SHV Genes Research Articles

Distribution and Molecular Characterization of Antibiotic-Resistant Pseudomonas aeruginosa in Hospital Settings of Sulaymaniyah, Iraq.

Pseudomonas aeruginosa is a significant pathogen in hospital settings, notorious for its role in hospital-acquired infections and its ability to develop resistance to multiple antibiotics. This study investigates the prevalence, distribution, and antibiotic resistance gene profiles of P. aeruginosa in seven hospitals in Sulaymaniyah City. A total of 300 samples were collected from various hospital surfaces including mops, sinks, medical equipment, beds, desks, and floors. Using bacteriological, biochemical, and molecular methods, 66 isolates were confirmed as Pseudomonas species, with 26 identified as P. aeruginosa. Antibiotic susceptibility testing revealed resistance rates of 23.3% to streptomycin, 13.6% to tobramycin, 22.7% to moxifloxacin, 21.2% to levofloxacin, and 22.7% to norfloxacin. Furthermore, the antibiotic resistance gene detection showed the presence of the bla CTX-M, bla SHV, qnrB, and bla ACC-1 genes among the isolates. The study highlights a 22% contamination rate of hospital surfaces with Pseudomonas species, emphasizing the urgent need for enhanced infection control measures and targeted antimicrobial stewardship to manage and reduce the spread of multidrug-resistant P. aeruginosa.

A Study on Extended Spectrum β-Lactamase Producing (ESBL) and Fluoroquinolone-Resistant Escherichia coli in the Japanese Quails

Antimicrobial resistance (AMR) is a growing public health threat worldwide, and members of the Enterobacteriaceae family are among the clinically important bacteria rapidly developing resistance to available antibacterial agents. The development of extended-spectrum β-lactamase (ESBL) enzymes is the primary cause of resistance to third-generation cephalosporins among Enterobacteriaceae. Quails also represent a source of animal protein deficiency in developing countries including India. This study aimed to detect the presence of extended-spectrum β-lactamases (ESBLs) and fluoroquinolone-resistant Escherichia coli from fecal samples of quails in Puducherry. A total of 36 fecal samples of quails from six different retail outlets were collected. E. coli could be isolated from 27 fecal samples and confirmed as E. coli by polymerase chain reaction by targeting the uspA gene. The isolates were preliminarily screened for ESBL production by indicator antimicrobial disc and phenotypic ESBL production was confirmed by combination disc method. The isolates were subjected to genotypic detection of ESBL (bla CTX-M, bla TEM, and bla SHV) by PCR. Out of 27 isolates, 16 isolates were positive for the presence of ESBL-producing genes, in which, 9 harbored bla CTX-M, 2 harbored bla TEM, and 5 harboured bla SHV genes. Similarly, out of 27 isolates 13 isolates were found positive for fluoroquinolone resistance in which 13 (48.18%) isolates harbored the qnrS gene, and 11 (40.74%) isolates harbored the qnrB gene. Antimicrobial susceptibility test showed that the isolates exhibited a high level of resistance against Cefpodoxime (90%), Amoxycillin/clavulanic acid (70%), Ceftriaxone (60%), Cefotaxime (55%), Ceftazidime (45%), Ciprofloxacin (40%) and Aztreonam (30%).

Distribution of TEM and SHV Genes in ESBL-Producing Klebsiella pneumoniae Strains Isolated From Various Clinical Samples in Al-Basrah Province, Iraq

Klebsiella pneumonia is an opportunistic pathogen causes several diseases including sepsis, pneumonia, and wound infections. Antimicrobial resistance (AMR) in MDR K.pneumonia carrying genes for extended-spectrum β-lactams (ESBLs) has been considered a risk for health problems around the world, which has greatly limited treatment options. The current study relied on n=26 Klebsiella pneumoniae isolates that were previously isolated by the studies of master’s student “Intithar Mahdi Mozan" between October 2022 and December 2022,the samples were collected from three hospitals in Al-Basrah province,Iraq. K. pneumoniae isolates reactivated and re-identified by cultivating isolates on MacConkey agar and Klebsiella pneumoniae chromogenic media and then re-identified K. pneumoniae isolates through diagnostic gene 16S rDNA by PCR technique, the results revealed that all n= 26 isolates of K. pneumoniae had a molecular weight of approximately 130 bp at a 100%. Double disk approximation test (DAM) was used to detected produced extended-spectrum β-lactams (ESBLs), the result of current study was showed positive results for the ESBLs. Extended-spectrum β-lactamses (ESBLs) genes was detected by using three specific primers: blaTEM, blaSHV and, blaCTX-M. Out of n=26 K. pneumoniae isolates results was shown the 23 (88.46%) K. pneumoniae isolates had gave positive results for the blaTEM gene. While 3 (11.538%) K. pneumoniae isolates revealed negative results for blaTEM gene. On other hand results were shown that only 5 (19.23%) K. pneumoniae isolates had positive results for blaSHV gene and 21 (80.769%) K. pneumoniae isolates had negative results for the blaSHV gene. Also the results shown the 5(19.23%)K. pneumoniae isolates had positive results for blaTEM and blaSHV genes.

