Extended-spectrum Beta-lactamase Research Articles

Extended-spectrum beta-lactamases in clinical isolates of Escherichia coli and Klebsiella pneumoniae recovered from patients at the Tamale Teaching Hospital, Ghana.

Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae are pathogens of significant public health interest for which new antibiotics are urgently needed. To determine the prevalence of ESBLs in E. coli and K. pneumoniae isolates from patients attending the Tamale Teaching Hospital (TTH) in Ghana. The study was a cross-sectional study involving convenience sampling of E. coli and K. pneumoniae isolates from consenting patients' clinical specimens, between April and June 2015. Antimicrobial susceptibility test was performed, and ESBL-producer phenotypes were further screened for BlaTEM, BlaSHV, and BlaCTX-M genes. Patients' clinical data were additionally collected using a structured questionnaire. Of the 150 non-duplicate E. coli and K. pneumoniae isolates identified, 140 were confirmed as E. coli (84%, n = 117) and K. pneumoniae (16%, n = 23). Of these, sixty-two (44%) [E. coli (84%; n = 52); K. pneumoniae (16%; n = 10)] phenotypically expressed ESBLs. The proportion of ESBL-producing isolates was higher in adults (15-65 years) than in neonates (< 28 days) (p = 0.14). Most of the isolates showed a high percentage resistance to ampicillin (96%) and tetracycline (89%), but a relatively lower resistance to amikacin (36%). No isolate was resistant to meropenem. More ESBL producers were multidrug resistant compared to non-ESBL-producers [23% (14/62) versus 18% (14/78); p = 0.573]. Overall, 74% (n = 46) of the ESBL genotypes expressed BlaCTX-M-1 genes, followed by 63% (n = 39) BlaTEM, and 16% (n = 10) BlaSHV. The study showed a high prevalence of ESBL-positive E. coli and K. pneumoniae, mostly CTX-M-1 producers at TTH. Routine laboratory ESBL screening is warranted to inform patient management.

Effect of poultry litter soil amendment on antibiotic-resistant Escherichia coli.

Given the high cost and non-renewability of mineral-based fertilizers, there is increasing interest in the innovative use of manure-based materials, such as poultry litter (PL). However, manure-based fertilizers add both nutrients and microbes to the soil, including antibiotic-resistant Escherichia coli (AREc). PL soil amendment impact on AREc in corn fields was evaluated in a randomized field experiment (May-October 2017). Two winter cropping systems (fallow and cover crop) were assigned to whole plots, with three spring-applied fertilizer treatments (untreated control [UC], PL, and commercial fertilizer [CF]) assigned to subplots. Soil was collected from 0 to 15cm on days 0, 7, 28, 70, 98, and 172 post-treatment applications. Samples were cultured for the enumeration and prevalence of generic, tetracycline-resistant (TETr), third-generation cephalosporin-resistant (3GCr) E. coli isolates, and extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae. PL soil amendment significantly (p < 0.05) increased the levels of generic E. coli, TETr E. coli, and 3GCr E. coli on days 7 and 28 compared to UC or CF. Beyond day 28, AREc did not significantly (p>0.05) differ by fertilizer treatment and returned to baseline on day 70. ESBL-producing Enterobacteriaceae were detected from 16 samples, mostly on day 70. Cover crop significantly decreased TETr E. coli concentration on day 28, with no significant effects on the prevalence of 3GCr E. coli and ESBL-producing Enterobacteriaceae compared to no cover crop. All ESBL-producing Enterobacteriaceae and 79% of the 3GCr E. coli isolates were positive for blaCTX-M gene by polymerase chain reaction. Results show that PL soil amendment transiently increases the levels of AREc compared to mineral fertilizer.

Molecular Characterization of Klebsiella pneumoniae Clinical Isolates Through Whole-Genome Sequencing: A Comprehensive Analysis of Serotypes, Sequence Types, and Antimicrobial and Virulence Genes.

