Lactamase Genes Research Articles

Prevalence of some Metallo β-lactamase enzymes genes in P. aeruginosa isolated from different clinical sample in Baghdad, Iraq

The increasing challenge of carbapenem antibiotics resistance caused by Pseudomonas aeruginosa is one of the global healthcare problems. P. aeruginosa is a significant opportunistic infection. For epidemiological reasons, identifying resistance genes is essential, and it is also desirable to quickly identify the techniques for producing carbapenemase enzymes. So, the study aims to determine the prevalence of beta-lactamase encoding genes (blaIMP, blaVIM, bla GIM, and bla SPM) phenotypically and genotypically in P. aeruginosa isolates to address the epidemiological spread of these genes in Baghdad city. The study identified P. aeruginosa isolates from various clinical sources by chemical characterization and VITEK 2 system, the antibiogram test, phenotypic screening carbapenem resistance by Combined disk synergy test (CDST), and conventional PCR were used to detect presence of VIM, IMP, SPM, and GIM genes. Bacterial susceptibility testing revealed (40%) of isolates were resistant to Imipenem and (85%) of them positive to CDST. Genotypic screening on phenotypic Metallo-β-lactamase isolates showed that (100%) isolates contained blaVIM genes, blaGIM genes (88%), whereas (65%) isolates carried blaSPM genes. In this investigation, there was no evidence of blaIMP among carbapenem-resistant isolates. The study appeared high prevalence of multi-drug resistance P. aeruginosa isolates that produce carbapenemase enzymes and having β- lactamase genes in local hospitalized patients compared to global ratio. Expanding the sample size and types of enzymes screening in MDR P. aeruginosa should be the main focus in the future research.

Epidemiological pattern and genomic insights into multidrug-resistant ST491 Acinetobacter baumannii BD20 isolated from an infected wound in Bangladesh: Concerning co-occurrence of three classes of beta lactamase genes

ObjectiveMultidrug-resistant (MDR) Acinetobacter baumannii is a major issue within healthcare facilities in Bangladesh due to its frequent association with hospital-acquired infections. In this study we report on a carbapenem-resistant draft genome sequence of an A. baumannii BD20 sample isolated from an infected wound in Bangladesh. MethodsA. baumannii BD20 was isolated from an infected burn wound. Whole-genome sequencing was carried out and annotated using PGAP and Prokka. Sequence type, antimicrobial resistance genes, virulence factor genes, and metal resistance genes were investigated. Core genome multilocus sequence typing–based phylogenomic analysis between A. baumannii BD20 and 213 A. baumannii strains retrieved from the NCBI GenBank database was performed using the BacWGSTdb 2.0 server. ResultsA. baumannii BD20 (MLST 491) was resistant to all the antibiotics tested, except for colistin and polymyxin B. Along with many other antibiotic resistance genes, the isolate harbored three classes of beta lactamase–producing genes: blaGES-11 (class A), blaOXA-69 (class D), blaADC-10 (class C), and blaADC-11 (class C). Additionally, the strain carried several virulence genes and metal resistance determinants, which may contribute to its increased virulence. Core genome MLST–based phylogenomic analysis revealed that A. baumannii BD20 was closely related to another ST491 strain isolated from Singapore. ConclusionsThe findings of this study underscore the growing challenge of MDR A. baumannii, emphasizing the need for vigilant surveillance and infection-control measures in healthcare settings in order to address these emerging threats effectively.

Prevalence, antimicrobial susceptibility profiles and resistant gene identification of bovine subclinical mastitis pathogens in Bangladesh

Subclinical mastitis (SCM), a silent threat in the dairy sector of Bangladesh poses a significant economic impact and serves as a potential source of infection for healthy cows, hindering efforts to achieve milk self-sufficiency. Despite the importance of this issue, limited research has been conducted on mastitis in Sylhet region of Bangladesh. This study aimed to investigate the molecular prevalence, antimicrobial susceptibility profile and resistant genes detection on pathogens (Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae) causing SCM. In a cross-sectional study utilizing convenience sampling, 325 milk samples were collected from apparently healthy dairy cows. Initially, SCM was detected using the modified Whiteside test (MWST) method. Suspected positive samples were then subjected to bacteriological culture and standard biochemical assays, followed by molecular identification through polymerase chain reaction (PCR). Finally, antimicrobial susceptibility testing was conducted on all PCR-positive samples using the Kirby-Bauer disk diffusion method on Mueller-Hinton agar. In the Sylhet district, the prevalence of SCM was 64.92 % (211 out of 325) at the individual animal level and 82.69 % (43 out of 52) at the herd level. Among the SCM-positive animals, S.aureus was found in 31.75 % (67 out of 211) of cases, E.coli in 81.99 % (173 out of 211), and K.pneumoniae in 66.82 % (141 out of 211). K.pneumoniae had the highest prevalence at 60 % in Zakiganj, S.aureus at 45 % in Zakiganj, and E.coli at 72 % in Bishwanath Upazila. Extended spectrum beta lactamase (ESBL) genes blaTEM, blaOXA, blaCTX-M1,blaCTX-M2,MultiCaseDHA were identified. Additionally, antibiotic resistance genes tet(A), AAC(3)-iv, and Sul1 were also detected. The pathogens exhibited resistance to Penicillins (ampicillin, amoxicillin), Cephems (cefuroxime, ceftazidime), and Tetracyclines (tetracycline). However, all three bacteria were highly sensitive to meropenem, amikacin, gentamicin, ciprofloxacin, and sulfamethoxazole-trimethoprim. These findings highlight the critical need for targeted antimicrobial stewardship and effective control measures to mitigate the impact of SCM on dairy production and animal health in the Sylhet region of Bangladesh.

