Detection Of Resistance Genes Research Articles

Deteksi Gen Resistensi Aeromonas hydrophila yang Diisolasi dari Budidaya Ikan Lele Di Jambi

Aeromonas hydrophila is one of the causative agents for Motile Aeromonas Septicemia (MAS) in catfish. MAS treatment is generally carried out by administering antibiotics. This study aimed to determine the antibiotic resistance profile of A. hydrophila and detect the presence of resistance genes. The samples are archived A. hydrophila isolates isolated from catfish ponds in the district of Muara Jambi, Bungo, Jambi Province, and Merangin total of 18 isolates. Testing the antibiotic resistance profile was carried out using the disk diffusion method against tetracycline, oxytetracycline, ampicillin, amoxicillin, ciprofloxacin, enrofloxacin, and streptomycin. The detected resistance genes were ampC, qnrA, qnrB, and qnrS. The results showed A. hydrophila isolates were resistant to tetracycline (100%), oxytetracycline (94.42%), ampicillin (77.70%), and amoxicillin (77.72%) and still sensitive to ciprofloxacin (83.32%), enrofloxacin (100 %), and streptomycin (100%). The ampC gene was found in 8 out of 14 isolates resistant to beta-lactam antibiotics (57.14%). The qnrA and qnrS genes were found in 3 out of 4 isolates resistant to quinolone antibiotics (75%), while qnrB was found in 1 isolate (25%). A. hydrophila isolated from catfish ponds in Jambi develops resistance to antibiotics and resistance genes found in isolates resistant to antibiotics.

Outbreak of colistin and carbapenem-resistant Klebsiella pneumoniae ST16 co-producing NDM-1 and OXA-48 isolates in an Iranian hospital

BackgroundColistin and carbapenem-resistant Klebsiella pneumoniae (Col-CRKP) represent a significant and constantly growing threat to global public health. We report here an outbreak of Col-CRKP infections during the fifth wave of COVID-19 pandemic.MethodsThe outbreak occurred in an intensive care unit with 22 beds at a teaching university hospital, Isfahan, Iran. We collected eight Col-CRKP strains from seven patients and characterized these strains for their antimicrobial susceptibility, determination of hypermucoviscous phenotype, capsular serotyping, molecular detection of virulence and resistance genes. Clonal relatedness of the isolates was performed using MLST.ResultsThe COVID-19 patients were aged 24–75 years with at least 50% pulmonary involvement and were admitted to the intensive care unit. They all had superinfection caused by Col-CRKP, and poor responses to antibiotic treatment and died. With the exception of one isolate that belonged to the ST11, all seven representative Col-CRKP strains belonged to the ST16. Of these eight isolates, one ST16 isolate carried the iucA and ybtS genes was identified as serotype K20 hypervirulent Col-CRKP. The blaSHV and blaNDM-1 genes were the most prevalent resistance genes, followed by blaOXA-48 and blaCTX-M-15 and blaTEM genes. Mobilized colistin-resistance genes were not detected in the isolates.ConclusionsThe continual emergence of ST16 Col-CRKP strains is a major threat to public health worldwide due to multidrug-resistant and highly transmissible characteristics. It seems that the potential dissemination of these clones highlights the importance of appropriate monitoring and strict infection control measures to prevent the spread of resistant bacteria in hospitals.

Development of a rapid detection method for the macrolide resistance gene in Mycobacterium avium using the amplification refractory mutation system-loop-mediated isothermal amplification method.

