Multiple drug resistance of Listeria monocytogenes isolated from aborted women by using serological and molecular techniques in Diwaniyah city/Iraq. - Related Documents




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234701.0000Multiple drug resistance of Listeria monocytogenes isolated from aborted women by using serological and molecular techniques in Diwaniyah city/Iraq. BACKGROUND AND OBJECTIVES: The study was sought to detect the effect of Listeria monocytogenes on pregnant Iraqi women at Al-Diwaniya hospitals and determination of virulence genes and antimicrobial susceptibility of isolates. MATERIALS AND METHODS: 360 specimens including blood, urine, vaginal and endocervical were collected from 90 patients with spontaneous abortions. Blood samples were displayed to immunological study and remaining specimens were subjected to bacteriological diagnosis. PCR was used to determine the virulence factors and antimicrobial resistance genes. RESULTS: Fifteen positive samples (16.6%) of patients and thirteen isolates (14.5%) from patients were recognized based on ELISA and PCR assay respectively. The general isolation of L. monocytogenes strains in cases of abortive women was 13/270 (4.8%). L. monocytogenes strains were highly virulent because of presence of virulence factors associated genes, namely actA, hlyA, plcA and prfA in all strains. Multiple drug resistance (MAR) index values of 15.4% of isolates were >0.2. CONCLUSION: It is necessary for conducting susceptibility testing and to select the suitable antibiotics and avoid the effects of these bacteria in pregnant women.202032994901
234610.9997Antibiotic resistance genes and molecular typing of Streptococcus agalactiae isolated from pregnant women. BACKGROUND: The antibiotic resistance of genital tract colonizing Streptococcus agalactiae in pregnant women is increasing. We aimed to determine the antibiotic resistance genes of different clonal types of this bacterium in pregnant women. METHODS: Four hundred twenty non-repeated vaginal and rectal specimens were collected from pregnant women and were transferred to the laboratory using Todd Hewitt Broth. The samples were cultured on a selective medium, and the grown bacteria were identified by standard microbiological and biochemical tests. Antimicrobial resistance pattern and inducible clindamycin resistance of the isolates were determined using the disk agar diffusion method. The genomic DNAs of S. agalactiae strains were extracted using an extraction kit, and the antibiotic resistance genes and RAPD types were detected using the PCR method. RESULTS: The average age of the participants was 30.74 ± 5.25 years. There was a significant relationship between the weeks of pregnancy and the number of positive bacterial cultures (P-value < 0.05). Moreover, 31 pregnant women had a history of abortion, and 18 had a history of membrane rupture. Among 420 specimens, 106 S. agalactiae isolates were detected. The highest antibiotic resistance rate was found against tetracycline (94.33%), and all isolates were susceptible to linezolid. Moreover, 15, 15, 42, and 7 isolates showed an iMLS(B), M-, cMLS(B), and L-phenotype. The ermB was the most prevalent resistance gene in the present study, while 38 (35.84%), 8 (7.54%), 79 (74.52%), 37 (34.9%), and 20 (18.86%) isolates were contained the ermTR, mefA/E, tetM, tetO, and aphA3 gene, respectively. CONCLUSIONS: The high-level antibiotic resistance and prevalence of resistance genes may be due to the arbitrarily use, livestock industry consumption, and the preventive use of antibiotics in pregnant women. Thus, the need to re-considering this problem seems to be necessary.202336658541
234920.9997DETECTION OF MECA AND NUC GENES OF MULTI-DRUG RESISTANT STAPHYLOCOCCUS AUREUS ISOLATED FROM DIFFERENT CLINICAL SAMPLES. BACKGROUND: During this study, six isolates of multiple antibiotic resistant Staphylococcus aureus bacteria were obtained from different clinical specimens (burn swabs, urinary tract infections, wound swabs): three isolates from burns, two isolates from urinary tract infections, and one isolate from wound swabs. They were obtained from private laboratories in Baghdad from 1/1/2023 to 3/15/2023. METHOD: The diagnosis of these isolates was confirmed using the Vitek2 device. A susceptibility test was conducted on ten antibiotics, and S. aureus bacteria showed resistance to most antibiotics, polymerase chain reaction was done to mecA and Nuc gene by conventional PCR. RESULTS: The results of the molecular detection of the MecA gene showed that all isolates of multi-drug-resistant S. aureus possess this gene. In contrast, the results of the molecular detection of the nuc gene showed that only isolates No. 1 and No. 4 carry this gene, while the rest of the isolates do not carry this gene. CONCLUSION: S. aureus are resistant to antibiotics because they possess resistance genes such as the mecA gene.202439724880
