scholarly journals Molecular Characterization of ESBL-Producing Enterobacteriaceae in Northern Portugal

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Rúben Fernandes ◽  
Paula Amador ◽  
Carla Oliveira ◽  
Cristina Prudêncio
Keyword(s):  
Klebsiella Oxytoca ◽  
Enterobacter Aerogenes ◽  
Generation Cephalosporin ◽  
Fourth Generation ◽  
Chain Reaction ◽  
E Coli ◽  
The North ◽  
Genetic Patterns ◽  
Polymerase Chain

Extended-spectrumβ-lactamases (ESBLs) prevalence was studied in the north of Portugal, among 193 clinical isolates belonging to citizens in a district in the boundaries between this country and Spain from a total of 7529 clinical strains. In the present study we recovered some members of Enterobacteriaceae family, producing ESBL enzymes, includingEscherichia coli(67.9%),Klebsiella pneumoniae(30.6%),Klebsiella oxytoca(0.5%),Enterobacter aerogenes(0.5%), andCitrobacter freundii(0.5%).β-lactamases genes blaTEM, blaSHV, and blaCTX-M were screened by polymerase chain reaction (PCR) and sequencing approaches. TEM enzymes were among the most prevalent types (40.9%) followed by CTX-M (37.3%) and SHV (23.3%). Among our sample of 193 ESBL-producing strains 99.0% were resistant to the fourth-generation cephalosporin cefepime. Of the 193 isolates 81.3% presented transferable plasmids harboringblaESBLgenes. Clonal studies were performed by PCR for the enterobacterial repetitive intragenic consensus (ERIC) sequences. This study reports a high diversity of genetic patterns. Ten clusters were found forE. coliisolates and five clusters forK. pneumoniaestrains by means of ERIC analysis. In conclusion, in this country, the most prevalent type is still the TEM-type, but CTX-M is growing rapidly.

scholarly journals IDENTIFICATION OF AMPC Β-LACTAMASE-PRODUCING CLINICAL ISOLATES OF ESCHERICHIA COLI

2017 ◽  
Vol 10 (12) ◽  
pp. 357
Author(s):  
Tanushree Barua Gupta ◽  
Malini Shariff ◽  
Thukral Ss ◽  
S.s Thukral
Keyword(s):  
Escherichia Coli ◽  
Detection Method ◽  
Clinical Isolates ◽  
Confirmatory Test ◽  
Chain Reaction ◽  
E Coli ◽  
Resistance To Penicillin ◽  
Polymerase Chain ◽  
Third Generation Cephalosporins

  Objective: Indiscriminate use of β-lactam antibiotics has resulted in the emergence of β-lactamase enzymes. AmpC β-lactamases, in particular, confer resistance to penicillin, first-, second-, and third-generation cephalosporins as well as monobactams and are responsible for antibiotic resistance in nosocomial pathogens. Therefore, this study was undertaken to screen nosocomial Escherichia coli isolates for the presence and characterization of AmpC β-lactamases. The study also envisaged on the detection of inducible AmpC β-lactamases and extended-spectrum β-lactamases (ESBLs) in AmpC β-lactamase-producing E. coli.Methods: A total of 102 clinical isolates of E. coli, were subjected to cefoxitin screening, and screen-positive isolates were further subjected to inhibitor-based detection method, phenotypic confirmatory test, disc antagonism test, polymerase chain reaction (PCR), and isoelectric focusing (IEF).Results: In this study, 33% of E. coli were resistant to cefoxitin, of which 35% were found to be positive for AmpC β-lactamase by inhibitor-based phenotypic test. Of the AmpC-positive isolates, 83% were positive for ESBLs, whereas 25% were producing inducible AmpC β-lactamases. PCR and IEF showed CIT and EBC types of AmpC β-lactamases present in the tested isolates.Conclusion: Our study showed the presence of inducible AmpC enzymes and ESBLs in E. coli isolates and PCR identified more isolates to be AmpC producers.

scholarly journals Genotypical characterization of thermotolerant coliforms isolated from food produced by a Solidarity Economic Venture of Bahia (Brazil)

