Colonization of residents and staff of a long-term-care facility and adjacent acute-care hospital geriatric unit by multiresistant bacteria. - Related Documents




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93901.0000Colonization of residents and staff of a long-term-care facility and adjacent acute-care hospital geriatric unit by multiresistant bacteria. Long-term-care facilities (LTCFs) are reservoirs of resistant bacteria. We undertook a point-prevalence survey and risk factor analysis for specific resistance types among residents and staff of a Bolzano LTCF and among geriatric unit patients in the associated acute-care hospital. Urine samples and rectal, inguinal, oropharyngeal and nasal swabs were plated on chromogenic agar; isolates were typed by pulsed-field gel electrophoresis; resistance genes and links to insertion sequences were sought by PCR; plasmids were analysed by PCR, restriction fragment length polymorphism and incompatibility grouping. Demographic data were collected. Of the LTCF residents, 74.8% were colonized with ≥1 resistant organism, 64% with extended-spectrum β-lactamase (ESBL) producers, 38.7% with methicillin-resistant Staphylococcus aureus (MRSA), 6.3% with metallo-β-lactamase (MBL) producers, and 2.7% with vancomycin-resistant enterococci. Corresponding rates for LTCF staff were 27.5%, 14.5%, 14.5%, 1.5% and 0%, respectively. Colonization frequencies for geriatric unit patients were lower than for those in the LTCF. Both clonal spread and plasmid transfer were implicated in the dissemination of MBL producers that harboured IncN plasmids bearing bla(VIM-1), qnrS, and bla(SHV-12). Most (44/45) ESBL-producing Escherichia coli isolates had bla(CTX-M) genes of group 1; a few had bla(CTX-M) genes of group 9 or bla(SHV-5); those with bla(CTX-M-15) or bla(SHV-5) were clonal. Risk factors for colonization of LTCF residents with resistant bacteria included age ≥86 years, antibiotic treatment in the previous 3 months, indwelling devices, chronic obstructive pulmonary disease, physical disability, and the particular LTCF unit; those for geriatric unit patients were age and dementia. In conclusion, ESBL-producing and MBL-producing Enterobacteriaceae and MRSA were prevalent among the LTCF residents and staff, but less so in the hospital geriatric unit. Education of LTCF employees and better infection control are proposed to minimize the spread of resistant bacteria in the facility.201019686277
94510.9997Extended Spectrum Beta Lactamase (ESBL), bla(TEM),bla(SHV) and bla(CTX-M), Resistance Genes in Community and Healthcare Associated Gram Negative Bacteria from Osun State, Nigeria. BACKGROUND: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the bla(TEM),bla(SHV) and bla(CTX-M) genes. The prevalence of ESBL producing bacteria has been on the increase globally, especially its upsurge among isolates from community-acquired infections has been observed. AIM: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. MATERIAL AND METHODS: A cross-sectional study was carried out from August 2016 - July 2017 in Osun State, Nigeria. Three hundred and sixty Gram-negative bacteria recovered from clinical samples obtained from both community and healthcare-associated infections were tested. They included 147 Escherichia coli (40.8%), 116 Klebsiella spp (32.2%), 44 Pseudomonas aeruginosa (12.2%) and 23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). RESULTS: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). The overall prevalence of ESBL producers was 41.4% with Klebsiella spp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs. 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the bla(CTX-M) gene predominating (47.0%) followed by bla(TEM) (30.9%) and bla(SHV) gene was the least, 22.1%. The bla(CTX-M) gene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. CONCLUSION: A high prevalence of ESBL producing gram-negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control the spread of these pathogens should be addressed.202132729432
94020.9996Acquisition of multidrug-resistant bacteria and colistin resistance genes in French medical students on internships abroad. BACKGROUND: Acquisition of multidrug resistant bacteria (MDR) and colistin resistance genes by international travellers has been demonstrated. Studies conducted in medical students during internships abroad are scant. METHODS: Nasopharyngeal, rectal, and vaginal swabs samples were collected from 382 French medical students before and after travel to investigate the acquisition of MDR bacteria. The bacterial diversity in the samples was assessed by culture on selective media. We also genetically characterised the isolates of MDR bacteria including Extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E), methicillin-resistant Staphylococcus aureus (MRSA), and Carbapenemase-producing Enterobacteriacae (CPE) using the real-time polymerase chain reaction method. The samples were collected from 293 students and were investigated for mcr colistin-resistance genes using RT-PCR directly on the samples, followed