Coli Strains Research Articles

Emerging antibiotic resistance pattern of urinary tract infection (UTI) due to Escherichia Coli (E.Coli).

Objective: To determine the emerging antibiotic resistance pattern of adult patients of MDR UTI cases caused by E.Coli. Study Design: Retrospective Data. Setting: Departments of Medicine and Obstetrics & Gynecology, Madinah Teaching Hospital / University Medical and Dental College, Faisalabad. Period: August 2016 to Sep 2018. Material & Methods: This retrospective study included 187 patients meeting the operational definition of multi-drug resistance cases of urinary tract infection caused by E. coli. All cases either male or female of age above 15 years with positive reports of urine culture and sensitivity for E.coli were included in the study. Along with the demographic variables urine WBC’s, results of urine culture and sensitivity were noted. Antibiotic sensitivity pattern of all the E.coli resistant UTI cases was analyzed. After identification of the microbe, antimicrobial sensitivity was accessed using Kirby–Bauer disc diffusion method on the Mueller–Hinton agar. The antimicrobial susceptibility test was performed against E. coli strains by using the following antibiotics: Piperacilin, Amoxicilin-Calvulanic, Pipmedic Acid, Cefipime, Ceftrixone, Cefuroxime, Cefoperazone-sulbactam, Ceftazidime, Cephradine, Ciprofloxacin, Ofloxacin, Levofloxacin, ofloxacin, Nitrofurantoin, Amikacin, Tombramycin, Gentamicin, Linezolid, Doxycycline, Sulphamethoxazole, Imipenem, Meropenem, Aztreonam in order to obtain an antibiogram. Results: In our data, 66.8% (n=125) were MDR UTI, and more than half of these cases were 63.2% (n=79) were females. Among the total 187 cases, 97.3% patients were resistant to most of the beta-lactam antibiotics, 95.7% were resistant to most of the quinolones and 68.4% were resistant to most of the aminoglycosides. Conclusion: This increasing antibiotic resistance is very alarming and steps should be taken to reduce the misuse of antibiotics. Studies should be done to control the risk factors leading to E. coli UTI. By following proper pharmacological guidelines, principles to use antibiotics and manage outpatient cases of UTIs, the incidence of MDR UTI can be controlled.

Basics of the Development of Microwave Intensification of Upstream on the Example of <i>Escherichia coli</i>

Introduction. E. coli strains are the main microorganisms used for the production of a number of important biopharmaceutical products. There are no natural sources of microwave radiation on Earth, as it is absorbed by the upper atmosphere. No one doubts the importance of studying the biological effect of microwave radiation. The number of publications devoted to this problem is growing every year, and new ideas for the use of microwaves in drug production technology are emerging.Aim. Reveal the main effects of microwave irradiation and develop a technology for microwave intensification of E. coli culture growth.Materials and methods. This study presents the results of atomic force microscopy, refractometry, NMR relaxometry, turbidimetry, and lumimetry, demonstrating the possibility of microwave intensification of the cultivation process.Results and discussion. It was found that microwave irradiation leads to changes in the mobility of protons and the adsorption of water molecules on biopolymers and cells. These are the main links in the mechanism of "non-thermal" microwave action. A single microwave irradiation, depending on a number of parameters, can decrease or increase the growth of biomass. Studies of the bioluminescence of the E. coli strain with the lux-operon have shown that the optimal processing conditions do not negatively affect the luciferase production and metabolic activity of cells. Conclusion. The intensification procedure using microwave radiation can be considered a promising method and can provide new ideas for various applications in biotechnology.

Transcriptome analysis of biofilm formation under aerobic and microaerobic conditions in clinical isolates of Campylobacter spp.