Molecular characterization of extended spectrum beta lactamase (ESβL) and quinolone (QNR) resistance genes in Escherichia Coli isolated from cloaca swabs of wild birds and poultry droppings in Jos Metropolis

Molecular characterization of extended spectrum beta lactamase (ESβL) and quinolone (QNR) resistance genes in Escherichia Coli isolated from cloaca swabs of wild birds and poultry droppings in Jos Metropolis

Ceftazidime-Avibactam resistance in clinical isolates of carbapenem-resistant Klebsiella pneumoniae: A phenotypic and genotypic analysis

ObjectiveTo find the prevalence of Ceftazidime-Avibactam (CAZ-AVI) resistant Klebsiella pneumoniae in clinical isolates and to determine the genes responsible for Ceftazidime-Avibactam resistance using PCR. MethodsA total of 89 carbapenem resistant Klebsiella pneumoniae from various clinical samples were included in the study. CAZ-AVI resistance was tested using E-test. CAZ-AVI resistant strains were subjected to conventional PCR for detection of carbapenamase genes blaNDM- 1, blaOXA-48, blaVIM, blaIMP, blaKPC. ResultsOf the 89 isolates screened for CAZ-AVI resistance, 45(50.5%) isolates were found to be resistant. 42 isolates were subjected to PCR for detection of β lactamase genes.34 isolates were positive for blaNDM-1 and all 42 isolates were positive for blaOXA-48. Co-expression of NDM-1 and OXA-48 was seen in 34 isolates. Sensitivity of mCIM test to identify a carbapenamse compared to PCR was 61.9%. Sensitivity of eCIM test to identify NDM-1 was 80%. ConclusionCAZ-AVI was effective in vitro in 49.4% of the isolates. Indicating that CAZ-AVI is a promising addition to antibiotics against CRE as well as a carbapenem sparing drug in ESBL producing organisms. β-Lactamase-related mutations are the main mechanism leading to CAZ-AVI resistance.

Simultaneous detection of phylogroups and ESBL genes in E. coli using Multiplex PCR

Background: Escherichia coli (E. coli) strains harbor various allelic versions of beta lactamase genes and their identification using conventional phenotypic tests is a tedious and time consuming task. In the present study, multiplex PCR is performed for the simulatanoues detection of E. coli phylogroups and extended-spectrum β-lactamase- (ESBL) genes.Methods: A total of 128 E. coli isolates from urine samples were screened for antibiotic resistance and expression of ESBL activity using phenotypic and genotypic methods. Uniplex and multiplex PCRs were used to detect E. coli phylogroup detrminants and blaCTX-M-15, blaOXA-1 and TEM1 genes. Chi Square test of independence was employed for evaluating significant levels at P value < 0.05. Results: Phylogroup B2 was detected as the predominant group (36%) followed by group D (30%), A (25%) and B1 (9%). The highest resistance was seen against nalidixic acid (100%) and the lowest against amoxicillin-clavulanic acid (55%). Significant P values were observed for resistance against cefotaxime and cefepime in the phylogroup B2 while resistance against cefoxitin, sulfamethoxazole and fosfomycin was significantly associated with group D. Combination disc diffusion test (CDDT) showed ESBL activity in 42% E. coli isolates. A significant association of blaCTX-M-15 gene was observed for phylogroup B2 (P = 0.007). Moreover, a combination genotype of blaCTX-M-15 and TEM1 was also found statistically prevalent in phylogroup B2 (P = 0.006).Conclusion: The study highlights the alarming rise in antibiotic resistance and delineates B2 a predominant phylogoup with a high prevalence of blaCTX-M-15 and TEM1 genes in urinary E. coli isolates.Keywords: UTI; E. Coli; Phylogenetic groups; ESBL genes; Multiplex PCR

Chromosomal and plasmid-encoded virulence and multidrug resistance of Escherichia coli ST58/24 infecting a 2-year-old sickle cell patient with sepsis in Kampala Uganda, East Africa