266730.9997Prevalence, virulence and antimicrobial resistance patterns of Aeromonas spp. isolated from children with diarrhea. BACKGROUND: Aeromonas spp. cause various intestinal and extraintestinal diseases. These bacteria are usually isolated from fecal samples, especially in children under five years old. The aim of this study was to assess the prevalence of Aeromonas spp. and their antimicrobial resistance profile in children with diarrhea referred to the Children Medical Center in Tehran, between 2013 and 2014. METHODS: A total number of 391 stool samples were collected from children with ages between 1 day and 14 years old, with diarrhea (acute or chronic), referred to the Children Hospital, Tehran, Iran, between 2013 and 2014. Samples were enriched in alkaline peptone water broth for 24 hours at 37 °C and then cultured. Suspicious colonies were analyzed through biochemical tests. Furthermore, antimicrobial susceptibility tests were carried out for the isolates. Isolates were further studied for act, ast, alt, aerA and hlyA virulence genes using polymerase chain reaction. RESULTS: In total, 12 isolates (3.1%) were identified as Aeromonas spp.; all were confirmed using the API-20E test. Of these isolates, five A. caviae (42%), four A. veronii (33%) and three A. hydrophila (25%) were identified in cases with gastroenteritis. Second to ampicillin (which was included in the growth medium used), the highest rate of antimicrobial resistance was seen against nalidixic acid and trimethoprim-sulfamethoxazole (5 isolates each, 41.6%) and the lowest rate of antimicrobial resistance was seen against gentamicin, amikacin and cefepime (none of the isolates). Results included 76.4% act, 64.7% ast, 71.5% alt, 83.3% aerA and 11.7% hlyA genes. CONCLUSION: Aeromonas spp. are important due to their role in diarrhea in children; therefore, isolation and identification of these fecal pathogens should seriously be considered in medical laboratories. Since virulence genes play a significant role in gastroenteritis symptoms caused by these bacteria, Aeromonas species that include virulence genes are potentially suspected to cause severe infections. Moreover, bacterial antimicrobial resistance is increasing, especially against trimethoprim-sulfamethoxazole and nalidixic acid.201627622161
236040.9997Evaluating the antibiotic resistance and frequency of adhesion markers among Escherichia coli isolated from type 2 diabetes patients with urinary tract infection and its association with common polymorphism of mannose-binding lectin gene. The present paper aims to determine the frequency and antibiotic resistance patterns of pathogenic bacteria, the virulence factor profile of Escherichia coli and mannose-binding lectin (MBL) gene polymorphism in individuals with diabetes mellitus (DM) and urinary tract infection (UTI). The population under study was 130 individuals with type 2 diabetes mellitus (T2DM) and UTI. The patients' clinical characteristics and urine and blood samples (5 mL) were collected. Antibiotic resistance was determined using a disc diffusion method, and the results were interpreted according to CLSI. The presence of virulence genes was detected by multiplex PCR. To detect the MBL gene polymorphism, PCR and restriction fragment length polymorphism methods were applied. The predominant Gram-negative and Gram-positive bacteria included E. coli and Streptococcus spp.viridans group, respectively. Women were more susceptible to the incidence of UTI than men. The E. coli isolates showed a high level of resistance to amoxicillin-clavulanic acid (87.35%), and nitrofurantoin and ceftizoxime were the most effective antimicrobial agents for E. coli. Cefotaxime and ceftizoxime were the most effective antimicrobial agents for Enterobacter spp., norfloxacin and ciprofloxacin were the most effective antimicrobial agents for Staphylococcus epidermidis and Staphylococcus saprophyticus. papGII (52.87%) and papEF (1.14%) had the highest and lowest frequency among examined genes in E. coli isolates, respectively. The GG genotype had the highest frequency among patients with T2DM and UTI. Results showed that the detection of E. coli in individuals with an AA genotype, codon 54 of the MBL gene, can play an important role in the molecular diagnosis and timely treatment of bacterial infections in individuals with diabetes.202033364032