It has been well documented that Campylobacter is the leading cause of foodborne infections and bacterial enteritis in high-income countries. The gastrointestinal tract of most warm-blooded animals, such as mammals and poultry, is prone to this pathogen. Infections caused by this bacterium in humans have usually been associated with the consumption of contaminated poultry meat. The important point about Campylobacter is that this bacterium has adapted to harsh environmental conditions along the food chain (poultry digestive tract to the consumer's plate) and developed an adapted mechanism to those conditions. This study aimed to compare the ability of Campylobacter jejuni and Campylobacter coli strains to form biofilms under aerobic and microaerobic conditions. The presence and expression of flab, FliS, DnaK, luxs, CsrA, Cj0688, and cosR genes involved in biofilm formation were investigated. Finally, the correlation between the biofilm forming ability of Campylobacter isolates and the presence/expression of selected genes has been explored. A significant correlation was observed between the presence and expression of some genes and the degree of biofilm formation in C. jejuni and C. coli isolates. A strong biofilm production was detected in strains harboring all selected genes with greater expression levels. The ability of C. jejuni and C. coli strains in biofilm formation is associated with the coordinated function and convergent expression of the selected genes. Seemingly, stress response- and motility-related genes have the most involvement in biofilm formation of C. jejuni and C. coli strains, while other genes have an accessory role in this phenomenon.

Dependence of the state of the large intestine parietal microflora on the activity of Helicobacter pylori associated chronic gastritis

Introduction. The aim of the study was to detect the dependence of the composition of the parietal microflora of the colon on the activity of Helicobacter pylori-associated chronic gastritis and the degree of Helicobacter pylori contamination of the gastric mucosa. Materials and methods. The study was carried out on 50 patients aged 24 to 72 years, who were diagnosed with HP-associated chronic gastritis in the phase of exacerbation based on the results of FEGDS and histological examination of biopsies of the mucous membrane of the antrum of the stomach in combination with a rapid urease test. A series of dilutions 10¹-10⁹ was prepared from a homogenized biopsy specimen of the mucous membrane of the sigmoid colon in 0.1 ml of isotonic sodium chloride solution, and 0.1 ml of a solution of the corresponding dilution was inoculated onto the surface of the nutrient medium. The study of mucosal microflora was carried out by means of bacteriological examination of biopsies of the mucous membrane of the sigmoid and cecum. The degree of H. pylori contamination of the gastric mucosa was assessed in the course of histological examination by a semi-quantitative method: low degree of contamination — up to 20 microbial bodies, average — 20-50 microbial bodies, strong — over 50 microbial bodies in the field of view. Results. The study demonstrated that the indicators of the average concentrations of bifidobacteria, lactobacilli and typical strains of E. coli progressively decreased with the transition to a higher degree of HP infection. This tendency was maximum in relation to bifidobacteria — 6.1-1.57-1.09 x 10⁸ CFU/g, with a difference between the maximum (I degree) and minimum (III degree) 5.59 times. A similar relationship, but much less pronounced, was observed for Lactobacterium spp. and E. coli. As the severity of HP infiltration of the gastric mucosa increases, there is a decrease in the concentration of the above microorganisms in the parietal mucin - the correlation coefficient is r = -0.74. Discussion. Since the mucous microflora is more stable and functionally significant than the cavity microflora, the data obtained allow us to consider Helicobacter pylori as an etiological factor of colonic dysbiosis. Conclusion. The probability of excessive growth of conditionally pathogenic fraction of intestinal parietal microflora has a direct dependence on the severity of Helicobacter pylori infestation, being maximum at degree III contamination. Content of bifidobacteria, lactobacilli and typical E. coli strains in the intestinal biotope of the large intestine is inversely related to the degree of histological activity of HP-associated gastritis.

Detecting Glucose Fluctuations in the Campylobacter jejuni N-Glycan Structure.

Campylobacter jejuni is a significant cause of human gastroenteritis worldwide, and all strains express an N-glycan that is added to at least 80 different proteins. We characterized 98 C. jejuni isolates from infants from 7 low- and middle-income countries and identified 4 isolates unreactive with our N-glycan-specific antiserum that was raised against the C. jejuni heptasaccharide composed of GalNAc-GalNAc-GalNAc(Glc)-GalNAc-GalNAc-diNAcBac. Mass spectrometric analyses indicated these isolates express a hexasaccharide lacking the glucose branch. Although all 4 strains encode the PglI glucosyltransferase (GlcTF), one aspartate in the DXDD motif was missing, an alteration also present in ∼4% of all available PglI sequences. Deleting this residue from an active PglI resulted in a nonfunctional GlcTF when the protein glycosylation system was reconstituted in E. coli, while replacement with Glu/Ala was not deleterious. Molecular modeling proposed a mechanism for how the DXDD residues and the structure/length beyond the motif influence activity. Mouse vaccination with an E. coli strain expressing the full-length heptasaccharide produced N-glycan-specific antibodies and a corresponding reduction in Campylobacter colonization and weight loss following challenge. However, the antibodies did not recognize the hexasaccharide and were unable to opsonize C. jejuni isolates lacking glucose, suggesting this should be considered when designing N-glycan-based vaccines to prevent campylobacteriosis.