Sepsis and drug resistance represent a complex of the most common global causes of mortality in intensive care units (ICUs) especially among patients with comorbidities. Extraintestinal pathogenic Escherichia coli (ExPEC) strains are highly implicated in systemic infections, with multidrug resistance exacerbating the risk of chronic conditions and patient mortality. The diversity of virulence and evolution of multidrug resistance are yet to be fully deciphered. In this work, we aimed at unveiling the pathogens and their genomic determinants of virulence and drug resistance relevant to increased sepsis in a sickle cell child admitted to ICU. From a rectal swab, we isolated a strain of E. coli from the patient and phenotypically tested it against a panel of selected beta lactams, fluoroquinolones, macrolides, aminoglycosides and colistin. We then sequenced the entire genome and integrated multiple bioinformatic pipelines to divulge the virulence and multidrug resistance profiles of the isolate. Our results revealed that the isolate belongs to the sequence type (ST) 58/24, which (ST58), is a known ExPEC. With the use of PathogenFinder, we were able to confirm that this isolate is a human pathogen (p = 0.936). The assembled chromosome and two plasmids encode virulence factors related to capsule (antiphagocytosis), serum survival and resistance, type 6 secretion system (T6SS), multiple siderophores (iron acquisition), and biosynthetic gene clusters for polyketides and nonribosomal peptides exhibiting host cell damaging activity in silico. The genome also harbors multidrug resistance genotypes including extended spectrum beta lactamase (ESBL) genes such as blaTEM-1A/B, sulfonamide resistance genes sul1/2, fluoroquinolone resistance genes dfrA5 and nonsynonymous mutations of the gene pmrB, conferring intrinsic colistin resistance. Conclusively, this pathogen holds the potential to cause systemic infection and might exacerbate sickle cell anemia in the patient. The virulence and multidrug resistance profiles are encoded by both the chromosome and plasmids. Genomic surveillance of pathogens with multidrug resistance among patients with comorbidities is crucial for effective disease management.

Extended Spectrum Beta Lactamase Escherichia Coli in Bagmati River, Kathmandu Valley.

Antimicrobial resistance organisms in the peripheral communities of an environment can be predicted by the presence of extended-spectrum beta-lactamase Escherichia coli in that environment. The close connectivity between humans and water sources can facilitate the entry of antimicrobial resistant organisms into the human ecosystem. The aim of this study was to assess beta lactamase producing Escherichia coli from Bagmati river within Kathmandu valley. In the year 2020, a cross-sectional study was conducted on water samples collected from 66 locations along the Bagmati River. Coliforms were isolated by five tubes dilution method and identified by cultural and biochemical tests. Further Escherichia coli was isolated in eosin methylene blue agar at 44.5 ⁰C. Antibiotic susceptibility test was performed by Kirby Bauer disk diffusion methods. Beta lactamase gene types were detected by using conventional multiplex polymerase chain reaction. A total of 615 bacterial isolates were identified among which 39 % (n=241) were Escherichia coli. Extended spectrum beta lactamase producing Escherichia coli was confirmed in 16.6 % (40/241) of total Escherichia coli isolates. Among 66 sites this isolate was detected in 26 (40 %) sampling sites excluding upstream regions. All the Escherichia coli isolates were multidrug resistance showing higher percentage (>99 %) of resistant for penicillin, tetracycline and erythromycin antibiotics. There were significant differences in resistance rate for cefotaxime and ceftazidime by extended spectrum beta lactamase producing and non-producing Escherichia coli (p<0.05). Presence of multidrug resistance extended spectrum beta lactamase producing Escherichia coli in river streams suggests the chances of circulating within river system and hence transmitting in human community. Bagmati river; drug resistance; escherichia coli; human.

Identification of Extended Spectrum Beta Lactamases Producing Pseudomonas aeruginosa from Subclinical Mastitis Milk Samples in an Organized Dairy Farm of SVVU, Andhra Pradesh

The present study was conducted during August 2021 at Buffalo Research Station, Venkatarammanagudem of West Godavari District, Andhra Pradesh, India and was aimed to detect the presence of extended-spectrum beta-lactamases (ESBL) producing P. aeruginosa from sub-clinical mastitis that are apparently healthy Murrah buffaloes. Inflammation of the udder by microbial infection is one of the leading economic disease in the dairy sector. Most of the pathogens especially Pseudomonas aeruginosa are refractory to the antibiotic theraphy. Nowadays, the antimicrobial resistance owed by the bacteria is at an alarming rate. Pseudomonas aeruginosa, a multi drug resistant pathogen carry the extended spectrum beta lactamase genes in its plasmids and thus become resistant to antibiotic therapy. A total of 276 milk samples were collected from 69 milch animals in the month of August, 2021 comprising from all the four quarters of each buffalo. Nine (n=9) (3.26%) P. aeruginosa isolates were identified from the sub-clinical mastitis milk samples from the total 276 samples collected in the study. The isolates on morphological analysis were typical to P. aeruginosa. Seven isolates were positive for phenotypical beta-lactamases production. A total of 1 and 2 isolates were found reactive of the blaSHV and blaOXA genes, and two isolates harboured both blaSHV and blaOXA genes, respectively on PCR assay. No blaTEM gene was found in the isolates. Antibiogram of the ESBL producing P. aeruginosa isolates (n=7) possessed 100% resistance against most of the commonly used antimicrobials like ampicillin, amoxycillin, clindamycin, co-trimoxazole oxytetracycline, streptomycin and ceftriaxone (85%). The P. aeruginosa isolates were sensitive to amikacin (100%), ciprofloxacin (87.4%), gentamicin (84%), norfloxacin (84.8%) and enrofloxacin (82%).

