Antibiotic Sensitivity Testing Research Articles

Bacteriological Study of Post-Operative Wound Infection and its Antibiograms

INTRODUCTIONPostoperative wound infections are a significant complication of surgical procedures, which increases morbidity, mortality, prolonged hospital stay, and increased healthcare costs. The study was undertaken with an aim to investigate the etiological agents of postoperative wounds and their sensitivity to commonly used antibiotics. MATERIAL AND METHODSThis study was a descriptive cross-sectional prospective study, carried out at Universal College Of Medical Sciences-Teaching Hospital, Bhairahawa, Nepal, on 100 postoperatively wounded infected cases, during April 2023 – August 2023, Non Probability sampling method (Purposive sampling) was used, and specimens were collected by sterile cotton swabs and pus was aspirated with a sterile needle and syringe and processed using standard bacteriological techniques. Antibiotic sensitivity test was conducted by disc diffusion (Kirby-Bauer) method. RESULTSPostoperative wound infections caused by Gram negative bacilli were more frequent and predominant pathogens involved were Pseudomonas aeruginosa, followed by Klebsiella spp and Enterobacter aerogenes. Among Gram positive bacteria Staphylococcus aureus was predominant. Gram positive cocci were highly sensitive to Ampicillin, and Gram negative bacilli were highly sensitive to Ceftazidime, Gentamicin, and Amikacin. Coagulase negative staphylococci were resistant to all drugs tested. CONCLUSIONPseudomonas aeruginosa was the commonest etiological agent for postoperative wound infections. Gram positive cocci and E.coli were highly sensitive to Ampicillin and Gram negative bacilli were highly sensitive to Ceftazidime, Gentamicin, Amikacin. Staphylococcus aureus was resistant to vancomycin and Teicoplanin and coagulase staphylococci were resistant to all drugs tested. Gram negative bacilli were resistant to Polymyxin B and Tigecycline. Increasing resistance to commonly used antibiotics warrants the judicious use of antibiotics and establishment of antibiotic policy in the hospital.

PHENOTYPIC DETECTION OF ESBL AND AMPC BETA LACTAMASES AMONG GRAM-NEGATIVE ISOLATES FROM CLINICAL SAMPLES IN A TERTIARY CARE HOSPITAL

Objectives: Drug-resistant Gram-negative bacilli expressing extended-spectrum beta-lactamases (ESBLs) and AmpC pose a serious therapeutic threat in nosocomial infections. Cost-effective screening methods are a boon to patients. This study aims to detect gram-negative bacilli and their antibiotic sensitivity patterns, as well as detect the ESBL and AmpC-producing isolates among Gram-negative bacilli. Methods: A prospective study was conducted with 150 samples. Gram-negative bacilli were isolated, and their antibiotic sensitivity tests were performed by the Kirby–Bauer disk diffusion method. Potential ESBL producers were screened using Ceftazidime disc, and AmpC producers were screened by Cefoxitin discs by the disc diffusion method. ESBL producers were confirmed by the combined disc diffusion assay method using ceftazidime and ceftazidime/Clavulanic acid disc. AmpC producers were confirmed by the Cefoxitin Cloxacillin Double Disc Synergy Test. Results: About 38% of 150 samples were gram-negative bacilli, of which 40.35% were Escherichia coli, followed by Pseudomonas aeruginosa (35.08%). Maximum sensitivity by E. coli was found toward imipenem, meropenem, and cotrimoxazole. P. aeruginosa showed maximum sensitivity toward piperacillin/tazobactam, imipenem, meropenem, and ceftazidime. 28.07% of Gram-negative isolates were ESBL producers, with E. coli (11 isolates) being the maximum, and 15.78% were AmpC producers, with E. coli (four isolates) being the maximum. Seven isolates were both ESBL and AmpC producers. Conclusion: Routine screening and timely reporting of ESBL and AmpC producers help in preventing the spread of multidrug-resistant strains. Antibiotic resistance surveillance helps in the implementation of strict infection control and prevention practices.