215250.9997Immunological and molecular detection of biofilm formation and antibiotic resistance genes of Pseudomonas aeruginosa isolated from urinary tract. BACKGROUND AND OBJECTIVES: Pseudomonas aeruginosa (P. aeruginosa) is one of the most common causes of hospital-acquired infections. It is associated with high morbidity and healthcare costs, especially when appropriate antibiotic treatment is delayed. Antibiotic selection for patients with P. aeruginosa infections is challenging due to the bacteria's inherent resistance to many commercially available antibiotics. This study investigated antibiotic-resistance genes in isolated bacteria, which play a key role in disease pathogenesis. MATERIALS AND METHODS: 100 samples out of the 140 samples collected from urinary tract infections (UTIs) cases between December 15(th), 2022, and April 15(th), 2023, were included in the study. Identification of bacterial isolates was based on colony morphology, microscopic examination, biochemical tests, and the Vitek-2 system. Antibiotic resistance genes; Aph(3)-llla, ParC, Tet/tet(M), and aac(6´)-Ib-cr were tested by polymerase chain reaction (PCR). RESULTS: The obtained results were based on bacterial identifications of 81 clinical samples. Only 26 (32%) of these isolates were P. aeruginosa, 21 (26%) were Escherichia coli, and 18 (22.2%) were other bacteria. These isolates were used to detect four genes including tet(M), Aph(3)-llla, Par-c, and aac(6´)-Ib-cr. Four types of primers were used for PCR detection. The results showed that 11/14 (78.57%) carried the tet(M) gene, 10/14 (71.42%) carried the Aph(3)-llla gene, 14/14 (100%) carried the Par-c gene, and 10/14 (71.42%) of the isolates carried the aac(6´)-Ib-cr gene. The biofilm formation examining the esp gene, showed that 9 (64.28) isolates carried this gene. CONCLUSION: The inability of antibiotics to penetrate biofilms is an important factor contributing to the antibiotic tolerance of bacterial biofilms.202540612720
215360.9997Molecular Characterization and Epidemiology of Antibiotic Resistance Genes of β-Lactamase Producing Bacterial Pathogens Causing Septicemia from Tertiary Care Hospitals. Septicemia is a systematic inflammatory response and can be a consequence of abdominal, urinary tract and lung infections. Keeping in view the importance of Gram-negative bacteria as one of the leading causes of septicemia, the current study was designed with the aim to determine the antibiotic susceptibility pattern, the molecular basis for antibiotic resistance and the mutations in selected genes of bacterial isolates. In this study, clinical samples (n = 3389) were collected from potentially infected male (n = 1898) and female (n = 1491) patients. A total of 443 (13.07%) patients were found to be positive for bacterial growth, of whom 181 (40.8%) were Gram-positive and 262 (59.1%) were Gram-negative. The infected patients included 238 males, who made up 12.5% of the total number tested, and 205 females, who made up 13.7%. The identification of bacterial isolates revealed that 184 patients (41.5%) were infected with Escherichia coli and 78 (17.6%) with Pseudomonas aeruginosa. The clinical isolates were identified using Gram staining biochemical tests and were confirmed using polymerase chain reaction (PCR), with specific primers for E. coli (USP) and P. aeruginosa (oprL). Most of the isolates were resistant to aztreonam (ATM), cefotaxime (CTX), ampicillin (AMP) and trimethoprim/sulfamethoxazole (SXT), and were sensitive to tigecycline (TGC), meropenem (MEM) and imipenem (IPM), as revealed by high minimum inhibitory concentration (MIC) values. Among the antibiotic-resistant bacteria, 126 (28.4%) samples were positive for ESBL, 105 (23.7%) for AmpC β-lactamases and 45 (10.1%) for MBL. The sequencing and mutational analysis of antibiotic resistance genes revealed mutations in TEM, SHV and AAC genes. We conclude that antibiotic resistance is increasing; this requires the attention of health authorities and clinicians for proper management of the disease burden.202336978484