Virulence genes of Escherichia coli vaginal isolates associated with postpartum dysgalactia syndrome in sows

Objective: Identify the occurrence of certain virulence genes of Escherichia coli vaginal isolates associated with postpartum dysgalactia syndrome (PDS) in sows. Materials and methods: Two hundred and two sows from 5 Macedonian pig farms were clinically examined for PDS 12 to 24 hours after farrowing. Vaginal swabs for bacteriological testing were taken from PDS-affected (PDSA, n = 47) and PDS-unaffected (PDSU, n = 155) sows. In total, 74 isolates of E coli were tested by multiplex polymerase chain reaction for the presence of virulence genes related to specific pathogenic strains. Results: Genes associated with extra-intestinal pathogenic E coli (ExPEC) strains were the most prevalent among all tested E coli isolates. The most dominant gene among all E coli isolates was fimC. The iss gene was more prevalent in PDSA sows compared to PDSU sows (P = .02). Multivariable logistic regression showed that lower parity sows (P ≤ .001) and presence of the iss (P = .003) and astA genes (P = .03) were correlated with the occurrence of PDS. Implications: Lower parity sows vaginally infected with E coli associated with particular ExPEC strains are at higher risk of developing PDS. Positive vaginal swabs for E coli and iss gene found early after farrowing were associated with PDS in sows. Classification of E coli into specific ExPEC pathotype was not possible by virulence genotyping only.

Campylobacter coli isolated in Brazil typed by core genome Multilocus Sequence Typing shows high genomic diversity in a global context.

The current study aimed to determine the phylogenetic relationships among Campylobacter coli strains isolated in Brazil and to compare them with international Campylobacter isolates available in some public databases. A total of 63C. coli strains isolated in Brazil were studied. The MLST analysis showed 18 different STs including three STs not yet described in the PubMLST database. The cgMLST allocated the Brazilian strains studied into five main clusters and each cluster comprised groups of strains with nearly identical cgMLST profiles and with significant genetic distance observed among the distinct clusters. The comparison of the Brazilian strains with 3401 isolates from different countries showed a wide distribution of these strains isolated in this country. The results showed a high similarity among some strains studied and a wide distribution of the Brazilian strains when compared to isolates from different countries, which is an interesting data set since it showed a high genetic diversity of these strains from Brazil in a global context. This study contributed for a better genomic characterization of C. coli strains isolated in Brazil and provided important information about the diversity of this clinically-relevant pathogen.

Expression of a Tagless Single-Chain Variable Fragment (scFv) of Anti-TNF-α by a Salt Inducible System and its Purification and Characterization.

Anti-TNF-α scFv is gaining acceptance as an effective drug for various diseases, such as rheumatoid arthritis and Crohn's disease that involve elevated levels of TNF-α. The single-chain variable fragment (scFv) consists of variable regions of heavy and light chains of monoclonal antibodies (mAb). Due to its smaller size, it curbs the mAb's auto-antibody effects and their limitation of penetration into the tissues during the neutralization of TNF-α. In this work, a cDNA coding for anti-TNF-α scFv was successfully cloned into a pRSET-B vector and efficiently expressed in an E. coli strain GJ1158, a salt inducible system that uses sodium chloride instead of IPTG as an inducer. The protein was expressed in the form of inclusion bodies (IB), solubilized using urea, and refolded by pulse dilution. Further, the amino acid sequence coverage of scFv was confirmed by ESI-Q-TOF MS/MS and MALDI-TOF. Further studies on scaling up the production of scFv and its application of scFv are being carried out. The soluble fraction of anti-TNF-α scFv was then purified in a single chromatographic step using CM-Sephadex chromatography, a weak cation exchanger with a yield of 10.3 mg/L. The molecular weight of the scFv was found to be ~ 28 kDa by SDS PAGE, and its presence was confirmed by western blot analysis and mass spectrometry. Anti-TNF-α scFv has been successfully purified in a salt inducible system GJ1158. As per the best of our knowledge, this is the first report of purification of Anti-TNF-α scFv in a salt inducible system from soluble fractions as well as inclusion bodies.