Isolation and Molecular Detection of Bacteria Causing Omphalitis in Poultry with Look on Antibiotic and Disinfectant Resistance

Antimicrobial resistance is an international concern and creates critical health issues for both humans and animals. This research aimed to ascertain the prevalence of isolates, antibiotic, and disinfectant resistance patterns from omphalitis suspected chicks in Dinajpur, Bangladesh. A total of 24 yolk swab samples were collected from different hatcheries for microbiological analysis and antibiotic sensitivity tests. The PCR employing 16s and 23s RNA genes was conducted to detect Escherichia coli and invA genes were used to detect Salmonella spp. and 23S rRNA genes were applied for Staphylococcus spp. A total of 35 isolates were identified where 14 (40%) E.coli, 10 (28.58%) Salmonella spp., and 11 (31.42%) Staphylococcus spp. from day 1-8, respectively. The largest amount of bacteria on day 5, including day 7 (20%), followed by days 3, 4, and 6, including 6 (17.14%) observed, respectively. The PCR band of E. coli was detected at 232 bp, Salmonella spp. 284 bp and Staphylococcus spp. 1267 bp respectively. E. coli was highly resistant to Amoxicillin (100%), followed by Tetracycline (83.33%), whereas highly sensitive to cefotaxime (100%), Gentamicin and Co-trimoxazole (83.33%). Salmonella spp. indicates high susceptibility to Cefotaxime, Co-trimoxazole, and Ceftazidime (83.33%), followed by Gentamicin (66.67%) respectively, whereas Staphylococcus spp. was found to be highly resistant to Methicillin (100%) and Ampicillin (100%) followed by Gentamicin and Tetracycline (83.33%) respectively. The MAR index calculation of isolated E. coli, Salmonella spp., and Staphylococcus spp. from different sources of poultry hatcheries was measured at 0.77, 0.79, and 0.77, which affect the newly hatched chicks in poultry industries. Therefore, an urgent surveillance program is needed to fight antimicrobial resistance in poultry production sectors in Bangladesh.

Multicenter Investigation of Drug-Resistance in Burkholderia Cepacia Bloodstream Infections in Hebei Province, China, from 2016 to 2021.

To compare the epidemiological characteristics and drug resistance of Burkholderia cepacia isolated from blood cultures, and to provide data support and a scientific basis for the clinical treatment and detection of hospital infections. The Hebei Province Antimicrobial Surveillance Network received 349 B. cepacia strains isolated from blood cultures reported by 83 hospitals, from 2016 to 2021. These strains were identified by MALDI-TOF MS and, the antibiotic sensitivity tests were carried out using the VITEK 2 COMPACT system. The 2023 Institute of Clinical and Laboratory Standardization drug-susceptibility breakpoints were used for drug susceptibility testing and the data were analyzed using WHONET5.6 software. A total of 349 B. cepacia strains were isolated from 2016 to 2021, including 68 strains from secondary hospitals and 281 strains from tertiary hospitals. The ratios of male: female patients with B. cepacia bloodstream infections in all hospitals, secondary hospitals, and tertiary hospitals were 1.49:1 (209/140), 2.09:1 (46/22), and 1.38:1 (163/118), respectively. Most B. cepacia strains were isolated in intensive care units (ICUs), followed by internal medicine departments, accounting for 49.57% (173/349) and 22.92% (80/349), respectively. Regarding the age distribution, most patients were elderly (>65 years, 57.59%, 201/349), with numbers of patients gradually declining with decreasing of age. The resistance rates for levofloxacin, ceftazidime, and sulfamethoxazole decreased over the 6-year period (P<0.05), while there were no significant changes in the resistance rates for meropenem, chloramphenicol, and minocycline (P>0.05). There was no significant difference in drug-resistance rates between secondary and tertiary hospitals (P>0.05). Attention should be paid to bloodstream infections caused by B. cepacia, especially elderly patients and patients admitted to the ICU. The difficult treatment characteristics of B. cepacia bloodstream infections mean that laboratories and clinicians should pay careful attention to drug resistance to provide a basis for their prevention and empirical treatment.