Macrolide antibiotics such as clarithromycin (CLR) and azithromycin are the key drugs used in multidrug therapy for Mycobacterium avium complex (MAC) diseases. For these antibacterial drugs, drug susceptibility has been correlated with clinical response in MAC diseases. We have previously demonstrated the correlation between drug susceptibility and mutations in the 23S rRNA gene, which confers resistance to macrolides. Herein, we developed a rapid detection method using the amplification refractory mutation system (ARMS)-loop-mediated isothermal amplification (LAMP) technique to identify mutations in the 23S rRNA gene of M. avium. We examined the applicability of the ARMS-LAMP method to genomic DNA extracted from six genotypes of M. avium clinical isolates. The M. avium isolates were classified into 21 CLR-resistant and 9 CLR-susceptible strains based on the results of drug susceptibility tests; the 23S rRNA genes of these strains were sequenced and analyzed using the ARMS-LAMP method. Sequence analysis revealed that the 9 CLR-sensitive strains were wild-type strains, whereas the 21 CLR-resistant strains comprised 20 mutant-type strains and one wild-type strain. Using ARMS-LAMP, no amplification from genomic DNAs of the 10 wild-type strains was observed using the mutant-type mismatch primer sets (MTPSs); however, amplification from the 20 mutant-type strain DNAs was observed using the MTPSs. The rapid detection method developed by us integrates ARMS-LAMP with a real-time turbidimeter, which can help determine drug resistance in a few hours. In conclusion, ARMS-LAMP might be a new clinically beneficial technology for rapid detection of mutations.IMPORTANCEMultidrug therapy for pulmonary Mycobacterium avium complex disease is centered on the macrolide antibiotics clarithromycin and azithromycin, and resistance to macrolides is an important prognosticator for clinical aggravation. Therefore, it is important to develop a quick and easy method for detecting resistance to macrolides. Drug resistance is known to be correlated with mutations in macrolide resistance genes. We developed a rapid detection method using amplification refractory mutation system (ARMS)-loop-mediated isothermal amplification (LAMP) to identify a mutation in the 23S rRNA gene, which is a macrolide resistance gene. Furthermore, we examined the applicability of this method using M. avium clinical isolates. The rapid method developed by us for detection of the macrolide resistance gene by integrating ARMS-LAMP and a real-time turbidimeter can help in detection of drug resistance within a few hours. Since this method does not require expensive equipment or special techniques and shows high analytical speed, it would be very useful in clinical practice.

Identification and Genomic Analyses of a Multidrug Resistant Avian Pathogenic Escherichia coli Coharboring mcr-1, blaTEM-176 and blaCTX-M-14 Genes

The emergence and transmission of the colistin-resistance gene mcr and extended-spectrum β-lactamase (ESBL) encoding genes pose a significant threat to global public health. In recent years, it has been reported that mcr-1 and ESBL genes can coexist in single bacteria strain. The objective of this study was to characterize a multidrug-resistant (MDR) avian pathogenic Escherichia coli (APEC) isolate carrying mcr and ESBL encoding genes in China. A total of 200 APEC isolates were collected for antimicrobial susceptibility testing by Kirby–Bauer (K–B) disk method. The MDR strain EC012 were then further analyzed for minimum inhibitory concentrations, antimicrobials resistance genes (ARGs) detection, conjugation, and whole-genome sequencing (WGS). Among all APEC isolates determined by K–B disk method, strain EC012 was resistant to almost all the antimicrobials, including polymyxin B, cefotaxime, and ceftazidime. Moreover, EC012 harbored ARGs mcr-1, blaTEM-176, and blaCTX-M-14. WGS analysis revealed that EC012 belonged to epidemic APEC serotype O1:H16 and multilocus sequence type ST295. EC012 consisted of one chromosome and six plasmids, encoding a broad ARGs. The blaCTX-M-14, mcr-1 or blaTEM-176 genes were located on conjugative plasmids pEC012-1 or pEC012-5, respectively. These plasmids were successfully transferred to transconjugants and resulted in the resistance to polymyxin B, cefotaxime, and ceftazidime. This study indicated that APEC was a potential reservoir of colistin-resistance gene mcr-1 and ESBL encoding genes, and highlighted the necessity for enhanced monitoring of ARGs dissemination among bacteria from different origins.

Rapid detection of mexX in Pseudomonas aeruginosa based on CRISPR-Cas13a coupled with recombinase polymerase amplification.

