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218900.9980High prevalence of Panton-Valentine Leucocidin (PVL) toxin carrying MRSA and multidrug resistant gram negative bacteria in late onset neonatal sepsis indicate nosocomial spread in a Pakistani tertiary care hospital. BACKGROUND: Neonatal sepsis has high incidence with significant mortality and morbidity rates in Pakistan. We investigated common etiological patterns of neonatal sepsis at a tertiary care setup. METHODS: 90 pus and blood, gram negative and gram positive bacterial isolates were analyzed for virulence and antibiotic resistance gene profiling using PCR and disc diffusion methods. RESULTS: Staphylococcus aureus showed strong association with neonatal sepsis (43 %) followed by Citrobacter freundii (21 %), Pseudomonas aeruginosa (13 %), Escherichia coli (15 %) and Salmonella enterica (8 %). Molecular typing of E. coli isolates depicted high prevalence of the virulent F and B2 phylogroups, with 4 hypervirulent phylogroup G isolates. 76.9 % S. aureus isolates showed presence of Luk-PV, encoding for Panton-valentine leucocidin (PVL) toxin with majority also carrying MecA gene and classified as methicillin resistant S. aureus (MRSA). ecpA, papC, fimH and traT virulence genes were detected in E. coli and Salmonella isolates. 47 % Citrobacter freundii isolates carried the shiga like toxin SltII B. Antimicrobial resistance profiling depicted common resistance to cephalosporins, beta lactams and fluoroquinolones. CONCLUSION: Presence of PVL carrying MRSA and multidrug resistant gram negative bacteria, all isolated from late onset sepsis neonates indicate a predominant nosocomial transmission pattern which may complicate management of the disease in NICU setups.202336621204
133510.9979Prevalence of virulence factor, antibiotic resistance, and serotype genes of Pasteurella multocida strains isolated from pigs in Vietnam. AIM: The study was conducted to determine the prevalence and characterization of the Pasteurella multocida isolates from suspected pigs in Vietnam. MATERIALS AND METHODS: A total of 83 P. multocida strains were isolated from lung samples and nasal swabs collected from pigs associated with pneumonia, progressive atrophic rhinitis, or reproductive and respiratory symptoms. Isolates were subjected to multiplex polymerase chain reaction (PCR) for capsular typing, detection of virulence-associated genes and antibiotic resistance genes by PCR. The antimicrobial sensitivity profiles of the isolates were tested by disk diffusion method. RESULTS: All the isolates 83/83 (100%) were identified as P. multocida by PCR: serogroup A was obtained from 40/83 (48.19%), serogroup D was detected from 24/83 strains (28.91%), and serogroup B was found in 19/83 (22.35%) isolates. The presence of 14 virulence genes was reported including adhesins group (ptfA - 93.97%, pfhA - 93.97%, and fimA - 90.36%), iron acquisition (exbB - 100%, and exbD - 85.54%), hyaluronidase (pmHAS - 84.33%), and protectins (ompA - 56.62%, ompH 68.67%, and oma87 - 100%). The dermonecrotoxin toxA had low prevalence (19.28%). The antimicrobial susceptibility testing revealed that cephalexin, cefotaxime, ceftriaxone, ofloxacin, pefloxacin, ciprofloxacin, and enrofloxacin were the drugs most likely active against P. multocida while amoxicillin and tetracycline were inactive. The usage of PCR revealed that 63/83 isolates were carrying at least one of the drug resistance genes. CONCLUSION: Unlike other parts of the word, serotype B was prevalent among Vietnamese porcine P. multocida strains. The high antibiotic resistance detected among these isolates gives us an alert about the current state of imprudent antibiotic usage in controlling the pathogenic bacteria.202032636585
125820.9979Occurrence of antimicrobial resistance and antimicrobial resistance genes in methicillin-resistant Staphylococcus aureus isolated from healthy rabbits. BACKGROUND AND AIM: Methicillin-resistant globally, Staphylococcus aureus (MRSA) is a major cause of disease in both humans and animals. Several studies have documented the presence of MRSA in healthy and infected animals. However, there is less information on MRSA occurrence in exotic pets, especially healthy rabbits. This study aimed to look into the antimicrobial resistance profile, hidden antimicrobial-resistant genes in isolated bacteria, and to estimate prevalence of MRSA in healthy rabbits. MATERIALS AND METHODS: Two-hundreds and eighteen samples, including 42 eyes, 44 ears, 44 oral, 44 ventral thoracic, and 44 perineal swabs, were taken from 44 healthy rabbits that visited the Prasu-Arthorn Animal Hospital, in Nakornpathom, Thailand, from January 2015 to March 2016. The traditional