Extensively Drug-Resistant Klebsiella pneumoniae Associated with Complicated Urinary Tract Infection in Northern India.

Klebsiella pneumoniae (Kp), which is associated with hospital-acquired infections, is extensively drug-resistant (XDR), making treatment difficult. Understanding the genetic epidemiology of XDR-Kp can help determine its potential to be hypervirulent (hv) through the presence of siderophores. We characterized the genomes of 18 colistin-resistant XDR-Kp isolated from 14 patients with complicated tract infection at an Indian healthcare facility. The 18 organisms comprised the following sequence types (STs): ST14 (n = 9), ST147 (n = 5), ST231 (n = 2), ST2096 (n = 1), and ST25 (n = 1). Many patients in each ward were infected with the same ST, suggesting a common source of infection. Some patients had recurrent infections with multiple STs circulating in the ward, providing evidence of hospital transmission. β-lactamase genes (blaCTX-M-1, blaSHV, and blaampH) were present in all isolates. blaNDM-1 was present in 15 isolates, blaOXA-1 in 16 isolates, blaTEM-1D in 13 isolates, and blaOXA-48 in 3 isolates. Disruption of mgrB by various insertion sequences was responsible for colistin resistance in 6 isolates. The most common K-type among isolates was K2 (n = 10). One XDR convergent hvKp ST2096 mutation (iuc+ybt+blaOXA-1+blaOXA-48) was associated with prolonged hospitalization. Convergent XDR-hvKp has outbreak potential, warranting effective antimicrobial stewardship and infection control.

Genomic landscape of NDM-1 producing multidrug-resistant Providencia stuartii causing burn wound infections in Bangladesh

The increasing antimicrobial resistance in Providencia stuartii (P. stuartii) worldwide, particularly concerning for immunocompromised and burn patients, has raised concern in Bangladesh, where the significance of this infectious opportunistic pathogen had been previously overlooked, prompting a need for investigation. The two strains of P. stuartii (P. stuartii SHNIBPS63 and P. stuartii SHNIBPS71) isolated from wound swab of two critically injured burn patients were found to be multidrug-resistant and P. stuartii SHNIBPS63 showed resistance to all the 22 antibiotics tested as well as revealed the co-existence of blaVEB-6 (Class A), blaNDM-1 (Class B), blaOXA-10 (Class D) beta lactamase genes. Complete resistance to carbapenems through the production of NDM-1, is indicative of an alarming situation as carbapenems are considered to be the last line antibiotic to combat this pathogen. Both isolates displayed strong biofilm-forming abilities and exhibited resistance to copper, zinc, and iron, in addition to carrying multiple genes associated with metal resistance and the formation of biofilms. The study also encompassed a pangenome analysis utilizing a dataset of eighty-six publicly available P. stuartii genomes (n = 86), revealing evidence of an open or expanding pangenome for P. stuartii. Also, an extensive genome-wide analysis of all the P. stuartii genomes revealed a concerning global prevalence of diverse antimicrobial resistance genes, with a particular alarm raised over the abundance of carbapenem resistance gene blaNDM-1. Additionally, this study highlighted the notable genetic diversity within P. stuartii, significant informations about phylogenomic relationships and ancestry, as well as potential for cross-species transmission, raising important implications for public health and microbial adaptation across different environments.

Molecular Detection of Beta Lactamase Genes from Salmonella Typhimurium Isolated from Poultry Droppings in Nyanya, Abuja