Rapid Identification of Escherichia coli Colistin-Resistant Strains by MALDI-TOF Mass Spectrometry.

Colistin resistance is one of the major threats for global public health, requiring reliable and rapid susceptibility testing methods. The aim of this study was the evaluation of a MALDI-TOF mass spectrometry (MS) peak-based assay to distinguish colistin resistant (colR) from susceptible (colS) Escherichia coli strains. To this end, a classifying algorithm model (CAM) was developed, testing three different algorithms: Genetic Algorithm (GA), Supervised Neural Network (SNN) and Quick Classifier (QC). Among them, the SNN- and GA-based CAMs showed the best performances: recognition capability (RC) of 100% each one, and cross validation (CV) of 97.62% and 100%, respectively. Even if both algorithms shared similar RC and CV values, the SNN-based CAM was the best performing one, correctly identifying 67/71 (94.4%) of the E. coli strains collected: in point of fact, it correctly identified the greatest number of colS strains (42/43; 97.7%), despite its lower ability in identifying the colR strains (15/18; 83.3%). In conclusion, although broth microdilution remains the gold standard method for testing colistin susceptibility, the CAM represents a useful tool to rapidly screen colR and colS strains in clinical practice.

Genome and Methylome analysis of a phylogenetic novel Campylobacter coli cluster with C. jejuni introgression.

The intriguing recent discovery of Campylobacter coli strains, especially of clade 1, that (i) possess mosaic C. coli / C. jejuni alleles, (ii) demonstrate mixed multilocus sequence types (MLSTs) and (iii) have undergone genome-wide introgression has led to the speculation that these two species may be involved in an accelerated rate of horizontal gene transfer that is progressively leading to the merging of both species in a process coined ‘despeciation’. In an MLST-based neighbour-joining tree of a number of C. coli and C. jejuni isolates of different clades, three prominent Campylobacter isolates formed a seemingly separate cluster besides the previously described C. coli and C. jejuni clades. In the light of the suspected, ongoing genetic introgression between the C. coli and C. jejuni species, this cluster of Campylobacter isolates is proposed to present one of the hybrid clonal complexes in the despeciation process of the genus. Specific DNA methylation as well as restriction modification systems are known to be involved in selective uptake of external DNA and their role in such genetic introgression remains to be further investigated. In this study, the phylogeny and DNA methylation of these putative C. coli / C. jejuni hybrid strains were explored, their genomic mosaic structure caused by C. jejuni introgression was demonstrated and basic phenotypic assays were used to characterize these isolates. The genomes of the three hybrid Campylobacter strains were sequenced using PacBio SMRT sequencing, followed by methylome analysis by Restriction-Modification Finder and genome analysis by Parsnp, Smash++ and blast. Additionally, the strains were phenotypically characterized with respect to growth behaviour, motility, eukaryotic cell invasion and adhesion, autoagglutination, biofilm formation, and water survival ability. Our analyses show that the three hybrid Campylobacter strains are clade 1 C . coli strains, which have acquired between 8.1 and 9.1 % of their genome from C. jejuni . The C. jejuni genomic segments acquired are distributed over the entire genome and do not form a coherent cluster. Most of the genes originating from C. jejuni are involved in chemotaxis and motility, membrane transport, cell signalling, or the resistance to toxic compounds such as bile acids. Interspecies gene transfer from C. jejuni has contributed 8.1–9.1% to the genome of three C. coli isolates and initiated the despeciation between C. jejuni and C. coli . Based on their functional annotation, the genes originating from C. jejuni enable the adaptation of the three strains to an intra-intestinal habitat. The transfer of a fused type II restriction-modification system that recognizes the CAYNNNNNCTC/GAGNNNNNRTG motif seems to be the key for the recombination of the C. jejuni genetic material with C. coli genomes.