2020 ◽  
Author(s):  
J. N. Silva ◽  
M. D. Baliza ◽  
F. Freitas ◽  
E. S Cruz ◽  
V. M. A. Camilo ◽  
...  
Keyword(s):  
Virulence Genes ◽  
Food Contamination ◽  
Microbiological Analysis ◽  
Chain Reaction ◽  
E Coli ◽  
Family Farmers ◽  
Two Samples ◽  
Thermotolerant Coliforms ◽  
Polymerase Chain

Abstract Many Solidarity Economic Venture (SEV) are family farmers who seek to add value to production through artisanal processing, which can lead to food contamination. Thus, this study aimed to genotypically characterize thermotolerant coliforms (TtC) strains from food produced by local agribusinesses of SEV during January to April 2019. Samples from thirteen production units (PU) from the SEV were submitted to a microbiological analysis of thermotolerant coliforms (AFNOR 3M1/2 – 09/89), using a fast count method in Petrifilm™ dishes. The Polymerase Chain Reaction (PCR) technique was used to verify the following virulence genes (VGs) associated with Escherichia coli: stx, typical from enterohemorrhagic E. coli (EHEC); bfpA typical from entheropathogenic E. coli (EPEC) and elt and slt, typical from entherotoxigenic E. coli (ETEC). The results showed that two samples of queijadinha (typical Brazilian candy made with eggs and coconut) and one sample of cassava cake presented characteristic colonies TtC. This way, three strains were isolated in order to perform the PCR technique. However, the genes used in the reaction were not detected in the isolated strains. Therefore, it is suggested that the isolated strains are from E. coli pathotypes with different virulence genes than the ones analyzed belong other types of TtC, such as Enterobacter and Klebsiella. Although the virulence of genes has not been confirmed, the presence of TtC on food indicates hygiene flaws during production and, therefore, measurements to control and prevent contamination should be taken.

scholarly journals Prevalence and characteristics ofEscherichia coilserotype O157:H7 and other verotoxin-producingE. coliin healthy cattle

1996 ◽  
Vol 117 (2) ◽  
pp. 251-257 ◽  
Author(s):  
M. Blanco ◽  
J. E. Blanco ◽  
J. Blanco ◽  
E. A. Gonzalez ◽  
A. Mora ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Gene Sequence ◽  
Heterogeneous Population ◽  
Chain Reaction ◽  
E Coli ◽  
Healthy Cattle ◽  
Faecal Samples ◽  
Polymerase Chain

SummaryFrom February to July of 1994, 328 faecal samples from 32 herds were collected and verotoxin-producingEscherichia coli(VTEC) found on 84% of the farms. The proportion of animals infected varied from 0–63%. VTEC were recovered from 52 (20%) of 257 cows and from 16 (23%) of 71 calves. Although the VTEC belonged to 25 different serogroups, 7 (O8. O20, O22, O77, O113, O126 and O162) accounted for 46% of strains. Nearly 45% of the 83 bovine VTEC strains belonged to serogroups associated with haemorrhagic colitis and haemolytic uraernic syndrome in humans. However, only 2 (2%) of 83 VTEC strains isolated from cattle belonged to enterohaemorrhagicE. coli(EHEC) serotypes (O26:H11 and O157:H7), and only 8 (10%) were positive for the attaching and effacingE. coli (eae)gene sequence. Polymerase chain reaction (PCR) showed that 17 (20%) of VTEC strains carried VT1 genes. 43 (52%) possessed VT2 genes, and 23 (28%) carried both VT1 and VT2 genes. Characterization of VTEC isolates revelated a heterogeneous population in terms of serogroup and toxin type in the positive herds. This study confirms that healthy cattle are a reservoir of VTEC, but, the absence ofeaegenes in most bovine VTEC strains suggests that they may be less virulent for humans thaneae-positive EHEC.