Introduction Antimicrobial resistance (AMR) has become a menace, spreading among bacterial species globally. AMR is now recognized as a silent pandemic responsible for treatment failures. Therefore, an effective surveillance mechanism is warranted to understand the bacterial species isolated from human clinical specimens. The present study employed next-generation sequencing (NGS) or whole-genome sequencing (WGS) to identify the resistance andvirulence genes,sequence type, and serotypes. Methods This study included 18 multidrug-resistant (MDR) Klebsiella pneumoniae (K. pneumoniae) isolates obtained from patients suffering from different infections attending the Prathima Institute of Medical Sciences, Karimnagar, India. All isolates were identified, and antimicrobial susceptibility profiles were determined through conventional microbiological techniques and confirmed by automated systems. All the isolates were investigated using NGSor WGSto identify the genes coding for resistance, such as extended-spectrum beta-lactamases (ESBLs), metallo-beta-lactamases, and virulence genes. Multilocus sequence typing (MLST) was conducted to identify the sequence types, and Kleborate analysis was performed to confirm the species, genes for AMR, and virulence and evaluate the capsular polysaccharide (KL) and cell wall/lipopolysaccharide (O) serotypes carried by the isolates. Results The mean age of the patients was 46.11±20.35 years. Among the patients included, 12 (66.66%) were males and 6 (33.33%) were females. A high percentage (>50%) of hypervirulent K. pneumoniae (hvKp) strains that had genes coding for AMR and plasmids having the potential to carryblaNDMand resistance genes were observed. Among the isolates, 16 (88.88%) revealed the presence of multiple antibiotic-resistant genes with evidence of at least one gene coding for beta-lactamase resistance. There was a high prevalence of blaSHV (17/18; 94.44%) and blaCTX-M-15 (16/18; 88.88%) AMR genes. Other AMR genes identified included blaTEM (83.33%; 15/18)and blaOXA (14/18; 77.77%). Two (11.11%) strains each showed the presence of blaNDM-1 and blaNDM-5 genes. The virulence genes identified includedgapA, infB, mdh, pgi, phoE, rpoB, tonB, and ybt. The most frequent K. pneumoniae serotypes found were KL51:O1v2 (3/18, 16.66%), KL17:O1v1 (3/18, 16.66%), and KL64:O2v1 (3/18, 16.66%). KL64 (4/18; 22.22%) was the most common capsular serotype identified among the isolates. The most frequent MLST-based sequence type(ST) identified included ST-147 (5/18, 27.77%), followed by ST-231 (3/18, 16.66%) and ST-101 (2/18, 11.11%). Conclusions The molecular analysis ofK. pneumoniaeisolates revealedmultipleAMR, plasmid,and virulence genes. Additionally, many global STs were noticed by MLST. The results noted a high prevalence of hvKp strains. Molecular characterization of bacterial strains using NGS/WGS is important to understand the epidemiology of bacterial strains and the antibiotic resistance and virulence genes they are potentially carrying. The data obtained from this studymay be utilized to devise careful antibiotic-prescribing approaches and improvepatient management practices.

Antimicrobial resistance in surgical infections: experience from open tibia fracture-related infection intraoperative tissue specimens

Background Antimicrobial resistance in surgical infections is known to have significant morbidity and mortality in extreme cases on top of increased costs in medical expenditure. This study aimed to analyze antimicrobial resistance patterns of microbes in open tibia fracture-related infection (FRI) using intraoperative tissue samples. Methods Between January and June 2022, a descriptive cross-sectional observational study was conducted involving patients aged 18 and above with open tibial shaft fractures. Microbiological culture and sensitivity of three intraoperative deep tissue and bone samples taken by sterile techniques using separate instruments in patients with infection. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method along with biomechanical tests for microbial identification. Results Fifty-seven deep tissue samples were taken from 19 patients with FRI who underwent surgery during the study period were analyzed. Almost all (96.5%) intraoperative tissue samples were culture-positive identifying 73 bacterial isolates out of which 57.9% were polymicrobial. The most frequent isolated bacteria were Gram-negative bacteria 56 (76.7%) with a predominance of Proteus spp 13 (17.8%). Methicilin Resistance Staphyaureus comprised 12 (16.4%) of the isolates whereas Inducible Clindamycin resistance was identified in 8 (57.1%) of Staphyloccus aureus isolates. More than half of the isolates 30 (53.6%) comprised of Extended Spectrum Beta Lactamase producing bacteria. There was high resistance to Cephalosporins and Amoxyclavulanic acid as the efficacy was low ranging from 28.2, 27.8, and 30.0% for cefotaxime, ceftazidime and amoxicillin/clavulanic acid, respectively. Almost all Gram-negative bacteria 47 (83.9%) had multidrug resistance pattern except for Pseudomonas aureginosa that had 77.8 to 88.9% susceptibility to all tested antibiotics. Conclusion High resistance to cephalosporins in patients with delayed or late infection in open tibia FRI is alarming and further studies to establish causes are indicated.