Prevalence of blaOXA-10, blaCTX-M-3 and SHV Genes among ESBL-Producing Escherichia coli and Klebsiella pneumoniae Isolated from Clinical samples in Basra city, Iraq

The current study was conducted to determine the prevalence of extended-spectrum β-lactamase (ESBL) in 78 drug-resistant clinical isolates (25 Klebsiella pneumoniae and 53 Escherichia coli strains) using phenotypic and molecular methods. The phenotypic method was performed using a double-disk synergy test (DDST), while the genotypic method screened for the blaSHV, blaCTX-M13U, and blaOXA-10 genes using specific primers. The phenotypic results showed that out of 53 tested strains of E. coli, 17 (32.07%) produced ESBL. Similarly, out of 25 tested strains of K. pneumoniae, 8 (32%) produced ESBL. Genotypic detection showed that in E. coli, the most abundant gene was SHV, present in 24 strains (45.28%), followed by blaOXA-10 in 23 strains (43.39%) and CTX-M-3 in 8 strains (15.09%). In K. pneumoniae, SHV was detected in 12 strains (48%), followed by OXA-10 and CTX-M-3, each found in 5 strains (20%).

Prevalence of CTX-M, TEM, SHV and OXA genes among single and multi-β-lactamase producing uropathogens in Katihar, Bihar, India

Prevalence of CTX-M, TEM, SHV and OXA genes among single and multi-β-lactamase producing uropathogens in Katihar, Bihar, India

Molecular Characterization and Evaluation of Antibiotic Resistance Gene of Bacteria Associated with Gills of Arius heudelotii and Clarias gariepinus

This study was aimed at the isolation and molecular characterization of bacteria associated with catfish from culture and wild environments. Samples of gills of Arius heudelotii and Claris gariepinus fishes of economic value from the New Calabar River Choba and earthen ponds from Aluu in Rivers State were collected and analysed. Characterization of isolates from the gills of the catfish was done using the 16S rRNA gene sequencing method to identify their relationship. The presence of a resistance gene was examined by polymerase chain reaction followed by agarose gel electrophoresis of the amplification products. The mean total heterotrophic bacterial counts was 6.5995±0.947 LogCFU/g for the gills’ samples obtained from the earthen pond Aluu. New Calabar River had 4.5041±1.054 LogCFU/g for the gills’ samples. A total of 12 bacterial flora were isolated from the gills of the sampled catfish: Escherichia coli, Klebsiella spp., Enterobacter spp., Citrobacter spp., Bacillus subtilis, Proteus vulgaris, Streptococcus spp., Shigella spp. Staphylococcus spp., Micrococcus spp., Vibrio spp. and Acinetobacter spp. Vibrio azureus, Aeromonas dhakensis, Proteus vulgaris, Aeromonas hydrophila. Vibrio azureus and Providencia alcalifaciens were found to be carriers of the TEM gene, while Providencia alcalifaciens carried the CTX-M gene. SHV gene was found in Vibrio azureus, Providencia alcalifacians, Proteus vulgaris, Aeromonas hydrophila, Providencia alcalifacians. However, the Oxa gene was not identified in the bacterial genera. The high probability of transferring them to other microbes to cause antibiotic pollution has become a serious public health challenge. Microbial loads with high or low presence of some potential pathogens call for public concern.