Characterization and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from chicken and pork

The prevalence and antimicrobial resistance (AMR) profile were investigated in Campylobacter jejuni and Campylobacter coli in chicken and pork in Fukuoka, Japan in 2019. Their AMR profiles were compared with those of C. jejuni and C. coli strains isolated in 2013. A total of 53 chicken and 14 pork samples were collected from different supermarkets in Fukuoka in 2019. Campylobacter spp. were isolated by conventional method and characterized by PCR and MALDI-TOF MS. Among 53 chicken samples tested in 2019, 24.5% and 5.7% were positive for C. jejuni and C. coli, respectively, and three (21.4%) of 14 pork samples were positive for C. coli, but not C. jejuni. From the positive samples, 13 and six strains of C. jejuni and C. coli were isolated, respectively. Antimicrobial susceptibility test against 12 different antimicrobials were performed on 48 isolates (43 C. jejuni and five C. coli) from chicken in 2013 and 19 isolates (13 C. jejuni from chicken, three C. coli from chicken and three C. coli from pork) in 2019 using the disk diffusion method. All the C. jejuni and C. coli isolated in 2013 and 2019 were highly resistant to cefazolin and sulfamethoxazole/trimethoprim. Among the C. jejuni isolates from chickens, 25.6% of 2013 isolates were resistant to nalidixic acid, ciprofloxacin, and levofloxacin, and 7% to ampicillin and minocycline, while 30.8% of the isolates were resistant to minocycline, 23.1% to nalidixic acid, ciprofloxacin, and levofloxacin, and 15.4% to ampicillin in 2019. Among the C. coli isolates, 80% of isolates from chickens in 2013, and 33.3% from chicken and 100% from pork in 2019 were resistant to nalidixic acid, ciprofloxacin, and levofloxacin. The frequency of multi-drug resistant (MDR) C. jejuni and C. coli strains from chickens in 2019 were 30.8% and 33.3%, respectively, which were lower than those isolated in 2013 (37.2% and 100%, respectively). One C. jejuni and two C. coli isolates from 2013 were resistant to six antibiotics. However, two C. jejuni and one C. coli isolate from chickens in 2019 were resistant to seven and five antibiotics, respectively. All the C. coli isolates from pork in 2019 were resistant to five antibiotics. The high frequency of AMR strains in C. coli isolates from pork suggests that appropriate use of antimicrobials is required in swine husbandry.

Anti-Bacterial Action of Plasma Multi-Jets in the Context of Chronic Wound Healing

This work is a contribution to the development and implementation of non-thermal plasma technology for decontamination in the perspective of nosocomial and chronic wound innovative therapies. Multi jets devices based on Plasma Gun® technology in static and scanning operation modes and bacterial lawns inoculated with resistant and non-resistant bacterial strains were designed and used. A pilot toxicity study exploring plasma treatment of wound bearing patients, performed with a low voltage plasma applicator, is documented as a first step for the translation of in vitro experiments to clinical care. Bacterial inactivation was demonstrated for Staphylococcus aureus, Pseudomonas aeruginosa and drug resistant S. aureus, P. aeruginosa and Escherichia Coli strains collected from patient wounds at Orleans (France) hospital. A few square centimeter large contaminated samples were inactivated following a single plasma exposure as short as one minute. Samples inoculated with a single but also a mix of three resistant pathogens were successfully inactivated not only right after their contamination but for mature lawns as well. Similar bactericidal action was demonstrated for antibiotic-resistant and non-resistant P. aeruginosa. The time exposure dependent increase of the inhibition spots, following multi jets exposure, is discussed as either the accumulation of reactive species or the likely combinatory action of both the reactive species and transient electric field delivery on inoculated samples.