Salmonella infections are one of the major global public health problems. During the last decade, antibiotic resistance and multiresistance of S. typhimurium. This study aimed at molecular detection of Beta Lactamase genes from S. typhimurium Isolated from poultry droppings in Nyanya, Abuja. Fecal droppings samples were collected from two different Poultry farms using sterile screw tubes. S. Typhimurium was isolated and identified using standard microbiology methods. Antimicrobial susceptibility was carried out using Clinical and Laboratory Standards Institute method. Out of 180 samples collected the overall occurrence of S. typhimurium was 20 (11.1%). S. typhimurium isolated were higher in Farm A 9(10.0%) than in farm B 11(12.2%). The S. typhimurium isolates from Farm A were highly resistant to Ampicillin with 77.7% but less resistant to gentamicin with 33.3%, ofloxacin, ciprofloxacin and Sulphamethoxazole/trimethoprim with 44.4%. Isolates from Farm B were highly resistant to Ampicillin and Sulphamethoxazole/trimethoprim 81.8%, but less resistant to Cefuroxime 36.3%. the most common phenotypic resistant observed were C-CIP-CAZ-CRO-AUG- OFL -SXT- CEF (22.2%) from Farm A. CIP-CRO-CAZ- AMP -AUG- OFL -SXT- CEF (18.1%) from Farm B, The commonest MAR index observed in this study was 0.8(33.3%) in farm A and (36.3%) in farm B. The order of occurrences of the 4 Beta Lactamase genes detected was: blaCTX-M-9 and blaSHV (75.0%) &gt; bla CMY2 and Bla CTX-M (50.0%). The resistance observed in this study indicates it will be difficult to use commonly use antibiotics to treat infection caused by S. typhimurium isolated in the study location

Detection Sequencing NDM and blaOXA Genes in Metallo-$\beta$-Lactamase Producing Klebsiella Pneumoniae

The prevalence of Carbapenem-resistance in Enterobacteriaceae is rising worldwide and poses a treatment challenge for healthcare facilities. This study aims to detect a sequence of New Delhi metallo- -lactamase (NDM) and bla OXA-48 gene in metalo-beta-lactamase-producing Klebsiella pneumoniae isolated from a few hospitals in the city of Baghdad. 74 Klebsiella pneumoniae were isolated from 320 clinical samples that were routinely submitted to the diagnostic laboratory of the chosen hospital. These samples included sputum, tissue swabs, folly tips, urine, ear swabs, bloodstream, pus, wound swabs, endotracheal tubes, and body fluids. In this study, 74 isolates were examined for expression of carbapenemase by phenotyping method. The modified Hodge Test (MHT) is utilized for phenotypic detection. MHT was positive for 24 carbapenem-resistant Klebsiella pneumoniae. Genotyping was accomplished by using polymerase chain reaction (PCR) to amplify target genes by employing particular primers. The NDM gene was detected in 19/24 isolates, and the blaOXA-48 gene was detected in 5/24 isolates. The presence of bla NDM and the dangerously high frequency of MBL in the Klebsiella pneumoniae strain are both concerning, and it has been demonstrated to be challenging to treat and inhibit infection.

Molecular prevalence of MecA and Blaz Genes with phenotypic analysis of Antibiotic Sensitivity Pattern for S.aureus Isolated From Dermal lesions of Sheep Breeders In Diyala Governorate – Iraq

Background: S. aureus is one of the dominant bacterial pathogens among dermal infections in human and animals, which have resistance to different antimicrobial drugs.&#x0D; Objective: Isolation and identification of S.aureus from skin lesions among sheep breeders by traditional methods, Vitek 2 system and PCR using Staur 4, 6 specific pri-mers for validation, Detection of genes (methicillin resistant (mecA), Beta- lac-tamase gene (blaZ) by conventional PCR ,determine antibiotic sensitivity pat-tern by kirby bauer disc diffusion method.&#x0D; Patients and Methods: A total of 44 swaps were collected from sheep breeders suffered from variety of infected skin lesions to detect methicillin sensitive and resistant s. aureus by employing traditional methods in addition to confirmatory techniques through fast rapid VETEK2 system , detection of genes (methicillin resistant (mecA), Be-ta- lactamase gene (blaZ) by conventional PCR , and determine antibiotic sensi-tivity pattern by kirby bauer disc diffusion method.&#x0D; Results: S.aureus was isolated from 15/44,( 34.09%) of skin lesion of sheep breeders . A total of 6/15 ,(40%) were methicillin resistant S.aureus (MRSA) ,which represent ( 13.63%) from total samples .Beta lactamase gene primers was detected in all S.aureus isolates. A 6/15,(40%) of S.aureus have resistance for members of antibiotics classes, penicillins, polypeptides, fluoroquinolones, macrolide which include the following : methicillin that was confirmed early by detection of me-cA gene , levofloxacin , ofloxacin , erythromycin and vancomycin. A 6/15,(40%) of S.aureus have multidrug resistant trait for Penicillins, Polypep-tides, Fluoroquinolones, Macrolide antibiotics. Non multidrug resistant S.aureus was reported for Penicillins (oxacillin,4/15, 26%) and Polypeptides antibiotics (vancomycin 8/15, 53.33%) of S.aureus . Absolute sensitivity was reported for gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol.&#x0D; Conclusion: Methicillin sensitive S.aureus was more common compared with MRSA isolated from dermal infections of sheep breeders. Blaz gene was predominantly ex-pressed by S.aureus isolates followed by Mec A gene. S.aureus have resistance for penicillins, polypeptides, fluoroquinolones, macrolide which include the fol-lowing : methicillin, levofloxacin , ofloxacin , erythromycin and vancomycin . S.aureus have multidrug resistant trait for Penicillins, Polypeptides, Fluoroquin-olones, Macrolide antibiotics. S.aureus was absolutely sensitive to gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol.