Etiologies, risk factors, and antibiotic pattern of lower respiratory tract infection in patients coming to Deccan College of Medical Sciences, Hyderabad, South India

Background. Lower Respiratory Tract Infections (LRTI) impose a huge problem on society due to their tenacious and unescapable health problems, and they were the reasons for consultation and hospitalization. Patients with LRTI were present with a wide spectrum of diseases that range from life-threatening infections to minor self-limiting illnesses. Materials and methods. Sputum samples from 310 patients with complaints of lower respiratory tract infections were collected and analyzed. After microscopic analysis, 30 were excluded from the study. Of the 280 that were included in the study, 171 were males and 109 were females. The age group between 56 and 65 years comprised the highest number of patients in the study. Different antibiotics were used to check antibiotic sensitivity patterns against isolated Gram-positive and negative bacteria. Results. Of the 280 samples processed, 122 (43.5%) were culture-positive. Among the 122 bacterial isolates, the predominant organism was Klebsiella pneumoniae 55 (45%). The most common predisposing factor identified in 115 (37%) patients was smoking. Using the Double Disk Synergy Test, 11 (20%) out of a total of 55 K. pneumoniae isolates and 4 (30.7%) out of 13 Pseudomonas isolates showed Extended Spectrum Beta Lactamases (ESBL). The Klebsiella and Pseudomonas isolates showed resistance to 2nd and 3rd generation Cephalosporins. Conclusion. Because of different geographical regions and conditions, the etiology and the antibiotic sensitivity patterns of LRTI vary, so the etiology, predisposing factors, and the antibiotic sensitivity pattern of LRTI need to be updated regularly. The diagnostic facilities for early and rapid identification of LRTI also need to be improved.

Microbiological and Antimicrobial Susceptibility Pattern of Asymptomatic Bacteriuria in Pregnant Women Attending SQUH.

Our research aimed to study the microbiology and antimicrobial resistance in asymptomatic bacteriuria (ASB) among Omani pregnant women. We conducted a retrospective study that included data from 196 Omani pregnant women with ASB who received care at Sultan Qaboos University Hospital from 2010-2019. Data were obtained from the patients' electronic medical records including demographics, clinical details, isolated organisms, antimicrobial susceptibility results, and prescribed antibiotics. ASB was detected in 56.1% of cases during the third trimester. Klebsiella pneumoniae(32.1%) was the most frequently isolated organism, followed by Escherichia coli (29.6%). Twenty-one (10.7%) isolates were extended-spectrum beta-lactamase (ESBL)-producing organisms. The overall microbiological susceptibility pattern showed that organisms have a high susceptibility rate to nitrofurantoin reaching 82.8%, followed to a lesser extent by cefuroxime and augmentin. The susceptibility of E. coli and K. pneumoniae to cefuroxime was 74.1% and 71.4%, respectively. Only 52.4% of all isolated ESBL-producing organisms were susceptible to nitrofurantoin. K. pneumoniae and E. coli were the most frequently isolated bacteria in ASB, representing 60.7% of total isolates. A high prevalence of ESBL-producing organisms, 10.7% of the total isolates, was observed. Cefuroxime is an appropriate empirical antibacterial therapy for ASB and urinary tract infection in pregnant women. Nitrofurantoin should be considered for empirical antibiotic therapy in settings of high prevalence of ESBL-producing organisms.

Extended Spectrum Beta Lactamase Escherichia Coli in Bagmati River, Kathmandu Valley.

Antimicrobial resistance organisms in the peripheral communities of an environment can be predicted by the presence of extended-spectrum beta-lactamase Escherichia coli in that environment. The close connectivity between humans and water sources can facilitate the entry of antimicrobial resistant organisms into the human ecosystem. The aim of this study was to assess beta lactamase producing Escherichia coli from Bagmati river within Kathmandu valley. In the year 2020, a cross-sectional study was conducted on water samples collected from 66 locations along the Bagmati River. Coliforms were isolated by five tubes dilution method and identified by cultural and biochemical tests. Further Escherichia coli was isolated in eosin methylene blue agar at 44.5 ⁰C. Antibiotic susceptibility test was performed by Kirby Bauer disk diffusion methods. Beta lactamase gene types were detected by using conventional multiplex polymerase chain reaction. A total of 615 bacterial isolates were identified among which 39 % (n=241) were Escherichia coli. Extended spectrum beta lactamase producing Escherichia coli was confirmed in 16.6 % (40/241) of total Escherichia coli isolates. Among 66 sites this isolate was detected in 26 (40 %) sampling sites excluding upstream regions. All the Escherichia coli isolates were multidrug resistance showing higher percentage (>99 %) of resistant for penicillin, tetracycline and erythromycin antibiotics. There were significant differences in resistance rate for cefotaxime and ceftazidime by extended spectrum beta lactamase producing and non-producing Escherichia coli (p<0.05). Presence of multidrug resistance extended spectrum beta lactamase producing Escherichia coli in river streams suggests the chances of circulating within river system and hence transmitting in human community. Bagmati river; drug resistance; escherichia coli; human.

