Impact

ABSTRACTResistance to extended-spectrum cephalosporins (ESCs) is an important health concern. Here, we studied the impact of the administration of a long-acting form of ceftiofur on the pig gut microbiota and ESC resistance inEscherichia coli. Pigs were orally inoculated with an ESC-resistantE. coliM63 strain harboring a conjugative plasmid carrying a gene conferring resistance,blaCTX-M-1. On the same day, they were given or not a unique injection of ceftiofur. Fecal microbiota were studied using quantitative PCR analysis of the main bacterial groups and quantification of short-chain fatty acids.E. coliand ESC-resistantE. coliwere determined by culture methods, and the ESC-resistantE. coliisolates were characterized. The copies of theblaCTX-M-1gene were quantified. After ceftiofur injection, the main change in gut microbiota was the significant but transitory decrease in theE. colipopulation. Acetate and butyrate levels were significantly lower in the treated group. In all inoculated groups,E. coliM63 persisted in most pigs, and theblaCTX-M-1gene was transferred to otherE. coli. Culture and PCR results showed that the ceftiofur-treated group shed significantly more resistant strains 1 and 3 days after ESC injection. Thereafter, on most dates, there were no differences between the groups, but notably, one pig in the nontreated group regularly excreted very high numbers of ESC-resistantE. coli, probably leading to a higher contamination level in its pen. In conclusion, the use of ESCs, and also the presence of high-shedding animals, are important features in the spread of ESC resistance.

A novel peroxiredoxin of the plant Sedum lineare is a homologue of Escherichia coli bacterioferritin co-migratory protein (Bcp)

2000 ◽  
Vol 351 (1) ◽  
pp. 107-114 ◽  
Author(s):  
Wei KONG ◽  
Susumu SHIOTA ◽  
Yixin SHI ◽  
Hiroaki NAKAYAMA ◽  
Koji NAKAYAMA
Keyword(s):  
Escherichia Coli ◽  
Peroxidase Activity ◽  
Discrete Group ◽  
Sequence Data ◽  
Accession Number ◽  
Gene Encoding ◽  
Reaction Cycle ◽  
Mutant Proteins ◽  
E Coli

We cloned a gene encoding a 17-kDa protein from a cDNA library of the plant Sedum lineare and found that its deduced amino acid sequence showed similarities to those of Escherichia coli bacterioferritin co-migratory protein (Bcp) and its homologues, which comprise a discrete group associated with the peroxiredoxin (Prx) family. Studies of the recombinant 17-kDa protein produced in E. coli cells revealed that it actually had a thioredoxin-dependent peroxidase activity, the hallmark of the Prx family. PrxQ, as we now designate the 17-kDa protein, had two cysteine residues (Cys-44 and Cys-49) well conserved among proteins of the Bcp group. These two cysteines were demonstrated to be essential for the thioredoxin-dependent peroxidase activity by analysis of mutant proteins, suggesting that these residues are involved in the formation of an intramolecular disulphide bond as an intermediate in the reaction cycle. Expression of PrxQ suppressed the hypersensitivity of an E. coli bcp mutant to peroxides, indicating that it might exert an antioxidant activity in vivo. The sequence data presented have been deposited in the GenBank/EMBL/DDBJ nucleotide sequence databases under the accession number AB037598.

Multiplex Real-Time PCR Detection of Heat-Labile and Heat-Stable Toxin Genes in Enterotoxigenic Escherichia coli †

2006 ◽  
Vol 69 (2) ◽  
pp. 412-416 ◽  
Author(s):  
MICHAEL A. GRANT ◽  
JINXIN HU ◽  
KAREN C. JINNEMAN
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Membrane Filtration ◽  
Enterotoxigenic Escherichia Coli ◽  
Pcr Analysis ◽  
E Coli ◽  
Toxin Genes ◽  
Heat Stable ◽  
Heat Labile

A multiplex real-time PCR method was developed for detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli. Approximately 10 CFU per reaction mixture could be detected in rinsates from produce samples. Several foods representative of varieties previously shown to have caused enterotoxigenic E. coli outbreaks were spiked and enriched for 4 or 6 h. Both heat-labile and heat-stable toxin genes could be detected in the foods tested, with the exception of hot sauce, with threshold cycle values ranging from 25.2 to 41.1. A procedure using membrane filtration which would allow enumeration of the enterotoxigenic E. coli population in a food sample in less than 28 h by real-time PCR analysis of colonies picked from media highly selective for E. coli was also developed.

scholarly journals Unexpected Stress-Reducing Effect of PhaP, a Poly(3-Hydroxybutyrate) Granule-Associated Protein, in Escherichia coli

2011 ◽  
Vol 77 (18) ◽  
pp. 6622-6629 ◽  
Author(s):  
Alejandra de Almeida ◽  
Mariela V. Catone ◽  
Virgil A. Rhodius ◽  
Carol A. Gross ◽  
M. Julia Pettinari
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Stress Proteins ◽  
Protective Role ◽  
Pcr Analysis ◽  
Content Type ◽  
E Coli ◽  
Protein Levels ◽  
Stress Related Genes ◽  
Heat Stress Proteins

ABSTRACTPhasins (PhaP) are proteins normally associated with granules of poly(3-hydroxybutyrate) (PHB), a biodegradable polymer accumulated by many bacteria as a reserve molecule. These proteins enhance growth and polymer production in natural and recombinant PHB producers. It has been shown that the production of PHB causes stress in recombinantEscherichia coli, revealed by an increase in the concentrations of several heat stress proteins. In this work, quantitative reverse transcription (qRT)-PCR analysis was used to study the effect of PHB accumulation, and that of PhaP fromAzotobactersp. strain FA8, on the expression of stress-related genes in PHB-producingE. coli. While PHB accumulation was found to increase the transcription ofdnaKandibpA, the expression of these genes and ofgroES,groEL,rpoH,dps, andyfiDwas reduced, when PhaP was coexpressed, to levels even lower than those detected in the non-PHB-accumulating control. These results demonstrated the protective role of PhaP in PHB-synthesizingE. coliand linked the effects of the protein to the expression of stress-related genes, especiallyibpA. The effect of PhaP was also analyzed in non-PHB-synthesizing strains, showing that expression of this heterologous protein has an unexpected protective effect inE. coli, under both normal and stress conditions, resulting in increased growth and higher resistance to both heat shock and superoxide stress by paraquat. In addition, PhaP expression was shown to reduce RpoH protein levels during heat shock, probably by reducing or titrating the levels of misfolded proteins.

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