by conventional PCR and sequencing. RESULTS: A proportion of 29.3% (112/382) of the participants had acquired ESBL-E and 2.6% (10/382) had acquired CPE. The most common species and ESBL-E encoding gene were Escherichia coli (125/127 isolates, 98.4%) and bla(CTX-M-A) (121/127, 95.3%), respectively. A proportion of 6.8% (20/293) of the participants had acquired mcr-1 genes, followed by mcr-3 (1/293, 0.3%) and mcr-8 (1/293, 0.3%). We found that taking part in humanitarian missions to orphanages (aRR = 2.01, p < 0.0001), being in contact with children during travel (aRR = 1.78, p = 0.006), the primary destination of travel being Vietnam (aRR = 2.15, p < 0.0001) and north India (aRR = 2.41, p = 0.001), using antibiotics during travel (aRR = 1.77, p = 0.01), and studying in 2018 (aRR = 1.55, p = 0.03) were associated with the acquisition of ESBL-E. When the primary destination of travel was Vietnam (aRR = 2.74, p < 0.0001) and the year of study was 2018 (aRR = 1.93, p < 0.002), this was associated with acquisition of colistin resistance genes. CONCLUSION: Medical students are at a potential risk of acquiring ESBL-E, CPE and colistin resistance genes. A number of risk factors have been identified, which may be used to develop targeted preventive measures.202133248262
144630.9996One-Day Prevalence of Extended-Spectrum β-Lactamase (ESBL) and Carbapenemase-Producing Bacteria in Fecal Samples from Surgical Patients: A Concerning Trend of Antibiotic Resistance. PURPOSE: Extended-spectrum β-lactamase (ESBL) and carbapenemase producing bacteria are of increasing concern due to their multidrug resistance and infection potential. This study determines the one-day prevalence of faecal carriage of ESBL and carbapenemase producing Gram-negative bacilli. METHODS: Fecal samples were collected from 30 post-surgery patients (hospitalized for at least 48 hours) in each of the four hospitals involved in the study and were analyzed for antibiotic-resistant bacteria. Identification was done using Maldi Tof mass spectrometry, and antibiotic susceptibility was tested using disk diffusion and specialized tests for ESBL (double disk synergy technique) and carbapenem (NG-TEST CARBA 5) resistance detection. PCR was conducted on isolates to detect betalactam resistance genes, carbapenemase genes and quinolone resistance genes. FINDINGS: Out of the 120 patients enrolled, 38.33% (n = 46) and 49.16.33% (n = 59) were found to carry ESBL- and carbapenemase-producing bacteria, respectively, in their fecal samples. Among the isolates, 51.08% (n = 47) exhibited ESBL production, with Escherichia coli (44.56%) being the most common species. The identification of bacteria with resistance to carbapenems showed a predominance of the species Escherichia coli (44.45%) followed by the species Klebsiella pneumoniae (16.06%) and Acinetobacter baumanii (13.58%). The study of the association of variables shows a high degree of association (p < 0.05) for the factors independent walking and use of a wheelchair with ESBL production. The most frequently detected genes among ESBL producing bacteria were bla(CTXM-1) (91.49%), qnrB (70.21%) and qnrs (63.82%). bla(NDM) (54.68%) was the most detected carbapenemase genes among carbapenemase producing isolates. CONCLUSION: This study demonstrates, for the first time, a significant prevalence of ESBL and carbapenemase producing gram-negative bacteria among surgical patients in Benin, with multiple resistance genes detected. Findings should be interpreted in light of the cross-sectional design and >48-hour hospitalization criterion.202540635768
105040.9996Antibiotic resistance and β-lactam resistant genes among bacterial isolates from clinical, river water and poultry samples from Kathmandu, Nepal. OBJECTIVE: To assess the antibiotic resistance and beta-lactam resistance genes among bacterial isolates from clinical, river water and poultry samples. METHODS: Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were isolated from clinical, poultry and river water samples collected during 2020-22. They were subjected to antimicrobial susceptibility tests following the CLSI guidelines. The bacteria were screened for β-lactam resistance genes bla (TEM), mcr-1, mecA and bla(NDM-1) . RESULTS: Among 2835 clinical samples, E. coli was the most frequently isolated bacterium (10.3%, 292), followed by S. aureus (6.0%, 169) and P. aeruginosa (4.0%, 143). Of the E. coli isolates, 64.4% exhibited multidrug resistance (MDR) and 43.8% were extended-spectrum β-lactamase (ESBL) producers, with 44.5% and 16.4% harbouring the blaTEM and mcr-1 genes, respectively. Among S. aureus isolates, 80.9% of methicillin-resistant strains (MRSA) carried the mecA gene, while 30.1% of metallo-β-lactamase (MBL)-producing P. aeruginosa were positive for the blaNDM-1 gene. In poultry samples, 30.4% of E. coli isolates harboured the blaTEM gene among 128 ESBL producers, and the prevalence of colistin-resistant isolates carrying mcr-1 was higher than in clinical samples. In contrast, the occurrence of ESBL-producing E. coli and MRSA, along with their associated resistance genes, was lower in water samples. CONCLUSIONS: This study demonstrated widespread multidrug resistance (MDR) and ESBL production among clinical, poultry and river water bacterial isolates in the Kathmandu valley. Colistin-resistant E. coli carrying the mcr-1 gene, methicillin-resistant S. aureus (MRSA) with mecA and metallo-β-lactamase (MBL)-producing P. aeruginosa harboring blaNDM-1 were detected across sources. These findings emphasize an urgent One Health approach to curb the growing threat of antimicrobial resistance in the region.202541113068
