An important chance of nosocomial acquired infections are caused by the opportunistic bacterium Klebsiella pneumoniae. Urine, wound, sputum, and blood samples were collected from all patients. This study aimed to detect the antibiotic resistance profile, the frequency of MDR, XDR, PDR, and detection of efflux pump and outer membrane permeability genes in K. pneumoniae isolates. One hundred twenty samples were collected from patients who were admitted to the Ramadi Teaching Hospitals in Al-Anbar Governorate. Fifty five of K. pneumoniae strains were collected from patients. The VITEK®2 Compact B System was used to detect the antibiotic resistance pattern of studied bacteria. The isolates were classified as MDR, XDR, or PDR based on established guidelines. The data were analyzed using Clinical and Laboratory Standards Institute (CLSI) breakpoints. PCR was used to detect the efflux pumps and porins genes. Out of the 120 samples studied, 45.83 % (55) tested positive for K. pneumoniae. The isolates displayed the greatest amount of resistance to cefazolin, ceftriaxone (98.2 %), ampicillin (100 %), and ceftazidime, cefepime (90.9 %). 20 % of the isolates were found to produce metallo-lactamases, and 41.81 % tested positive for extended-spectrum beta-lactamases. Overall, the rates of multi-drug resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) isolates were 57.2 %, 10.9 %, and 9.09 %, respectively. Additionally, the prevalence of efflux pump genes acrAB, mdtK, and tolC were 94.54 %, 14.54 %, and 89.09 %, respectively, while the porin-encoding genes ompK35 and ompK36 were found in 96.36 % and 98.18 % of the isolates. This investigation concluded that the study isolates had a high degree of antibiotic resistance heterogenicity. High frequencies of resistance to ampicillin, cefazolin, and ceftriaxone are present in study isolates. Most strains were categorized as MDR strains, with six being XDR strains and five being PDR strains. One of the main routes of antibiotic resistance in multidrug-resistant K. pneumoniae strains is through the acrAB efflux system. The high prevalence of the acrAB, tolC, ompk35, and ompK36 genes were increases the ability of these isolates combat antimicrobial treatments.

There is still no agreement on the gold standard technique for diagnosing of H. Pylori in Iraq, as well as a paucity of data on the validity of different diagnostic techniques. This study aimed to investigate the prevalence of this bacterium with different methods and compare them to the quantitative polymerase chain reaction (qPCR) as a golden standard technique among Iraqi patients. In total, 115 Iraqi patients strongly suspected of H. pylori infection were enrolled in the current study. Invasive techniques including rapid urease testing (RUT) and gastric tissue culture in addition to non-invasive techniques including ¹⁴C-Urea breath test (¹⁴C-UBT), stool antigen test (SAT), CagA-IgG serology, and qPCR were performed to confirm the H. pylori infection. The qPCR was used as the gold standard to estimate the sensitivity, specificity, positive and negative predictive values for the studied diagnostic tests. Overall, the prevalence rate of H. pylori in Iraqi patients was ranged from 47.8 to 70.4% using different methods. The positive results for each test were as follows: qPCR 81, (70.4%) UBT 79 (68.7%), SAT 77 (67%), RUT 76 (66.1%), Cag-IgG 61 (53%), and culture 55 (47.8%). The ¹⁴C-UBT showed the highest overall performance with 97.5% sensitivity, 97% specificity, and total accuracy of 97.3% followed by SAT, RUT, Cag-IgG, and culture method. Based on the accuracy of the studied methods for H. pylori detection, they can be arranged in order as follows: qPCR > UBT > SAT > RUT> CagA IgG > culture. Since a universal gold standard assay for the diagnosis of H. pylori has not yet been established in Iraq, the UBT may be recommended as first choice due to its higher performance compared to other methods.

To evaluate the potential probiotic properties ofLactobacillus strains isolated from feces of infants and also to determine their antimicrobial activity against some enteropathogenic bacteria. The Fecal samples were prepared from 120 infants aged less than 24 months. In total,105 Lactobacillus strains were identified by phenotypic tests. Thirty isolates were randomly selected to study their potential probiotic properties. These isolates were examined for resistance to acid (pH: 2.5, 2 h) and bile (oxgall 0.3%, 8 h), adhesion to HT-29 cells, antibiotic susceptibility, and antimicrobial activities. On basis of 16S rRNA sequencing, 30 isolates identified as Lactobacillus fermentum (n = 11; 36.7%), Lactobacillus plantarum (n = 9; 30%), Lactobacillus rhamnosus (n = 6; 20%), and Lactobacillus paracasei (n = 4; 13.3%). All tested strains survived at acid and bile conditions. Six Lactobacillus strains revealed high adherence to HT-29 cells. Three strains including the L. fermentum (N2, N7), and the L. plantarum (N20) showed good probiotic potential and inhibited the growth of Yersinia enterocolitica ATCC 23715, Shigella flexneri ATCC 12022, Salmonella enterica ATCC 9270, and enteropathogenic Escherichia coli (EPEC) ATCC 43887. The antibiotic resistance test showed that all the isolates were susceptible to tetracycline, and chloramphenicol. Lactobacillus strains like L. fermentum (N2, N7), and the L. plantarum (N20), could be potential probiotic, but further in vitro and in vivo studies on these probiotic strains are still required.