Molecular prevalence of MecA and Blaz Genes with phenotypic analysis of Antibiotic Sensitivity Pattern for S.aureus Isolated From Dermal lesions of Sheep Breeders In Diyala Governorate – Iraq

Background: S. aureus is one of the dominant bacterial pathogens among dermal infections in human and animals, which have resistance to different antimicrobial drugs. Objective: Isolation and identification of S.aureus from skin lesions among sheep breeders by traditional methods, Vitek 2 system and PCR using Staur 4, 6 specific pri-mers for validation, Detection of genes (methicillin resistant (mecA), Beta- lac-tamase gene (blaZ) by conventional PCR ,determine antibiotic sensitivity pat-tern by kirby bauer disc diffusion method. Patients and Methods: A total of 44 swaps were collected from sheep breeders suffered from variety of infected skin lesions to detect methicillin sensitive and resistant s. aureus by employing traditional methods in addition to confirmatory techniques through fast rapid VETEK2 system , detection of genes (methicillin resistant (mecA), Be-ta- lactamase gene (blaZ) by conventional PCR , and determine antibiotic sensi-tivity pattern by kirby bauer disc diffusion method. Results: S.aureus was isolated from 15/44,( 34.09%) of skin lesion of sheep breeders . A total of 6/15 ,(40%) were methicillin resistant S.aureus (MRSA) ,which represent ( 13.63%) from total samples .Beta lactamase gene primers was detected in all S.aureus isolates. A 6/15,(40%) of S.aureus have resistance for members of antibiotics classes, penicillins, polypeptides, fluoroquinolones, macrolide which include the following : methicillin that was confirmed early by detection of me-cA gene , levofloxacin , ofloxacin , erythromycin and vancomycin. A 6/15,(40%) of S.aureus have multidrug resistant trait for Penicillins, Polypep-tides, Fluoroquinolones, Macrolide antibiotics. Non multidrug resistant S.aureus was reported for Penicillins (oxacillin,4/15, 26%) and Polypeptides antibiotics (vancomycin 8/15, 53.33%) of S.aureus . Absolute sensitivity was reported for gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol. Conclusion: Methicillin sensitive S.aureus was more common compared with MRSA isolated from dermal infections of sheep breeders. Blaz gene was predominantly ex-pressed by S.aureus isolates followed by Mec A gene. S.aureus have resistance for penicillins, polypeptides, fluoroquinolones, macrolide which include the fol-lowing : methicillin, levofloxacin , ofloxacin , erythromycin and vancomycin . S.aureus have multidrug resistant trait for Penicillins, Polypeptides, Fluoroquin-olones, Macrolide antibiotics. S.aureus was absolutely sensitive to gentamycin, tetracycline, rifampicin, imipenem and chloramphenicol.

Abundance and prevalence of ESBL coding genes in patients undergoing first line eradication therapy for Helicobacter pylori.

The spread of extended-spectrum beta-lactamases (ESBLs) in nosocomial and community-acquired enterobacteria is an important challenge for clinicians due to the limited therapeutic options for infections that are caused by these organisms. Here, we developed a panel of ESBL coding genes, evaluated the abundance and prevalence of ESBL encoding genes in patients undergoing H. pylori eradication therapy, and summarized the effects of eradication therapy on functional profiles of the gut microbiome. To assess the repertoire of known beta lactamase (BL) genes, they were divided into clusters according to their evolutionary relation. Primers were designed for amplification of cluster marker regions, and the efficiency of this amplification panel was assessed in 120 fecal samples acquired from 60 patients undergoing H. pylori eradication therapy. In addition, fecal samples from an additional 30 patients were used to validate the detection efficiency of the developed ESBL panel. The presence for majority of targeted clusters was confirmed by NGS of amplification products. Metagenomic sequencing revealed that the abundance of ESBL genes within the pool of microorganisms was very low. The global relative abundances of the ESBL-coding gene clusters did not differ significantly among treatment states. However, at the level of each cluster, classical ESBL producers such as Klebsiella sp. for blaOXY (p = 0.0076), Acinetobacter sp. for blaADC (p = 0.02297) and others, differed significantly with a tendency to decrease compared to the pre- and post-eradication states. Only 13 clusters were common across all three datasets, suggesting a patient-specific distribution profile of ESBL-coding genes. The number of AMR genes detected in the post-eradication state was higher than that in the pre-eradication state, which could be attributed, at least in part, to the therapy. This study demonstrated that the ESBL screening panel was effective in targeting ESBL-coding gene clusters from bacterial DNA and that minor differences exist in the abundance and prevalence of ESBL-coding gene levels before and after eradication therapy.