Enhanced expression and purification of nucleotide-specific ribonucleases MC1 and Cusativin

Combinations of ribonucleases (RNases) are commonly used to digest RNA into oligoribonucleotide fragments prior to liquid chromatography-mass spectrometry (LC-MS) analysis. The distribution of the RNase target sequences or nucleobase sites within an RNA molecule is critical for achieving a high mapping coverage. Cusativin and MC1 are nucleotide-specific endoribonucleases encoded in the cucumber and bitter melon genomes, respectively. Their high specificity for cytidine (Cusativin) and uridine (MC1) make them ideal molecular biology tools for RNA modification mapping. However, heterogenous recombinant expression of either enzyme has been challenging because of their high toxicity to expression hosts and the requirement of posttranslational modifications. Here, we present two highly efficient and time-saving protocols that overcome these hurdles and enhance the expression and purification of these RNases. We first purified MC1 and Cusativin from bacteria by expressing and shuttling both enzymes to the periplasm as MBP-fusion proteins in T7 Express lysY/IqE. coli strain at low temperature. The RNases were enriched using amylose affinity chromatography, followed by a subsequent purification via a C-terminal 6xHIS tag. This fast, two-step purification allows for the purification of highly active recombinant RNases significantly surpassing yields reported in previous studies. In addition, we expressed and purified a Cusativin-CBD fusion enzyme in P. pastoris using chitin magnetic beads. Both Cusativin variants exhibited a similar sequence preference, suggesting that neither posttranslational modifications nor the epitope-tags have a substantial effect on the sequence specificity of the enzyme.

The Prevalence Of Uropathogenic Escherichia Coli Strains Among Outpatients With Urinary Tract Infection In Zakho City, Iraq

This study involved the prevalence of uropathogenic Escherichia coli (UPEC) among outpatients of UTI attending three major hospitals in Zakho city. Four hundred urine samples were collected from patients of UTI of both sexes and different ages (≤ 1 year to over 50 years), during the period from July 2018 until January 2019. All urine samples were analyzed by conventional bacteriological method for the presence of Escherichia coli (E. coli), while molecular method was used for the presence of species-specific uidA gene in the isolated E. coli. Out of 400 samples, 141 (35.25%) were infected with UPEC from enrolled patients. The rate was higher in females than males (90.78% vs 9.22%), respectively. In both sexes, the age group 41-50 years in both sexes showed the highest rate (46.67%) of infection, and statistically this rate of infection was significant (p< 0.013) among both sexes and various age groups. Furthermore, in all ages, married patients showed slightly higher prevalence than un-married one (38% vs 32.5%), but this difference was statistically non-significant (p>0.05%). The rate of UTI was higher among urban inhabitants (40.56%) than others. During the months of the year, the peak (90.48%) in both sexes was during December while the lowest rates (13.64%) was during January.

Antimicrobial resistance and mcr-1 gene in Escherichia coli isolated from poultry samples submitted to a bacteriology laboratory in South Africa.

Background and Aim:Antimicrobial resistance (AMR) and recently mobilized colistin resistance (mcr-1) associated colistin resistance among Escherichia coli isolates have been attributed to the overuse of antimicrobials in livestock production. E. coli remains an important pathogen, often associated with mortality and low carcass weight in poultry medicine; therefore, the need to use antimicrobials is common. The study aimed to determine the AMR profile and presence of mcr-1 and mcr-2 genes in avian pathogenic E. coli from poultry samples tested at a bacteriology laboratory for routine diagnosis. This is a first step in understanding the effectiveness of mitigation strategies.Materials and Methods:Fifty E. coli strains were assessed for resistance against ten antimicrobial drugs using broth microdilution. All isolates with a colistin minimum inhibitory concentration (MIC) of 2 μg/mL were analyzed for the presence of mcr-1 and mcr-2 genes by employing the polymerase chain reaction. For each isolate, the following farm information was obtained: farm location, type of farm, and on-farm use of colistin.Results:Sixty-eight percent of the strains were resistant to at least one antimicrobial; 44% were multiple drug-resistant (MDR). Most E. coli isolates were resistant to doxycycline (44%), trimethoprim-sulfamethoxazole (38%), ampicillin (32%), and enrofloxacin (32%). None of the E. coli strains was resistant to colistin sulfate (MIC90 of 2 μg/mL). Only one E. coli isolate held the mcr-1 gene; none carried the mcr-2 gene.Conclusion:Resistance among E. coli isolates in this study was fairly high. Resistance to commonly used antimicrobials was observed, such as doxycycline, trimethoprim-sulfamethoxazole, and enrofloxacin. Only a single E. coli strain carried the mcr-1 gene, suggesting that mcr-1 and mcr-2 genes are common among isolates in this study. The prevalence of AMR, however, suggests that farmers must implement standard biosecurity measures to reduce E. coli burden, and antimicrobial use to prolong the efficacy life span of some of these drugs.