methods of Gram stain, mannitol fermentation, hemolysis on blood agar, catalase test, and coagulase production were used to confirm the presence of Staphylococcus aureus in all specimens. All bacterial isolates were determined by antimicrobial susceptibility test by the disk diffusion method. The polymerase chain reaction was used to identify the antimicrobial-resistant genes (blaZ, mecA, aacA-aphD, msrA, tetK, gyrA, grlA, and dfrG) in isolates of MRSA with a cefoxitin-resistant phenotype. RESULTS: From 218 specimens, 185 S. aureus were isolated, with the majority of these being found in the oral cavity (29.73%) and ventral thoracic area (22.7%), respectively. Forty-seven (25.41%) MRSAs were found in S. aureus isolates, with the majority of these being found in the perineum (16, 34.04%) and ventral thoracic area (13, 27.66%) specimens. Among MRSAs, 29 (61.7%) isolates were multidrug-resistant (MDR) strains. Most of MRSA isolates were resistant to penicillin (100%), followed by ceftriaxone (44.68%) and azithromycin (44.68%). In addition, these bacteria contained the most drug-resistance genes, blaZ (47.83%), followed by gyrA (36.17%) and tetK (23.4%). CONCLUSION: This study revealed that MRSA could be found even in healthy rabbits. Some MRSAs strains were MDR-MRSA, which means that when an infection occurs, the available antibiotics were not effective in treating it. To prevent the spread of MDR-MRSA from pets to owners, it may be helpful to educate owners about effective prevention and hygiene measures.202236590129
147230.9979Incidence and antibiotic susceptibility profile of uropathogenic Escherichia coli positive for extended spectrum β-lactamase among HIV/AIDS patients in Awka metropolis, Nigeria. BACKGROUND AND OBJECTIVES: This study investigated the incidence and antibiotic susceptibility profile of extended spectrum β-lactamase (ESBL) producing uropathogenic Escherichia coli recovered from HIV/AIDS patients in Awka metropolis, Nigeria. MATERIALS AND METHODS: A total of 363 urine samples were bacteriologically analyzed for the isolation of E. coli isolates which were further characterized using standard microbiology techniques. The isolated uropathogenic E. coli was tested for susceptibility to a range of clinically important antibiotics using the modified disk diffusion technique. All E. coli isolates were phenotypically screened for ESBL production using the combined disk technique, and strains which were positive were further confirmed for the presence of ESBL genes using PCR technique. RESULTS: A total 160 (44.1%) non-duplicate isolates were bacteriologically confirmed to be uropathogenic E. coli (UPEC). The E. coli isolates showed reduced susceptibility to important antibiotics including ceftazidime (76.88%), cefuroxime (77.5%), cefixime (61.88%), amoxicillin-clavulanic (32.5%) and ciprofloxacin (34.38%). Twenty-seven of the UPEC isolates were phenotypically confirmed to be ESBL producers. PCR test confirmed some important genes mediating ESBL production in Gram negative bacteria including bla (TEM) (5.0%) and bla (CTX-M-15) (6.9%) genes. CONCLUSION: We report a high prevalence of ESBL producers among HIV/AIDS patients in Awka, Nigeria. This result is important as antibiotic resistance (ABR) particularly those mediated by multidrug resistant bacteria as reported in this current study could complicate treatment outcome, worsen the individual's health, and even increase cost of treatment and hospitalization. It is therefore important to lookout for ESBL positive UPEC amongst HIV/AIDS patients in Nigeria.202237124857
127640.9978Should we leave the paper currency? A microbiological examination. OBJECTIVE: Pathogens can be transmitted to banknotes due to the personal unhygienic habits. The aim of study was to find the possible pathogens on the banknotes circulating in the market and also to present their antibacterial resistance and their various virulence factors using genotypic and phenotypic methods. METHODS: A total of 150 samples of bank-notes were randomly collected between August 2017 and March 2018. VITEK systems were used for identification and antimicrobial susceptibility testing respectively. Antimicrobial resistance genes (mecA, van, extended-spectrum β-lactamase [ESBL] and carbapenemases) and staphyloccoccal virulence genes (staphyloccoccal enterotoxins [SEs], pvl, and tsst-1) were determined using with real-time PCR. RESULTS: Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Enterococcus spp., Gram-negative enteric bacteria, non-fermentative Gram-negative bacteria and Candida spp. were detected 48%, 54.7%, 56%, 21.3%, 18.7%, and 4%, respectively. Methicillin-resistant S. aureus, vancomycin-resistant enterococci and