Carriage of antimicrobial-resistant Enterobacterales among pregnant women and newborns in Amhara, Ethiopia

ObjectivesInfections are one of the most common causes of neonatal mortality, and maternal colonization has been associated with neonatal infection. In this study, we sought to quantify carriage prevalence of extended-spectrum-beta-lactamase (ESBL) -producing and carbapenem-resistant Enterobacterales (CRE) among pregnant women and their neonates and to characterize risk factors for carriage in rural Amhara, Ethiopia. MethodsWe conducted a prospective cohort study nested in the Birhan field site. We collected rectal and vaginal samples from 211 pregnant women in their third trimester and/or during labor/delivery and perirectal or stool samples from 159 of their neonates in the first week of life. ResultsWe found that carriage of ESBL-producing organisms was fairly common (women: 22.3%, 95% CI: 16.8-28.5; neonates: 24.5%, 95% CI: 18.1-32.0), while carriage of CRE (women: 0.9%, 95% CI: 0.1-3.4; neonates: 2.5%, 95% CI: 0.7-6.3) was rare. Neonates whose mothers tested positive for ESBL-producing organisms were nearly twice as likely to also test positive for ESBL-producing organisms (38.7% vs 21.1%, P-value = 0.06). Carriage of ESBL-producing organisms was also associated with Woreda (district) of sample collection and recent antibiotic use. ConclusionUnderstanding carriage patterns of potential pathogens and antibiotic susceptibility among pregnant women and newborns will inform local, data-driven recommendations to prevent and treat neonatal infections.

Discerning the dissemination mechanisms of antibiotic resistance genes through whole genome sequencing of extended-spectrum beta-lactamase (ESBL)-producing E. coli isolated from veterinary clinics and farms in South Korea

Extended-spectrum beta-lactamase (ESBL)-producing bacteria are resistant to most beta-lactams, including third-generation cephalosporins, limiting the treatment methods against the infections they cause. In this study, we performed whole genome sequencing of ESBL-producing E. coli to determine the mechanisms underlying the dissemination of antibiotic resistance genes. We analyzed 141 ESBL-producing isolates which had been collected from 16 veterinary clinics and 16 farms in South Korea. Long- and short-read sequencing platforms were used to obtain high-quality assemblies. The results showed that blaCTX-M is the dominant ESBL gene type found in South Korea. The spread of blaCTX-M appears to have been facilitated by both clonal spread between different host species and conjugation. Most blaCTX-M genes were found associated with diverse mobile genetic elements that may contribute to the chromosomal integration of the genes. Diverse incompatibility groups of blaCTX-M-harboring plasmids were also observed, which allows their spread among a variety of bacteria. Comprehensive whole genome sequence analysis was useful for the identification of the most prevalent types of ESBL genes and their dissemination mechanisms. The results of this study suggest that the propagation of ESBL genes can occur through clonal spread and plasmid-mediated dissemination, and that suitable action plans should be developed to prevent further propagation of these genes.

What Is the Rate of Antimicrobial Resistance of a Prosthetic Joint Infection in a Major Orthopaedic Centre?

Periprosthetic joint infections (PJIs) are important factors in decreasing the success of hip and knee arthroplasties. It is a necessity to explore the epidemiological data and develop applications for rational antibiotic use, to address future infection control concerns. We aimed to investigate the microorganisms that were responsible and the related antibiograms in 121 patients with PJI, who were managed by two-stage revision surgery. Patients' data records, demographics, comorbidities, sites of arthroplasty, synovial fluid and deep tissue culture results and antibiotic treatment were summarized on a standardized case report form. There were 43 (35.5%) culture-negative PJI cases and 12 (9.9%) polymicrobial growths. The causative pathogens included Gram-positive (50.4%) and Gram-negative microorganisms (23.1%) and fungi (0.8%). Methicillin resistance was 64.3% for S. aureus and 89.5% for coagulase-negative staphylococcus (CoNS). The extended spectrum beta lactamase (ESBL) rate for Enterobacteriaceae was 68.4%. This study shows that antibiotic resistance is encountered in more than half of the cases, which is valid for all microorganisms most common in PJI. The success of treatment decreases significantly in cases where antibiotic-resistant microorganisms are isolated or in cases where the culture is negative.

