The emergence of carbapenemase-producing Enterobacteriaceae has limited therapeutic options and presents a major public health problem. Resistances against carbapenems in this bacterial family are mostly conveyed by metallo-beta-lactamases (MBL) including VIM, which are often encoded on resistance plasmids. We characterized four VIM-positive isolates that were obtained as part of a routine diagnostic screening from two laboratories in north-eastern Germany be-tween June and August 2020. Whole-genome sequences were generated to address a) their phylo-genetic properties, b) their plasmid content, and c) the underlying mechanisms of resistance and virulence. In addition, we performed phenotypic antibiotic and mercury resistance analyses. The genomic analysis revealed three different bacterial species including C. freundii, E. coli and K. oxytoca with four different sequence types. All isolates were geno- and phenotypically multi-drug-resistant (MDR). Three isolates of four carried identical VIM-1-resistance plasmids, which – in addition – encoded for a mercury resistance operon and showed some similarity to two ex-ternal plasmid sequences. The distribution of virulence features among the genomes was limited. Our results highlight the circulation of an identical VIM-1-resistance plasmid in three of four bacterial isolates of different Enterobacteriaceae in north-eastern Germany.

The incidence of carbapenem-resistant Enterobacteriaceae (CRE) has been increasing considerably annually since being first reported in 2008. Gyeonggi Province has the largest number of healthcare facilities in South Korea, which contribute to its regional vulnerability to CRE infections. Thus, there is a need to obtain up to date information on the occurrence of CRE infections in the Gyeonggi Province to take corrective measures. Therefore, in this study, we aimed to investigate the overall trends of CRE occurrence in Gyeonggi Province over the past 4 years and the epidemiological characteristics of CRE-related mortality. Patients admitted to a healthcare facility in Gyeonggi Province from January 2018 to December 2021 with positive blood cultures to CRE were included in the study and investigated. Additionally, the risk factors for CRE-related death were determined using data from CRE-positive patients who died within 30 days of last blood sampling. Older adults aged 70 and over accounted for the highest percentage of both survivors and non-survivors with CRE bacteremia. The number of non-survivors was higher in tertiary hospitals and among patients in intensive care unit. There were no significant differences in the mortality risk according to the use of antibiotics. Moreover, non-survivors were predominant among patients with hypertension, malignant tumor, and those having two or more underlying diseases. Klebsiella pneumoniae was the most common CRE strain in both survivors and non-survivors. Among the survivors, carbapenemase-producing Enterobacteriaceae was detected in 151 cases (71.2%) and 49 cases (23.1%) in the KPC and NDM groups, respectively. Among the non-survivors, it was detected in 25 cases (86.2%) and 5 cases (17.2%) in the KPC and NDM groups, respectively. The endemicity of CRE in South Korea has been proposed for several years; our results have suggested endemicity of CRE in the Gyeonggi Province. Despite the strengthened infection control policies in healthcare facilities during the pandemic, there has been an increasing trend in the isolation of CRE strains in South Korean long-term care hospitals and hospitals within Gyeonggi Province. Factors such as history of hospitalization and underlying diseases have been identified as risk factors for CRE-related deaths. However, to address this issue strictly, it is important to establish infection control infrastructure tailored based on each type of healthcare facility, and elaborate specific government support measures, accordingly.

This study aimed to isolate, identify enteric bacterial species from the migratory seagulls migrated to Libyan cost and to assess the antibiotic resistance of these bacteria. A total of 50 fecal samples were collected from slender-billed gulls in January, 2023 at Farwa Island near the city of Zwara. Bacteria were isolated by conventional culturing method, identified by using the Enterosystem 18R, and antibiotic susceptibility testing was conducted on the isolated bacteria. Only 32 of 46 bacteria were identified using biochemical tests. These identified bacteria belong to six species of Enterobacteriacae namely Citrobacter freundii, Pantoea agglomerans, E. coli, Enterobacter cloacae, Serratia liquifaciens, and Proteus mirabilis, with percentages of (17) 53.125%, (10) 31.25%, (2) 6.25%, (1) 3.125%, (1) 3.125%, and (1) 3.125%, respectively. All isolated bacterial species in this study were 100% sensitive to gentamicin and ciprofloxacin. The highest resistance rate was observed against the antibiotic cefoxitin. The results indicated that C. freundii was the most antibiotic-resistant bacterial species isolated in this study. In conclusion, slender-billed gulls carry multi-drug resistant bacteria. The study recommends implementation of a national program to survey antibiotic-resistant bacteria and expanding scientific research on migratory birds.

