scholarly journals Prevalence of Antibiotic-Resistant Escherichia coli Isolated from Swine Faeces and Lagoons in Bulgaria

Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 940
Author(s):  
Lyudmila Dimitrova ◽  
Mila Kaleva ◽  
Maya M. Zaharieva ◽  
Christina Stoykova ◽  
Iva Tsvetkova ◽  
...  

Antimicrobial resistance (AMR) is a worldwide health problem affecting humans, animals, and the environment within the framework of the “One Health” concept. The aim of our study was to evaluate the prevalence of pathogenic strains of the species Escherichia coli (E. coli), their AMR profile, and biofilm-forming potential. The isolated strains from three swine faeces and free lagoons (ISO 16654:2001/Amd 1:2017) were confirmed using Phoenix M50 and 16S rDNA PCR. The antibiotic sensitivity to 34 clinically applied antibiotics was determined by Phoenix M50 and the disc diffusion method, according to the protocols of the CLSI and EUCAST. We confirmed the presence of 16 E. coli isolates, of which 87.5% were multi-drug-resistant and 31.25% performed strong biofilms. The possibility for the carrying and transmission of antibiotic-resistance genes to quinolones (qnr), aminoglycosides (aac(3)), β-lactamase-producing plasmid genes ampC, and blaSHV/blaTEM was investigated. We confirmed the carrying of blaSHV/blaTEM in one and ampC in seven isolates. The strains were negative for the virulence genes (ETEC (LT, STa, and F4), EPEC (eae), and STEC/VTEC (stx and stx2all)). The results should contribute to the development of effective measures for limitation and control on the use of antibiotics, which is a key point in the WHO action plan.

2020 ◽  
Vol 28 (2) ◽  
pp. 81
Author(s):  
Raouia Ben Rhouma ◽  
Ahlem Jouini ◽  
Amira Klibi ◽  
Safa Hamrouni ◽  
Aziza Boubaker ◽  
...  

The purpose of this study was to identify <em>Escherichia coli</em> isolates in diarrhoeic and healthy rabbits in Tunisia and characterise their virulence and antibiotic resistance genes. In the 2014-2015 period, 60 faecal samples from diarrhoeic and healthy rabbits were collected from different breeding farms in Tunisia. Susceptibility to 14 antimicrobial agents was tested by disc diffusion method and the mechanisms of gene resistance were evaluated using polymerase chain reaction and sequencing methods. Forty <em>E. coli</em> isolates were recovered in selective media. High frequency of resistance to tetracycline (95%) was detected, followed by different levels of resistance to sulphonamide (72.5%), streptomycin (62.5%), trimethoprim-sulfamethoxazole (60%), nalidixic acid (32.5%), ampicillin (37.5%) and ticarcillin (35%). <em>E. coli</em> strains were susceptible to cefotaxime, ceftazidime and imipenem. Different variants of bla<sub>TEM</sub>, <em>tet</em>, <em>sul</em> genes were detected in most of the strains resistant to ampicillin, tetracycline and sulphonamide, respectively. The presence of class 1 integron was studied in 29 sulphonamide-resistant <em>E. coli</em> strains from which 15 harboured class 1 integron with four different arrangements of gene cassettes, <em>dfrA17</em>+<em>aadA5</em> (n=9), <em>dfrA1</em> + <em>aadA1</em> (n=4), <em>dfrA12</em> + <em>addA2</em> (n=1), <em>dfrA12</em>+<em>orf</em>+<em>addA2</em> (n=1). The <em>qnrB</em> gene was detected in six strains out of 13 quinolone-resistant <em>E. coli</em> strains. Seventeen <em>E. coli</em> isolates from diarrhoeic rabbits harboured the enteropathogenic eae genes associated with different virulence genes tested (<em>fimA</em>, <em>cnf1</em>, <em>aer</em>), and affiliated to B2 (n=8) and D (n=9) phylogroups. Isolated <em>E. coli</em> strains from healthy rabbit were harbouring <em>fim A</em> and/or <em>cnf1</em> genes and affiliated to A and B1 phylogroups. This study showed that <em>E. coli</em> strains from the intestinal tract of rabbits are resistant to the widely prescribed antibiotics in medicine. Therefore, they constitute a reservoir of antimicrobial-resistant genes, which may play a significant role in the spread of antimicrobial resistance. In addition, the eae virulence gene seemed to be implicated in diarrhoea in breeder rabbits in Tunisia.