144750.9996Molecular detection of β-lactamase and integron genes in clinical strains of Klebsiella pneumoniae by multiplex polymerase chain reaction. INTRODUCTION: Infections caused by β-lactamase-producing gram-negative bacteria, such as Klebsiella pneumoniae, are increasing globally with high morbidity and mortality. The aim of the current study was to determine antimicrobial susceptibility patterns and the prevalence of antibiotic resistance genes (β-lactamase and integron genes) using multiplex PCR. METHODS: One-hundred K. pneumoniae isolates were collected from different clinical samples. Antibiotic susceptibility testing was performed with thirteen different antibiotics. Multiplex-PCR was used to detect β-lactamase (bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC) and integron genes (int I, int II, and int III). RESULTS: The highest and lowest rate of resistance was exhibited against amikacin (93%) and imipenem (8%), respectively. The frequency of β-lactamase-positive K. pneumoniae was 37%, and the prevalence of the bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC genes was 38%, 24%, 19%, 12%, 6%, 11%, 33%, 0%, 28%, and 23%, respectively. Of the 100 isolates, eight (8%) were positive for class I integrons; however, class II and III integrons were not detected in any of the strains. CONCLUSIONS: These results indicate co-carriage of a number of β-lactamase genes and antibiotic resistance integrons on the same plasmids harboring multi-drug resistance genes. It seems that these properties help to decrease treatment complications due to resistant bacterial infections by rapid detection, infection-control programs and prevention of transmission of drug resistance.201728700049
94460.9996Fecal shedding of extended-spectrum beta-lactamase-producing Enterobacterales in cats admitted to an animal shelter. OBJECTIVES: The objective of this study was to evaluate shedding of extended-spectrum beta-lactamase (ESBL)-producing bacteria in cats admitted to an animal shelter. METHODS: Fecal samples were collected from cats admitted to an animal shelter between 12 June and 23 August 2018. Selective enrichment culture for ESBL-producing bacteria was performed and isolates were speciated and tested for selected ESBL genes using PCR. RESULTS: ESBL-producing Enterobacterales were identified in fecal samples from 2/87 (2.3%; 95% confidence interval 0.6-8.0) cats. One isolate was an Escherichia coli that possessed bla(CTX-M-1), bla(CMY-2) and bla(TEM) genes. The other was Enterobacter cloacae possessing bla(CTX-M-1) and bla(CMY-2). CONCLUSIONS AND RELEVANCE: While the study sample size and prevalence rate for ESBL-producing bacteria were low, these data document that cats admitted to similar shelters could harbor these agents. The risk posed by ESBL-producing bacterium shedding in cats, both to cats and other species, is currently unclear. However, these findings support the need for more investigation of interspecies transmission of ESBL-producing bacteria and ESBL genes, as well as the importance of antimicrobial stewardship and routine infection control measures.202235133182
107170.9996Characterization of Beta-Lactamase and Fluoroquinolone Resistance Determinants in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa Isolates from a Tertiary Hospital in Yola, Nigeria. Infections due to antimicrobial resistant gram-negative bacteria cause significant morbidity and mortality in sub-Saharan Africa. To elucidate the molecular epidemiology of antimicrobial resistance in gram-negative bacteria, we characterized beta-lactam and fluoroquinolone resistance determinants in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa isolates collected from November 2017 to February 2018 (Period 1) and October 2021 to January 2022 (Period 2) in a tertiary medical center in north-eastern Nigeria. Whole genome sequencing (WGS) was used to identify sequence types and resistance determinants in 52 non-duplicate, phenotypically resistant isolates. Antimicrobial susceptibility was determined using broth microdilution and modified Kirby-Bauer disk diffusion methods. Twenty sequence types (STs) were identified among isolates from both periods using WGS, with increased strain diversity observed in Period 2. Common ESBL genes identified included bla(CTX-M), bla(SHV,) and bla(TEM) in both E. coli and K. pneumoniae. Notably, 50% of the E. coli in Period 2 harbored either bla(CTX-M-15) or bla(CTX-M-1 4) and phenotypically produced ESBLs. The bla(NDM-7) and