Coexistence Of Blaoxa-48, Blavim, And Blashv Genes In Klebsiella Pneumoniae And Escherichia Coli Isolated From Urinary Tract Infections: Microbiological And Epidemiological Analysis.

To investigate antimicrobial resistance mechanisms of isolated bacterialstrains, and their correlation with virulence profile. The cross-sectional study was conducted in January 2020 at outpatient health centres in Kafrelsheikh Governorate of Egypt, and comprised urine samples from patients regardless of age and gender. Midstream samples were collected into sterile swaps which were kept in ice-cooled boxes until transported to the laboratory within 5h. Antimicrobial resistance profile of the isolated Enterobacteriaceae was done using Kirby-Bauer disk diffusion method and was confirmed withVitek compact 2. The phenotypic of carbapenemases and extended-spectrum beta lactamase was determined, and polymerase chain reaction was used, as appropriate. Data was analysed using SPSS 20. Of the 199 patients, 101(50.7%) were females and 98(49.3%) were males. The majority 73(36.6%) were aged 30-50 years. Urinary tract infection was found in 68(34.2%) patients. In 28(41.2%) of these patients, there were 32 isolates of Enterobacterales; 21(65.62%) Klebsiella pneumoniae, 7(21.87%) Escherichia coli and 4(12.5%) Enterobacter cloacae. Of the 28(41.2%) patients, 24(85.7%) were infected with a single strain; 17(70.8%) Klebsiella pneumoniae, 4(16.7%) Escherichia coli and 3(12.5%) Enterobacter cloacae. In 3(10.7%) cases, there was co-infection with Escherichia coli and Klebsiella pneumoniae, and 1(3.6%)sample had mixed infection with Klebsiella pneumoniae and Enterobacter cloacae. The other 40(58.8%) patients had other causative agents. Housewives, agricultural workers and those aged >50 years had a higher risk of urinary tract infections(p<0.05) Among Klebsiella pneumonia isolates, 6(28.5%) possessed carbapenemase-related genes and 4(19.1%) extended-spectrum beta lactamase-related genes. The carbapenemase related genes were bla-Verona integron-encoded metallo beta lactamase 6(100%) bla-New Delhi metallo beta lactamase-1 4(66.6%) and bla-oxacillinase-48 2(33.3%). The 4(19.1%) cases of extended-spectrum beta lactamase related genes had bla-temoneira gene 3(75%) and bla-sulfhydryl variable gene 4(100%). In Escherichia coli isolates, bla-oxacillinase-48 and bla-Cefotaximase genes were observed in 2(28.5%) cases.Virulence genes uridine diphosphate glucose 4-epimerase, fimbrial adhesion and mannose-resistance adhesin of Klebsiella spp genes in Klebsiella pneumoniae isolates were positive in in 16(76.2%), 14(66.7%) and 10(47.6%) cases, respectively. All 21(100%) isolates of Klebsiella pneumoniae were negative for mucoviscosity-associated gene A. There was evidence of the coexistence of bla- oxacillinase-48, bla-Verona integron-encoded metallo beta lactamase and bla-sulfhydryl variable genes in Klebsiella pneumoniae and Escherichia coli isolates from mixed urinary tract infection samples.

Phylogenetic analysis of zoonotic Acinetobacter spp. isolated from Geoffroy's bat (Myotis emarginatus), Northern Iraq

The current study was conducted to isolate and evaluate the phylogenetic status of the zoonotic Acinetobacter spp. from the Geoffroy's bat (Myotis emarginatus). The sample collection included 35 bats captured in some caves in Northern Iraq. Intestine parts and swabs were taken from each bat. The specimens were subjected to bacterial cultivation processes and 16S rRNA gene and beta lactamase (bla TEM-1) dependent polymerase chain reaction (PCR) methods to detect the genus level and the virulence activity, respectively. The phylogenetic tree was built utilizing the 16S rRNA gene. The findings of the bacterial cultivation revealed the presence of the bacterium in 23 (65.7%) of the collected bats; however, the 16S rRNA-PCR showed that only 10 (28.57%) of the bats demonstrated the incidence of this microorganism in their intestines. The bla TEM-1-PCR reported that 4 (40%) isolates of the 16S rRNA-PCR positive bats carried the β-lactamase gene in their genetic materials. The phylogenetic tree showed that the genetic similarity of the current study isolates was closely related to those from Egypt and China. The present data show that Acinetobacter spp. is present in the intestine of the Geoffroy's bat (Myotis emarginatus) located in some caves from Northern Iraq, and some isolates have virulence potential represented by the composition of the beta lactamase gene.