Antimicrobial Resistance Profile and Extended Spectrum Beta-lactamase Resistance Genes in Escherichia coli from Patients in General Hospital, Karshi, Abuja, Nigeria

Aims: This study investigated the antimicrobial resistance profile and extended spectrum beta-lactamase resistance genes in Escherichia coli of from urine of patients sourced from General Hospital, Karshi, Abuja, Nigeria. Study Design: Cross-sectional study. Place and Duration of Study: Department of Microbiology, Nasarawa State University, Keffi, between August 2022 and February 2023. Methodology: A total of 120 samples were collected from patients. Escherichia coli was isolated from the samples using standard microbiological methods. Antibiotic susceptibility testing was evaluated as described by the Clinical and Laboratory Standards Institute (CLSI). The detection of ESBL production in E. coli isolates was carried out using double disc synergy test. In addition, molecular detection of ESBL genes was carried out using Polymerase Chain Reaction (PCR) method. Results: The prevalence of E. coli was 17.5% (21/120), out of which two (2) of the positive isolates (9.5%) were male, and 19 (90.5%) female. Antibiotic resistances in the isolates in decreasing order were as follows: sulphamethoxazole / trimethoprim (SXT: 81.0%), amoxicillin/clavulanic acid (AMC: 61.9%), ofloxacin (OFX: 66.7%), cefotaxime (CTX: 53.4%), gentamicin (CN: 42.9%), ceftriaxone (CRO: 33.3%), imipenem (IPM: 33.3%), meropenem (MOR: 42.4%), nitrofurantoin (NET: 20.3%) and ciprofloxacin (CIP: 23.8%). The commonest antibiotic resistant resistance phenotype was CIP-OFX-SXT (23.8%). Multiple antibiotic resistance (MAR) was observed in 90.5% (19/21) of the isolates, with the common MAR index being 0.3 (23.8%). Six of the twenty one beta-lactam resistant isolates (28.5%) were confirmed ESBL producers. The 6 ESBL positive isolates carried bla genes as follows: blaTEM (1/6, 16.7%) and blaCTX-M (1/6, 16.7%). blaSHV only was not found in any of the isolates. Conclusion: The E. coli isolates from urine of patients in General Hospital, Karshi, Abuja, Nigeria was less resistant to ciprofloxacin, nitrofurantoin, meropenem and imipenem. This implies that the antibiotics are useful in the treatment of infection caused by E. coli. Also, ESBL-positive E. coli isolates harbored ESBL genes, with blaCTX-M and blaTEM as the most common.

THE USE OF A POLYVALENT KLEBSIELLA BACTERIOPHAGE IN INFECTION AFTER RECONSTRUCTIVE PLASTIC SURGERY ON THE SPINE IN A CHILD (A CLINICAL CASE)

The incidence of infection after reconstructive plastic surgery on the spine (in particular, spondylodesis) in children remains at a fairly high level and ranges from 0.4% to 8.7%. These complications are often caused by polyresistant bacterial pathogens; this accentuates the need for new effective therapeutic approaches. Bacteriophage preparations in combination with antibiotics can optimize existing strategies for the treatment of infectious diseases, since they are able to selectively destroy bacteria, including biofilm-forming ones. The aim of the study was to draw the attention of practitioners to modern problems of treating polyresistant infections on the example of a case of purulent complication after spinal surgery in a child. Materials and methods. On the basis of the FSBI "Federal Center for Traumatology, Orthopedics and Endoprosthetics" under the Health Ministry of the Russian Federation (the town of Cheboksary) a retrospective analysis of a clinical case of treating early infection after reconstructive plastic surgery on the spine caused by a polyresistant strain of Klebsiella pneumoniae was conducted. Research results. Patient P., 2.2 years old, with congenital pathology of the musculoskeletal system, who repeatedly received medical care in various medical organizations, underwent surgical spine stabilization. In the early postoperative period, an infectious complication developed caused by a polyresistant strain of Klebsiella pneumoniae (a producer of extended-spectrum beta-lactamases and carbapenemases). Along with surgical debridement of the postoperative wound, a combined etiotropic antibacterial therapy was performed (12 weeks). However, complete healing of the postoperative wound was achieved in combination with the administration of a polyvalent klebsiella bacteriophage (10 weeks). Conclusions. This clinical case demonstrates that the use of a polyvalent klebsiella bacteriophage can increase the effectiveness of antibacterial pharmacotherapy in surgical treatment with preservation of metal structures.

Unusual and complicated peritonitis: Your questions answered.