Enterobacteriaceae can contaminate meat during various stages of processing, including slaughter, evisceration, and butchering, causing foodborne illnesses. The purpose of the study was to investigate the prevalence of Enterobacteriaceae associated with carcasses obtained from slaughterhouses and raw cut meat collected from butcher shops. A total of 120 samples of camels and cattle were identified using biochemistry and PCR testing. Total viable count (TVC) was range from 4.91 to 5.37 Log10 CFU/g in slaughterhouses and butcher shops. E. coli was predominating with 84 (70%) among all samples, where the camels had the highest with 100% and sheep the lowest with 30%. Salmonella spp. was confirmed in 40% of camel samples, 47.5% of cattle samples, and 32.5% in sheep samples. Twenty-five Enterobacteriaceae genera were confirmed using PCR. Where sheep’s samples had the highest occurrence of Enterobacteriaceae with 15 different genera followed by camels and cattle samples with 14 different genera. The prevalence of Enterobacteriaceae among camel, cattle, and sheep carcasses raises significant concerns regarding food safety. Adherence to good hygiene practices throughout animal slaughtering is crucial to minimize the risk of infection and transmission and ensure food safety.

Beta-lactam resistant bacteria, commonly resident in tertiary hospitals, have emerged as a worldwide health problem because of ready-to-eat vegetable intake. We aimed to characterize the genes providing resistance to beta-lactam antibiotics in Enterobacteriaceae, isolated from five commercial salad brands for human consumption in Mexico City. 25 samples were collected, grow in blood agar plates, the bacteria were biochemistry identified and antimicrobial susceptibility testing was done, the carried family genes were identified by endpoint PCR and the specific genes were confirmed with WGS by NGS. 12 positive cultures were identified and their microbiological distribution was as follows, 8.3% for Enterobacter aerogene (n=1), 8.3% for Serratia fonricola (n=1), 16.7% for Serratia marcesens (n=2), 16.7% for Klebsiella pneumoniae (n=2), and 50% (n=6) for Enterobacter cloacae. The endpoint PCR results showed 11 colonies positive for blaBIL (91.7%), 11 for blaSHV (91.7%), 11 for blaCTX (97.7%), 12 for blaDHA (100%),4 for blaVIM (33.3%), 2 for blaOXA (16.7%), 2 for blaIMP (16.7%), 1 for blaKPC (8.3%) and 1 for blaTEM (8.3%) gene, all samples were negative blaROB, blaCMY, blaP, blaCFX and blaLAP gene. The sequencing analysis revels a specific genotypes for Enterobacter cloacae (blaSHV-12, blaCTX-M-15, blaDHA-1, blaKPC-2); Serratia marcescens (blaSHV-1, blaCTX-M-3, blaDHA-1, blaVIM-2); Klebsiella pneumoniae (blaSHV-12, blaCTX-M-15, blaDHA-1); Serratia fonticola (blaSHV-12, blaVIM-1, blaDHA-1) and Enterobacter aerogene (blaSHV-1, blaCTX-M-1, blaDHA-1, blaVIM-2, blaOXA-9). Our results indicate that beta-lactam resistant bacteria have acquired integrons with a different number of genes that providing panresistance to beta-lactam antibiotics, including penicillins, oxacillins, cefalosporins, monobactams, carbapenems and imipenems.

In South Western Indian ocean (IO), Extended-Spectrum β-Lactamase producing Enterobacteriaceae (ESBL) are a main public health issue. In livestock, ESBL burden was unknown. The aim of this study was estimating the prevalence of ESBL on commercial farms in Reunion, Mayotte and Madagascar and genes involved. Secondly, risk factors of ESBL occurrence in broiler, beef cattle and pig farms were explored. In 2016-2017, commercial farms were sampled using boot swabs and samples stored at 4°C before microbiological analysis for phenotypical ESBL and gene characterization. A semi-directive questionnaire was performed. Prevalences observed in all production types and territories were elevated, except for beef cattle in Reunion which differed significantly. The most common ESBL gene was the CTX-M-1 subtype. Generalized linear models explaining ESBL occurrence varied between livestock production sectors and allowed identifying main protective (e.g., water quality control and detergent use for cleaning) and risk factors (e.g., recent antibiotic use, other farmers visiting the exploitation, pet presence). This study is the first to explore tools for antibiotic resistance management in IO farms. It provides interesting hypothesis to explore about antibiotic use in IO and ESBL transmission between pig, beef cattle and humans in Madagascar.