Tracking the Phylogenetic Origins of Proteus mirabilis and P. vulgaris in Iraq

Abstract Background: Common gram-negative rod-shaped bacteria called Proteus spp. are widely known for their swarming motility and urease activity. It is the main pathogen that causes complicated urethral tract infections. Objectives: Proteus mirabilis and P. vulgaris isolates were isolated, and the positions of the bla TEM, bla CTX-M, and bla SHV genes on the P. mirabilis isolates’ genomes were identified. The MEGA X10.2.4 software tool was employed to achieve this, and the genetic trees of P. mirabilis were created. Additionally, the isolates that represent the prevalence and degree of pathogenicity of these bacteria in Iraq were used to produce biofilms. Materials and Methods: A total of 210 urine samples from various patients who were hospitalized in general hospitals in Iraq between September 2021 and July 2022 were collected and yielded forty isolates of P. mirabilis and P. vulgaris. Traditional methods and the VITEK 2 compact were used to identify the isolates, which were given the names PM1 to PV40. Results: The results showed that all isolates were biofilm-forming, with the following ratios: 5 (15%) were weak, 31 (5.77%) were medium, and 4 (5.7%) were strong. The matching of this study isolates with NCBI-Gen bank global strains found that four isolates (PM1, PM2, PM3, and PM4) were originated in India, two isolates (PM5 and PM8) were originated in Iraq, and one (PM6) isolate was originated in Egypt. Conclusion: According to the results of this investigation, P. mirabilis and P. vulgaris isolates from various geographical areas show varying degrees of virulence factors. Therefore, additional researches are required to create a genetic diversity map of P. mirabilis and P. vulgaris due to variable frequencies in the sequencing of bla TEM, bla CTX-M, and bla SHV genes of the isolates.

CTX-M, TEM, and SHV Genes in Escherichia coli, Klebsiella pneumoniae, and Enterobacter spp Isolated from Hematologic Cancer Patients with Bacteremia in Uganda.

We determined the phenotypic resistance to third-generation cephalosporins, phenotypic extended spectrum beta-lactamase (ESBL) prevalence, and genotypic prevalence of ESBL-encoding genes blaCTX-M, blaTEM, and blaSHV in Enterobacteriaceae isolated from hematologic cancer patients with febrile neutropenia and bacteremia at the Uganda Cancer Institute (UCI). Blood cultures from hematologic cancer patients with febrile neutropenia were processed in BACTEC 9120. E. coli, K. pneumoniae, and Enterobacter spp. isolates were identified using conventional biochemical methods. Antimicrobial susceptibility tests, phenotypic ESBL characterization, and genotypic characterization of the ESBL-encoding genes blaCTX-M, blaTEM, and blaSHV were determined for pure isolates of E. coli, K. pneumoniae, and Enterobacter spp. Two hundred and two patients were included in the study. Median age of patients was 19 years (IQR: 10-30 years). Majority (N=119, 59%) were male patients. Sixty (30%) of the participants had at least one febrile episode due to Enterobacteriaceae. Eighty-three organisms were isolated with E. coli being predominant (45, 54%). Seventy-nine (95%) Enterobacteriaceae were multidrug resistant. The ESBL phenotype was detected in 54/73 (74%) of Enterobacteriaceae that were resistant to third-generation cephalosporins. A higher proportion of Enterobacteriaceae with ESBL-positive phenotype were resistant to piperacillin-tazobactam (p=0.024), gentamicin (p=0.000), ciprofloxacin (p=0.000), and cotrimoxazole (p=0.000) compared to Enterobacteriaceae, which were sensitive to third-generation cephalosporins. The organisms were more susceptible to carbapenems and chloramphenicol than resistant. ESBL-encoding genes (blaCTX-M, blaTEM, and blaSHV) were detected in 55 (75%) of the 73 Enterobacteriaceae that were resistant to third-generation cephalosporins. BlaCTX-M, was the most common ESBL-encoding gene identified with 50 (91%). ESBL-producing Enterobacteriaceae are a predominant cause of bacteremia in hematologic cancer patients at UCI. The most common ESBL-encoding gene identified in the ESBL-PE was blaCTX-M. Resistance to imipenem and meropenem was low.

Epidemiological and Molecular Characteristics of Hypermucoviscous and Hypervirulent Klebsiella pneumoniae Isolates in Community Patients in Shanghai, China.