220470.9997Multidrug resistance pattern of bacterial agents isolated from patient with chronic sinusitis. BACKGROUND: Treatment of chronic sinusitis is complicated due to increase of antibiotic-resistant bacteria. The aim of this study was to determine the multidrug resistance (MDR) pattern of the bacteria causing chronic sinusitis in north of Iran. METHODS: This cross-sectional study was carried out on patients with chronic sinusitis. Bacterial susceptibility to antimicrobial agents was determined according to the CLSI 2013 standards. Double-disk synergy (DDS) test was performed for the detection of extended-spectrum beta-lactamase (ESBL) producing bacteria; also methicillin-resistant Staphylococcus (MRSA) strains were identified by MRSA screen agar. The MDR isolates were defined as resistant to 3 or more antibiotics. Data were analyzed using SPSS 17 software. Descriptive statistics was used to describe the features of the data in this study. RESULTS: The rate of ESBL-producing bacteria was 28.75-37.03% among enterobacteriaceae and the rate of MRSA was 42.75%-60% among Staphylococcus strains. The most detectable rate of the MDR bacterial isolates was Gram-negative bacteria 39 (76.47%) and Enterobacter spp. 19(70.37%) was the most multidrug resistant isolate among Gram negative bacteria. Also 36 (73.46%) of the gram positive bacterial isolated were multidrug resistance and Staphylococcus aureus 9(90%) was the most MDR among Gram positive bacteria. CONCLUSION: Antimicrobial resistance is increasing in chronic bacterial sinusitis. The emergence of MRSA and ESBL bacteria causing chronic sinusitis is increasing.201627386063
215780.9997Prevalence and antibiotic resistance pattern of bacteria isolated from urinary tract infections in Northern Iran. BACKGROUND: This study aimed to investigate the bacteria associated with urinary tract infection (UTI) and antibiotic susceptibility pattern of the isolates during 2013-2015 in Northern Iran. MATERIALS AND METHODS: Overall 3798 patients with clinical symptoms of UTI were subjected as samples, and they were cultured and pure isolated bacteria were identified using biochemical tests and subjected to antibiogram assessment using disc diffusion method. RESULTS: Totally, 568 (14.96%) from 3798 patients had positive UTI. Four hundred and ninety-seven (87.5%) from 568 isolated bacteria were resistant to at least one antibiotic. Escherichia coli, Staphylococcus spp., and Pseudomonas spp. were the most prevalent bacteria. Isolated bacteria indicated the highest antibiotic resistance to methicillin (76.06%) and ampicillin (89.29%) and also revealed the most sensitivity to imipenem (99.1%) and amikacin (91.57%). Statistical analysis of the resistance pattern trend during 3 years indicated the insignificant increase (P > 0.05) in antibiotic resistance of the isolates. CONCLUSION: The results of this study revealed a great concern for emerging UTI-related multidrug-resistant strains of bacteria causing UTI in Iran.201729026424
235990.9996Virulence Factor Genes and Antimicrobial Susceptibility of Staphylococcus aureus Strains Isolated from Blood and Chronic Wounds. Staphylococcus aureus is one of the predominant bacteria isolated from skin and soft tissue infections and a common cause of bloodstream infections. The aim of this study was to compare the rate of resistance to various antimicrobial agents and virulence patterns in a total of 200 S. aureus strains isolated from patients with bacteremia and chronic wounds. Disk diffusion assay and in the case of vancomycin and teicoplanin-microdilution assay, were performed to study the antimicrobial susceptibility of the isolates. The prevalence of genes encoding six enterotoxins, two exfoliative toxins, the Panton-Valentine leukocidin and the toxic shock syndrome toxin was determined by PCR. Of the 100 blood strains tested, the highest percentage (85.0%, 31.0%, and 29.0%) were resistant to benzylpenicillin, erythromycin and clindamycin, respectively. Out of the 100 chronic wound strains, the highest percentage (86.0%, 32.0%, 31.0%, 31.0%, 30.0%, and 29.0%) were confirmed as resistant to benzylpenicillin, tobramycin, amikacin, norfloxacin, erythromycin, and clindamycin, respectively. A significantly higher prevalence of resistance to amikacin, gentamicin, and tobramycin was noted in strains obtained from chronic wounds. Moreover, a significant difference in the distribution of sea and sei genes was found. These genes were detected in 6.0%, 46.0% of blood strains and in 19.0%, and 61.0% of wound strains, respectively. Our results suggest that S. aureus strains obtained from chronic wounds seem to be more often resistant to antibiotics and harbor more virulence genes compared to strains isolated from blood.202134357963