2017 ◽  
Vol 14 (2) ◽  
pp. 289-295
Author(s):  
M. M. Islam ◽  
S. Ahamed ◽  
M. Y. Arafat ◽  
I. Hasan ◽  
M. Rahman ◽  
...  

This study was designed to determine the shiga toxin producing genes and to investigate antibiotic sensitivity or resistant patterns of the Escherichia coli isolated from diarrheic children at Mymensingh Medical College Hospital, Bangladesh. A total of 83 stool samples were collected and screened for the detection of E. coli on the basis of cultural, staining and biochemical properties followed by molecular detection by Polymerase Chain Reaction (PCR) using genus specific 16SrRNA primers. Antimicrobial susceptibility pattern of E. coli was determined by disc diffusion method against 9 antimicrobial agents. In this study, 27 (32.53%) out of 83 samples, were confirmed as E. coli. Overall prevalence of shiga toxin producing E. coli (STEC) among the examined children was 1.20% (n=1/83).  Further, 27 E. coli isolates were analyzed for the presence of Stx-1 and Stx-2 genes by duplex-PCR.  The STEC isolate was confirmed to be positive for the presence of the Stx-2 gene only. Highest susceptibility of the E. coli isolates was found against Gentamicin (92.59%), followed by Ciprofloxacin (48.14%) and Moxifloxacin (33.33%). More than 77.78% of the isolates were resistant to more than three antibiotics thus defined as multi-drug resistant (MDR). In conclusion, Gentamicin and Ciprofloxacin can be recommended as the effective drugs successful treatment of STEC infections in children.


2021 ◽  
Author(s):  
Rene Dembele ◽  
Wendpoulomdé A.D. Kaboré ◽  
Issiaka Soulama ◽  
Oumar Traoré ◽  
Nafissatou Ouédraogo ◽  
...  

Abstract Background The aim of this study was to determine the resistance of diarrheagenic Escherichia coli strains to β-lactams antibiotics and to perform the molecular characterization of Extended Spectrum β-lactamases (ESBL) and integrons genes. Methods This study was carried out from August 2013 to October 2015 and involved 31 DEC strains isolated from diarrheal stools samples collected from children less than five years of age. The identification and characterization of DEC strains was done through the standard biochemical tests those were confirmed using API 20E and Polymerase Chain Reaction (PCR). The determination of antimicrobial resistance was realized by the disk diffusion method then an amplification of the β-lactamase resistance genes and integrons by PCR was done. Results Out of the 419 E. coli strains identified, 31 isolates (7.4%) harbored the DEC virulence genes. From these DEC, 21 (67.7%) were ESBL-producing E. coli. Susceptibility to ESBL-producing E. coli showed that the majority of isolates were highly resistant to amoxicillin (77.4%), amoxicillin clavulanic acid (77.4%) and piperacillin (64.5%). The following antibiotic resistance genes and integron were identified from the 31 DEC isolates: blaTEM (6.5%), blaSHV (19.4%), blaOXA (38.7%) blaCTX−M (9.7%), Int1 (58.1%) and Int3 (19.4%). No class 2 integrons (Int2) was characterized. Conclusions Because of the high prevalence of multidrug-resistant ESBL organisms found in this study among pediatric patients, there is a need of stringent pediatric infection control measures.


2021 ◽  
Vol 8 (1) ◽  
pp. 24-27
Author(s):  
Wanikda Eh Chuan ◽  
Akrimah Yusof ◽  
Aweng Eh Rak ◽  
Shareef Mohd Hafiz Mani ◽  
Seong Wei Lee