The occurrence and dissemination of hypermucoviscous and hypervirulent Klebsiella pneumoniae (hm-hvKp) isolates in clinical settings are a critical public health problem in the world. However, the data on these isolates in community populations are limited. This study aims to understand the prevalence and molecular characteristics of hm-hvKp isolates in community patients in Shanghai, China. In 2018, an active surveillance system focused on hm-hvKp in community diarrhoeal cases was implemented in Pudong New Area, Shanghai, China, involving 12 sentinel hospitals. The antimicrobial susceptibility of hm-hvKp isolates from fecal samples was tested, and whole-genome sequencing (WGS) was performed to predict the serotypes and sequence types and to identify antimicrobial resistance determinants, virulence determinants, and phylogenetic clusters. The overall prevalence of hm K. pneumoniae isolates was 2.48% (31/1252), with the proportions of 1.76% (22/1252) for hm-hvKp and 0.72% (9/1252) for hm not hv K. pneumoniae. The prevalence of hm-hvKp isolates among different age groups and different months was statistically significant. All the 22 hm-hvKp isolates were susceptible to 20 antimicrobial agents and only carried bla SHV gene, and KL1 and KL2 accounted for eight (36.36%) cases and seven (31.82%) cases, respectively. The eight ST23/KL1 isolates belonged to the predominant CG23-I clade, which typically possessed the virulence determinants profile of rmpA/rmpA2-iro-iuc-ybt-irp-clb. The five ST86/KL2 isolates were assigned to the global clusters ST86/KL2-1 (n=2), ST86/KL2-2 (n=2), ST86/KL2-3 (n=1), all lack of the clb gene. Shanghai ST23/KL1 and ST86/KL2 isolates were closely related to the global isolates from liver abscesses, blood, and urine. Hm-hvKp is carried by the community population of Shanghai, with ST23/KL1 and ST86/KL2 isolates predominant. Hm-hvKp isolates of different continents, different sources, and different virulence levels were closely related. Ongoing surveillance of hm-hvKp isolates in the community population is warranted.

Phylogenetic Characterization of Resistant Salmonella Strains in Typhoid Fever Patients in Nigeria.

Salmonella species are Enterobacteriaceae associated with typhoid fever. In this study, the distribution of broad-spectrum β-lactamase regulatory genes and genetic relatedness of isolates was determined. Stool samples (400) were collected from patients with fever in Dalhatu Araf Specialist Hospital (DASH), Lafia, Nigeria, between March 2020 and April 2021. Salmonella species were isolated and extended-spectrum β-lactamase distribution was determined among resistant isolates using polymerase chain reaction (PCR). Genetic relatedness of Salmonella species resistant to the 10 first-line antibiotics administered was determined among S typhi isolated. Of the 60 isolates that were confirmed to belong to the genus Salmonella, 12 (20.0%) isolates with bla SHV genes were the most prevalent, blaOXA-1 and blaCTX-M-9 were present in 5 isolates each, while blaCTX-M-4 and blaTEM genes with a prevalence of 1.7% each were the least obtained in the isolates. Two isolates had a multidrug-resistant index (MDRI) of 1, and 2 others were positive with the S typhi staG gene. Sequencing to determine their diversity showed that isolates ST36 and ST138, respectively, had MDRI = 1 and are clustered in a group with a similarity coefficient of 0.00634. The 2 isolates had the highest genetic similarity, which indicates that the genetic diversity between the isolates is low, while Salmonella strain ST313L2 had a high level of genetic distance from the other isolates. The most resistant isolates are closely related which calls for concern.

Frequency of Fecal Carriage of ESBL Resistance Genes in Multidrug-Resistant Pseudomonas aeruginosa Isolates from Cancer Patients at Laquintinie Hospital, Douala, Littoral Region, Cameroon.