The principal pathogen responsible for chronic urinary tract infections, immunocompromised hosts, and cystic fibrosis patients is Pseudomonas aeruginosa, which is difficult to eradicate. Due to the extensive use of antibiotics, multidrug-resistant P. aeruginosa has evolved, complicating clinical therapy. Therefore, a rapid and efficient approach for detecting P. aeruginosa strains and their resistance genes is necessary for early clinical diagnosis and appropriate treatment. This study combines recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats-association protein 13a (CRISPR-Cas13a) to establish a one-tube and two-step reaction systems for detecting the mexX gene in P. aeruginosa. The test times for one-tube and two-step RPA-Cas13a methods were 5 and 40 min (including a 30 min RPA amplification reaction), respectively. Both methods outperform Quantitative Real-time Polymerase Chain Reactions (qRT-PCR) and traditional PCR. The limit of detection (LoD) of P. aeruginosa genome in one-tube and two-step RPA-Cas13a is 10 aM and 1 aM, respectively. Meanwhile, the designed primers have a high specificity for P. aeruginosa mexX gene. These two methods were also verified with actual samples isolated from industrial settings and demonstrated great accuracy. Furthermore, the results of the two-step RPA-Cas13a assay could also be visualized using a commercial lateral flow dipstick with a LoD of 10 fM, which is a useful adjunt to the gold-standard qRT-PCR assay in field detection. Taken together, the procedure developed in this study using RPA and CRISPR-Cas13a provides a simple and fast way for detecting resistance genes.

Cost-effective multiplex PCR assay for simultaneous detection of bacterial leaf blight, blast and brown planthopper resistance genes in rice

Cost-effective multiplex PCR assay for simultaneous detection of bacterial leaf blight, blast and brown planthopper resistance genes in rice

A microfluidic card-based electrochemical assay for the detection of sulfonamide resistance genes

Most electroanalytical detection schemes for DNA markers require considerable time and effort from expert personnel to thoroughly follow the analysis and obtain reliable outcomes. This work aims to present an electrochemical assay performed inside a small card-based platform powered by microfluidic manipulation, requiring minimal human intervention and consumables. The assay couples a sample/signal dual amplification and DNA-modified magnetic particles for the detection of DNA amplification products. Particularly, the sul1 and sul4 genes involved in the resistance against sulfonamide antibiotics were analyzed. As recognized by the World Health Organization, antimicrobial resistance threatens global public health by hampering medication efficacy against infections. Consequently, analytical methods for the determination of such genes in environmental and clinical matrices are imperative. Herein, the resistance genes were extracted from E. coli cells and amplified using an enzyme-assisted isothermal amplification at 37 °C. The amplification products were analyzed in an easily-produced, low-cost, card-based set-up implementing a microfluidic system, demanding limited manual work and small sample volumes. The target amplicon was thus captured and isolated using versatile DNA-modified magnetic beads injected into the microchannel and exposed to the various reagents in a continuously controlled microfluidic flow. After the optimization of the efficiency of each phase of the assay, the platform achieved limits of detections of 44.2 pmol L−1 for sul1 and 48.5 pmol L−1 for sul4, and was able to detect down to ≥500-fold diluted amplification products of sul1 extracted from E. coli living cells in around 1 h, thus enabling numerous end-point analyses with a single amplification reaction.

Molecular Characteristics of Staphylococcus aureus Isolates form Food-Poisoning Outbreaks (2011-2022) in Sichuan, China.

Staphylococcal food poisoning (SFP) is one of the most common foodborne diseases in the world. This study aimed to investigate the molecular epidemiological characteristics of Staphylococcus aureus isolated from SFP. A total of 103 S. aureus isolates were obtained during 2011-2022 in Sichuan, southwest China. All isolates were tested for the genomic characteristics and phylogenetic analysis by performing whole-genome sequencing. Multilocus sequence typing analysis showed 17 multilocus sequence types (STs), ST7 (23.30%), ST5 (22.33%), and ST6 (16.50%) being the most common. A total of 45 virulence genes were detected, 22 of which were staphylococcal enterotoxin (SE) genes. Among the identified SE genes, selX exhibited the highest prevalence (86.4%). All isolates carried at least one SE gene. The results of the antimicrobial resistance (AMR) gene detection revealed 41 AMR genes of 12 classes. β-lactam resistance genes (blal, blaR1, blaZ) and tetracycline resistance gene (tet(38)) exhibited a higher prevalence rate. Core genome single nucleotide polymorphism showed phylogenetic clustering of the isolates with the same region, year, and ST. The results indicated that the SFP isolates in southwest of China harbored multiple toxin and resistance genes, with a high prevalence of new SEs. Therefore, it is important to monitor the antimicrobial susceptibility and SE of S. aureus to reduce the potential risks to public health.