ESBL producing Gram-negative were found 46.8%, 1.3%, and 28.7%, respectively. Pvl, tsst-1, and SEs genes were found in a 2.8/4.9%, 1.4/1.2%, and 100/ 87.8% of the S. aureus/CoNS strains, respectively. The sea gene was found the most common enterotoxigenic gene. blaTEM, blaSHV, blaCTX-M-2, blaCTX-M-1, blaKPC, and blaOXA-48 were found 55.8%, 46.5%, 41.2%, 18.6%, 18.6%, and 18.6%, respectively in Gram-negative strains. CONCLUSIONS: These results is very important to highlight hygienic status of paper currencies. This can be considered as an indication that banknotes may contribute to the spread of pathogens and antimicrobial resistance. Therefore, we may need to start using alternative products instead of banknotes.202032066229
218550.9978Isolation of multidrug-resistant Escherichia coli, Staphylococcus spp., and Streptococcus spp. from dogs in Chattogram Metropolitan Area, Bangladesh. OBJECTIVES: Antibacterial resistance is a great concern in human and food animal medicine, and it poses a significant concern in pet animals like dogs. This cross-sectional study was conducted to evaluate the antimicrobial resistance pattern of Escherichia coli, Staphylococcus spp., and Streptococcus spp. along with the carryover of some resistance genes in E. coli from dogs in the Chattogram metropolitan area, Bangladesh. MATERIALS AND METHODS: Rectal swab (n = 50), nasal swab (n = 50), and skin swab (n = 50) samples were collected from dogs having respiratory infections, skin infections, and/or enteritis, respectively. Three types of bacteria were identified and isolated by conventional bacteriological techniques and biochemical tests. Antimicrobial susceptibility testing was carried out against 12 antimicrobials by disk diffusion methods. Six resistance genes, namely bla (TEM), bla (CTX-M), tetA, tetB, Sul-I, and Sul-II, were screened for phenotypically resistant E. coli isolates by the polymerase chain reaction. RESULTS: A total of 39 (78%) E. coli, 25 (50%) Staphylococcus spp., and 24 (48%) Streptococcus spp. isolates were isolated from the rectal swab, nasal swab, and skin swab samples, respectively. In the cultural sensitivity test, the E. coli isolates showed resistance to ceftriaxone (79%) and sulfamethoxazole/trimethoprim (64%). Doxycycline (80%) demonstrated the highest resistance among Staphylococcus isolates, followed by sulfamethoxazole/trimethoprim (60%). Streptococcus isolates showed the highest resistance to penicillin (63%), followed by ceftriaxone (54%), while no isolate showed resistance to gentamycin. The prevalence of bla (TEM), bla (CTX-M), tetA, tetB, Sul-I, and Sul-II genes in phenotypically resistant E. coli isolates were 100%, 61.29%, 100%, 8.33%, 56%, and 72%, respectively. CONCLUSIONS: Spillover of such multidrug-resistant bacteria and resistance genes from pet dogs pose a serious public health risk.202033409311
211460.9978Clinical, phenotypic, and genotypic characteristics of ESBL-producing Salmonella enterica bloodstream infections from Qatar. BACKGROUND: Resistant Salmonella infections are a major global public health challenge particularly for multidrug-resistant (MDR) isolates manifesting as bloodstream infections (BSIs). OBJECTIVES: To evaluate clinical, phenotypic, and genotypic characteristics of extended-spectrum beta-lactamase (ESBL) producing Salmonella enterica BSIs from Qatar. METHODS: Phenotypic ESBL Salmonella enterica from adult patients presenting with positive BSIs were collected between January 2019 to May 2020. Microbiological identification and characterization were performed using standard methods while genetic characteristics were examined through whole genome sequencing studies. RESULTS: Of 151 episodes of Salmonella enterica BSI, 15 (10%) phenotypic ESBL isolates were collected. Recent travel was recorded in most cases (80%) with recent exposure to antimicrobials (27%). High-level resistance to quinolines, aminoglycosides, and cephalosporins was recorded (80-100%) while meropenem, tigecycline and colistin demonstrated universal susceptibility. Genomic evaluation demonstrated dominance of serotype Salmonella Typhi sequence type 1 (93%) while antimicrobial resistance genes revealed dominance of aminoglycoside resistance (100%), qnrS1 quinolones resistance (80%), bla(CTX-M-15) ESBLs (86.7%), and paucity of AmpC resistance genes (6.7%). CONCLUSIONS: Invasive MDR Salmonella enterica is mainly imported, connected to patients from high prevalent regions with recent travel and antimicrobial use caused by specific resistant clones. In suspected cases of multidrug resistance, carbapenem therapy is recommended.202438742235