Distribution of blaTEM Gene among Escherichia Coli Strains Isolated from Different Clinical Samples in Erbil City

Background: Emerging antibiotic resistance and Extended-Spectrum Beta-Lactamase (ESBL) producing Escherichia coli causing different nosocomial infections are rapidly increasing at alarming levels and it poses a major health burden in the 21st century globally. Objective: The aim of this study is to determine the distribution of blaTEM gene ESBLproducing E. coli from clinical specimens in different hospitals in Erbil city. Methods: A total of 200 samples were collected from (sputum, urine, wound) from patients attended public hospitals in Erbil city. The isolation and identification of Escherichia coliand antimicrobial susceptibilitywere performed by using Vitek 2 compact -lactamase production in E.coli was confirmed by using both Double disk diffusion and Standard disk diffusion techniques. Moreover, PCR technique was used for genotypic detection of an ESBL gene blaTEMaccording to the standard protocol. Results: Out of 200 samples 60(30%) of E. coli isolates and the highest rate of E. coli isolates were obtained from the urine samples (20%) and the lowest number was isolated from sputum specimens (2.5%). The ESBL-producing Escherichia coli isolates were detected using double disk synergy test (76.7%) in comparison to standard disk diffusion test (80%). Genotypic screening results confirmed that all ESBL-producing E. coli isolates (66.7%) were carried blaTEM gene (700 bp) in clinical specimens (50% urine, 13.33% wound and 3.33% sputum). All ESBL-positive E. coli isolates showed high rates of susceptibility to Carbapenems antibiotic group including Imipenem (83.3%), Meropenem (81.7%), and Ertapenem (80.0%). Conclusions: -lactamase gene in ESBL-producing E. coli observed in this study for the first time is considered as alarming because there is a limited treatment options remained for infections. Attempts to reduce the dissemination of multi-drug resistant E. coli through compliance with strict hospital infection control and prevention standards are imperative. Findings of this study may help clinicians selecting appropriate antimicrobial therapy in patients with different infections caused by ESBLproducing E. coli.

Effects of common Gram-negative pathogens causing male genitourinary-tract infections on human sperm functions

Male genitourinary tract (MGT) bacterial infections are considered responsible for 15% of male infertility, but the mechanisms underlying decreased semen quality are poorly known. We evaluated in vitro the effect of strains of Gram-negative uropathogenic species (two E.coli strains, three K. pneumoniae strains, P. aeruginosa and E. cloacae) on motility, viability, mitochondrial oxidative status, DNA fragmentation and caspase activity of human spermatozoa. All strains, except P. aeruginosa, reduced significantly sperm motility, with variable effects. Sperm Immobilizing Factor (SIF) was largely responsible for deteriorating effects on sperm motility of E. coli strains since they were completely reverted by knockout of SIF coding recX gene. Sequence alignment for RecX showed the presence of high homologous sequences in K. pneumoniae and E. cloacae but not in P. aeruginosa. These results suggest that, in addition to E.coli, other common uropathogenic Gram-negative bacteria affect sperm motility through RecX products. In addition to sperm motility, the E. coli strain ATCC 35218 also affected sperm viability, and induced caspase activity, oxidative stress and DNA fragmentation suggesting an interspecies variability in the amount and/or type of the produced spermatotoxic factors. In general, our results highlight the need for a careful evaluation of semen infections in the diagnostic process of the infertile man.