Background. Opportunistic infections are the second cause of death among cancer patients. This study aimed at determining the antimicrobial profile and the prevalence of extended-spectrum beta-lactamase (ESBL)-gene carriage of Pseudomonas aeruginosa isolates among cancer patients at the Douala Laquintinie Hospital, Littoral Region of Cameroon. Between October 2021 and March 2023, 507 study participants were recruited among whom 307 (60.55%) were cancer patients, compared to 200 (39.45%) noncancer patients. Fifty-eight P. aeruginosa isolates were isolated from fecal samples of forty-five cancer patients and thirteen noncancer patients using Cetrimide agar. The antimicrobial resistance profile of the isolates was determined using the Kirby-Bauer disk diffusion method. The polymerase chain reaction was used to detect the presence of extended-spectrum beta-lactamase genes among P. aeruginosa isolates. P. aeruginosa showed significant resistance rates in cancer patients compared to noncancer patients to imipenem, cefotaxime, and ceftazidime, piperacillin-tazobactam, ticarcillin-clavulanic acid, and ciprofloxacin. The multidrug resistance (MDR) rate was significantly (p < 0.05) higher in cancer patients than in noncancer patients. The frequency of beta-lactamase genes in the 58 ESBL-producing P. aeruginosa isolates was determined as 72.41% for bla TEM, 37.93% for bla OXA, 74.14% for blaCTX-M, and 44.83% for bla SHV genes. The study revealed an alarmingly high prevalence of fecal carriage of ESBL-producing P. aeruginosa with a high rate of MDR among cancer patients. It indicates that regular monitoring and surveillance of ESBL-producing P. aeruginosa among cancer patients are needed to improve the management of patients.

SURVEY ON MULTI-DRUG RESISTANT AND EXTENDED SPECTRUM BETA LACTAMASES PRODUCING BACTERIA ON CONTACT SURFACES AT MURTALA MUHAMMAD SPECIALIST HOSPITAL, MAKKAH EYE CLINIC AND UMC ZHAHIR HOSPITAL, KANO STATE

The contaminated environmental surfaces are not only potential reservoirs for spread of microbial agents inside hospital but also in community. The development and evolution of antimicrobial resistance in pathogen has been reported to be one of the major issues confronting the global health community. Members of family Enterobacteriacae able to produce extended spectrum of beta lactamase which is responsible to hydrolyze the third generation of cephalosporin group antibiotics resulting treatment failure. The use of beta-lactams has increased considerably since its discovery so also resistant genes leading to Extended-Spectrum Beta-Lactamases (ESBL) mediated by the presence of blaCTX-M, blaTEM, and blaSHV genes present in most Gram-negative bacteria. Out of 300 swab samples collected, 218 (72.7%) bacteria were isolated. 158 (72.5%) was gram positive and 60(27.5%) was gram negative. A total of 96(44.0%) isolates were multidrug resistant. Antibiotics susceptibility test was performed by Kirby Bauer technique according to CLSI guidelines. Organisms that tested positive phenotypically for ESBL were subjected to PCR for molecular analysis. ESBL had a percentage rate of 11.3%. The coexistence of bla CTX-M, bla TEM and bla SHV gene was detected

Molecular Screening of Carbapenem-Resistant K. pneumoniae (CRKP) Clinical Isolates for Concomitant Occurrence of Beta-Lactam Genes (CTX-M, TEM, and SHV) in the Kingdom of Bahrain.

The emergence of extended-spectrum β-lactamase-producing Klebsiella pneumoniae, including CRKP infections, has resulted in significant morbidity and mortality worldwide. We aimed to explore the presence of bla genes (CTX-M, TEM, and SHV) in CRKP isolates. A total of 24 CRKP isolates were randomly selected from the Salmaniya Medical Complex Microbiology Laboratory. These isolates, which were positive for carbapenemases, were further explored for CTX-M, TEM, and SHV genes using PCR. All the CTX-M PCR amplicons were sent for sequencing. To determine genetic relatedness, molecular typing by ERIC-PCR was performed. The bla gene testing demonstrated that a significant proportion of these isolates harbored SHV, CTX-M, and TEM genes (100%, 91.6%, and 45.8%), respectively. Bioinformatic analyses confirmed CTX-M-15 in these isolates. ERIC-PCR analysis showed three clusters demonstrating genetic relatedness. The study findings reveal the concomitant carriage of the SHV and CTX-M-15 and a comparatively lower carriage of TEM genes in CRKP isolates. Our findings highlight the significance of routinely reporting the presence of antibiotic resistance genes along with regular antibiotic sensitivity reports, as this will aid clinicians in prescribing appropriate antibiotics.