In the present study, antibiogram of Escherichia coli isolated from farmed Asian clam, Corbiculafluminea was characterised. Asian clam or locally known as ‘etak’ is processed to became smokedclam and consumed as snack by Kelantanese. However, there are many diarrhoea cases afterconsuming smoked clam. Furthermore, there are also insufficient information about the E.coli levelin Asian clam farm and effective antibiotic in controlling the bacteria in the litereature. Hence, thisstudy was carried out to provide information of antibiogram of E.coli to be reference in the future.Eosin Methylene Blue agar (EMB) was used to isolate E. coli. A total of 100 isolated bacteria weresubjected to antibiotic sensitivity test using disk diffusion method. A total of 18 types of antibioticsnamely novobiocin (30µg/disk), fosfomycin (50 µg/disk), tetracycline (30µg/disk), lincomycin(15µg/disk), flumequine (30µg/disk), sulphamethoxazole (25 µg/disk), amoxycillin (25 µg/disk),chloramphenicol (30 µg/disk), oleandomycin (15 µg/disk), spiramycin (100 µg/disk), ampicillin(10 µg/disk), oxytetracycline (30 µg/disk), doxycycline (30 µg/disk), nalidixic acid (30 µg/disk),florfenicol (30 µg/disk), erythromycin (15 µg/disk), kanamycin (30 µg/disk) and oxolinic acid (2µg/disk). The findings of the present study showed total plate count of E. coli was 6.45 x 103 colonyforming unit (CFU/100g) of sampled Asian clam. Hence, the clam is needed to be under cleansingtreatment before can consider safe for human consumption. Antibiotic results showed 51 % wasrecorded as antibiotic resistance case, 44 % antibiotic sensitive case and 5 % as antibioticintermediary sensitive case. None of the tested antibiotics was successfully inhibited the growth ofthe present bacterial isolates indicating more antibiotics are needed to be screen in the future studyto find out the most effective antibiotic in controlling isolated E. coli.


2021 ◽  
Vol 12 ◽  
Author(s):  
Akiyo Nakano ◽  
Ryuichi Nakano ◽  
Ryuji Nishisouzu ◽  
Yuki Suzuki ◽  
Saori Horiuchi ◽  
...  

Colistin is used to treat infectious diseases in humans and livestock; it has also been used as a feed additive for livestock for approximately 50 years. Since the mcr-1 plasmid-mediated colistin resistance gene was discovered in China in 2015, it has been detected worldwide, mainly in livestock. In this study, we investigated the prevalence and characteristics of mcr-mediated colistin-resistant Escherichia coli in livestock and farmers in Japan. We collected fecal samples from 295 healthy livestock (202 cattle and 93 swine) and 62 healthy farmers from 72 livestock farms (58 cattle farms and 14 swine farms) between 2013 and 2015. Twenty-eight mcr-1-harboring E. coli strains were isolated from 25 livestock (six cattle and 19 swine) and three farmers (two cattle farmers and one swine farmer). The prevalence rates of mcr-1-harboring E. coli in livestock and farmers were 8.47 and 4.84%, respectively. Of the 28 strains, the resistance genes of three were transferable via the mcr-1-coding plasmids to E. coli J53 at low frequencies (10−7–10−8). Six strains coharbored mcr-1 with CTX-M β-lactamases (CTX-M-14, CTX-M-27, or CTX-M-156). Of the isolates obtained from livestock and farmers in four farms (farms C, I, N, and P), nine strains had the same genotypical characteristics (sequence types and pulsed-field gel electrophoresis band patterns), plasmid characteristics (incompatibility group and plasmid transferability), and minimum inhibitory concentrations. Thus, the findings suggested that clonal strains could spread among livestock and farmers within farms. To our knowledge, this is the first study to detect clonal relatedness of mcr-1-mediated colistin-resistant E. coli in livestock and farmers. It is suggested that farmers are at a higher risk of acquiring mcr-1-harboring strains, calling for our attention based on the One Health concept.


2018 ◽  
Vol 4 (1) ◽  
pp. 36-43 ◽  
Author(s):  
Md Rashed Kamal ◽  
Md Fakhruzzaman ◽  
Mir Rowshan Akter ◽  
Md Atiqul Haque