133270.9978First study on capsular serotypes and virulence factors of Pasteurella multocida isolates from Phan Rang sheep in Vietnam. BACKGROUND AND AIM: Pasteurella multocida is considered as a main factor mediating pneumonic pasteurellosis in ruminants, including sheep. It is also a current threat to Phan Rang sheep in Vietnam. This study aimed to characterize P. multocida isolated from Phan Rang sheep, their antibiotic resistance profile, and the prevalence of some virulence-associated genes of these strains. MATERIALS AND METHODS: Bacteria were isolated on brain heart infusion, 10% sheep blood agar plates, and screened by biochemical tests. The polymerase chain reaction technique was used with specific primers to identify P. multocida, the presence of virulence-associated genes, and serotypes of isolates. Antimicrobial susceptibility and biofilm formation of isolates were examined using the disk diffusion method and crystal violet-based method, respectively. RESULTS: A total of 41 P. multocida strains were isolated from 485 samples from clinically sick and healthy sheep. Of the isolates, 58.53% were serotype A, 9.75% were serotype B, and 31.71% were serotype D. Healthy animals were infected with serotype D only. All 15 virulence genes were identified in all strains isolated from clinically sick sheep, while strains isolated from healthy sheep carried 11/15 virulence genes tested. Among virulence-associated genes exbB, exbD, tonB, ompA, oma87, fimA, hgbA, and nanB were detected in over 90% of isolates, whereas hgbB, nanH, tbpA and pfhA were less frequent. Interestingly, pmHAS and tadD were highly prevalent in capsular type A strains, whereas the toxA gene was detected in capsular type D strains only. All of the isolated strains were fully susceptible to enrofloxacin, ciprofloxacin, neomycin, and ofloxacin. About 92.68% were susceptible to chloramphenicol and 90.24% to amikacin, but there was high resistance to erythromycin, tetracycline, and amoxicillin. Our results reveal that 53.65% of 41 isolates could produce biofilm, whereas 46.34% could not. CONCLUSION: Pasteurella multocida from Phan Rang sheep possess many virulence genes and resistance to several common antibiotics such as erythromycin, tetracycline, and amoxicillin. The results are an important warning regarding antibiotic resistance of P. multocida.202337042011
145780.9978Detection of TEM and CTX-M Genes in Escherichia coli Isolated from Clinical Specimens at Tertiary Care Heart Hospital, Kathmandu, Nepal. BACKGROUND: Antimicrobial resistance (AMR) among Gram-negative pathogens, predominantly ESBL-producing clinical isolates, are increasing worldwide. The main aim of this study was to determine the prevalence of ESBL-producing clinical isolates, their antibiogram, and the frequency of ESBL genes (bla(TEM) and bla(CTX-M)) in the clinical samples from patients. METHODS: A total of 1065 clinical specimens from patients suspected of heart infections were collected between February and August 2019. Bacterial isolates were identified on colony morphology and biochemical properties. Thus, obtained clinical isolates were screened for antimicrobial susceptibility testing (AST) using modified Kirby-Bauer disk diffusion method, while ESBL producers were identified by using a combination disk diffusion method. ESBL positive isolates were further assessed using conventional polymerase chain reaction (PCR) to detect the ESBL genes bla(TEM) and bla(CTX-M). RESULTS: Out of 1065 clinical specimens, 17.8% (190/1065) showed bacterial growth. Among 190 bacterial isolates, 57.4% (109/190) were Gram-negative bacteria. Among 109 Gram-negative bacteria, 40.3% (44/109) were E. coli, and 30.2% (33/109) were K. pneumoniae. In AST, 57.7% (n = 63) Gram-negative bacterial isolates were resistant to ampicillin and 47.7% (n = 52) were resistant to nalidixic acid. Over half of the isolates (51.3%; 56/109) were multidrug resistant (MDR). Of 44 E. coli, 27.3% (12/44) were ESBL producers. Among ESBL producer E. coli isolates, 58.4% (7/12) tested positive for the bla(CTX-M) gene and 41.6% (5/12) tested positive for the bla(TEM) gene. CONCLUSION: Half of the Gram-negative bacteria in our study were MDR. Routine identification of an infectious agent followed by AST is critical to optimize the treatment and prevent antimicrobial resistance.202133562276