Baseline susceptibility of Anopheles gambiae to clothianidin in northern Ghana

BackgroundClothianidin, an insecticide with a novel mode of action, has been deployed in the annual indoor residual spraying programme in northern Ghana since March 2021. To inform pragmatic management strategies and guide future studies, baseline data on local Anopheles gambiae sensu lato (s.l.) susceptibility to the clothianidin insecticide were collected in Kpalsogu, a village in the Northern region, Ghana.MethodsPhenotypic susceptibility of An. gambiae mosquitoes to clothianidin was assessed using the World Health Organization (WHO) insecticide resistance monitoring bioassay. The WHO cone bioassays were conducted on mud and cement walls sprayed with Sumishield 50 wettable granules (WG) (with clothianidin active ingredient). Daily mortalities were recorded for up to 7 days to observe for delayed mortalities. Polymerase chain reaction (PCR) technique was used to differentiate the sibling species of the An. gambiae complex and also for the detection of knock down resistance genes (kdr) and the insensitive acetylcholinesterase mutation (ace-1).ResultsThe WHO susceptibility bioassay revealed a delayed killing effect of clothianidin. Mosquitoes exposed to the cone bioassays for 5 min died 120 h after exposure. Slightly higher mortalities were observed in mosquitoes exposed to clothianidin-treated cement wall surfaces than mosquitoes exposed to mud wall surfaces. The kdr target-site mutation L1014F occurred at very high frequencies (0.89–0.94) across all vector species identified whereas the ace-1 mutation occurred at moderate levels (0.32–0.44). Anopheles gambiae sensu stricto was the most abundant species observed at 63%, whereas Anopheles arabiensis was the least observed at 9%.ConclusionsAnopheles gambiae s.l. mosquitoes in northern Ghana were susceptible to clothianidin. They harboured kdr mutations at high frequencies. The ace-1 mutation occurred in moderation. The results of this study confirm that clothianidin is an effective active ingredient and should be utilized in malaria vector control interventions.

Pathogenicity assessment of Arcobacter butzleri isolated from Canadian agricultural surface water

BackgroundWater is considered a source for the transmission of Arcobacter species to both humans and animals. This study was conducted to assess the prevalence, distribution, and pathogenicity of A. butzleri strains, which can potentially pose health risks to humans and animals. Cultures were isolated from surface waters of a mixed-use but predominately agricultural watershed in eastern Ontario, Canada. The detection of antimicrobial resistance (AMR) and virulence-associated genes (VAGs), as well as enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) assays were performed on 913 A. butzleri strains isolated from 11 agricultural sampling sites.ResultsAll strains were resistant to one or more antimicrobial agents, with a high rate of resistance to clindamycin (99%) and chloramphenicol (77%), followed by azithromycin (48%) and nalidixic acid (49%). However, isolates showed a significantly (p < 0.05) high rate of susceptibility to tetracycline (1%), gentamycin (2%), ciprofloxacin (4%), and erythromycin (5%). Of the eight VAGs tested, ciaB, mviN, tlyA, and pldA were detected at high frequency (> 85%) compared to irgA (25%), hecB (19%), hecA (15%), and cj1349 (12%) genes. Co-occurrence analysis showed A. butzleri strains resistant to clindamycin, chloramphenicol, nalidixic acid, and azithromycin were positive for ciaB, tlyA, mviN and pldA VAGs. ERIC-PCR fingerprint analysis revealed high genetic similarity among strains isolated from three sites, and the genotypes were significantly associated with AMR and VAGs results, which highlight their potential environmental ubiquity and potential as pathogenic.ConclusionsThe study results show that agricultural activities likely contribute to the contamination of A. butzleri in surface water. The findings underscore the importance of farm management practices in controlling the potential spread of A. butzleri and its associated health risks to humans and animals through contaminated water.