bla(VIM-5) metallo-beta-lactamase genes were dominant in E. coli and P. aeruginosa in Period 1, but in Period 2, only K. pneumoniae contained bla(NDM-7), while bla(NDM-1) was predominant in P. aeruginosa. The overall rate of fluoroquinolone resistance was 77% in Period 1 but decreased to 47.8% in Period 2. Various plasmid-mediated quinolone resistance (PMQR) genes were identified in both periods, including aac(6')-Ib-cr, oqxA/oqxB, qnrA1, qnrB1, qnrB6, qnrB18, qnrVC1, as well as mutations in the chromosomal gyrA, parC and parE genes. One E. coli isolate in Period 2, which was phenotypically multidrug resistant, had ESBL bla(CTX-M-15,) the serine carbapenemase, bla(OXA-181) and mutations in the gyrA gene. The co-existence of beta-lactam and fluoroquinolone resistance markers observed in this study is consistent with widespread use of these antimicrobial agents in Nigeria. The presence of multidrug resistant isolates is concerning and highlights the importance of continued surveillance to support antimicrobial stewardship programs and curb the spread of antimicrobial resistance.202337999619
94680.9996Identification and Characterization of Multidrug-Resistant Extended-Spectrum Beta-Lactamase-Producing Bacteria from Healthy and Diseased Dogs and Cats Admitted to a Veterinary Hospital in Brazil. The objective of this study was to identify the main extended-spectrum beta-lactamase (ESBL)-producing bacteria and to detect the frequency of the major genes responsible to trigger this resistance in hospitalized animals. We collected 106 rectal swabs from cats (n = 25) and dogs (n = 81) to detect ESBL-producing isolates. ESBL-positive samples were submitted to the antimicrobial susceptibility test, and polymerase chain reaction was performed to detect TEM, SHV, and CTX-M genes from different groups. We observed that 44.34% of these samples (11 cats and 36 dogs) were positive for ESBL-producing bacteria. Thirteen animals (27.66%-seven cats and six dogs) were hospitalized for elective castration (healthy animals). Only a single animal was positive for ESBL-producing bacteria at hospital admission (the animal also showed an ESBL-positive isolate after leaving the hospital), whereas 11 were positive only at the hospital discharge. Of the 73 ESBL-producing isolates, 13 were isolated from cats (8 sick and 7 healthy) and 60 from dogs (53 sick and 7 healthy). Escherichia coli was the major ESBL-producing bacterium isolated (53.42%), followed by Pseudomonas aeruginosa (15.07%), Salmonella sp., and Proteus mirabilis (5.48% each one). Antimicrobial resistance profile of ESBL-producing isolates showed that 67 isolates (91.78%) were resistant to 3 or more antibiotic classes, while 13 of them (17.81%-2 healthy cats and 11 sick dogs) were resistant to all tested antimicrobial classes. The bla(TEM) gene exhibited the highest frequency in ESBL-producing isolates, followed by the bla(CTX-M) group 8/25, bla(CTX-M) group 1 and bla(CTX-M) group 9 genes. These results are useful to assess the predominance of ESBL-producing isolates recovered from dogs and in cats in Brazil. Consequently, we draw attention to these animals, as they can act as reservoirs for these microorganisms, which are the major pathogens of nosocomial infections worldwide.202133185513
144290.9996Superbugs in the supermarket? Assessing the rate of contamination with third-generation cephalosporin-resistant gram-negative bacteria in fresh Australian pork and chicken. BACKGROUND: Antibiotic misuse in food-producing animals is potentially associated with human acquisition of multidrug-resistant (MDR; resistance to ≥ 3 drug classes) bacteria via the food chain. We aimed to determine if MDR Gram-negative (GNB) organisms are present in fresh Australian chicken and pork products. METHODS: We sampled raw, chicken drumsticks (CD) and pork ribs (PR) from 30 local supermarkets/butchers across Melbourne on two occasions. Specimens were sub-cultured onto selective media for third-generation cephalosporin-resistant (3GCR) GNBs, with species identification and antibiotic susceptibility determined for all unique colonies. Isolates were assessed by PCR for SHV, TEM, CTX-M, AmpC and carbapenemase genes (encoding IMP, VIM, KPC, OXA-48, NDM). RESULTS: From 120 specimens (60 CD, 60 PR), 112 (93%) grew a 3GCR-GNB (n = 164 isolates; 86 CD, 78 PR); common species were Acinetobacter baumannii (37%), Pseudomonas aeruginosa (13%) and Serratia fonticola (12%), but only one E. coli isolate. Fifty-nine (36%) had evidence of 3GCR alone, 93/163 (57%) displayed 3GCR plus resistance to one additional antibiotic class, and 9/163 (6%) were 3GCR plus resistance to two additional classes. Of 158 DNA specimens, all were negative for ESBL/carbapenemase genes, except 23 (15%) which were positive for AmpC, with 22/23 considered to be inherently chromosomal, but the sole E. coli isolate contained a plasmid-mediated CMY-2 AmpC. CONCLUSIONS: We found low rates of MDR-GNBs in Australian chicken and pork meat, but potential 3GCR-GNBs are common (93% specimens). Testing programs that only assess for E. coli are likely to severely underestimate the diversity of 3GCR organisms in fresh meat.201829484175