104490.9978Molecular Characterization of Antimicrobial Resistance and Virulence Genes of Bacterial Pathogens from Bovine and Caprine Mastitis in Northern Lebanon. Mastitis is an infectious disease encountered in dairy animals worldwide that is currently a growing concern in Lebanon. This study aimed at investigating the etiology of the main mastitis-causing pathogens in Northern Lebanon, determining their antimicrobial susceptibility profiles, and identifying their antimicrobial resistance (AMR) genes. A total of 101 quarter milk samples were collected from 77 cows and 11 goats presenting symptoms of mastitis on 45 dairy farms. Bacterial identification was carried out through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibility was tested by disc diffusion and broth microdilution methods. Molecular characterization included polymerase chain reaction (PCR) screening for genes encoding extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC among Enterobacterales isolates, and virulence factors among Staphylococcus isolates. Escherichia coli isolates were subjected to phylogenetic typing by a quadruplex PCR method. The most frequently identified species were Streptococcus uberis (19.2%), Streptococcus agalactiae (15.1%), E. coli (12.3%), and Staphylococcus aureus (10.96%). Gram-positive bacteria were resistant to macrolides and tetracycline, whereas gram-negative bacteria displayed resistance to ampicillin and tetracycline. Two ESBL genes, bla(TEM) (83.3%) and bla(OXA) (16.7%), and one AmpC beta-lactamase gene, bla(CMY-II) (16.7%), were detected among six E. coli isolates, which mainly belonged to phylogenetic group B1. Among Staphylococcus spp., the mecA gene was present in three isolates. Furthermore, four isolates contained at least one toxin gene, and all S. aureus isolates carried the ica operon. These findings revealed the alarming risk of AMR in the Lebanese dairy chain and the importance of monitoring antimicrobial usage.202134071800
2156100.9977Antimicrobial 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
1470110.9977Occurrence of extended-spectrum beta-lactamase (ESBL) in Gram-negative bacterial isolates from high vaginal swabs in a teaching hospital in Nigeria. OBJECTIVE: This study aims to determine the antibiotic susceptibility pattern and incidence of extended-spectrum beta-lactamase (ESBL) genes in isolates from vaginal discharge of symptomatic female patients. STUDY DESIGN: Cross-sectional study. PARTICIPANT: Pregnant and non-pregnant women between 18 and 50 years who presented with genital tract infection and had not received antimicrobial therapy in the two weeks prior. INTERVENTIONS: The study determines the prevalence of bacteria in the vaginal discharge of female patients of reproductive age, the antibiotic susceptibility pattern of the isolates and the incidence of ESBL genes in Gram-negative isolates from the sample. RESULTS: Bacteria were found in 74 (80.4%) and 88 (81.5%) samples from pregnant and non-pregnant women, respectively. Escherichia coli (n=48; 27.6%) occurred mostly in the samples, followed by Staphylococcus aureus (n=38; 21.8%). Among the Gram-positive, all Streptococcus. pneumoniae and Staphylococcus. epidermidis were sensitive to imipenem and meropenem (100%). S. aureus was the most resistant to cephalexin (71.4%), cefoxitin (60.5%) carbenicillin (60.5%) and ceftazidime (57.9%). Escherichia coli was highly resistant to carbenicillin (85.4%), cephalexin (64.6%) and cefotaxime (56.3%). Klebsiella pneumoniae showed the highest level of imipenem resistance (31.6%), followed by E. coli (29.2%). The prevalence of ESBL genes in Gram-negative isolates from pregnant women was 25.6% (11/43), compared to 30.3% (23/76) in non-pregnant women. Both bla (TEM) and bla (SHV) had the highest occurrence of 14.3% (17/119) of the isolates. CONCLUSION: This study found Gram-negative pathogens isolated from the vaginal tract of both pregnant and non-pregnant women to be resistant to multiple antibiotics and have ESBL genes. FUNDING: None declared.202440585516
1469120.9977Investigation of Bacterial Infections and Antibiotic Resistance Patterns Among Clinical Isolates in the Center of Iran. Introduction: Bacterial infection is a considerable problem in hospitals. Thus, this study was executed to appraise the rampancy of bacterial infections, antimicrobial susceptibility patterns, and molecular characterization of isolates among patients in Bafgh Hospital in Yazd, Iran, in 2020. Methods: In the current study, we surveyed 103 isolates of 400 clinical specimens from early March 2020 to September 2020 in Bafgh Hospital. We assessed phenotypic traits and antibiotic resistance with standard microbiological methods. Phenotypic methods were also performed to identify extended-spectrum beta-lactamases (ESBLs) in Gram-negative bacilli, inducible clindamycin resistance, and methicillin resistance in Staphylococcus according to CLSI guidelines. Molecular identification of isolates was done by conventional PCR 16S rRNA gene sequencing. Furthermore, we investigated the prevalence of resistant genes including bla (TEM), bla (PER-2), bla (CTX-M), bla (SHV), and bla (VEB-1) in