Effective treatment of infections is a growing challenge owing to antimicrobial resistance. Peritoneal dialysis (PD) patients experience more frequent hospitalisations than the general population and have greater exposure to antibiotics, making them particularly vulnerable to this threat. Over the last decade, we have noted a surge in cases of complicated peritoneal dialysis-associated peritonitis (PD peritonitis) caused by antimicrobial-resistant organisms, including extended-spectrum beta-lactamase (ESBL), AmpC beta-lactamase-producing Enterobacterales, Pseudomonas aeruginosa and fungi. Practitioners must be alert to these organisms, seek early recognition of these resistance patterns and make timely adjustments in order to avoid delay in treatment that may increase risk of PD catheter removal and technique failure. We present a case of successful treatment of ESBL peritonitis, highlight its challenges, while providing guidance on management of other unusual and complicated PD peritonitis.

Connectivity at the human-wildlife interface: starling movements relate to carriage of E. coli

Synanthropic bird species in human, poultry or livestock environments can increase the spread of pathogens and antibiotic-resistant bacteria between wild and domestic animals. We present the first telemetry-based spatial networks for a small songbird. We quantified landscape connectivity exerted by spotless starling movements, and aimed to determine if connectivity patterns were related to carriage of potential pathogens. We captured 28 starlings on a partridge farm in 2020 and tested them for Avian influenza virus, West Nile virus WNV, Avian orthoavulavirus 1, Coronavirus, Salmonella spp. and Escherichia coli. We did not detect any viruses or Salmonella, but one individual had antibodies against WNV or cross-reacting Flaviviruses. We found E. coli in 61 % (17 of 28) of starlings, 76 % (13 of 17) of which were resistant to gentamicin, 12 % (2 of 17) to cefotaxime/enrofloxacin and 6 % (1 of 17) were phenotypic extended spectrum beta-lactamase (ESBL) carriers. We GPS-tracked 17 starlings and constructed spatial networks showing how their movements (i.e. links) connect different farms with nearby urban and natural habitats (i.e. nodes with different attributes). Using E. coli carriage as a proxy for acquisition/dispersal of bacteria, we found differences across spatial networks constructed for E. coli positive (n = 7) and E. coli negative (n = 9) starlings. We used Exponential Random Graph Models to reveal significant differences between networks. In particular, an urban roost was more connected to other sites by movements of E. coli positive than by movements of E. coli negative starlings. Furthermore, an open pine forest used mainly for roosting was more connected to other sites by movements of E. coli negative than by movements of E. coli positive starlings. Using E. coli as a proxy for a potential pathogen carried by starlings, we reveal the pathways of spread that starlings could provide between farms, urban and natural habitats.

Cervical cancer screening practices in HIV positive females - a missing link in health care delivery in Pakistan.

To probe cervical cancer screening practices in local women positive for human immunodeficiency virus, and to determine the cervical cytological changes in them. The serial cross-sectional study was conducted at the Jinnah Hospital and Services Hospital, Lahore, Pakistan, from April 2019 to October 2020, and comprised female patients aged 18-45 years who were positive for human immunodeficiency virus or acquired immunodeficiency syndrome and were registered with the relevant programme being run by the provincial government in Punjab. Blood samples of all the patients were collected for the determination of human immunodeficiency virus viral load and cluster of differentiation 4+ count. Cervical smears were taken for cytopathological analysis, while the swabs were analysed for culture sensitivity. The same individuals were subjected to the same testing one year later, and the status of the disease and clinical stability or disease progression was explored. Data was analysed using SPSS 25. There were 150 women with mean age 32.08±7.13 years (range: 21-45 years). Age at marriage/sexual activity was 17.33±4.73 years in 15(10%) subjects. Cytological examination showed atypical squamous cells of undetermined significance in 6(4%) of the cases whereas 3(2%) cases showed atypical squamous cells, which cannot rule out high grade squamous intraepithelial lesion on cytology, while the rest were classified as negative for intraepithelial lesion or malignancy. Cervical microbial changes revealed methicillin-resistant staphylococcus aureus infection in 9(6%) cases, extended-spectrum beta-lactamase in 15(10%) cases, whereas fungal infection and trichomonas vaginalis infection were found in 30(20%) smears. There was a significant association between cluster of differentiation 4+ cell count and stability of high-risk patients (p<0.001). After one year, 84(56%) patients remained clinically stable, while 51(34%) developed some chronic illness. There was a significant association between cluster of differentiation 4+ cell count <200/mm3 and the risk of developing a chronic illness (p<0.001). There was a dire need to educate healthcare workers to offer regular cervical screening to patients with high-risk sexually-transmitted infections to prevent them from the morbidity and mortality related to cervical cancer.