992100.9996Phenotypic and genotypic evaluation of beta-lactamases (ESBL and KPC) among enterobacteria isolated from community-acquired monomicrobial urinary tract infections. Beta-lactamases enzymes such as extended-spectrum beta-lactamases (ESBL) and carbapenemase type beta-lactamases (KPC) confer resistance to beta-lactam drugs among Gram-negative rods, mainly Enterobacteriaceae, as those frequently related to urinary tract infections (UTI). The aim of this study was to evaluate ESBL and KPC among enterobacteria isolated from monomicrobial UTI and to establish correlations between the presence of genetic markers and the phenotypic resistance to beta-lactam antibiotics. Out of 12 304 urine samples collected during 2009, 93 enterobacteria showing an ESBL phenotype were recovered. Imipenem was used for KPC screening and modified disk approximation assay was used for detection of ESBL phenotype. Polymerase chain reaction was used for screening of bla(SHV), bla(TEM), bla(CTX-M), and bla(KPC). Considering the isolated bacteria showing ESBL phenotype 56% of the isolates were positive for two genes. The bla(TEM) was the most frequent (87·1%). Neither KPC phenotype nor bla(KPC)-harboring bacteria were observed. Monitoring the antimicrobial resistance is extremely important to sustain empirical therapy of community-acquired urinary tract infections (Co-UTI).201424621159
951110.9996Analyses of Extended-Spectrum-β-Lactamase, Metallo-β-Lactamase, and AmpC-β-Lactamase Producing Enterobacteriaceae from the Dairy Value Chain in India. The consumption of milk contaminated with antibiotic-resistant bacteria poses a significant health threat to humans. This study aimed to investigate the prevalence of Enterobacteriaceae producing β-lactamases (ESBL, MBL, and AmpC) in cow and buffalo milk samples from two Indian states, Haryana and Assam. A total of 401 milk samples were collected from dairy farmers and vendors in the specified districts. Microbiological assays, antibiotic susceptibility testing, and PCR-based genotyping were employed to analyze 421 Gram-negative bacterial isolates. The overall prevalence of β-lactamase genes was 10% (confidence interval (CI) (7-13)), with higher rates in Haryana (13%, CI (9-19)) compared to Assam (7%, CI (4-11)). The identified β-lactamase genes in isolates were bla(CMY), bla(MOX), bla(FOX), bla(EBC), and bla(DHA), associated with AmpC production. Additionally, bla(CTX-M1), bla(SHV), and bla(TEM) were detected as ESBL producers, while bla(VIM), bla(IMP), bla(SPM), bla(SIM), and bla(GIM) were identified as MBL producers. Notably, Shigella spp. were the dominant β-lactamase producers among identified Enterobacteriaceae. This study highlights the presence of various prevalent β-lactamase genes in milk isolates, indicating the potential risk of antimicrobial-resistant bacteria in dairy products. The presence of β-lactam resistance raises concern as this could restrict antibiotic options for treatment. The discordance between genotypic and phenotypic methods emphasizes the necessity for comprehensive approaches that integrate both techniques to accurately assess antibiotic resistance. Urgent collaborative action incorporating rational and regulated use of antibiotics across the dairy value chain is required to address the global challenge of β-lactam resistance.202337760745
956120.9996Detection of Extended-Spectrum Beta-Lactamase-Producing and Carbapenem-Resistant Bacteria from Mink Feces and Feed in the United States. Antibiotic-resistant infections caused by extended-spectrum β-lactamases (ESBLs) and carbapenemases are increasing worldwide. Bacteria resistant to extended-spectrum cephalosporins and last resort carbapenems have been reported from food animals and their environments. Other concentrated nonfood-producing animals such as mink farming can be a reservoir of bacteria resistant to these critically important antibiotics. The objective of this study was to determine the prevalence of ESBL-producing bacteria and carbapenem-resistant (CR) bacteria from mink fecal (n = 42) and feed (n = 8) samples obtained from a commercial mink farm in the United States. The most prevalent ESBL-producing bacteria identified from the fecal samples were Escherichia coli (93%), Klebsiella pneumoniae (76%), and Proteus species (88%). E. coli (100%) and K. pneumoniae (75%) were also the most prevalent ESBL-producing bacteria identified from feed samples. All ESBL E. coli isolates were resistant to penicillin and most cephem beta-lactam antibiotics. Among the ESBL E. coli isolates, co-resistance was observed to ciprofloxacin (33%) and gentamicin (28%) indicating multidrug resistance. ESBL E. coli isolates predominantly carried bla(CTX-M-14) and bla(CTX-M-15) genes. Although all feed K. pneumoniae isolates carried bla(CTX-M-9), all fecal K. pneumoniae isolates carried bla(SHV). CR Pseudomonas species (7%), Hafnia alvei (24%), and Myroides odoratimimus (9.5%) were detected from fecal samples. H. alvei (37.5%) was the only CR bacteria detected from the feed samples. All CR isolates were polymerase chain reaction negative for the tested carbapenemases that are commonly reported, which may indicate intrinsic rather than acquired resistance. This study indicates that mink production can be a reservoir for bacteria resistant to the highest priority critically important antibiotics for human health.202133978469