Enterovirulent Escherichia coli remain as an important etiological agent of goat diarrhoea in Bangladesh. The present study was designed with a view to isolate and identifies E. coli from field cases. For this purpose, a total of 135 faecal samples (85 from diarrhoeic and 50 from apparently healthy goat) were collected during the period from January 2012 to July 2012 from different areas in Rangpur District. It was found that the prevalence of E. coli was higher (18.82 %) in diarrhoeic goats while it was lower (14.00 %) in non diarrhoeic goats. Age wise distribution of E. coli isolates were 26.42% in day old to 1 year, 10.53% in 1-2 years and 11.36% in above 2 years age of goat respectively. All the isolates of E. coli revealed greenish black colony with metallic sheen in Eosine methylene blue agar, bright pink color smooth transparent colony in MacConkey agar and slight pinkish smooth colony in Salmonella-Shigella agar. Gram stain and hanging drop techniques were performed with the cultured bacteria. Biochemical properties of the isolates were studied, and antibiotic sensitivity test was done by agar disk diffusion method. In Gram stain, the organisms revealed Gram negative, small rod shaped, occurs singly or paired. Biochemically, all of the isolates showed fermentation of dextrose, sucrose and maltose with the production of acid and gas, negative result to Voges-Proskauer test, positive result to Methylred test and differential result to Indol test. All the isolates of E. coli were highly sensitive to ciprofloxacin and gentamicin while moderately sensitive to colistin, livofloxacin and azithromycin and less sensitive to ceftraexon and tetracyclin and resistant to amoxycillin, ampicillin, erythromycin, and neomycin. Therefore, ciprofloxacin and gentamicin may be the antibiotics of first choice, and colistin, livofloxacin and azithromycin may be the second choice among the test antibiotics for the treatment of illness caused by these bacteria.Asian J. Med. Biol. Res. March 2018, 4(1): 36-43


2021 ◽  
Vol 14 ◽  
pp. 117863612110168
Author(s):  
Sanjida Akter ◽  
A. M. Masudul Azad Chowdhury ◽  
Sohana Akter Mina

In developing countries, the occurrence of antibiotic resistance is increasing day by day and antibiotic resistant microorganisms are being found in almost every environmental setting. Plasmids are considered as the main vector in the procurement and propagation of antibiotic resistance in many microorganisms such as Escherichia coli ( E. coli). The goal of this study was to examine the antibiotic resistance and screening of plasmid in E. coli strains which were previously identified from human sewage samples. During this study antibiotic susceptibility of E. coli isolates were determined by Kirby-Bauer disk diffusion method against 5 antibiotics (ampicilin, ceftriaxone, amoxicillin, ciprofloxacin, azithromycin). Furthermore, plasmid extraction of each isolate was done according to the protocol of FavorPrepTMPlasmid Mini Kit and plasmid profiling was done by agarose gel electrophoresis. In antibiotic sensitivity test, all E. coli strains showed resistance to ampicilin, amoxicillin, and ceftriaxone. In the plasmid profiling, it was revealed that all the isolates of E. coli harbored plasmids. The plasmid sizes ranged from approximately 1.5 to 15 kb. The findings of this study prove the consequences of antibiotic resistance as well as relationship of plasmid with antibiotic resistance which necessitates proper surveillance on antibiotic usage in the developing countries.


Author(s):  
Mojtaba Bonyadian ◽  
Sara Barati ◽  
Mohammad Reza Mahzounieh

Background and Objectives: Escherichia coli is a common enteric pathogen of human and livevestock. Antibiotic resis- tance is the main concern of public health. The aim of this study was to detect this bacterium in stool samples of diarrheal patients and identify the phenotypic and genotypic characterizations of antibiotic-resistant isolates such as dfrA1, sul1, citm, tetA, qnr, aac(3)-IV in Shahrekord. Materials and Methods: Two hundred fifty diarrheal stool samples from patients were collected. Microbiological and biochemical examinations were done to detect E. coli. Phenotypic and genotypic antibiotic resistance of the isolates were determined using dick diffusion method and polymerase chain reaction (PCR), respectively. Results: Among 114 E. coli isolates, the least resistance was for gentamicin (0%) and the most resistance was for trimetho- prim (79.8%). The resistance to sulfamethoxazole, ciprofloxacin, ampicillin, and tetracycline were 71.05%, 10.5%, 52.63%, and 3.5% respectively. The results of PCR assay revealed that 10 isolates contain sul1, 49 isolates harbor citm, 8 isolates tetA, 36 isolates dfrA1, 11 isolates qnr genes but there was no isolate with aac(3)-IV gene. In comparison between phenotypic and genotypic of the isolates revealed that citm, tetA, dfrA1, qnr, sul1, aac(3)-IV genes covered 42.98%, 7.01%, 31.57%, 9.64%, 8.7%, 0% of the antibiotic resistance, respectively. Conclusion: Our results revealed that all isolates harbor one or more antibiotic resistance genes and that the PCR is a fast practical and appropriate method to determine the presence of antibiotic resistance genes.