Multiple-Drug Resistant Shiga Toxin-Producing E. coli in Raw Milk of Dairy Bovine.

Raw milk may contain pathogenic microorganisms harmful to humans, e.g., multidrug-resistant Escherichia coli non-O157:H7, which can cause severe colitis, hemolytic uremia, and meningitis in children. No studies are available on the prevalence of Shiga toxin-producing E. coli (STEC O157:H7) in sick or healthy dairy animals in the Khyber Pakhtunkhwa Province of Pakistan. This study aimed to isolate, characterize, and detect antibiotic resistance in STEC non-O157:H7 from unpasteurized milk of dairy bovines in this province. We collected raw milk samples (n = 800) from dairy farms, street vendors, and milk shops from different parts of the Khyber Pakhtunkhwa Province. E. coli was isolated from these samples followed by latex agglutination tests for serotyping. The detection of STEC was conducted phenotypically and confirmed by the detection of virulence genes genotypically. An antibiogram of STEC isolates was performed against 12 antibiotics using the disc diffusion method. A total of 321 (40.12%) samples were found to be positive for E. coli in this study. These samples were processed for the presence of four virulence genes (Stx1, Stx2, ehxA, eae). Forty samples (5.0%) were STEC-positive. Of these, 38%, 25%, 19%, and 18% were positive for Stx1, Stx2, ehxA, and eae, respectively. Genotypically, we found that 1.37% of STEC isolates produced extended-spectrum beta-lactamase (ESBL) and contained the blaCTX M gene. Resistance to various antibiotics ranged from 18% to 77%. This study highlights the risk of virulent and multidrug-resistant STEC non-O157:H7 in raw milk and the need for proper quality surveillance and assurance plans to mitigate the potential public health threat.

Bacterial resistome in different stages of wastewater treatment plant is highly impacted by the abundance of the Pseudomonadota community

Wastewater treatment plants (WWTPs) are significant sources of antimicrobial resistance (AMR). We studied bacterial communities and antibiotic-resistance genes (ARGs) in different stages of a WWTP in the UAE. We found high levels of extended-spectrum beta-lactamase bacteria in the returned activated sludge (RAS) stage, including Escherichia coli and Aeromonas sobria, resistant to ESBL, ampicillin, and fosfomycin. Bacterial richness was highest in the primary effluent (PE) stage, with distinct community structures influenced by environmental factors. Pseudomonadota dominated across all stages, with Bacteroidota and Bacillota in PE, and Actinomycetota and Pseudomonadota in AS and RAS. Acidovorax sp. showed strong connections with ARGs in PE and RAS, while Delftia acidovorans had the most associations with ARGs in AS. These findings underscore the role of WWTP stages in shaping bacterial communities and ARG abundance, highlighting the potential of certain bacteria in AMR development and dissemination.

Detection of Extended-Spectrum Beta-Lactamase (ESBL)-Producing Enterobacteriaceae from Diseased Broiler Chickens in Lusaka District, Zambia.

Poultry products in Zambia form an integral part of the human diet in many households, as they are cheap and easy to produce. The burden of poultry diseases has, however, remained a major challenge. Growing consumer demand for poultry products in Zambia has resulted in non-prudent antimicrobial use on farms, intending to prevent and treat poultry diseases for growth optimisation and maximising profits. This cross-sectional study aimed to identify the different types of bacteria causing diseases in chickens in Lusaka and to detect the extended-spectrum lactamase (ESBL)-encoding genes. We collected 215 samples from 91 diseased chickens at three post-mortem facilities and screened them for Gram-negative bacteria. Of these samples, 103 tested positive for various clinically relevant Enterobacteriaceae, including Enterobacter (43/103, 41.7%), Escherichia coli (20/103, 19.4%), Salmonella (10/103, 9.7%), and Shigella (8/103, 7.8%). Other isolated bacteria included Yersinia, Morganella, Proteus, and Klebsiella, which accounted for 21.4%. E. coli, Enterobacter, Salmonella, and Shigella were subjected to antimicrobial susceptibility testing. The results revealed that E. coli, Enterobacter, and Shigella were highly resistant to tetracycline, ampicillin, amoxicillin, and trimethoprim-sulfamethoxazole, while Salmonella showed complete susceptibility to all tested antibiotics. The observed resistance patterns correlated with antimicrobial usage estimated from sales data from a large-scale wholesale and retail company. Six (6/14, 42.9%) E. coli isolates tested positive for blaCTX-M, whilst eight (8/14, 57.1%) Enterobacter samples tested positive for blaTEM. Interestingly, four (4/6, 66.7%) of the E. coli isolates carrying blaCTX-M-positive strains were also positive for blaTEM. Sanger sequencing of the PCR products revealed that five (5/6, 83.3%) of the abovementioned isolates possessed the blaCTX-M-15 allele. The results suggest the presence of potentially pathogenic ESBL-producing Enterobacteriaceae in poultry, threatening public health.