2137100.9996High prevalence of antibiotic resistance and biofilm formation in Salmonella Gallinarum. BACKGROUND AND OBJECTIVES: Antibiotic resistance is an indicator of the passively acquired and circulating resistance genes. Salmonella Gallinarum significantly affects the poultry food industry. The present study is the first study of the S. Gallinarum biofilm in Iran, which is focused on the characterization of the S. Gallinarum serovars and their acquired antibiotic resistance genes circulating in poultry fields in central and northwestern Iran. MATERIALS AND METHODS: Sixty isolates of S. Gallinarum serovar were collected from feces of live poultry. The bacteria were isolated using biochemical tests and confirmed by Multiplex PCR. Biofilm formation ability and the antibacterial resistance were evaluated using both phenotypic and genotypic methods. The data were analyzed using SPSS software. RESULTS: According to Multiplex PCR for ratA, SteB, and rhs genes, all 60 S. Gallinarum serovars were Gallinarum biovars. In our study, the antibiotic resistance rate among isolated strains was as follows: Penicillin (100%), nitrofurantoin (80%), nalidixic acid (45%), cefoxitin (35%), neomycin sulfate (30%), chloramphenicol (20%), and ciprofloxacin (5%). All isolates were susceptible to imipenem, ertapenem, ceftriaxone, ceftazidime, and ceftazidime+clavulanic acid. All sixty isolates did not express the resistance genes IMP, VIM, NDM, DHA, bla(OXA48), and qnrA. On the other hand, they expressed GES (85%), qnrB (75%), Fox M (70%), SHV (60%), CITM (20%), KPC (15%), FOX (10%), MOXM (5%), and qnrS (5%). All S. Gallinarum isolates formed biofilm and expressed sdiA gene. CONCLUSION: Considering that the presence of this bacteria is equal to the death penalty to the herd, the distribution of resistance genes could be a critical alarm for pathogen monitoring programs in the region. This study showed a positive correlation between biofilm formation and 50% of tested resistance genes. Also, it was found that the most common circulating S. gallinarum biovars are multidrug-resistant.202337941876
2355110.9996Causative bacteria and antibiotic resistance in neonatal sepsis. BACKGROUND: Neonatal sepsis is characterised by bacteraemia and clinical symptoms caused by microorganisms and their toxic products. Gram negative bacteria are the commonest causes of neonatal Sepsis. The resistance to the commonly used antibiotics is alarmingly high. The major reason for emerging resistance against antibiotics is that doctors often do not take blood cultures before starting antibiotics. We have carried out this study to find out various bacteria causing neonatal sepsis and their susceptibility to antibiotics for better management of neonatal sepsis. METHODS: A total of 130 neonates with sepsis who were found to be blood culture positive were taken in this study. Culture/sensitivity was done, isolated organisms identified and their sensitivity/resistance was noted against different antibiotics. Data were arranged in terms of frequencies and percentage. RESULTS: Out of 130 culture proven cases of neonatal sepsis, gram negative bacteria were found in 71 (54.6%) cases and gram positive bacteria in 59 (45.4%) cases. Staphylococcus aureus was the most common bacteria found in 35 (26.9%) cases followed by Escherichia coli in 30 (23.1%) cases. Acinetobacter species, Staphylococcus epidermidis, Klebseila, Streptococci, Enterobacter cloacae and Morexella species were found in 17 (13.1%), 17 (13.1%), 13 (10%), 7 (5.4%), 6 (4.6%), and 5 (3.8%) cases respectively. In most of the cases causative organisms were found to be resistant to commonly used antibiotics like ampicillin, amoxicillin, cefotaxime, and ceftriaxone (77.7%, 81.5%, 63.1%, and 66.9% respectively). There was comparatively less (56.9%) resistance to ceftazidime. Gentamicin had resistance in 55.1% cases, while amikacin and tobramycin had relatively less resistance (17.4% and 34.8% cases respectively). Quinolones and imipenem had relatively less resistance. Vancomycin was found to be effective in 100% cases of Staphylococcus group. CONCLUSION: Staphylococcus aureus are the most common gram positive bacteria and Escherichia coli are the most common gram negative bacteria causing neonatal sepsis. Resistance to commonly used antibiotics is alarmingly increasing. Continued surveillance is mandatory to assess the resistance pattern at a certain level.201224669633