Resistance genes and plasmid profile of bacteria Isolated from selected water bodies in Owo Metropolis

Aquatic samples from three water bodies were collected following laid down procedures. The bacteria were isolated using, Eosin Methylene Blue agar (EMB), Mannitol salt agar (MSA), Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS) Agar, Muller Hinton (MH). The isolates were subjected to colonial, morphological and biochemical characterization. Pathogenicity test was carried out on the isolates using blood agar to determine their hemolytic activities. Antibiotic resistance status was also investigated against standard antibiotics using the Kirby Bauer disk diffusion method and the multiple antibiotic resistance indices of the isolates were also determined. The plasmid profiles were determined and the resistance genes were identified using molecular means. The bacteria species isolated were Veillonella Spp, Vibrio orientalis and Micrococcus luteus. Veillonella Spp showed Beta hemolysis while Micrococcus luteus showed alpha hemolysis and Vibrio orientalis showed gamma hemolysis. Antibiotic susceptibility test showed all isolates to be antibiotic resistant to varying degrees and multiple antibiotic resistance indices (MARi) depending on their level of resistance. Veillonella. spp. had MARi of 0.3 while M. luteus and V. orietalis had MARi of 0.5. Plasmid profiling showed all isolates to have plasmids with bandwidth of 10,000 bp. Vibrio spp and Veillonella Spp were positive for beta lactamase genes (bla SHV). Only Vibrio was positive for resistance to aminoglycosides (aac(3)-IV). Vibrio orientalis and Micrococcus luteus were found to have the resistance genes for quinolones (qnrA gene) while all the isolates possess genes against the sulphonamides (sul1 gene) used. The result of this work establishes the antibiotic resistance status of bacteria isolated from aquatic habitat and suggests their important role in the spread of antimicrobial resistance AMR, and antibiotic resistance genes (ARGs)

PREVALENCE OF MULTIDRUG RESISTANT ESCHERICHIA COLI AND KLEBSIELLA PNEUMONIAE SPECIES ISOLATED FROM URINE SAMPLES OF PATIENTS AT ALEX EKWUEME FEDERAL UNIVERSITY TEACHING HOSPITAL ABAKALIKI, EBONYI STATE, NIGERIA

Aim and Objective: In hospitals and community, morbidity and mortality are attributed to urinary tract infections (UTIs). This study investigated the prevalence of multi-drug resistant isolates of Escherichia coli and Klebsiella species from urine samples of patients visiting Alex Ekwueme Federal University Teaching Hospital Abakaliki (AE-FUTHA), Ebonyi State. Method: With the use of standard microbiological and biochemical techniques for analysis, a total of 300 mid-stream samples of urine were collected in sterile bottles. Phynotype screening for extended spectrum beta lactamase (ESBL) production was achieved by double disc synergy test (DDST). Disc diffusion method was used to check for ESBL producing bacteria susceptible to antibiotics. With specific primers, the presence of temoniera (TEM) and sulfhydryl variable (SHV) beta lactamases genes was determined using polymerase chain reaction (PCR). Results: 88 isolates of E coli and Klebsiella species were isolated from the urine samples; 29 were E coli and 59 were K. pneumonia species. 47 ESBL positive isolates were identified with E. coli, 19 (40%) and Klebsiella, 28 (60%). Escherichia coli isolated from GOPD patients, gynecology, and men's surgery ward were 100% resistant to ofloxacin, ceftazidime, amoxicillin, nitrofurantoin, chloramphenicol, and aztreonam, while Klebsiella species isolated from gynecology department, maternity ward, and psychiatric ward were 100% resistant to ofloxacin, Nitrofurantoin and chloramphenicol. Multiple antibiotics resistance index (MARI) of E. coli and Klebsiella species isolated recorded an average of 0.64 for E. coli and 0.41 for Klebsiella species.The molecular analysis revealed that 44.4% of SHV beta lactamase and 65.6% TEM-type β-lactamase genes were present in ESBL producing E. coli while 54.5% of SHV beta lactamase gene and 45.5% TEM-type β-lactamase genes were present in ESBL producing Klebsiella species. Conlusion: In conclusion, TEM and SHV-type β-lactamase genes are the primary cause of β-lactam antibiotic resistance in E. Coli and Klebsiella species resulting in increased infections caused by organisms harboring the ESBL gene. Nitrofurantoin, ceftazidime, cefalexin, and cefotaxime may be the antibiotic of choice in the treatment of UTIs. Peer Review History: Received: 1 January 2023; Revised: 14 February; Accepted: 7 March 2023, Available online: 15 March 2023 Academic Editor: Dr. Amany Mohamed Alboghdadly, Princess Nourah bint abdulrahman university, Riyadh, amalbgadley@pnu.edu.sa Received file: Reviewer's Comments: Average Peer review marks at initial stage: 5.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Prof. Hassan A.H. Al-Shamahy, Sana'a University, Yemen, shmahe@yemen.net.ye Dr. Tamer Elhabibi, Suez Canal University, Egypt, tamer_hassan@pharm.suez.edu.eg Similar Articles: ANALYSIS OF THE ANTIBIOGRAM PROFILES OF BIOFILM FORMING STAPHYLOCOCCUS AUREUS AND ESCHERICHIA COLI