Spectrum and antibiotic resistance in community- and hospital-acquired urinary tract infections among adults: Experience from a large tertiary care center in a developing country.

It is important to note that the causative agents and patterns of antibiotic resistance vary between urinary tract infections (UTIs) acquired in the community and those acquired in a hospital setting. Therefore, the aim of this study was to compare the types of organisms and patterns of antibiotic resistance in adult patients with community-acquired urinary tract infections (CA-UTIs) and hospital-acquired urinary tract infections (HA-UTIs). Retrospectively, we collected urine samples from patients at An-Najah National University Hospital who experienced nonrecurring urinary tract infections (UTIs) between January 2019 and December 2020. The data were subsequently analyzed using IBM-SPSS® 21.0 software to determine the distribution of microorganisms and evaluate the rates of antibiotic resistance. A total of 798 nonrepetitive UTI patients were included in our study, in which more than half of the UTIs occurred in female patients (472; 59.1%), and most of the UTIs were of community origin (611; 76.6%). Both community-acquired urinary tract infections (CA-UTIs) and hospital-acquired urinary tract infections (HA-UTIs) were more common in female patients (45.6% and 13.5%, respectively). Escherichia coli was the most commonly isolated urinary pathogen in both the CA-UTIs and HA-UTIs. The five most common isolated urinary pathogens were E. coli, Klebsiella pneumoniae, Enterococcus faecalis, E. faecium, and Pseudomonas aeruginosa. Among the isolated bacteria, 28.2% were extended-spectrum beta-lactamase (ESBL)-producing bacteria, 4.1% were carbapenem-resistant Enterobacterales (CRE), and 6.5% were vancomycin-resistant Enterococci. The most commonly isolated urinary pathogens from HA-UTIs showed higher resistance rates against all the tested antibiotics except for E. faecium, which showed greater resistance to tetracycline (42.5%) and quinupristin/dalfopristin (17.5%) in CA-UTIs. There are similarities in the etiological profiles of CA-UTI and HA-UTI, but the resistance rates are high, and urine culture is essential for the correct treatment of individual cases, even in primary care.

Identification of Extended Spectrum Beta Lactamases Producing Pseudomonas aeruginosa from Subclinical Mastitis Milk Samples in an Organized Dairy Farm of SVVU, Andhra Pradesh

The present study was conducted during August 2021 at Buffalo Research Station, Venkatarammanagudem of West Godavari District, Andhra Pradesh, India and was aimed to detect the presence of extended-spectrum beta-lactamases (ESBL) producing P. aeruginosa from sub-clinical mastitis that are apparently healthy Murrah buffaloes. Inflammation of the udder by microbial infection is one of the leading economic disease in the dairy sector. Most of the pathogens especially Pseudomonas aeruginosa are refractory to the antibiotic theraphy. Nowadays, the antimicrobial resistance owed by the bacteria is at an alarming rate. Pseudomonas aeruginosa, a multi drug resistant pathogen carry the extended spectrum beta lactamase genes in its plasmids and thus become resistant to antibiotic therapy. A total of 276 milk samples were collected from 69 milch animals in the month of August, 2021 comprising from all the four quarters of each buffalo. Nine (n=9) (3.26%) P. aeruginosa isolates were identified from the sub-clinical mastitis milk samples from the total 276 samples collected in the study. The isolates on morphological analysis were typical to P. aeruginosa. Seven isolates were positive for phenotypical beta-lactamases production. A total of 1 and 2 isolates were found reactive of the blaSHV and blaOXA genes, and two isolates harboured both blaSHV and blaOXA genes, respectively on PCR assay. No blaTEM gene was found in the isolates. Antibiogram of the ESBL producing P. aeruginosa isolates (n=7) possessed 100% resistance against most of the commonly used antimicrobials like ampicillin, amoxycillin, clindamycin, co-trimoxazole oxytetracycline, streptomycin and ceftriaxone (85%). The P. aeruginosa isolates were sensitive to amikacin (100%), ciprofloxacin (87.4%), gentamicin (84%), norfloxacin (84.8%) and enrofloxacin (82%).