2234120.9996Clinical relevance of molecular identification of microorganisms and detection of antimicrobial resistance genes in bloodstream infections of paediatric cancer patients. BACKGROUND: Bloodstream infections (BSIs) are the major cause of mortality in cancer patients. Molecular techniques are used for rapid diagnosis of BSI, allowing early therapy and improving survival. We aimed to establish whether real-time quantitative polymerase chain reaction (qPCR) could improve early diagnosis and therapy in paediatric cancer patients, and describe the predominant pathogens of BSI and their antimicrobial susceptibility. METHODS: Blood samples were processed by the BACTEC system and microbial identification and susceptibility tests were performed by the Phoenix system. All samples were screened by multiplex 16 s rDNA qPCR. Seventeen species were evaluated using sex-specific TaqMan probes and resistance genes blaSHV, blaTEM, blaCTX, blaKPC, blaIMP, blaSPM, blaVIM, vanA, vanB and mecA were screened by SYBR Green reactions. Therapeutic efficacy was evaluated at the time of positive blood culture and at final phenotypic identification and antimicrobial susceptibility results. RESULTS: We analyzed 69 episodes of BSI from 64 patients. Gram-positive bacteria were identified in 61 % of the samples, Gram-negative bacteria in 32 % and fungi in 7 %. There was 78.2 % of agreement between the phenotypic and molecular methods in final species identification. The mecA gene was detected in 81.4 % of Staphylococcus spp., and 91.6 % were concordant with the phenotypic method. Detection of vanA gene was 100 % concordant. The concordance for Gram-negative susceptibilities was 71.4 % for Enterobacteriaceae and 50 % for Pseudomonas aeruginosa. Therapy was more frequently inadequate in patients who died, and the molecular test was concordant with the phenotypic susceptibility test in 50 %. CONCLUSIONS: qPCR has potential indication for early identification of pathogens and antimicrobial resistance genes from BSI in paediatric cancer patients and may improve antimicrobial therapy.201627585633
2156130.9996Antimicrobial resistance in urinary pathogens and culture-independent detection of trimethoprim resistance in urine from patients with urinary tract infection. BACKGROUND: Although urinary tract infections (UTIs) are extremely common, isolation of causative uropathogens is not always routinely performed, with antibiotics frequently prescribed empirically. This study determined the susceptibility of urinary isolates from two Health and Social Care Trusts (HSCTs) in Northern Ireland to a range of antibiotics commonly used in the treatment of UTIs. Furthermore, we determined if detection of trimethoprim resistance genes (dfrA) could be used as a potential biomarker for rapid detection of phenotypic trimethoprim resistance in urinary pathogens and from urine without culture. METHODS: Susceptibility of E. coli and Klebsiella spp. isolates (n = 124) to trimethoprim, amoxicillin, ceftazidime, ciprofloxacin, co-amoxiclav and nitrofurantoin in addition to susceptibility of Proteus mirabilis (n = 61) and Staphylococcus saprophyticus (n = 17) to trimethoprim was determined by ETEST® and interpreted according to EUCAST breakpoints. PCR was used to detect dfrA genes in bacterial isolates (n = 202) and urine samples(n = 94). RESULTS: Resistance to trimethoprim was observed in 37/124 (29.8%) E. coli and Klebsiella spp. isolates with an MIC(90) > 32 mg/L. DfrA genes were detected in 29/37 (78.4%) trimethoprim-resistant isolates. Detection of dfrA was highly sensitive (93.6%) and specific (91.4%) in predicting phenotypic trimethoprim resistance among E. coli and Klebsiella spp. isolates. The dfrA genes analysed were detected using a culture-independent PCR method in 16/94 (17%) urine samples. Phenotypic trimethoprim resistance was apparent in isolates cultured from 15/16 (94%) dfrA-positive urine samples. There was a significant association (P < 0.0001) between the presence of dfrA and trimethoprim resistance in urine samples containing Gram-negative bacteria (Sensitivity = 75%; Specificity = 96.9%; PPV = 93.8%; NPV = 86.1%). CONCLUSIONS: This study demonstrates that molecular detection of dfrA genes is a good indicator of trimethoprim resistance without the need for culture and susceptibility testing.202235610571
2151140.9996Study of the Genomic Characterization of Antibiotic-Resistant Escherichia Coli Isolated From Iraqi Patients with Urinary Tract Infections. Urinary tract infection is one of the last diseases prevalent in humans, with various causative agents affecting 250 million people annually, This study analyzed UTIs in Iraqi patients caused by Escherichia coli. ESBL enzymes contribute to antibiotic resistance. The research aimed to analyze ESBL gene frequency, resistance patterns, and genetic diversity of E. coli strains; Between Dec 2020 and May 2021, 200 urine samples were collected, cultured on blood agar, EMB, and MacConkey's plates, samples incubated at 37 °C for 24 h. Positive samples (> 100 cfu/ml) underwent Kirby-Bauer and CLSI antibiotic susceptibility testing. PCR detected virulence genes, Beta-lactamase coding genes, and biofilm-associated resistance genes in E. coli isolates; Out of 200 isolates, 