844130.9996Whole Genome Sequencing of Extended Spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae Isolated from Hospitalized Patients in KwaZulu-Natal, South Africa. Extended spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae remain a critical clinical concern worldwide. The aim of this study was to characterize ESBL-producing K. pneumoniae detected within and between two hospitals in uMgungundlovu district, South Africa, using whole genome sequencing (WGS). An observational period prevalence study on antibiotic-resistant ESKAPE (i.e. Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) bacteria was carried out in hospitalized patients during a two-month period in 2017. Rectal swabs and clinical specimens were collected from patients hospitalized and were screened for ESBL-producing, Gram-negative ESKAPE bacteria using cefotaxime-containing MacConkey agar and ESBL combination disk tests. Nine confirmed ESBL-K. pneumoniae isolated from six patients and two hospitals were whole genome sequenced using an Illumina MiSeq platform. Genome sequences were screened for presence of integrons, insertion sequences, plasmid replicons, CRISPR regions, resistance genes and virulence genes using different software tools. Of the 159 resistant Gram-negative isolates collected, 31 (19.50%) were ESBL-producers, of which, nine (29.03%) were ESBL-K. pneumoniae. The nine K. pneumoniae isolates harboured several β-lactamase genes, including bla(CTX-M-15), bla(TEM-1b), bla(SHV-1), bla(OXA-1) concomitantly with many other resistance genes e.g. acc(6')-lb-cr, aadAI6, oqxA and oqxB that confer resistance to aminoglycosides and/or fluoroquinolones, respectively. Three replicon plasmid types were detected in both clinical and carriage isolates, namely ColRNAI, IncFIB(K), IncF(II). Sequence type ST152 was confirmed in two patients (one carriage isolate detected on admission and one isolate implicated in infection) in one hospital. In contrast, ST983 was confirmed in a clinical and a carriage isolate of two patients in two different hospitals. Our data indicate introduction of ESBL-producing K. pneumoniae isolates into hospitals from the community. We also found evidence of nosocomial transmission within a hospital and transmission between different hospitals. The Clustered Regularly Interspaced Palindromic Repeats (CRISPR)-associated cas3 genes were further detected in two of the nine ESBL-KP isolates. This study showed that both district and tertiary hospital in uMgungundlovu District were reservoirs for several resistance determinants and highlighted the necessity to efficiently and routinely screen patients, particularly those receiving extensive antibiotic treatment and long-term hospitalization stay. It also reinforced the importance of infection, prevention and control measures to reduce the dissemination of antibiotic resistance within the hospital referral system in this district.201931000772
957140.9996Occurrence, Typing, and Resistance Genes of ESBL/AmpC-Producing Enterobacterales in Fresh Vegetables Purchased in Central Israel. Beta-lactam resistance can lead to increased mortality, higher healthcare expenses, and limited therapeutic options. The primary mechanism of beta-lactam resistance is the production of extended-spectrum beta-lactamases (ESBL) and AmpC beta-lactamases. The spread of beta-lactamase-producing Enterobacterales via the food chain may create a resistance reservoir. The aims of this study were to determine the prevalence of ESBL/AmpC-producing Enterobacterales in vegetables, to examine the association between EBSL/AmpC-producing bacteria and types of vegetables, packaging, and markets, and to investigate the genetic features of ESBL-producing isolates. The antibiotic susceptibilities were determined using VITEK. Phenotypic ESBL/AmpC production was confirmed using disk diffusion. ESBL-producing isolates were subjected to Fourier-transform infrared (FT-IR) spectroscopy and to whole genome sequencing using Oxford Nanopore sequencing technology. Of the 301 vegetable samples, 20 (6.6%) were positive for ESBL producers (16 Klebsiella pneumoniae and 4 Escherichia coli), and 63 (20.9%) were positive for AmpC producers (56 Enterobacter cloacae complex, 4 Enterobacter aerogenes/cancerogenus, and 3 Pantoea spp., Aeromonas hydrophila, and Citrobacter braakii). The blaCTX-M and blaSHV genes were most common among ESBL-producing isolates. The beta-lactamase genes of the ESBL producers were mainly carried on plasmids. Multilocus sequence typing and FT-IR typing revealed high diversity among the ESBL producers. AmpC producers were significantly more common in leafy greens and ESBL producers were significantly less common in climbing vegetables. The presence of ESBL/AmpC-producing Enterobacterales in raw vegetables may contribute to the dissemination of resistance genes in the community.202337887229