Evaluation of thymol effect on antimicrobial susceptibility and biofilm production in clinical Enterobacter spp. producing ESBLs and KPC enzymes

ABSTRACT Background Multidrug-resistant Enterobacter strains that harbour Klebsiella pneumoniaecarbapenemase and Extended Spectrum β-Lactamases genes are a therapeuticchallenge in hospitals that have made infection control and antibiotic therapydifficult and use of natural compounds such as thymol is interesting in thetreatment of infections. Methods Sixty-seven Enterobacter spp. isolates were obtained from patients admittedto hospitals in Hamadan and Tehran. Antimicrobial susceptibility, ESBLs andKPC genes detection and biofilm production of isolates were measured. Findings The results showed that the highest and lowest antibiotic resistance wererelated to ceftazidime (82.1%) and tigecycline (3%), respectively. Also, 79.1% of the isolates were MDR. The highestpresence of genes was related to TEM (100%), SHV (82%), CTX-M(77%) and KPC (8.96%) genes. The MICand MBIC of thymol were obtained 8-31 μg/ml and16-62 μg/ml, respectively. Conclusion Use of thymol, as an aid in antibiotic therapy is recommended.

Genetic determinants of antibiotic resistance in enterobacteria isolated during microbiological monitoring in the perinatal center

Relevance. Currently, studies of the prevalence of antibiotic resistance and its genetic characteristics are focused primarily on the adult population, although infection with multiple drug infection has been registered as etiological agents of a general infection in obstetric and gynecological and pediatric institutions. The study of the prevalence of genetic determinants of antibiotic resistance is an important area of scientific research.Aim. To analyze the results of the studies carried out to identify the genetic determinants of antibiotic resistance of enterobacteria isolated during microbiological monitoring in the perinatal center.Materials and methods. The genetic profile of antibiotic resistance was studied in ESBL-producing strains isolated from 45 women and 35 children examined at the departments of the Federal State Budgetary Institution «NII OMM» of the Ministry of Health of Russia. To determine the determinants of antibiotic resistance, 80 non-duplicate strains of 7 species of the Enterobacteriaceae family were studied. DNA of bacterial cells was isolated from a daily culture of microorganisms using the PROBA-NK kit, detection of the tem, ctx-M-1, shv genes; oxa-40-like, oxa-48-like, oxa-23-like, oxa-51-like, imp, kpc, ges, ndm, vim were carried out using the diagnostic kit «BacResista GLA» on the detecting amplifier DT-48 (DNA -technology, Russia). To assess the statistical significance of differences in the frequency of occurrence of genes, Pearson's c2 test with Yates' correction was used.Results and discussion. When analyzing the results of studies on the molecular genetic detection of antibiotic resistance determinants, which we conducted in 2022, it was found that 8 genovariants were found in bacterial strains isolated from patients of the departments of the Research Institute of OMM in Yekaterinburg, providing resistance to beta-lactam antibiotics. The dominant genome, as in 2021, remains blaCTX-M-1, found in 29 cases. The blaTEM gene was identified both in association with other genes and as a single variant in Escherichiae coli and Klebsiella pneumoniae strains. Of the eight strains of K. pneumoniae, 4 were found to have three antibiotic resistance genes blaCTX-M, blaTEM, blaSHV, strains with a genetic profile of blaCTX-M, blaTEM, blaSHV, blaNDM were isolated once; and blaTEM, blaSHV, blaKPC. In one strain of K. pneumoniae, phenotypically showing resistance to AB, no genetic determinants of AB resistance were found. In addition to resistance to beta-lactam antibiotics, the strains demonstrate resistance to such groups of antibacterial drugs as fluoroquinolones, phosphonic acid derivatives (fosfomycin), and aminoglycosides. The data obtained indicate that the intestines of newborns during their stay at the stationary stage of nursing in some cases are colonized by strains of enterobacteria with multidrug resistance. Consequently, children are a reservoir of resistant microorganisms and can be sources of pathogens of infectious diseases in families and children's organized groups.

The Prevalence of Extended Spectrum Beta Lactamase Producing Citrrobacter Freundii and Pprovidencia Stuartii from Medical Laboratories in Parts of Lagos State, Nigeria