Gram-negative bacteria and the mecA gene in staphylococcal species. Results: From 400 different clinical specimens, 103 isolates of Gram-positive and Gram-negative bacteria were isolated. Based on phenotypic and molecular methods, most common isolates were Escherichia coli (53 isolates), followed by Klebsiella spp. (18 isolates), and Staphylococcus aureus (16 isolates). The highest resistance was found in Gram-positive bacteria to erythromycin (66.67%) and penicillin (55.56%), while considering Gram-negative bacteria, the most resistant was cefixime (49.41%) and trimethoprim-sulfamethoxazole (47.05%). In addition, out of 16 S. aureus isolates, 62.5% and 17.65% were resistant to methicillin and clindamycin, respectively. Among 83 Gram-negative isolates, 22.89% were ESBL-positive. The prevalence of bla (SHV), bla (PER2), bla (TEM), bla (CTX-M), and bla (VEB-1) genes was 78.31%, 59.03%, 40.96%, 30.12%, and 0%, respectively. Conclusions: The outbreak of bacterial infections is relatively high in hospitals. Recognizing risk agents for bacterial infections and restricting the administration of multidrug-resistant antibiotics is a substantial measure that must be taken to prevent patient mortality.202540822981
1468130.9977Phenotypic and Molecular Characterization of Multidrug Resistant Klebsiella pneumoniae Isolated from Different Clinical Sources in Al-Najaf Province-Iraq. BACKGROUND AND OBJECTIVE: Burns infections and urinary tract infections are the most important prevalent diseases in Asian countries, such as Iraq. Klebsiella pneumoniae is one of the most important bacteria cause this type of infections especially in hospitals. Therefore, the aim of this study was to investigate the prevalence of multi-drug resistance K. pneumoniae and extended-spectrum beta-lactamases producing K. pneumoniae isolates from inpatients with urinary tract infection and burns infections in Al-Kufa hospital in Al-Najaf province, Iraq. MATERIALS AND METHODS: A total of 285 clinical samples were collected from in-patients infected with urinary tract infection (141 urine samples) and burns infections (144 burns swabs). Fourteen different antibiotics were used by disc diffusion method and 13 antimicrobials resistance genes were used by PCR technique. RESULTS: A total of 43 K. pneumoniae strains were isolated. The highest resistance rate was observed for amoxicillin 25 μg and amoxicillin+clavulanic acid 20+10 μg (97.67%) while the lowest resistance rate was observed for imipenem 10 μg (9.30%). The most common resistance associated-genes were blaSHV (86.04%) and at lower prevalence were IMP (9.30%). CONCLUSION: Klebsiella pneumoniae strains isolated from burns infections were more virulent than those isolated from urinary tract infections.201729023034
1168140.9977Dairy Cattle and the Iconic Autochthonous Cattle in Northern Portugal Are Reservoirs of Multidrug-Resistant Escherichia coli. Background/Objectives: Animals destined for human consumption play a key role in potentially transmitting bacteria carrying antibiotic resistance genes. However, there is limited knowledge about the carriage of antibiotic-resistant bacteria in native breeds. We aimed to characterize the phenotypic profiles and antibiotic resistance genes in Escherichia coli isolated from bovines, including three native Portuguese bovine breeds. Methods: Forty-nine E. coli isolates were selected from 640 fecal samples pooled by age group (eight adult or eight calf samples) from each farm, representing both dairy cattle raised in intensive systems and meat cattle raised in extensive systems in Northern Portugal. The presumptive E. coli colonies plated onto MacConkey agar were confirmed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The antibiotic resistance profiles were screened by antimicrobial susceptibility testing (EUCAST/CLSI guidelines), and the antibiotic resistance genes by PCR. Results: Most isolates showed resistance to ampicillin (69%), tetracycline (57%), gentamicin (55%), and trimethoprim + sulfamethoxazole (53%), with no resistance to imipenem. Resistance to at least one antibiotic was found in 92% of isolates, while 59% exhibited multidrug resistance. Most calf isolates, including those from native breeds, showed a multidrug-resistant phenotype. Among the adults, this was only observed in Holstein-Friesian and Barrosã cattle. None of the Holstein-Friesian isolates were susceptible to all the tested antibiotics. ESBL-producing E. coli was identified in 39% of isolates, including those from Holstein-Friesian calves and adults, Cachena calves and Minhota adults. The sul2 gene was detected in 69% of isolates, followed by bla(CTX-M) (45%), aac(3')-IV (41%), and aac(6')-Ib-cr (31%), with a higher prevalence in adults. Conclusions: This pioneering study highlights the concerning presence of multidrug-resistant E. coli in native Portuguese cattle breeds.202439766598