2122150.9996Phenotypic and genotypic characteristics of carbapenemase- and extended spectrum β-lactamase-producing Klebsiella pneumoniae ozaenae clinical isolates within a hospital in Panama City. Klebsiella pneumoniae spp ozaenae is a versatile bacterial species able to acquire antimicrobial resistance; the species presents a higher antimicrobial resistance profile compared to Klebsiella pneumoniae spp pneumoniae. Carbapenemase and extended spectrum β-lactamase (ESBL)-producing bacteria commonly arise in clinical settings where antimicrobial stewardship is limited. Our study aims to report the phenotypical and genetic characteristics of nosocomial Klebsiella pneumoniae spp ozaenae isolates associated with mortality collected from a tertiary-level hospital in Panama City. In October 2020, 11 consecutive multidrug-resistant Gram-negative isolates were recovered from secretions and blood cultures from hospitalized patients. Nearly 90% (10/11) of these patients died, and bacteria was obtained from six patients for investigation. Biochemical evaluation of the six isolates revealed the presence of multidrug-resistant Klebsiella pneumoniae spp ozaenae. Phenotypic evaluation indicated resistance to carbapenemase and EBSL. In contrast, genetic evaluation by PCR showed that only 30% (2/6) were resistant to CTX-M-1 (CTX-M group 1), whereas 60.7% (4/6) presented carbapenemase resistance genes, and 33.3% (2/6) presented New Delhi metallo-β-lactamase (NDM) resistance genes. Klebsiella pneumoniae ST258 was identified in 83.3% (5/6) of the isolates. Phylogenetic analysis using 16S revealed low homology among the six isolates. These results suggest that antibiotic resistance genes may have been incorporated into these Klebsiella pneumoniae spp ozaenae isolates within the hospital environment. We recommend strengthening the antimicrobial stewardship program and antibiotic control policy, as well as heightened infection control and prevention measures, such as ward sanitation and increased hand washing frequency.202134733509
950160.9996Incidence of Extended Spectrum β-Lactamase Genes (ESBLs) among community and health care infection in Mansoura University Hospital, Egypt. BACKGROUND: Multidrug-resistant (MDR) Gram-negative bacteria pose a significant challenge due to their limited treatment options. The production of extended-spectrum β-lactamases (ESBLs) is an important mechanism of resistance. This study aimed to identify the incidence and characteristics of ESBL-encoding genes (bla(CTX-M), bla(TEM), bla(SHV), and bla(OXA)) in MDR isolates. MATERIALS AND METHODS: A cross-sectional study was conducted from September 2022 to May 2023. ESBL-producing isolates (n = 105) out of 412 were recovered from hospitalized and outpatient settings and analyzed. Standard microbiological methods were used for isolates identification, susceptibility testing, and phenotypic ESBL detection. Additionally, bla(CTX-M), bla(TEM), bla(SHV), and bla(OXA) genes were identified using conventional PCR. RESULTS: Molecular profiling of β-lactamase determinants was conducted via PCR targeting bla(CTX-M), bla(TEM), bla(SHV), and bla(OXA) genes. Among phenotypically confirmed (100%) ESBL producers, 98% harbored one or more target genes, with bla(CTX-M) predominant (81%), followed by bla(SHV) (70.4%), bla(TEM) (62%), and bla(OXA) (30.4%). Carbapenem resistance was higher in ESBL-producing strains compared to non-ESBL strains. Extensively drug-resistant (XDR) isolates were the most common across hospital departments and outpatients. DISCUSSION: This study highlights the significant prevalence of ESBL genes and multidrug resistance among Gram-negative bacteria. The dominance of bla(CTX-M) and the existence of multiple resistance genes raise concerns about limited treatment options. The findings emphasize the need for stricter antibiotic stewardship and infection control measures to curb the spread of MDR pathogens. CONCLUSION: This study provides valuable insights into the alarming incidence of ESBL genes and MDR in Mansoura, Egypt. Continuous surveillance and implementation of effective control strategies are crucial to combat this growing public health threat.202540405086