Molecular detection of Klebsiella pneumoniae producing extended-spectrum beta-lactamase isolated from bat feces from the Tanjung Ringgit bat cave, Lombok Island, Indonesia

Background and Aim: Bats are a reservoir for the pathogenic bacteria Klebsiella pneumoniae and can spread it through feces that fall in nests/caves, carried, and dropped while they fly near human settlements, and from their saliva. The emergence and spread of multidrug resistance (MDR) strains of K. pneumoniae indicate that resistant to antibiotics, especially extended-spectrum beta-lactamase (ESBL), is considered an important global health threat. The aim of this study was to determine the presence of the gene encoding extended beta-lactamase in K. pneumoniae isolated from fresh bat feces collected from the Tanjung Ringgit bat cave, East Lombok. Materials and Methods: In this study, 150 fresh fecal samples were analyzed using standard microbiological techniques for the presence of K. pneumoniae. K. pneumoniae-positive isolates were subjected to antibiotic sensitivity testing, followed by molecular detection using polymerase chain reaction. Results: This study showed that 14 (9.3%) of 150 samples were positive for K. pneumoniae. Ten of the 14 samples (71.4%) were MDR isolates and 6 (42.9%) had the blaSHV gene identified. Conclusion: The presence of K. pneumoniae isolated from fresh bat feces, which is MDR and has the blaSHV gene encoding ESBL indicates that bats can be a reservoir for the transmission of MDR and ESBL bacteria has an impact on public health in the study area. Keywords: bat, blaSHV, extended-spectrum beta-lactamase, Klebsiella pneumoniae, public health.

Detection and antibiogram of bacterial contaminants in some commonly consumed indigenous street vended foods in Kasua Central Market in Kaduna North, Kaduna State, Nigeria

Some indigenous ready-to-eat foods, due to its affordability and availability serve as a major alternative to restaurants and high profiled eateries in developing countries. The hygiene standard of these foods is a critical point that needs to be considered due to microbial contamination by potential pathogens. A triplicate of three different food samples: Fura, Nunu, and Tuwo were collected and analysed to determine their microbiological safety. Bacterial isolates were identified using standard methods while antibiotic sensitivity test and multiple drug resistance pattern were carried out. The isolated bacteria: Escherichia coli, Klebsiella pneumoniae, Salmonella 100yphi, Staphylococcus aureus, and Enterobacter sp. While fungal isolates include: Aspergillus niger, Aspergillus flavus, Rhizopus. oryzae and Penicillium sp. Salmonella sp. was observed to be resistant to 70% of the antibiotics while both Klebsiella sp. And Enterobacter sp. exhibited resistance to 60% of the tested antibiotics. S. aureus had the least resistance to about 30% of the tested Gram-positive antibiotics. Out of all the tested antibiotics, only Tarivid was observed to be effective against all the Gram-negative bacterial isolates, while Reflacine and Ciprofloxacine were effective against 80% of them. The presence of coliform bacteria in the sampled foods raise a great concern about the safety of the street vended foods in the sampled region in Nigeria. The microbiologically low quality of the vended ready-to-eat foods assayed in this study might have resulted from a relatively low level of hygiene observed by the vendors, as well as absence of regulatory body or hygiene codes to guide the street food vendors. From the results of this study, efforts should be intensified by all concerned to make sure that the safety of people remains paramount in the ways the vended foods are produced and distributed.

In Vitro Evaluation of Phytobiotic Mixture Antibacterial Potential against Enterococcus spp. Strains Isolated from Broiler Chicken.

