Investigation of Bacterial Persistence and Filaments Formation in Clinical Klebsiella pneumoniae: First Report from Iraq

Bacterial persistence is recognized as a major cause of antibiotic therapy failure, causing biofilms, and chronic intractable infections. The emergence of persisters in Klebsiella pneumoniae isolates has become a worldwide public health concern. The goal of the present study is to investigate the formation of persister cells beside filaments in Iraqi K. pneumoniae isolates. A total of fifty clinical K. pneumoniae isolates were collected from different clinical specimens and identified using the genotypic identification by using specific primer (rpoB gene) from housekeeping genes. Persister cells investigation is performed by exposure of stationary phase K. pneumoniae isolates to a high concentration of ciprofloxacin (×10 MIC) and counting the number of viable persister cells by CFU counts. Bacterial filament formation is detected and measured by light microscope scanning electron microscope. The results show the  bility of these pathogenic bacteria to form persister cells to survive the bactericidal antibiotics and to cause chronic infection.Furthermore, persistent isolates have the ability to change in shape and size extensively, about 4 times increase in cell length than their normal length. These phenomena are possibly the initial stages of bacterial resistance prevalence

Artificial Intelligence and the Silent Pandemic of Antimicrobial Resistance: A Comprehensive Exploration

The rise of antimicrobial resistance (AMR) in the 21st century has made it a worldwide disaster. Due to the fast spread of AMR illnesses and the lack of novel antimicrobials, the silent pandemic is well known. This issue requires a fast and meaningful response, not just speculation. To address this dilemma, deep learning (DL) and machine learning (ML) have become essential in many sectors. As a cornerstone of modern research, machine learning helps handle the many aspects of AMR. AI helps researchers construct clinical decision-support systems by collecting clinical data. These methods enable antimicrobial resistance monitoring and wise use. Additionally, AI applications help research new drugs. AI also excels at synergistic medicine combinations, providing new treatment methods. This paper summarizes our extensive study of AI and the silent epidemic of antibiotic resistance. Through deep learning and machine learning applications across multiple dimensions, we hope to contribute to the proactive management of AMR, moving away from its presentation as a future problem to present-day solutions

Synthesis and study of biological activity of some new Imidazole derivatives

In this work ester derivatives were synthesized by the reaction of imidazole derivatives (C1) with ethylchloroacetate in ethanol and NaOH to give the corresponding (C2) .While compound (C3) acetohydrazide was synthesized by the reaction of ester derivatives (C2) with hydrazine hydrat in ethanol. Compound (C3) from the reaction with different aromatic aldehydes in absolute ethanol gave the Schiff′s bases (C4,C5). The product compounds were characterized by FT-IR, U.V and 1HNMR spectra and the biological activities were studied as antibacterial

Unveiling the High Prevalence of Antibiotic Resistance and Quorum Sensing Genes in Uropathogenic Escherichia coli

Escherichia coli is considered one of the uropathogenic bacteria with different infection symptoms representing mild illness to acute sepsis. This study aims to detect E. coli in patients with urinary infection and investigate quorum sensing genes (lux S and motA) in multi-drug resistant isolates of E. coli.  200 urine samples were collected from patients with urinary tract infections from several hospitals in Baghdad. The antibiotics sensitivity test showed high resistance of isolates for Ampicillin (100%), Cefazolin (97%), Trimethoprim/ Sulfamethoxazole (83%), Ceftriaxone (77%), Ceftazidime and Ciprofloxacin (70% each of them), and moderate resistance of isolates for Levofloxacin (50%), Gentamicin (47%), Cefepime (40%), while low resistance Piperacillin/ Tazobactam (33%), Cefoxitin (30%), Nitrofurantoin (17%), Imipenem (10%), Ertapenem and Amikacin (7% each of them), and Tigecycline (3%). The results showed an increase percentage of infection in females was 30% in the ages 30-44 years, whereas in ages 15-29 and more than 45 years was 17%.  There was a high percentage (57.11%) of resistant isolates in females which are ages 30-44 years. While the ages more than 45 years were 66.4% and ages 15-29 were 34%. While, in males, the percentage was high in ages more than 45 years (35.25%) followed by age groups 30-44 years (31.5%) and 15-29 years (31%).  The prevalence percentage for luxS and motA genes in E. coli was 100%. In conclusion, E. coli isolates were multi-drug resistant due to all isolates had quorum sensing genes. Moreover, uropathogenic of E. coli in females was more frequent than in males due to the resistance of bacteria to antibiotics.    