Ultrasensitive Electrochemiluminescence Biosensor to Detect Ampicillin Resistance Gene (ARGAMP) Based on a Novel Near-Infrared Ruthenium Carbene Complex/TPrA/PEI Ternary ECL System.

The establishment of rapid target identification and analysis methods for antibiotic resistance genes (ARGs) is urgently needed. In this study, we unprecedently designed a target-catalyzed hairpin assembly (CHA) electrochemiluminescent (ECL) biosensor for the ultrasensitive detection of ampicillin resistance genes (ARGAMP) based on a novel, efficient near-infrared ruthenium carbene complex/TPrA/PEI ternary ECL system with low oxidation potential. The ternary NIR-ECL system illustrated in this work displayed double ECL intensity in comparison with their corresponding traditional binary ECL system. The as-prepared ECL biosensor illustrated in this work demonstrates highly selective and sensitive determination of ARGAMP from 1 fM to 1 nM and a low detection limit of 0.23 fM. Importantly, it also exhibits good accuracy and stabilities to identify ARGAMP in plasmid and bacterial genome DNA, which demonstrates its excellent reliability and great potential in detecting ARGAMP in real environmental samples.

Phenotypic and Molecular Characterization of Hypervirulent and Multidrug-Resistant Acinetobacter baumannii Isolated from ICU Respiratory Infections.

The emergence of hypervirulent strains of Acinetobacter baumannii poses a significant threat in intensive care units (ICU). This study aimed to molecularly characterize hypervirulent A. baumannii strains isolated from ICU patients with respiratory infections. Six strains were isolated from ICU patients over one month. Isolates were identified by phenotypic characterization biochemical properties and 16s RNA sequencing. Antibiotic susceptibility testing was conducted followed by resistance genes detection by PCR. MLST, and PFGE were employed to analyse clonal relationships among strains. Plasmid replicon typing and plasmid transmission frequencies were determined. The isolated strains exhibited diverse clinical manifestations, including acute respiratory distress syndrome (ARDS). Antibiotic susceptibility testing revealed multidrug-resistance phenotype. Molecular analysis revealed a complex genetic landscape of antibiotic resistance genes, including ESBLs and carbapenemases, as well as virulence genes such as ompA, csuE, and exoS. The multiple sequence types indicating genetic diversity among the strains as ST1512, ST622, and ST149 (each type two isolates). Plasmid characterization revealed the presence of diverse replicon types associated with multidrug resistance. This study provides comprehensive insights into the phenotypic, molecular, and epidemiological characteristics of hypervirulent A. baumannii outbark in ICU.

Whole-genome analysis of Escherichia coli isolated from wild Amur tiger (Panthera tigris altaica) and North China leopard (Panthera pardus japonensis).

Escherichia coli is an important intestinal flora, of which pathogenic E. coli is capable of causing many enteric and extra-intestinal diseases. Antibiotics are essential for the treatment of bacterial infections caused by pathogenic E. coli; however, with the widespread use of antibiotics, drug resistance in E. coli has become particularly serious, posing a global threat to human, animal, and environmental health. While the drug resistance and pathogenicity of E. coli carried by tigers and leopards in captivity have been studied intensively in recent years, there is an extreme lack of information on E. coli in these top predators in the wild environment. Whole genome sequencing data of 32 E. coli strains collected from the feces of wild Amur tiger (Panthera tigris altaica, n = 24) and North China leopard (Panthera pardus japonensis, n = 8) were analyzed in this article. The multi-locus sequence types, serotypes, virulence and resistance genotypes, plasmid replicon types, and core genomic SNPs phylogeny of these isolates were studied. Additionally, antimicrobial susceptibility testing (AST) was performed on these E. coli isolates. Among the E. coli isolates studied, 18 different sequence types were identified, with ST939 (21.9%), ST10 (15.6%), and ST3246 (9.4%) being the most prevalent. A total of 111 virulence genes were detected, averaging about 54 virulence genes per sample. They contribute to invasion, adherence, immune evasion, efflux pump, toxin, motility, stress adaption, and other virulence-related functions of E. coli. Sixty-eight AMR genes and point mutations were identified. Among the detected resistance genes, those belonging to the efflux pump family were the most abundant. Thirty-two E. coli isolates showed the highest rate of resistance to tetracycline (14/32; 43.8%), followed by imipenem (4/32; 12.5%), ciprofloxacin (3/32; 9.4%), doxycycline (2/32; 6.3%), and norfloxacin (1/32; 3.1%). Our results suggest that E. coli isolates carried by wild Amur tigers and North China leopards have potential pathogenicity and drug resistance.