Distribution ofEscherichia colistrains harbouring Shiga toxin-producingE. coli(STEC)-associated virulence factors (stx1, stx2, eae, ehxA) from very young calves in the North Island of New Zealand

2014 ◽  
Vol 142 (12) ◽  
pp. 2548-2558 ◽  
Author(s):  
H. IRSHAD ◽  
A. L. COOKSON ◽  
D. J. PRATTLEY ◽  
M. DUFOUR ◽  
N. P. FRENCH
Keyword(s):  
New Zealand ◽  
Shiga Toxin ◽  
Macconkey Agar ◽  
Chain Reaction ◽  
Potassium Tellurite ◽  
E Coli ◽  
The North ◽  
Virulence Markers ◽  
Polymerase Chain ◽  
Common Combination

SUMMARYThe objective of this study was to determine the distribution of Shiga toxin-producingEscherichia coli(STEC) virulence markers (stx1,stx2,eae,ehxA) inE. colistrains isolated from young calves aged fewer than 7 days (bobby calves). In total, 299 recto-anal mucosal swabs were collected from animals at two slaughter plants and inoculated onto tryptone bile X-glucuronide and sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Isolates were analysed using multiplex polymerase chain reaction to detectstx1,stx2,eaeandehxAgenes. The most common combination of virulence markers wereeae, ehxA(n = 35) followed byeae(n = 9). In total, STEC and atypical enteropathogenicE. coli(aEPEC) were isolated from 8/299 (2·6%) and 37/299 (12·3%) calves, respectively. All the isolates could be assigned to 15 genotype clusters with >70% similarity cut-off usingXbaI pulsed-field gel electrophoresis. It may be concluded that healthy calves from the dairy industry are asymptomatic carriers of a diverse population of STEC and aEPEC in New Zealand.

Development and characterization of polymorphic microsatellite markers for Spathoglottis plicata (Orchidaceae)

2018 ◽  
Vol 16 (6) ◽  
pp. 572-575
Author(s):  
Chiuan-Yu Li ◽  
Chi-Chun Huang ◽  
Chaur-Tzuhn Chen ◽  
Kuo-Hsiang Hung
Keyword(s):  
Microsatellite Markers ◽  
Microsatellite Primers ◽  
Wild Populations ◽  
Terrestrial Orchid ◽  
Chain Reaction ◽  
Expected Heterozygosity ◽  
Genetic Patterns ◽  
Polymerase Chain ◽  
Polymorphic Microsatellite

AbstractWe developed novel and polymorphic microsatellite primers for Spathoglottis plicata, a tropical and subtropical terrestrial orchid, to investigate the genetic patterns and population structure among wild populations, and also to identify the varieties and hybrids of S. plicata in horticultural industry. The 12 novel microsatellites from S. plicata were developed by using polymerase chain reaction (PCR)-based isolation of microsatellite arrays. These markers that were successfully PCR amplified exhibited polymorphisms in S. plicata. The number of alleles, observed heterozygosity, expected heterozygosity and polymorphism information content values across loci ranged from 2.000 to 8.000, 0.000 to 0.756, 0.208 to 0.813 and 0.405 to 0.805 in total populations, respectively. The newly developed microsatellite markers exhibited variation in S. plicata. These markers can be used as a tool to further investigate the genetic diversity, conservation genetics and variety/hybrid identification of S. plicata.

Molecular prevalence and genetic characterization of piroplasms in dogs from Tunisia

Parasitology ◽  
2016 ◽  
Vol 143 (12) ◽  
pp. 1622-1628 ◽  
Author(s):  
MOHAMED R. RJEIBI ◽  
SAFA AMAIRIA ◽  
MARIEM ROUATBI ◽  
FATMA BEN SALEM ◽  
MOEZ MABROUK ◽  
...  
Keyword(s):  
North Africa ◽  
Molecular Detection ◽  
Genetic Characterization ◽  
Chain Reaction ◽  
Companion Dogs ◽  
Central Tunisia ◽  
The North ◽  
Babesia Vogeli ◽  
Polymerase Chain