1079170.9996CTX-M-15-producing Escherichia coli clone B2-O25b-ST131 and Klebsiella spp. isolates in municipal wastewater treatment plant effluents. OBJECTIVES: The global occurrence of antibiotic resistance genes in bacteria in water environments is an increasing concern. Treated wastewater was sampled daily over a 45 day period from the outflow of a municipal wastewater treatment plant in Brno, Czech Republic, and examined for extended-spectrum β-lactamase (ESBL)-producing bacteria. METHODS: Water samples were cultivated on MacConkey agar with cefotaxime (2 mg/L) and individual colonies were examined for ESBL production. Phenotypic ESBL-positive bacteria identified as Escherichia coli or Klebsiella spp. were tested for the presence of antibiotic resistance genes, the virulence gene afa/dra and the bla(CTX-M) upstream region. Genetic relatedness was analysed by PFGE, multilocus sequence typing and plasmid analysis. RESULTS: A total of 68 ESBL-producing Enterobacteriaceae isolates were detected in 34 out of 45 wastewater samples. ESBL-producing isolates included 26 E. coli isolates, 4 Klebsiella pneumoniae isolates and 1 Klebsiella oxytoca isolate. The pandemic and multiresistant B2-O25b-ST131 clone was predominant, being detected among 19 E. coli isolates, and 17 of the B2-O25b-ST131 isolates were positive for the FIA replicon and the afa/dra operon and had an IS26 element flanking bla(CTX-M-15). Seventeen of the B2-O25b-ST131 isolates showed closely related PFGE profiles (defined by 84% band similarity) and belonged to identical clonal groups. CONCLUSIONS: The results highlight the inadequacy of the treatment process in removing multiresistant bacteria from municipal wastewater and point to a risk of transmission of clinically important multiresistant strains, such as the pandemic ST131 clone, to the environment. This is the first study demonstrating the pandemic ST131 clone in wastewater.201121954457
1049180.9996Multiple Antibiotic-Resistant, Extended Spectrum-β-Lactamase (ESBL)-Producing Enterobacteria in Fresh Seafood. Members of the family Enterobacteriaceae include several human pathogens that can be acquired through contaminated food and water. In this study, the incidence of extended spectrum β-lactamase (ESBL)-producing enterobacteria was investigated in fresh seafood sold in retail markets. The ESBL-positive phenotype was detected in 169 (78.60%) isolates, with Escherichia coli being the predominant species (53), followed by Klebsiella oxytoca (27), and K. pneumoniae (23). More than 90% of the isolates were resistant to third generation cephalosporins, cefotaxime, ceftazidime, and cefpodoxime. Sixty-five percent of the isolates were resistant to the monobactam drug aztreonam, 40.82% to ertapenem, and 31.36% to meropenem. Resistance to at least five antibiotics was observed in 38.46% of the isolates. Polymerase Chain Reaction (PCR) analysis of ESBL-encoding genes detected bla(CTX), bla(SHV), and bla(TEM) genes in 76.92%, 63.3%, and 44.37% of the isolates, respectively. Multiple ESBL genes were detected in majority of the isolates. The recently discovered New Delhi metallo-β-lactamase gene (bla(NDM-1)) was detected in two ESBL⁺ isolates. Our study shows that secondary contamination of fresh seafood with enteric bacteria resistant to multiple antibiotics may implicate seafood as a potential carrier of antibiotic resistant bacteria and emphasizes an urgent need to prevent environmental contamination and dissemination of such bacteria.201728867789
1443190.9995Wastewater Surveillance Detected Carbapenemase Enzymes in Clinically Relevant Gram-Negative Bacteria in Helsinki, Finland; 2011-2012. Antimicrobial resistance profiling of pathogens helps to identify the emergence of rare or new resistance threats and prioritize possible actions to be taken against them. The analysis of wastewater (WW) can reveal the circulation of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARG) among the catchment communities. Here, we analyzed WW influent samples to determine the prevalence of carbapenemase genes-carrying Gram-negative bacteria (Carba-GNB) in Helsinki, Finland. This study set important historical reference points from the very early stage of the carbapenemase era, during the period 2011-2012. A total of 405 bacterial isolates grown on CHROMagarKPC (n = 195) and CHROMagarESBL (n = 210) from WW influent samples were collected between October 2011 and August 2012 and were analyzed. The bacterial DNA from the isolates was extracted, and the prevalence of carbapenemases genes bla (KPC), bla (NDM), bla (GES), bla (OXA-48), bla (IMP), bla (IMI), and bla (VIM) were screened with multiplexed PCR. All carbapenemase-positive isolates were identified taxonomically to species or genus level with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The nucleic acid extraction was successful for 399 isolates, of which 59 (14.8%) were found to carry carbapenemase genes. A total of 89.8% of the carbapenemase positive isolates (53 out of 59) were obtained from CHROMagarKPC plates and only 10.2% (six out of 59) were obtained from CHROMagar ESBL plates. Among the Carba-GNB isolates, 86.4% were bla (GES) (51 out of 59), 10.2% were bla (KPC) (six out of 59), and 3.4% were bla (VIM) (two out of 59). The most common carba-gene, bla (GES), was carried by 10 different bacterial species, including Aeromonas spp., Enterobacter spp., and Kluyvera spp.; the bla (KPC) gene was carried by Escherichia coli, Klebsiella pneumoniae, and Kluyvera cryocescens; and the bla (VIM) gene was carried by Aeromonas hydrophila/caviae and Citrobacter amalonaticus. This study emphasizes that wastewater surveillance (WWS) can be an additional tool for monitoring antimicrobial resistance (AMR) at the population level.202235722284