80% comprised Gram-positive and Gram-negative bacteria. Specifically, 120 isolates (60%) were Gram-negative, while 40 isolates (20%) were Gram-positive. Among Gram-negative isolates, 20% were identified as E. coli. Remarkably, all E. coli strains showed resistance to all tested antibiotics, ranging from 80 to 95% resistance. The E. coli isolates harbored three identified resistance genes: blaTEM, blaSHV, and blaCTXM. Regarding biofilm production, 10% showed no formation, 12% weak formation, 62% moderate formation, and 16% strong formation; our study found that pathogenic E. coli caused 20% of UTIs. The majority of studied E. coli strains from UTI patients carried the identified virulence genes, which are vital for infection development and persistence.202439011020
2186150.9996Bacterial drug-resistance patterns and genetic diversity of bacteria-associated bacteriuria in diabetic patients in Ghana. OBJECTIVES: Our study aimed to determine the etiology of urinary tract infections (UTIs), resistance profiles of isolated bacteria, and virulence factors of Escherichia coli associated with bacteriuria in diabetic patients in Ghana. METHODS: Midstream urine samples from 982 diabetic patients were tested for uropathogens at the National Diabetes Management and Research Centre in Ghana, using standard bacteriological methods, with antibiogram testing of the isolates using the Kirby-Bauer disk diffusion, as per CLSI guidelines. Polymerase chain reaction (PCR) was used to investigate the phylogenetic groupings and virulence factor (VF) genes of isolated E. coli. RESULTS: The overall prevalence of UTIs was 9.2%, and the main uropathogens were Klebsiella spp. (55.6%) and Escherichia coli (31.3%). Age, duration of diabetes, and a previous history of UTIs were risk factors associated with UTI (p-value < 0.05). High levels of antibacterial resistance to cefuroxime (84%), ampicillin (80%), and gentamicin (70.7%) were observed. The distribution of VFs in each phylogenetic group revealed that sfa-iutA-KpsTMII-KpsTMIII genes were associated with group B2, and iutA-ibe were associated with group D. CONCLUSIONS: The isolated uropathogens were highly resistant, and the E. coli isolates possessed varying VFs. Continuous monitoring of bacteria associated with UTI in diabetics is highly recommended.202135757820
1166160.9996Yearly incidence of acute childhood gastroenteritis in Nigeria: Implicated pathogens predominantly harbor blaCTXM and blaTEM genes. BACKGROUND AND OBJECTIVES: Routine use of antibiotics for infectious diarrhea in children is associated with the risk of increasing antibiotic resistance in developing countries. This work aimed to study the predominant extended spectrum beta-lactamase (ESBL) genes among bacteria pathogens implicated in acute childhood gastroenteritis in a tertiary hospital in Nigeria. MATERIALS AND METHODS: The stool samples of children diagnosed with acute gastroenteritis were collected. Isolation and identification of bacterial pathogens from the stool samples using standard microbiological and molecular sequencing methods. Pure cultures of the probable bacteria pathogens were subjected to antibiotics susceptibility profiling using the Kirby-Bauer Disk Diffusion Method and also screened for ESBL and AmpC using the Modified Double Disc Synergy Test. Primers for 5 different ESBL genes associated with beta-lactam antibiotic resistance were amplified and sequenced. RESULTS: Out of the 62 isolates, the highest number of organisms identified within the isolates were Bacillus sp at 38.7% (24) followed by Alcaligenes sp at 37% (23). Resistance to cefepime and ceftazidime were recorded at 50.8% (30) each. Ceftriaxone and cefotaxime were resisted in 47.4% (28) of the isolates. Out of 34 isolates resistant to all the cephalosporins used, 41.2% (14) were ESBL-producing, of which blaCTXM-1 and blaCTXM-2 were detected in 85.7%, while blaTEM was seen in 64.3%. CONCLUSIONS: blaCTXM and blaTEM may be the predominant ESBL genes haboured in the bacteria pathogens implicated in the yearly incidence of acute childhood gastroenteritis in Nigeria. This may be due to the widespread use of antibiotics in treating this disease.202539977466