1456150.9977Resistance and Co-Resistance of Metallo-Beta-Lactamase Genes in Diarrheal and Urinary-Tract Pathogens in Bangladesh. Carbapenems are the antibiotics of choice for treating multidrug-resistant bacterial infections. Metallo-β-lactamases (MBLs) are carbapenemases capable of hydrolyzing nearly all therapeutically available beta-lactam antibiotics. Consequently, this research assessed the distribution of two MBL genes and three β-lactamases and their associated phenotypic resistance in diarrheal and urinary-tract infections (UTIs) to guide future policies. Samples were collected through a cross-sectional study, and β-lactamase genes were detected via PCR. A total of 228 diarrheal bacteria were isolated from 240 samples. The most predominant pathogens were Escherichia coli (32%) and Klebsiella spp. (7%). Phenotypic resistance to amoxicillin-clavulanic acid, aztreonam, cefuroxime, cefixime, cefepime, imipenem, meropenem, gentamicin, netilmicin, and amikacin was 50.4%, 65.6%, 66.8%, 80.5%, 54.4%, 41.6%, 25.7%, 41.2%, 37.2%, and 42.9%, respectively. A total of 142 UTI pathogens were identified from 150 urine samples. Klebsiella spp. (39%) and Escherichia coli (24%) were the major pathogens isolated. Phenotypic resistance to amoxicillin-clavulanic acid, aztreonam, cefuroxime, cefixime, cefepime, imipenem, meropenem, gentamicin, netilmicin, and amikacin was 93.7%, 75.0%, 91.5%, 93.7%, 88.0%, 72.5%, 13.6%, 44.4%, 71.1%, and 43%, respectively. Twenty-four diarrheal isolates carried blaNDM-1 or blaVIM genes. The overall MBL gene prevalence was 10.5%. Thirty-six UTI pathogens carried either blaNDM-1 or blaVIM genes (25.4%). Seven isolates carried both blaNDM-1 and blaVIM genes. MBL genes were strongly associated with phenotypic carbapenem and other β-lactam antibiotic resistance. blaOXA imparted significantly higher phenotypic resistance to β-lactam antibiotics. Active surveillance and stewardship programs are urgently needed to reduce carbapenem resistance in Bangladesh.202439203431
1129160.9977Genotypic and phenotypic profiles of antibiotic-resistant bacteria isolated from hospitalised patients in Bangladesh. OBJECTIVES: Characterisation of resistance phenotype and genotype is crucial to understanding the burden and transmission of antimicrobial resistance (AMR). This study aims to determine the spectrum of AMR and associated genes encoding aminoglycoside, macrolide and β-lactam classes of antimicrobials in bacteria isolated from hospitalised patients in Bangladesh. METHODS: 430 bacterial isolates from patients with respiratory, intestinal, wound infections and typhoid fever, presenting to clinical care from 2015 to 2019, were examined. They included Escherichia coli (n = 85); Staphylococcus aureus (n = 84); Salmonella typhi (n = 82); Klebsiella pneumoniae (n = 42); Streptococcus pneumoniae (n = 36); coagulase-negative staphylococci (n = 28); Enterococcus faecalis (n = 27); Pseudomonas aeruginosa (n = 26); and Acinetobacter baumannii (n = 20). Reconfirmation of these clinical isolates and antimicrobial susceptibility tests was performed. PCR amplification using resistance gene-specific primers was done, and the amplified products were confirmed by Sanger sequencing. RESULTS: 53% of isolates were multidrug-resistant (MDR), including 97% of Escherichia coli. There was a year-wise gradual increase in MDR isolates from 2015 to 2018, and there was an almost twofold increase in the number of MDR strains isolated in 2019 (P = 0.00058). Among the 5 extended-spectrum β-lactamases investigated, CTX-M-1 was the most prevalent (63%) followed by NDM-1 (22%); Escherichia coli was the major reservoir of these genes. The ermB (55%) and aac(6')-Ib (35%) genes were the most frequently detected macrolide and aminoglycoside resistance genes, respectively. CONCLUSION: MDR pathogens are highly prevalent in hospital settings of Bangladesh.202133838068