SUMMARYIn this study, the prevalence of piroplasms in dogs was assessed using polymerase chain reaction (PCR) to identifyBabesiaandTheileriaspecies in 200 dogs from Northern and Central Tunisia between spring and autumn 2014. The overall molecular prevalence for piroplasms was 14·5% ± 0·05 (29/200); PCR detected 2 species, namelyBabesia vogeliandTheileria annulatawith an overall prevalence of 12·5 ± 0·04 and 2% ± 0·02, respectively. No differences in the molecular prevalences ofB. vogeliwere revealed for age and sex (P> 0·05). The molecular prevalence ofB. vogeliwas significantly higher in central Tunisia (26·5% ± 0·01) compared with the North (9·6% ± 0·04) (P< 0·05). More working and companion dogs were infected byB. vogeli(25·8 ± 0·15 and 21·1% ± 0·13, respectively) in comparison with guarding dogs (1·8% ± 0·03) (P< 0·05). ConcerningT. annulata,no significant variation was observed for all studied risk factors (P> 0·05). Comparison of the partial sequences of18S rRNAandTams 1genes confirmed the presence of 2 novelB. vogeliandT. annulatagenotypes. This is the first molecular detection ofT. annulataand genetic characterization of dogs’ piroplasms in Tunisia. Further studies are needed to better assess the epidemiological feature of piroplasms infection in North Africa.

scholarly journals Prevalence of Escherichia coli adhesion-related genes in neonatal calf diarrhea in Uruguay

2016 ◽  
Vol 10 (05) ◽  
pp. 472-477 ◽  
Author(s):  
Ana Umpiérrez ◽  
Sofía Acquistapace ◽  
Sofía Fernández ◽  
Martín Oliver ◽  
Patricia Acuña ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Economic Losses ◽  
Chain Reaction ◽  
Beef Calves ◽  
E Coli ◽  
Neonatal Calf ◽  
Calf Diarrhea ◽  
Polymerase Chain ◽  
Neonatal Calf Diarrhea

Introduction: Neonatal calf diarrhea (NCD), one of the most important diseases of neonatal dairy and beef calves in Uruguay, has become relevant in association with intensive systems. This disease generates substantial economic losses every year worldwide as a result of increased morbidity and mortality. Escherichia coli, one of the pathogens associated with NCD, can express several fimbrial and afimbrial adhesins. The objective of this study was to assess the presence of clpG, f5, f17A, f17G(II), and f17G(I) genes that encode three important adhesins expressed in diarrheagenic E. coli: F5, F17 and CS31A, isolated from feces of calves in Uruguay. Methodology: Feces of 86 (70 diarrheic and 16 healthy) calves, from 15 animal facilities in Uruguay, were collected between 2012 and 2013. Biochemical and molecular identification were performed to finally obtain 298 E. coli isolates. Partial amplification of adhesion-related genes was performed by polymerase chain reaction. Results: The most prevalent gene was f17A (31.2%), followed by f17G(II), clpG, f17G(I) and f5 (25.8%, 17.5%, 3.7% and 0.7%, respectively). All genes were present in diarrheic and healthy animals except f5 and f17G(I); these genes were present only in affected calves, although in low numbers. Conclusions: This is the first report of the presence of F5, F17, and CS31A genes in E. coli strains from NCD cases in Uruguay. Prevalence values of the genes, except f5, were in accordance with regional findings. It is expected that further characterization of locally transmitted strains will contribute to control a problem of regional and international magnitude.

Molecular Characterization of Antimicrobial Drug Resistance in Escherichia coli Isolated from Clinical Samples

2017 ◽  
Vol 8 (01) ◽  
Author(s):  
Ibtisam Habeeb AL-Azawi ◽  
Aqeel Reheeum Hassan ◽  
Alaa Hamza Jaber
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Antimicrobial Drug Resistance ◽  
Biochemical Characteristics ◽  
E Coli ◽  
Medical City ◽  
Polymerase Chain

A total of 49 different clinical samples (urine n=30, stool n=10, and blood n=8) were collected from patient admitted to the Al-Sadder medical City in Al-Najaf Governorate-Iraq. The results demonstrated that 49 specimens (100%) were diagnosed as E. coli by cultural, biochemical characteristics and Vitek2® system. Polymerase Chain Reaction has been used to detect of some genes which coding antimicrobial resistance in E. coli isolates. Regarding genes that responsible for ESBL enzymes (blaCTX-M, blaOXA and blaTEM), the current results proved that blaTEM genes have highest rate (97.95%) followed by blaTEM and blaOXA (93.75%) for each.