Antibiotic Resistance Profile of Escherichia coli Isolated from Bovine Subclinical Mastitis of Dairy Farms in Algeria from 2017 to 2019

Mastitis in cows is a major problem in dairy farms leading to a decrease in the quantity and quality of milk. The aim of the present study was to examine the association between the presence of Escherichia coli (E. coli) in milk and the subclinical mastitis, and to characterize the antibiotic resistance profiles of the isolated E. coli. In the current study, a total of 360 cow raw milk samples from three dairy farms of the region of Algiers were analyzed. The analysis period lasted from Spring 2017 to Winter 2019. The California Mastitis Test (CMT) was applied to detect subclinical mastitis. The E. coli strains were isolated from milk using conventional bacteriological methods. The antibiotic resistance profile of the isolated E. coli strains to 12 different antibiotics was tested using the disk diffusion method. On β-lactamase-producing strains, a double diffusion test was applied to identify the Extended-spectrum β-lactamase (ESBL) phenotype. Finally, the ctXx-M genes were amplified by PCR. Two-thirds (66.4%) of the milk samples were positive for the CMT test. A total of 97 E. coli strains were isolated from the milk samples, their resistance to antibiotics was tested, and 3.1% of the strains were resistant to trimethoprim-sulfamethoxazole, 6.2% to chloramphenicol, 12.3% to gentamicin, 13.4% to colistin, 23.3% to amoxicillin/clavulanate, 31.9% to kanamycin, 39.2% to enrofloxacin, 51.5% to cefotaxime, 52% to tetracycline, 57.7% to ampicillin, 74.3% to nalidixic acid, and 75.3% to amoxicillin. Furthermore, most of the E. coli strains (92.8%) were resistant to more than one antibiotic with a Multiple Antibiotic Resistance index ranging from 0 to 0.8. The 50 strains resistant to cefotaxime were analyzed for an ESBL phenotype. 39 of them (78%) were positive to the double-disk synergy test. Among the 39 ESBL positive strains, 27 (69.2%) were confirmed for the presence of a CTX-M gene by PCR. The present study showed that multiple drug-resistant E. coli, including ESBL-carriers, were frequently isolated from the milk of dairy cows in Algeria. The results underlined that the use of antibiotics on farms must be reasoned to avoid the spread of resistant strains in animals and human populations.

Manufacturing Characteristics of Probiotic Esherichia Coli Strain M-17 After Immobilization in Gel Carriers and Storage at Various Low Temperatures

Manufacturing Characteristics of Probiotic Esherichia Coli Strain M-17 After Immobilization in Gel Carriers and Storage at Various Low Temperatures

Efficacy of Novel Bacteriophages against Escherichia coli Biofilms on Stainless Steel

Biofilm formation by E. coli is a serious threat to meat processing plants. Chemical disinfectants often fail to eliminate biofilms; thus, bacteriophages are a promising alternative to solve this problem, since they are widely distributed, environmentally friendly, and nontoxic to humans. In this study, the biofilm formation of 10 E. coli strains isolated from the meat industry and E. coli ATCC BAA-1430 and ATCC 11303 were evaluated. Three strains, isolated from the meat contact surfaces, showed adhesion ability and produced extracellular polymeric substances. Biofilms of these three strains were developed onto stainless steel (SS) surfaces and enumerated at 2, 12, 24, 48, and 120 h, and were visualized by scanning electron microscopy. Subsequently, three bacteriophages showing podovirus morphology were isolated from ground beef and poultry liver samples, which showed lytic activity against the abovementioned biofilm-forming strains. SS surfaces with biofilms of 2, 14, and 48 h maturity were treated with mixed and individual bacteriophages at 8 and 9 log10 PFU/mL for 1 h. The results showed reductions greater than 6 log10 CFU/cm2 as a result of exposing SS surfaces with biofilms of 24 h maturity to 9 log10 PFU/mL of bacteriophages; however, the E. coli and bacteriophage strains, phage concentration, and biofilm development stage had significant effects on biofilm reduction (p < 0.05). In conclusion, the isolated bacteriophages showed effectiveness at reducing biofilms of isolated E. coli; however, it is necessary to increase the libraries of phages with lytic activity against the strains isolated from production environments.