Carbapenem resistant Enterobacteriaceae (CRE) has been public health risk in several countries and recent reports indicate the emergence of CRE in food animals. This study was conducted to investigate the occurrence, resistance patterns, and phylogenetic diversity of CRE E.coli from chicken. Routine bacteriology, PCR detection of E.coli species, multiplex PCR to detect carbapenemase encoding genes and phylogeny of CRE E. coli were conducted. The results show that 24.36 % (19/78) were identified as CRE based on the phenotypic identifications of which 17 were positive for the tested carabanemase genes. The majority, 57.99% (11/19) of the isolates harbored multiple carbapenemase genes. Four isolates harbored all blaNDM blaOXA, blaIMP, five and two different isolates harbored blaNDM and blaOXA, and blaOXA and blaIMP respectively. The Meropenem, Imipenem and Ertapenem MIC values for the isolates ranged from 2g/mL to ≥256g/mL. Phylogenetic grouping showed that the CRE E.coli isolates belonged to five different groups; groups A, B1, C, D and unknown. The detection of carbapenem resistant E.coli in this study shows that CRE is has become an emerging problem in farm animals, particularly, in poultry farms. This also implies the potential public health risks posed by CRE from chicken to the consumers.

: 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 optimization 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 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 bla_CTX-M, whilst eight (8/14, 57.1%) Enterobacter samples tested positive for bla_TEM. Interestingly, four (4/6, 66.7%) of the E. coli isolates carrying bla_CTX-M-positive strains were also positive for bla_TEM. Sanger sequencing of the PCR products revealed that five (5/6,83.3%) of the abovementioned isolates possessed the bla_CTX-M-15 allele. The results suggest the presence of potentially pathogenic ESBL-producing Enterobacteriaceae in poultry, threatening public health. Keywords:. Enterobacteriaceae, bla_CTX-M, bla_TEM

Escherichia coli causes several infections like intestinal diarrhea, urinary tract infections, and pneumonia, which are difficult to treat due to the emergence of antimicrobial resistance. The World Health Organisation (WHO) declared carbapenem-resistant E. coli a critical pathogen and called for the development of new antimicrobial agents. Phage therapy, a century-old technique, has been revitalised recently with many successful applications against multi-drug resistant infections. As such, phage therapy is considered one of the most effective alternatives or adjuvants to antibiotics and is also effective in treating multidrug-resistant (MDR) bacterial infections. However, few phages that can kill pathogenic E. coli strains from Southeast Asia have been studied at a molecular level, although the burden of MDR, particularly carbapenem resistance, is high in this region. In this study, three closely related obligately virulent Escherichia phages (øEc_Makalu_001, øEc_Makalu_002, and øEc_Makalu_003) that could infect uropathogenic E. coli were isolated from sewage samples and characterized using morphological as well as sequence-based analysis. Further, a comparative genomic and phylogenetic analysis was done to predict the functional genes and evolutionary relationships. Electron microscopy and genome-based classification revealed that all three phages belonged Krischvirus genus of Straboviridae family. One-step growth curve showed that the latent period of the phages ranged from 15-20 min, the outbreak period was about 50 min, and the burst size ranged between 74-127 PFU/bacterium. Moreover, the phages could tolerate a pH range of 6 to 9 and a temperature range of 25-37°C for up to 180 minutes without significant loss of phage viability. Spotting the purified phages in different clinical isolates showed a broad host-range of isolated Escherichia phages and could lyse up to 30% of the tested pathogenic clinical isolates, including an intra-genus isolate of Klebsiella pneumoniae. The genome organization of all three phages was almost identical and similar to T4-like phage with a full-length genome of ~163 kb. Around 284 coding sequences (CDS) were predicted in each phage, of which only 40% were functional with an average gene length of 951 bp. The coding density in all phages was approximately 95%, while the gene density was 1.73 gene/kbp. Putative lysin, holin, endolysin, and spanin genes were found in the genomes of all three phages, resembling that of T4-like phages implying their phylogenetic relatedness. Furthermore, various in silico sequence analysis tools confirmed that all three phages were strictly virulent with functional lysis modules but devoid of any known virulence or toxin genes, integron-associated genes and antimicrobial-resistance (AMR) genes. In summary, pre-clinical experimental analysis and exploration of genomic features suggest that all three phages may be suitable candidates for therapeutic applications against uropathogenic E. coli.