2651170.9996Virulence and resistance determinants in Pseudomonas aeruginosa isolated from pericarditis in diseased broiler chickens in Egypt. OBJECTIVE: This study was performed to probe the antimicrobial resistance and virulence genes profiling in Pseudomonas aeruginosa recovered from the cases of pericarditis in broiler chickens. MATERIALS AND METHODS: The samples (n = 250) collected from the cases of pericarditis in broiler chickens were bacteriologically examined. Antimicrobial susceptibility was tested by disc diffusion technique. The isolates were genotypically studied for the presence of antimicrobial resistance and virulence gene traits. Finally, the nucleotide sequence of representative resistance gene (mexR gene) and virulence genes (toxA and lasI genes) was analyzed. RESULTS: P. aeruginosa was isolated from 45 samples (18%). Antimicrobial susceptibility testing revealed multidrug resistance in most of the recovered P. aeruginosa isolates, whereas colistin and imipenem were the furthermost in vitro-sensitive antibiotics. Antimicrobial resistance genes, such as bla (CTX), fox, and mexR, were prevalent in 100%, 80%, and 100% of the isolates, respectively. PCR confirmed virulence genes such as toxA, exoY, lasB, and lasI in 100%, 60%, 80%, and 80% of the isolates, respectively. Nucleotide sequence analysis of representative resistance gene (mexR gene) and virulence genes (toxA and lasI genes) revealed a high correlation between P. aeruginosa recovered from pericarditis in broiler chickens in the present study with PAO1 (reference strain) and with other sequences published on the GenBank representing different localities worldwide. CONCLUSION: It could be concluded that P. aeruginosa recovered from pericarditis in broiler chickens in the current study is highly virulent bacteria, resisting most of the therapeutic agents which not only bear hazards for poultry industry but also represent a public health concern.202033005671
1130180.9996The characteristic of antibiotic drug resistance of Salmonella Typhi isolated from tertiary care hospital in Faisalabad. Salmonella Typhi, a human-restricted pathogen, is demonstrating multi-drug resistance (MDR) due to widespread and inappropriate antibiotic use. This study aims to molecular identify the pattern of antibiotic resistance. Blood samples from 2456 suspected patients were assessed. Molecular identification of Salmonella Typhi was performed by amplifying the fliC gene. The Disc diffusion method was used to measure the susceptibility of antibiotics. 2456 patient samples, bacterial growth and Salmonella Typhi were 152 (6.2 %) positive. PCR analysis confirmed that all 152 isolated strains were Salmonella Typhi (100%) through the amplification of the fliC gene. Salmonella Typhi isolates showed resistance to trimethoprim (58%), ampicillin (63%), ciprofloxacin (79%) and chloramphenicol (58%). Fifty-eight percent of the isolates showed multi-drug resistance, whereas 26 percent had extensive drug resistance. Antibiotic resistance gene of quinolones was isolated as 44 (36.4%), whereas 88 (57.9 %) were positive for bla(CTX-M) gene were detected among cephalosporin-resistance bacteria 56 (36.8 %) resistance bla(IMP) and bla(OXA-48) were detected among carbapenem-resistance bacteria. For the azithromycin resistance, more genes were detected as a percentage 03 (50 %) from isolates. It concludes that several multidrug resistance and extensive drug-resistance Salmonella Typhi were found. The majority of isolates were sensitive to meropenem, Imipenem and Azithromycin.202540996203
2220190.9996Rapid detection and molecular survey of blaVIM, blaIMP and blaNDM genes among clinical isolates of Acinetobacter baumannii using new multiplex real-time PCR and melting curve analysis. BACKGROUND: Acinetobacter baumannii is a cosmopolitan bacterium that is frequently reported from hospitalized patients, especially those patients who admitted in the intensive care unit. Recently, multiplex real-time PCR has been introduced for rapid detection of the resistance genes in clinical isolates of bacteria. The current study aimed to develop and evaluate multiplex real-time PCR to detect common resistance genes among clinical isolates of A. baumannii. RESULTS: Multiplex real-time PCR based on melting curve analysis showed different T(m) corresponding to the amplified fragment consisted of 83.5 °C, 93.3 °C and 89.3 °C for blaIMP, blaVIM and blaNDM, respectively. Results of multiplex real-time PCR showed that the prevalence of blaIMP, blaVIM and blaNDM among the clinical isolates of A. baumannii were 5/128(3.9%), 9/128(7.03%) and 0/128(0%), respectively. Multiplex real-time PCR was able to simultaneously identify the resistance genes, while showed 100% concordance with the results of conventional PCR. CONCLUSIONS: The current study showed that blaVIM, was the most prevalent MBL gene among the clinical isolates of A. baumannii while no amplification of blaNDM was seen. Multiplex real-time PCR can be sensitive and reliable technique for rapid detection of resistance genes in clinical isolates.201931182026