Detection of Tetracycline Resistance Genes and Susceptibility Profile of Bacteria Isolated from Clinical Samples

The use of Antibiotics by humans for an extensive period of time has an impact not just on the disease being treated, but also on the microbial population in that environment. Antibiotic abuse in many developing nations have aided the emergence and widespread transmission of antibiotic resistant resistance genes in the environment. Tetracycline is a broad-spectrum antibiotic used to treat diseases in both humans and animals. Tetracycline is regulated by tet genes, which are engaged in active drug efflux, ribosome protection, or enzymatic drug modification. The use of Tetracycline’s as a treatment option for treating oral and systemic disorders haves been limited due to the existence of these genes. Hence, this study aimed to detect tetracycline tet (A) and tet (B) resistance genes in bacterial isolated from clinical samples and to determine their susceptibility profile. An experimental design was used for this study. A total of forty human clinical isolates from blood, urine, throat, wound, and sputum were randomly selected for this study. The isolates were collected and sub-cultured in nutrient broth for analysis and PCR was used for the detection of genes. The result showed that antibiotics resistant genes were present in the all of the isolates. However, tetB had the highest occurrence (91.6%) of than tetA (8.4%). All the isolates showed high resistant to the various antibiotics including ceftazidime, cefuroxime, gentamicin, ofloxacin, amoxicillin, nitrofurantoin, cefixime, and ciprofloxacin. This could be due to the frequent use of broad-spectrum antibiotics in clinical settings which have led to the development of antibiotic resistant strains among bacteria. Antibiotics resistant genes (tetA &amp; tetB) are present in bacteria isolated from clinical setting and may become a serious public health concern when these genes are transferred among organisms that affect both humans and animals. Hence, preventative measures should be employed to reduce the diffusion of these genes in the clinical setting since they present a serious public health concern when they are transferred among organisms that affect both humans and animals.

Molecular Detection of Resistance and Virulence Genes in Coagulase Negative Staphylococci Isolated from Blood Cultures at the University Teaching Hospital of Bouake

Molecular Detection of Resistance and Virulence Genes in Coagulase Negative Staphylococci Isolated from Blood Cultures at the University Teaching Hospital of Bouake

A multi-channel electrochemical biosensor based on polyadenine tetrahedra for the detection of multiple drug resistance genes.

Antimicrobial resistance poses a serious threat to human health due to the high morbidity and mortality caused by drug-resistant microbial infections. Therefore, the development of rapid, sensitive and selective identification methods is key to improving the survival rate of patients. In this paper, a sandwich-type electrochemical DNA biosensor based on a polyadenine-DNA tetrahedron probe was constructed. The key experimental conditions were optimized, including the length of polyadenine, the concentration of the polyadenine DNA tetrahedron, the concentration of the signal probe and the hybridization time. At the same time, poly-avidin-HRP80 was used to enhance the electrochemical detection signal. Finally, excellent biosensor performance was achieved, and the detection limit for the synthetic DNA target was as low as 1 fM. In addition, we verified the practicability of the system by analyzing E. coli with the MCR-1 plasmid and realized multi-channel detection of the drug resistance genes MCR-1, blaNDM, blaKPC and blaOXA. With the ideal electrochemical interface, the polyA-based biosensor exhibits excellent stability, which provides powerful technical support for the rapid detection of antibiotic-resistant strains in the field.