Enterococcus spp. are normal intestinal tract microflorae found in poultry. However, the last decades have shown that several species, e.g., Enterococcus cecorum, have become emerging pathogens in broilers and may cause numerous losses in flocks. In this study, two combinations (H1 and H2) of menthol, 1,8-cineol, linalool, methyl salicylate, γ-terpinene, p-cymene, trans-anethole, terpinen-4-ol and thymol were used in an in vitro model, analyzing its effectiveness against the strains E. cecorum, E. faecalis, E. faecium, E. hirae and E. gallinarum isolated from broiler chickens from industrial farms. To identify the isolated strains classical microbiological methods and VITEK 2 GP cards were used. Moreover for E. cecorum a PCR test was used.. Antibiotic sensitivity (MIC) tests were performed for all the strains. For the composition H1, the effective dilution for E. cecorum and E. hirae strains was 1:512, and for E. faecalis, E. faecium and E. gallinarum, 1:1024. The second mixture (H2) showed very similar results with an effectiveness at 1:512 for E. cecorum and E. hirae and 1:1024 for E. faecalis, E. faecium and E. gallinarum. The presented results suggest that the proposed composition is effective against selected strains of Enterococcus in an in vitro model, and its effect is comparable to classical antibiotics used to treat this pathogen in poultry. This may suggest that this product may also be effective in vivo and provide effective support in the management of enterococcosis in broiler chickens.

Antibiotic susceptibility of Streptococcus species that cause pharyngitis in children

Background: Group A Streptococcus (GAS) significantly impacts global health, especially among children aged 5–14 years, causing approximately 288 million episodes of sore throat annually and resulting in around 100,000 disability-adjusted life years (DALYs) each year. GAS infections are particularly prevalent in developing countries. Objective: To assess the patterns of antibiotic resistance of Streptococcus spp. that cause pharyngitis in children. Patients and Methods: It was collected 100 pediatric patients. It was taken throat swabs for Streptococcus species culture and antibiotic sensitivity test. In addition, it was recorded the data of patient’s age and gender. Results: It was shown in this study that an average age of 4.4 ± 3.7 years for pharyngitis in children. In this study was diagnosed that Streptococcus agalactiae (24%) and Streptococcus pyogenes (18%) as the common causes Streptococcus species causes pharyngitis in children. It was noted that the differences in bacterial species according to the patient’s ages. Antibiotic sensitivity testing determined that there are high sensitivity of S. pneumoniae and S. parasanguinis to multiple antibiotics. In addition, it was showed resistance of S. pyogenes to Metronidazole and Azithromycin. Conclusion: In this study, it was found that the risk age for pharyngitis in children is four years old. Streptococcus agalactiae and Streptococcus pyogenes are the common bacterial causes for this infection. In addition, it was shown that S. pneumoniae appeared a high sensitivity to Ceftriaxone, Ciprofloxacin, and Meropenem, and S. parasanguinis was sensitive to Azithromycin, Cefixime, and Cefotaxime, and S. pyogenes showed a high sensitivity to Meropenem and Imipenem.

Molecular Detection of Vancomycin Resistance Genes in Staph aureus Isolates from Different Clinical Specimens

The study screened the occurrence of VanA, VanB, Vanc genes in Staph aureus vancomycin-resistant isolates recovered in Hilla -hospitals. From various clinical samples, 160 isolates from patients between July and November of 2022 were included in the search. Among these samples are (65) burn samples, (55) urine samples, and (40) samples from Blood. 40 (or 25%) of the 160 isolates were Staph aureus. Characterization of the VanA, VanB, and VanC genes in Staph aureus isolates obtained from various specimens is the main goal of the work. The RESULTS of samples were obtained from a variety of clinical specimens, including 20 (40.625%) burn swabs, 10 (25%) urine samples, and 10 (34.375%) blood. Staph aureus was resistant to Cefepime (91.7%), Piperacillin (73.3%), and Ceftazidime (71.7%), although little relationship was detected with Aztreonam resistance (35%). 40 isolates were used for antibiotic sensitivity test (AST). Only 18.3% and 15% of isolates, respectively, were resistant to meropenem and imipenem. Fluoroquinolone resistance to lomefloxacin, ciprofloxacin, ofloxacin, and levofloxacin was 51.7%, 48.3%, 35%), and 30%, respectively. While the most prevalent vancomycin resistance genes were found in 40 isolates of S. aureus and were discovered by VanC 40 (100%), VanA 20 (100%), and VanB28 (70%), according to the current investigation. In Conclusion :The PCR-reaction method was discovered to be an easy-to-use tool for detecting staph aureus by pcr that can be regularly used for identifying staph aureus and detection the molecular resistance apparatuses to vancomycin in staph aureus by VanA, VanB, VanC), efflux systems plays To choose the best antibiotics, prevent the establishment of resistant strains, and create novel antimicrobial. medications, it is beneficial to research the particular resistance mechanisms vancomycin to staph aureus.