Антимікробна резистентність провідних збудників інфекцій сечових шляхів у Києві (Україна)

2017 ◽  
Vol 1 (2) ◽  
pp. 48-60
Author(s):  
A.G. Salmanov ◽  
A.V. Rudenko
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Staphylococcus Epidermidis ◽  
Klebsiella Oxytoca ◽  
Enterobacter Aerogenes ◽  
Enterobacter Cloacae ◽  
Proteus Vulgaris ◽  
Providencia Rettgeri ◽  
E Coli

Мета роботи — вивчити резистентність до антибіотиків бактеріальних збудників інфекцій сечових шляхів (ІСШ), виділених у пацієнтів урологічного стаціонару в м. Києві. Матеріали і методи. Досліджено 1612 штамів бактерій, виділених із сечі хворих з ІСШ (цистит, уретрит, пієлонефрит), госпіталізованих в урологічне відділення ДУ «Інститут урології НАМН України» у м. Києві протягом 2016 р. Серед пацієнтів переважали жінки — 1201 (74,5 %). Вік хворих становив від 17 до 74 років. Для збору даних використано медичну документацію лікарні. Мікробіологічні дослідження виконано у лабораторії мікробіології ДУ «Інститут урології НАМН України». Аналізували результати культурального дослідження зразків сечі, зібраних за наявності клінічних ознак ІСШ. Дослідження клінічного матеріалу та інтерпретацію отриманих результатів проводили загальноприйнятими методами. Вивчено чутливість уропатогенів до 31 антибіотика дискодифузійним методом відповідно до рекомендацій Інституту клінічних та лабораторних стандартів США (Clinical and Laboratory Standards Institute (CLSI)). Результати та обговорення. Аналіз мікробного спектра сечі виявив домінування серед уропатогенів штамів Escherichia coli (32,0 %), Enterococcus faecalis (19,5 %), Klebsiella pneumoniae (10,9 %), Staphylococcus epidermidis (8,9 %), S. haemolyticus (6,5 %) та Pseudomonas aeruginosa (6,4 %). Частка Enterococcus faecium, Enterobacter aerogenes і Streptococcus viridans становила відповідно 2,5, 2,2 і 1,6 %, Enterobacter cloacae, Klebsiella oxytoca, Acinetobacter baumannii, Proteus vulgaris та Providencia rettgeri — менше 1,0 %. У більшості випадків (69,7 %) мікроорганізми виділено у монокультурі, у решті випадків — у мікробних асоціа- ціях. Високу резистентність до тестованих антибіотиків виявили штами E. aerogenes (45,1 %), E. cloacae (45,7 %), E. faecium (40,9 %), E. faecalis (40,7 %), E. coli (39,9 %), P. aeruginosa (34,0 %), K. pneumoniae (28,6 %). Найбільш активними до уропатогенів були іміпенем (E. coli — 87,6 %, P. aeruginosa — 75,7 %, E. cloacae — 67,3 %, E. aerogenes — 72,6 %, K. pneumoniae — 93,2 %), меропенем (E. coli — 89,1 %, P. aeruginosa — 76,7 %, K. pneumoniae — 82,6 %), лефлоцин (E. coli — 74,5 %, ентерококи — 78,7 %, P. aeruginosa — 76,7 %, E. cloacae — 73,9 %, E. aerogenes — 80,4 %, K. pneumoniae — 83,5 %), амоксицилін/клавуланат (ентерококи — 84,6 %), фурагін (ентерококи — 82,6 %), цефоперазон (K. pneumoniae — 89,2 %, P. aeruginosa — 73,8 %), цефтріаксон (K. pneumoniae — 80,1 %). Висновки. Антибіотикорезистентність збудників ІСШ — важлива терапевтична проблема. Найбільшою активністю до уропатогенів характеризуються іміпенем, меропенем, лефлоцин, амоксицилін/ клавуланат, фурагін, цефоперазон, цефтріаксон, які можна розглядати як препарат вибору для призначення стартової терапії ІСШ. Необхідно здійснювати постійний моніторинг за резистентністю до дії антибіотиків. Політику використання антибіотиків у кожному стаціонарі слід визначати залежно від локальних даних щодо резистентності до протимікробних препаратів.

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