Multi-drug resistant (MDR) Gram-negative pathogenic bacteria isolated from poultry in the Noakhali region of Bangladesh.

Rapidly increasing antibiotic-resistant bacterial strains in Bangladesh's food and farm animals stem from the excessive and inappropriate use of antibiotics. To assess the prevalence of multi-drug resistant (MDR) Gram-negative bacteria in poultry chicks, we sought to isolate and identify strains carrying antimicrobial resistance genes. Isolation and identification involved biochemical tests, 16S rRNA sequencing, and PCR screening of species-specific genes. MDR patterns were evaluated using CLSI guidelines with seventeen antibiotics across twelve classes. Targeted gene sequences were amplified for the detection of Extended-spectrum β-Lactamase (ESBL), carbapenem, tetracycline, sulfonamide, and colistin resistance genes. Common isolates, such as Escherichia coli, Klebsiella pneumoniae, Proteus penneri, and Enterobacter hormaechei, exhibited average Multiple Antimicrobial Resistance (MAR) indices of 0.66, 0.76, 0.8, 0.84, and 0.81, 0.76, 0.84, 0.41 for broiler and layer chicken, respectively. Providencia stuartii and Salmonella enterica, exclusive to broiler samples, had MAR indices of 0.82 and 0.84, respectively. Additional isolates Morganella morganii, Aeromonas spp., and Wohlfahrtiimonas chitiniclastica were found in layers (Average MAR indices: 0.73, 0.71, and 0.91). Notably, M. morganii, E. hormaechei and W. chitiniclastica were identified for the first time in Bangladeshi poultry chicken, although their evolution is yet to be understood. In this study, Pan-drug resistance was observed in one P. stuartii (broiler) and one Aeromonas spp. (layer) with a MAR index 1, while all isolates exhibited MAR indices >0.2, indicating MDR. Antimicrobial resistance (AMR) gene screening identified blaTEM, blaSHV, tetA, and sul1 in a majority of the MDR strains. Interestingly, E. coli (lactose positive and negative) and E. hormaechei were exclusively found to possess the tetB gene. In addition, E. coli (lactose negative), Klebsiella pneumoniae, Enterobacter hormaechei, M. morganii, and P. stuartii were observed to carry the colistin-resistant mcr-1 gene, whereas sul2 was detected in E. coli (lactose positive and negative), E. hormaechei, P. stuartii, and P. penneri. These findings emphasize the health risk of our consumers of both broiler and layer chickens as they have turned into a potent reservoir of various AMR gene carrying MDR and Pan-drug resistant bacteria.

Evaluation of the BioFire FilmArray Pneumonia Panel Plus for Detection of Viral, Bacterial Pathogens and Antimicrobial Resistance Genes in Lower Respiratory Tract Specimens at Al-Zahra Hospital for Pediatrics in Najaf, Iraq

The study aimed to evaluate the BioFire FilmArray Pneumonia Panel (BFPP) in rapidly diagnosing pathogens and detecting antimicrobial resistance genes in pediatric respiratory specimens, with an aim to enhance clinical decision-making in a pediatric hospital in Najaf, Iraq. For a period from 24/11/2022 to 22/5/2023, 36 specimens from pediatric patients with lower respiratory tract infection of males and females of different ages less than six years were collected. All specimens met the inclusion criteria, where Procalcitonin (PCT) levels above 0.5 ng/ml and negative blood cultures. The most prevalent pathogen detected were Acinetobacter calcoaceticus-baumannii complex in 63.89% of samples. Staphylococcus aureus and Klebsiella pneumoniae were detected in 44.44% and 41.67% of specimens, respectively. Less frequent were Streptococcus pneumoniae and Escherichia coli at 19.44%, Pseudomonas aeruginosa at 16.67%, Haemophilus influenzae at 11.11%, and Moraxella catarrhalis and Serratia marcescens each at 5.56%. Adenovirus was the most frequently detected virus, in 33.33% of the total samples. Other viruses detected include Human Rhinovirus/Enterovirus 19.44%, Respiratory Syncytial Virus 16.66%, Parainfluenza 11.11%, Human Metapneumovirus and Influenza B account for 5.55% while Corona virus not detected at all specimens. VIM gene was found in 63.89% of specimens, underscoring extensive carbapenem resistance, while other resistance genes appeared sporadically like mecA/C in 13.88%, IMP in 5.55% and KPC in 2.77 while OXA-48 like not detected at all.