1334170.9977Intimin (eae) and virulence membrane protein pagC genes are associated with biofilm formation and multidrug resistance in Escherichia coli and Salmonella enterica isolates from calves with diarrhea. OBJECTIVES: This study aimed to evaluate the association of the intimin (eae) and pagC genes with biofilm formation and multidrug resistance (MDR) phenotype in Escherichia coli and Salmonella enterica collected from calves with diarrhea. RESULTS: Fecal samples (n: 150) were collected from calves with diarrhea. Of 150 fecal samples, 122 (81.3%) were culture positive and 115/122 (94.2%) were Gram-negative bacteria. Among them, E. coli (n = 64/115, 55.6%) was the most common isolate followed by S. enterica (n = 41/115, 35.6%). Also, 10 (8.6%) isolates were other Enterobacteriaceae bacteria including Klebsiella and Proteus species. Eighty-nine isolates (77.4%) from calf diarrhea, including 52 (81.3%) E. coli and 37 (90.2%) S. enterica were MDR. The eae and pagC genes were detected in 33 (51.5%) E. coli and 28 (68.3%) S. enterica isolates, respectively. There was a strong association between these genes and biofilm formation and MDR phenotype (P-value = 0.000). All E. coli isolates carrying the eae gene were biofilm producers and MDR. Also, all pagC-positive S. enterica isolates were MDR and 25 (89.3%) isolates of them produced biofilm.202236221149
1119180.9977Prevalence and molecular characterization of antibiotic resistance and associated genes in Klebsiella pneumoniae isolates: A clinical observational study in different hospitals in Chattogram, Bangladesh. OBJECTIVE: This study was performed to investigate the prevalence of multidrug resistance and molecular characterization of Klebsiella pneumoniae (KPN) from clinical isolates in the southern region of Bangladesh. Additional analysis of the prevalence of blaNDM-1, blaSHV-11, uge genes of KPN was also carried out among these clinical isolates. METHOD: The study was carried out using 1000 clinical isolates collected from two different hospitals of Chattogram. A drug susceptibility test was performed by the disk diffusion method to detect KPN's response to 16 antibiotics. The presence of antibiotic-resistant and (or) virulent genes blaNDM-1, blaSHV-11, uge were investigated using the PCR technique. Isolates having blaNDM-1, blaSHV-11, uge gene were further validated by sequencing followed by phylogenetic analysis. Phylogenetic relationships among these isolates were determined by Clustal omega and MEGA7. RESULT: A total of 79%, 77%, 74.9%, 71%, 66% and 65% isolates exhibited resistance against cefuroxime, cefixime, cefotaxime, ceftazidime, cefepime and ceftriaxone respectively. The frequency of resistance to other antibiotics varied from 26.5% to 61.8%. PCR analysis showed that 64% of strains harbored blaNDM-1 gene, and 38% strains harbored blaSHV-11 gene. Moreover, 47% of samples were carrying uge gene, and 19% of samples carried blaNDM-1, blaSHV-11, uge genes together. CONCLUSION: In this study, we've analysed the pattern of expression as well as prevalence of blaNDM-1, blaSHV-11, and uge genes in Klebsiella isolates. Upon molecular and statistical analysis, we found a high prevalence of multi-drug resistance KPN strains in the isolates. The Klebsiella isolates were confirmed to harbor multiple ESBL genes and 64% of the isolates were found to be producing NDM-1. As multidrug resistance is an alarming issue, continuous surveillance and routine clinical detection of resistant bacteria and plasmids are necessary to prevent catastrophic public health incidents.202134506611
1123190.9977Molecular detection of blaSHV gene in multidrug resistance of Klebsiella pneumoniae isolated from chicken egg shell swab from a traditional market in Surabaya. BACKGROUND: Contamination with Klebsiella pneumoniae in food ingredients, including eggs, causes various dangers because it threatens public health, because it acts as a multidrug resistance (MDR) bacteria, especially the extended-spectrum beta-lactamase (ESBL) strain. The ESBL blaSHV gene is part of a broad-spectrum ESBL that is often found in Gram-negative bacteria. AIM: This study aimed to identify the ESBL blaSHV gene in K. pneumoniae MDR from chicken eggshells. METHODS: This study used 160 samples of chicken eggshell swabs isolated on 1% BPW media from 10 traditional Surabaya markets. Samples were isolated using MCA media and were identified using Gram staining and biochemical tests. Detection of MDR using Muller-Hinton Agar. RESULTS: Confirmation of ESBL in multidrug-resistant (MDR) isolates was performed using polymerase chain reaction to detect ESBL genes. The results showed that the isolation and identification of K. pneumoniae bacteria were 25.62% (41/160). Amoxicillin antibiotics showed the highest level of resistance at a percentage of 100% (41/41), followed by antibiotic resistance to erythromycin (90.24% (37/41), Streptomycin antibiotics were 26.82% (11/41), ciprofloxacin (14.63% (6/41), and Tetracycline antibiotic resistance was 7.31% (3/41). The results of MDR from K. pneumoniae showed 34.14% (14/41) of the isolates were then tested by PCR, which showed positive results for the blaSHV gene of 71.42% (10/14). CONCLUSION: The data from this study confirm the existence of K. pneumoniae bacteria isolated from egg shell swabs carrying the blaSHV gene from MDR isolates.202540557075