Peptic ulcer disease, chronic gastritis, and stomach cancer are all caused by H. pylori. The most notable drug for the treatment is the antibiotic clarithromycin, which is currently the drug of choice. H. pylori clarithromycin resistance has been associated with point mutations in 23srRNA, the most prominent of which are A2143 and A2144G. In H. pylori bacteria, methylase synthesis, macrolide-inactivating enzyme activity, and active efflux have all been found to be resistance mechanisms. The goal of the study is to determine how resistant H. pylori is to clarithromycin and what the minimum inhibitory concentration is for various antimicrobials. Furthermore, gastro-endoscopy will be performed on Iraqi patients to detect the presence of A2143G and A2144G point mutations in Helicobacter pylori infections, as diagnosed from the pyloric region and other anatomical regions. One hundred fifteen samples were collected from patients strongly suspected of H. pylori infection presented for upper gastrointestinal endoscopy at Ramadi Teaching Hospitals and Private Clinics for the period from January 2020 until February 2021. Specimens were cultured on brain heart infusion agar containing various antibiotics and were incubated at 37 °C under microaerophilic conditions. For identification of H. pylori, isolates of the biochemical tests and RT-PCR assay were applied. The Epsilometer test was used in the antibiotic susceptibility testing as dependent on the CLSI standard. The Restriction Fragment Length Polymorphism technique was used to determine point mutations. In total, 55 (47.8%) Helicobacter pylori isolates were cultured from the 115 biopsy specimens, among which 16 (29.1%), 38 (69.1%), 20 (36.4%), and 40 (72.7%) revealed some degree of resistance to levofloxacin, clarithromycin, ciprofloxacin, and metronidazole, respectively. The frequency of A2144G and A2143 point mutations were 23 (60.5%) and 19 (50%), respectively. According to our results, Helicobacter pylori showed high resistance to clarithromycin. Our results demonstrate the requirement for antibiotic susceptibility testing and molecular methods in selecting drug regimens.

Antimicrobial resistance, with the production of extended-spectrum β-lactamases (ESBL) and carbapenemases, is common in the opportunistic pathogen, Klebsiella pneumoniae. This organism has a genome that can contain clustered regularly interspaced short palindromic repeats (CRISPRs), which operate as a defense mechanism against external invaders such as plasmids and viruses. This study aims to determine the association of the CRISPR/Cas systems with antibiotic resistance in K. pneumoniae isolates from Iraqi patients. A total of 100 K. pneumoniae isolates were collected and characterized according to their susceptibility to different antimicrobial agents. The CRISPR/Cas systems were detected via PCR. The phenotypic detection of ESBLs and carbapenemases was performed. The production of ESBL was detected in 71% of the isolates. Carbapenem-resistance was detected in 15% of the isolates, while only 14% were susceptible to all antimicrobial agents. Furthermore, the bacteria were classified into multidrug (77%), extensively drug-resistant (11.0%) and pandrug-resistant (4.0%). There was an inverse association between the presence of the CRISPR/Cas systems and antibiotic resistance, as resistance was higher in the absence of the CRISPR/Cas system. Multidrug resistance in ESBL-producing and carbapenem-resistant K. pneumoniae occurred more frequently in strains negative for the CRISPR/Cas system. Thus, we conclude that genes for exogenous antibiotic resistance can be acquired in the absence of the CRISPR/Cas modules that can protect the bacteria against acquiring foreign DNA.

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology has emerged as an adaptable instrument for several uses. The CRISPR–Cas system employs Cas proteins and programmable RNA molecules to guide the recognition and cleavage of specific DNA regions, permitting accurate genome editing. It is derived from the bacterial immune system and allows for accurate and efficient modification of DNA sequences. This technique provides unparalleled gene editing, control, and precise alteration opportunities. This review aims to offer a comprehensive update of the core concepts of the CRISPR–Cas system and recent progress, while also providing an overview of the significant applications in diverse fields such as microbiology and medicine. The CRISPR–Cas9 gene editing technique has facilitated substantial advancements in comprehending gene function, simulating diseases, and creating innovative therapeutics. CRISPR-based therapeutics present a hopeful prospect for addressing intricate ailments, including genetic disorders, malignancies, and infectious diseases, as they serve as viable substitutes for conventional pharmaceuticals. In microbiology, this method serves as a diagnostic and therapeutic tool that proves highly efficient in eliminating bacteria that have developed resistance to various antibiotics. Despite its significant potential, CRISPR encounters ethical, safety, and regulatory obstacles that necessitate meticulous deliberation. Concerns regarding off-target effects, poor delivery to target tissues, and unwanted side effects emphasize the necessity to thoroughly examine the technology. It is necessary to balance the advantages and difficulties CRISPR presents. Consequently, more rigorous preclinical and clinical experiments are essential before using it in humans.