Surveillance of antimicrobial resistance in Streptococcus agalactiae isolated from pregnant women and newborns in 2010–2022

Objective. To study the dynamics and molecular mechanisms of resistance of Streptococcus agalactiae strains isolated from pregnant women and newborns in St. Petersburg in 2010–2022. Materials and Methods. The strains of S. agalactiae (n = 248) were obtained by analyzing the clinical material of pregnant women and newborns who were hospitalized at Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott in 2010–2011 and 2020–2022. Determination of susceptibility to antibacterial drugs was evaluated by disk diffusion method. The detection of resistance genes to macrolides and lincosamides (mef and ermB) was performed by real-time PCR using reagents «Resistom MLSB-Streptococcus» (Litech, Russia). Results. All isolates S. agalactiae (n = 248) were sensitive to penicillin and vancomycin. Compared to 2010–2011, the resistance of S. agalactiae to erythromycin increased from 18.4% to 42.1% and to clindamycin from 15.6% to 39.3%. Among S. agalactiae strains (n = 71) resistant to erythromycin and clindamycin, cMLSB phenotype was predominant (59.1%); iMLSB (31%) and M-phenotype (9.9%) were less common. The results of detection of genes encoding resistance to erythromycin and clindamycin showed that in 38 (53.5%) and 9 (12.7%) isolates of S. agalactiae resistance genes ermB and mefA were detected as one resistance determinant, respectively, association of two ermB/mefA genes was detected in 2 (2.8%) strains; resistance genes were not detected in 22 (31%) isolates. Conclusions. Penicillin and vancomycin are effective drugs for the treatment and prevention of infections caused by S. agalactiae. The increase in resistance to macrolides and lincosamides reflects the global trend of increasing resistance to these antibiotics. Regular monitoring of antibiotic resistance of the pathogen is necessary to update the clinical recommendations.

Genetic detection of Resistance Genes among Enterobacter cloacae and Citrobacter spp. isolates

Internationally, there is an increasing prevalence of diseases caused by germs that are resistant to many drugs, which is often a reason for worry. The objective of the present investigation was to identify specific genes linked to the resistance of beta-lactam, macrolide, and sulfa medications in Enterobacter cloacae and Citrobacter spp. isolates. A total of 364 patients were included in the study, with 132 (36.26%) being male and 232 (63.74%) being female. The results indicated that 135 samples, accounting for 37.08% of the total, exhibited positive bacterial growth. The findings indicated that Enterobacter cloacae was present in 25 (6.86%) of the patients with urinary tract infections, whereas Citrobacter spp. was found in only 2 (0.54%) cases (consisting of 1 (0.27%) C. freundii isolate and 1 (0.27%) C. farmer isolate). The PCR findings indicated that the sul-1 gene was present in 23 out of 25 (92%) E. cloacae isolates and in 2 out of 2 (100%) Citrobacter spp. isolates. On the other hand, the sul-2 gene was detected in 22 out of 25 (88%) E. cloacae isolates and in 2 out of 2 (100%) Citrobacter spp. isolates. The mph(A) gene was present in 80% (20/25) of E. cloacae isolates and in 100% (2/2) of Citrobacter spp. isolates. The findings showed that 96% of the E. cloacae isolates and 100% of the Citrobacter spp. isolates tested positive for both ctx-m and ctx-m-10 genes. The prevalence of the veb gene in E. cloacae isolates was 76% (19/25), but in Citrobacter spp. isolates it was 100% (2/2). The investigation did not detect the gene ctx-m-14 in any of the local isolates examined. Genes of sul-3 and ctx-m-14 were no recorded in this study.