Citrobacter freundii and Providencia stuartii are members of the family Enterobacteriaceae which are emerging as important Extended Spectrum Beta Lactamase (ESBL) producing pathogens. This study was aimed to determine the prevalence of ESBL and carbapenemase production in C. freundii and P. stuartii. from laboratories in the northwest of Lagos. A total of 134 isolates were obtained from laboratories. Isolate identification, antibiotics susceptibility testing, and phenotypic detection of ESBL production were done using standardized procedures. Multiplex polymerase chain reaction (PCR) was performed on ESBL producing isolates to detect blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM, blaIMP and blaOXA. Out of 134 isolates 15 (11.2%) were Citrobacter freundii and only 2 (1.5%) were of the Providencia genus both of which were Providencia rettgeri and Providencia stuartii. A total of 9 (60%) tested positive for the production of Extended Spectrum Beta Lactamase (ESBL) enzyme while only Providencia stuartii of the Providencia genus also produced ESBL. Also, 8 (47%) of the initial Citrobacter and Providencia isolates produced carbapenemase Molecular characterization showed that 4 (44.4%) and 1 (100%) of the Citrobacter freundii and Providencia stuartii isolates respectively, displayed the presence of CTX-M gene while 2 (20%) of all ESBL producing isolates also possessed SHV gene including 1 (11.1%) of ESBL producing Citrobacter freundii and 1 (100%) of ESBL producing Providencia stuartii. Antibiotic susceptibility testing showed that Citrobacter freundii displayed the highest resistance to Ceftazidime and Cefotaxime at 71.4% and the least resistance to Amoxicillin-Clavulanate at 28.6%. Providencia stuartii also displayed absolute resistance (100%), to cefotaxime and ciprofloxacin and absolute susceptibility (100%) to Ceftazidime. Also, Citrobacter freundii showed the same resistance patterns with 57.1% resistance to Gentamicin, Cefuroxime and Ceftriaxone antibiotics while Providencia stuartii showed varied resistance patterns with absolute resistance (100%), absolute susceptibility (100%) and absolute resistance (100%) to Gentamicin, Cefuroxime and Ceftriaxone antibiotics respectively. In conclusion, the Multiple Antibiotic Resistance Index values of both bacterial genera showed that 70.6% were from High-Risk sources indicating that the majority of bacteria analysed were from environments where they have been subjected to a high burden of antibiotic exposure.

Genetic Profiling of Pathogens Associated with Pyroxia of Unknown Origin Treated for Plasmodiasis in Bayelsa State, Nigeria

Antimicrobial resistance of Salmonella is an increasing problem and has become a public health issue worldwide. Moreover the evidences on hazard of therapeutic failure due to the increasing incidence of antimicrobial resistance among typhoidal antibodies and malaria are on the increase. This study was aimed at the genetic profiling of pathogens associated with Pyroxia of unknown origin treated for Plasmodiasis in Bayelsa State, Nigeria. A total of 1200 samples (200 Stool and 1000 blood) were collected from consented subjects in Federal Medical Center and Niger Delta University Teaching Hospital Bayelsa State, Nigeria. Blood samples were used for detection of Malaria parasites and Typhoidal antibodies while the stool samples were used for culturing on Salmonella-Shigella Agar. Antibiotic sensitivity of the pure isolates were carried out using sensitivity disc, biochemical analysis using API 20E was carried out to actually differentiate bacterial from each isolates and finally by genetic profiling of the resistant genes of the pure isolates using polymerase chain reaction. Antibiogram of the isolates showed high resistance to Oxacillin (73.6 %), Ampicillin (66.6%), Tetracycline (60.0%), and Erythromycin (53.3%) while Gentamycin, 22(73.3%), Chloramphenicol 18(60%) and Azithromycin 18(60%) had the highest sensitivity. The distribution of resistant genes from the genetic profiling showed that out of 20 amplified isolates, NDM resistant genes were 17(85%) followed by Bla TEM genes 14(70%) followed by CTXM and SHV genes as 11(55%) and 10(50%) respectively. The bacterial isolates showed high AMR and MDR to the commonly used antimicrobial drugs and the distribution of ESBL genes revealed a high level of resistant genes in Bayelsa State.

Evaluation of presence of clone ST131 and biofilm formation in ESBL producing and non-producing Escherichia coli strains.