Bacterial Filaments Induced by Antibiotic Minimal Inhibitory Concentrations in Persister Cells

Background: The ability of minor subpopulations among clonal populations to survive antibiotics is referred to as bacterial persistence. It is believed that persisters come from latent cells, where antibiotic target areas are less active and incapable of being affected. Objective: 112 clinical Escherichia coli isolates were acquired out of diverse medical samples and genetically identified using the uspA gene, which is part of the housekeeping genes. Methods: The examination of persister cells was carried out by subjecting isolates of E. coli in the exponential phase with high dose of ciprofloxacin (20 fold MIC) and calculating the surviving persister cells using CFU (colony forming units) counts. The detection and measurement of bacterial filament production was done using scanning electron and light microscopy. Results: Results showed that the bacterial filament cells kept on lengthen but cease to divide (no septa formation) at sub-minimal inhibitory doses of ciprofloxacin. Persistent isolates were shown to exhibit a wide range of form and size variations, with cells up to 4.5 times longer than usual. Conclusions: The results showed the importance of antibiotic stress on persisted cells that result in the production of filaments as a means of survival and the need to examine these rare phenotypic variations. These occurrences may be the beginning of the spread of bacterial resistance

Using the Lactobacillus gasseri filtrate to protect the mice from the pathogenic bacteria Aeromonas spp.

The protective effect of the Probiotic bacteria (Lactobacillus gasseri) against the pathogenic Bacteria (Aeromonas spp) was studied in vivo and in vitro, the inhibitory effect of the Probiotic bacteria filtrate was tested with the Well diffusion method, the filtrate showed a clear inhibiting efficiency toward the pathogenic bacteria and the diameter of the inhibition zone was 16 mm. A group of mice were injected intraperitoneally with (0.25 ml) of the filtrate for 10 days, then they were injected with (0.2 ml) of the Aeromonas spp bacteria living cells ( cell\ml) intraperitoneally, while the control group were injected with 0.25 ml of PBS. The mice were killed after 12 hours of injection with the pathogenic bacteria, they were injected with (5ml) of PBS intraperitoneally, the contents of the periton, Liver and the Spleen were taken after killing the mice, then a dilutions of the Periton contents were made and 0.1 ml was streaking on the media to calculate the number of the growing bacterial colonies, then a part of the Spleen was homogenized and streaking on the media, then the growing colonies were calculated and compared with the control mice, also the macrophage in the contents of the periton were counted, and a samples of the organs were taken directly after the killing and were putted in the formalin solution (10%) for study the histopathological changes. The results shows that the mice who were injected with the probiotic bacteria wasn't effected when they were exposed to the pathogenic bacteria unlike the group that were only exposed to the pathogenic bacteria without being injected with the probiotoic bacteria, and that shows the protective effect of the (Lactobacillus gasseri) against the pathogenic bacteria (Aeromonas spp), and the pathogenic changes shows an inflammatory response and that the tissues were appeared in their natural form

The Effect of pH Variation on Antibiotic Susceptibility of MDR Klebsiella pneumoniae Isolates

Background: Klebsiella pneumoniae is a significant opportunistic pathogen responsible for various nosocomial infections in humans. The emergence of multi-drug resistant strains poses a significant challenge in clinical settings, necessitating a deeper understanding of factors influencing antimicrobial resistance. Objective: This research aimed to investigate the impact of pH variation on the resistance patterns of multi-resistant K. pneumoniae isolated from Iraqi patients with urinary tract infections and wound infections against different antibiotics. Methods: Forty K. pneumoniae isolates were obtained from urine samples and wound swabs, and their identification was confirmed using the VITEK ® 2 compact system and molecular identification of the rpoB housekeeping gene. Antibiotic susceptibility testing was performed using the Kirby Bauer’s disk diffusion method under varying pH conditions (pH 5, 7, 9, and 11) at 37°C for 18 to 24 hours. Results: The study findings indicated that K. pneumoniae isolates exhibited differential susceptibility to antibiotics based on pH conditions. Cefotaxime demonstrated increased efficacy under alkaline pH, while tetracycline showed optimal efficacy under acidic conditions. However, ciprofloxacin displayed resistant phenotypes at acidic pH 5 and either resistant or intermediate phenotypes at alkaline pH 9. Conclusions: The results suggest a potential influence of pH on the antibiotic susceptibility profiles of K. pneumoniae isolates. Understanding the role of pH in antimicrobial resistance can inform strategies for better managing infections caused by multi-resistant pathogens. Further research is warranted to elucidate the underlying mechanisms and implications for clinical practice