2018 ◽  
Vol 55 (4) ◽  
pp. 390-396 ◽  
Author(s):  
Roger Lafontaine Mesquita TABORDA ◽  
Luiz Antônio da SILVA ◽  
Patricia Puccinelli ORLANDI ◽  
Flávia Serrano BATISTA ◽  
Renata Santos RODRIGUES ◽  
...  

ABSTRACT BACKGROUND: Enteroaggregative Escherichia coli (EAEC) is one of the main acute and chronic diarrhea causes both in children and adults, mainly in developing countries. OBJECTIVE: The aim of the present study is to characterize EAEC strains isolated from faecal samples and to identify genes potentially contributing to virulence, biofilm production and antimicrobial resistance in children admitted to a pediatric hospital in Porto Velho, Rondônia State. METHODS: The total of 1,625 E. coli specimens were isolated from 591 children in the age group 6 years or younger who were hospitalized in Cosme and Damião Children Hospital in Porto Velho, between February 2010 and February 2012, with acute gastroenteritis. Colonies suggestive of E. coli were subjected to polymerase chain reaction testing in order to identify the virulence factors. The in vitro adhesion assays using HEp-2 adherence were tests. Biofilm detection through spectrophotometry and antimicrobial susceptibility tests were conducted in the disk diffusion method. RESULTS: The mentioned study examined 591 stool samples from children with diarrhea. Diarrheogenic E. coli was found in 27.4% (162/591) of the children. EAEC was the diarreagenic E. coli most frequently associated with diarrhea 52.4% (85/162), which was followed by enteropathogenic E. coli 43.8% (71/162), enterotoxigenic E. coli 2.4% (4/162), and enterohemorrhagic E. coli 1.2% (2/162). The aggR gene was detected in 63.5% (54/85) of EAEC isolates; moreover, statistically significant correlation was observed among typical EAEC (aggR) and aatA (P<0.0001), irp2 (P=0.0357) and shf (P=0.0328). It was recorded that 69% (59/85) of the 85 analyzed EAEC strains were biofilm producers; 73% (43/59) of the biofilm producers carried the aggR gene versus 42.3% (11/26) of non-producers (P=0.0135). In addition, there was association between the aatA gene and biofilm production; 61% (36/59) of the samples presented producer strains, versus 19.2% (5/26) of non-producers (P<0.0004). Antibiotic sensitivity test evidenced that most EAEC were ampicillin 70.6% (60/85), sulfamethoxazole 60% (51/85), tetracycline 44.7% (38/85) and cefotaxime 22.4% (19/85) resistant. CONCLUSION: As far as it is known, the present study is pioneer in Northern Brazil to investigate EAEC virulence factors and to show the antimicrobial susceptibility of EAEC strains isolated from children with diarrhea.


2021 ◽  
Vol 9 (2) ◽  
pp. 308
Author(s):  
Michaela Kubelová ◽  
Ivana Koláčková ◽  
Tereza Gelbíčová ◽  
Martina Florianová ◽  
Alžběta Kalová ◽  
...  

The great plasticity and diversity of the Escherichia coli genome, together with the ubiquitous occurrence, make E. coli a bacterium of world-wide concern. Of particular interest are pathogenic strains and strains harboring antimicrobial resistance genes. Overlapping virulence-associated traits between avian-source E. coli and human extraintestinal pathogenic E. coli (ExPEC) suggest zoonotic potential and safety threat of poultry food products. We analyzed whole-genome sequencing (WGS) data of 46 mcr-1-positive E. coli strains isolated from retail raw meat purchased in the Czech Republic. The investigated strains were characterized by their phylogroup—B1 (43%), A (30%), D (11%), E (7%), F (4%), B2 (2%), C (2%), MLST type, and serotype. A total of 30 multilocus sequence types (STs), of which ST744 was the most common (11%), were identified, with O8 and O89 as the most prevalent serogroups. Using the VirulenceFinder tool, 3 to 26 virulence genes were detected in the examined strains and a total of 7 (15%) strains met the pathogenic criteria for ExPEC. Four strains were defined as UPEC (9%) and 18 (39%) E. coli strains could be classified as APEC. The WGS methods and available on-line tools for their evaluation enable a comprehensive approach to the diagnosis of virulent properties of E. coli strains and represent a suitable and comfortable platform for their detection. Our results show that poultry meat may serve as an important reservoir of strains carrying both virulence and antibiotic resistance genes for animal and human populations.


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