Escherichia coli ST131 is a pandemic clone associated with multidrug resistance, starting with beta-lactamase production and fluoroquinolone resistance in the first place, leading to significant systemic infections. Clones that develop due to the frequency of antimicrobial resistance and the rate of spread in our country are important issues that need to be investigated. This study aims to investigate the incidence of ST131which is a "high-risk pandemic clone E. coli" in ESBL-producing and non-ESBL-producing strains, as well as their biofilm-forming abilities and antibiotic resistance rates. A total of 86 E. coli isolates were used in the study. Bacterial identifications were performed by conventional and automated methods. The double disc synergy method was used to demonstrate the presence of ESBL. Molecular studies in all E. coli strains were performed by real-time PCR method. 86 strains were studied, of which 83.72% were urine, 6.98% were wound, 4.65% were blood, and 2.33% were tracheal aspirate and sputum. 79.07% of these strains were ESBL-positive. 58.1% of the strains were female, whereas 41.9% were male patients, and the average age was 46.2. Out of 86 strains, 38.72% were ST131 positive, the H30 subclone was detected in 27.27% of them, and the H30-Rx subclone was detected in all of the H30 subclone positive strains. The presence of the ESBL resistance gene was detected at the rate of TEM 41.86%, SHV 37.21%, CTX-M 36.04%, and OXA 4.65%. Most commonly SHV gene (54.54%) was seen in ST131 clone-positive samples. Finally, while it was found that 48.83% of the strains formed biofilm by any method, biofilm formation was detected in 69.7% of the samples that were positive for the ST131 clone. Our study can reveal the dramatic prevalence of the ESBL-producing E. coli strains along with the high-risk ST131 clone, the dominance of the H30Rx subclone of this risky clone, as well as the importance of the influence of resistance mechanisms along with resistance and biofilm.

Abundance and phenotypic-genotypic analysis of antibiotic-resistant Escherichia coli isolated from wastewater of the Zainoel Abidin Hospital, Banda Aceh, Indonesia

Abstract. Anelia N, Suhartono S, Hayati Z. 2023. Abundance and phenotypic-genotypic analysis of antibiotic-resistant Escherichia coli isolated from wastewater of the Zainoel Abidin Hospital, Banda Aceh, Indonesia. Biodiversitas 24: 2294-2301. Excessive and inappropriate use of antibiotics can initiate the emergence of antibiotic resistance in bacteria. Wastewater Treatment Plant (WWTP) is believed to be a reservoir for antibiotic-resistant bacteria, including Escherichia coli. This study aimed to perform the enumeration, isolation, characterization, and evaluation of resistance properties and antibiotic resistance-coding genes encoded by a plasmid of E. coli isolates from WWTP of the dr. Zainoel Abidin General Hospital (RSUDZA). The study was conducted through several stages, including the isolation and quantification of E. coli, antibiotic resistance testing, confirmation testing of ESBL-E. coli isolates, isolation of plasmid DNA, and amplification of TEM, SHV, and CTX-M genes. The results showed that the wastewater in the sum pit tank, aeration tank, and final tank of the WWTP contained E. coli at 7.5x104 CFU/mL, 3x102 CFU/mL, and 1x102 CFU/mL, respectively. A total of 22 isolates were obtained, with ten isolates from the sum pit tank, seven from the aeration tank, and five from the final tank. Escherichia coli isolates had resistance to eight types of antibiotics in the aeration tank and seven types in the final tank, compared to the sum pit tank of the WWTP. Therefore, 59% of the isolates were classified as Multi-drug Resistant (MDR) and 32% as Extended Spectrum ?-Lactamase E. coli (ESBL-EC). TEM, SHV, and CTX-M genes were detected in all locations at 63.6%, 22.7%, and 95.5%, respectively. In addition, CTX-M was the most common gene found in wastewater isolates at RSUDZA WWTP. The hospital wastewater treatment process must be improved to reduce the presence of antibiotic-resistant bacteria and the spread of antibiotic resistance genes.

Characteristic of extended substrate-spectrum beta-lactamases producing Enterobacteriales isolated from pork meat

Antibiotic resistance among bacteria is an ever-growing problem in both human and veterinary medicine. Livestock and meat from their slaughter are often sources of drug-resistant bacteria. The ability to produce extended-spectrum beta-lactamases (ESBLs) is common among Enterobacteriales, including commensal bacteria. These bacteria represent a largely ignored reservoir of ESBL genes. The aim of this study was to assess the drug susceptibility of non-pathogenic Enterobacteriales isolates obtained from meat used in food production. Among the strains tested, ESBL-type resistance was demonstrated - 13 of 18 (72%) isolates tested were found to be ESBL-producing strains. Of these, eight possessed the CTX-M gene, three the TEM gene and one the SHV gene.