The Prevalence of Microorganisms in H1N1 Patients Compared to Seasonal Influenza in a Sample of Iraqi Patients

This study provides valuable information on secondary microbial infections in H1N1 patients compared to Seasonal Influenza in Iraqi Patients. Nasopharynx  swabs were collected from  (12 ) patients  infected with Seasonal influenza (11  from Baghdad  and 1 Patient from south of Iraq) ,and ( 22 ) samples from patients with 2009 H1N1 ( 20 from Baghdad and  2 from  south of Iraq). The results show that the patients infected with 2009 H1N1 Virus were younger than healthy subjects and those infected with seasonal influenza. And the difference reached to the level of significance     (p< 0.01) compared with healthy subjects.Two cases infected with 2009 H1N1 virus (9.1%) were from south of the Iraq and remaining 20 cases were from Baghdad . Polymicrobial isolates from nasopharynx swab were observed in patients infected with 2009 H1N1 virus. Polybacterial infections (2-7 microorganisms) and fungal infection were reported in 21 out of 22 patients (95.5%) and 5 out of 22 (22.7 %) respectively.The predominant isolated microorganisms were Streptococcus pyogenes , Staphylococcus aureus and Streptococcus pneumoniae  were found in 95.2 %  , 95.2 % and 90.5 %  respectively .The results also show  that seven microorganisms were isolated from 10 (47.6 %) patients infected with 2009 H1N1 , no microorganism was isolated from patients infected with seasonal influenza or healthy persons. Key words: Seasonal Influenza , 2009 H1N1, Nasopharynx  swab

The unfinished agenda of communicable diseases among children and adolescents before the COVID-19 pandemic, 1990-2019: a systematic analysis of the Global Burden of Disease Study 2019

BACKGROUND: Communicable disease control has long been a focus of global health policy. There have been substantial reductions in the burden and mortality of communicable diseases among children younger than 5 years, but we know less about this burden in older children and adolescents, and it is unclear whether current programmes and policies remain aligned with targets for intervention. This knowledge is especially important for policy and programmes in the context of the COVID-19 pandemic. We aimed to use the Global Burden of Disease (GBD) Study 2019 to systematically characterise the burden of communicable diseases across childhood and adolescence. METHODS: In this systematic analysis of the GBD study from 1990 to 2019, all communicable diseases and their manifestations as modelled within GBD 2019 were included, categorised as 16 subgroups of common diseases or presentations. Data were reported for absolute count, prevalence, and incidence across measures of cause-specific mortality (deaths and years of life lost), disability (years lived with disability [YLDs]), and disease burden (disability-adjusted life-years [DALYs]) for children and adolescents aged 0-24 years. Data were reported across the Socio-demographic Index (SDI) and across time (1990-2019), and for 204 countries and territories. For HIV, we reported the mortality-to-incidence ratio (MIR) as a measure of health system performance. FINDINGS: In 2019, there were 3·0 million deaths and 30·0 million years of healthy life lost to disability (as measured by YLDs), corresponding to 288·4 million DALYs from communicable diseases among children and adolescents globally (57·3% of total communicable disease burden across all ages). Over time, there has been a shift in communicable disease burden from young children to older children and adolescents (largely driven by the considerable reductions in children younger than 5 years and slower progress elsewhere), although children younger than 5 years still accounted for most of the communicable disease burden in 2019. Disease burden and mortality were predominantly in low-SDI settings, with high and high-middle SDI settings also having an appreciable burden of communicable disease morbidity (4·0 million YLDs in 2019 alone). Three cause groups (enteric infections, lower-respiratory-tract infections, and malaria) accounted for 59·8% of the global communicable disease burden in children and adolescents, with tuberculosis and HIV both emerging as important causes during adolescence. HIV was the only cause for which disease burden increased over time, particularly in children and adolescents older than 5 years, and especially in females. Excess MIRs for HIV were observed for males aged 15-19 years in low-SDI settings. INTERPRETATION: Our analysis supports continued policy focus on enteric infections and lower-respiratory-tract infections, with orientation to children younger than 5 years in settings of low socioeconomic development. However, efforts should also be targeted to other conditions, particularly HIV, given its increased burden in older children and adolescents. Older children and adolescents also experience a large burden of communicable disease, further highlighting the need for efforts to extend beyond the first 5 years of life. Our analysis also identified substantial morbidity caused by communicable diseases affecting child and adolescent health across the world. FUNDING: The Australian National Health and Medical Research Council Centre for Research Excellence for Driving Investment in Global Adolescent Health and the Bill & Melinda Gates Foundation