Epidemiology of antibiotic consumption and resistance in Mauritius

IntroductionThis study aims at determining the pattern of antibiotic consumption and resistance in Mauritius, a tropical island in the Indian Ocean.MethodologyAntibiotic consumption was measured in kilograms of purchased antibiotics and also in defined daily dose (DDD) in different health institutions from 2015 to 2017. Data on antibiotic resistance was collected at the Central Health Laboratory (CHL) at Victoria Hospital and at Jeetoo Hospital Laboratory, where antibiotic sensitivity testing is done for all public health institutions. For this study, Escherichia coli, Klebsiella species, Acinetobacter species, and Pseudomonas aeruginosa isolates from blood samples of patients from 2015 to 2023 were included. The resistance rate and prevalence of multi-drug-resistant (MDR) organisms were calculated.ResultsThe amount of antibiotics (in kilograms) distributed to the human sector was between 11,000 to 13,000 kg, compared to only 700 to 1,500 kg in the animal sector. The DDD per 1,000 inhabitants per day was 20.9, 22.1, and 21.7 in 2015, 2016, and 2017, respectively, with a greater consumption of WATCH and RESERVE group antibiotics in the private sector. In public health institutions, health centers in the northern region had the highest DDD per 1,000 outpatients per day for beta-lactams penicillins and quinolones. Concerning antibiotic resistance, the proportion of MDR Acinetobacter baumannii and Pseudomonas aeruginosa has increased from 58% to 74% and from 33% to 45%, respectively, from 2015 to 2023. During the same period, the proportion of E. coli and K. pneumoniae isolates sensitive to ceftriaxone decreased from 55% to 39% and from 37% to 22%, respectively, while the proportion of E. coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa isolates sensitive to meropenem decreased from 98% to 94%, 83% to 53%, 45% to 28%, and 63% to 47%, respectively.ConclusionThis study provides valuable insights on antibiotic consumption and resistance in the country and emphasizes the significance of adopting a One Health approach to combat antimicrobial resistance (AMR) effectively. These findings will aid policymakers in formulating targeted strategies to address the challenge of AMR and should be integrated into the National Action Plan on AMR in Mauritius.

Phylogenomic analyses and comparative genomics of Pseudomonas syringae associated with almond (Prunus dulcis) in California.

We sequenced and comprehensively analysed the genomic architecture of 98 fluorescent pseudomonads isolated from different symptomatic and asymptomatic tissues of almond and a few other Prunus spp. Phylogenomic analyses, genome mining, field pathogenicity tests, and in vitro ice nucleation and antibiotic sensitivity tests were integrated to improve knowledge of the biology and management of bacterial blast and bacterial canker of almond. We identified Pseudomonas syringae pv. syringae, P. cerasi, and P. viridiflava as almond canker pathogens. P. syringae pv. syringae caused both canker and foliar (blast) symptoms. In contrast, P. cerasi and P. viridiflava only caused cankers, and P. viridiflava appeared to be a weak pathogen of almond. Isolates belonging to P. syringae pv. syringae were the most frequently isolated among the pathogenic species/pathovars, composing 75% of all pathogenic isolates. P. cerasi and P. viridiflava isolates composed 8.3 and 16.7% of the pathogenic isolates, respectively. Laboratory leaf infiltration bioassays produced results distinct from experiments in the field with both P. cerasi and P. syringae pv. syringae, causing significant necrosis and browning of detached leaves, whereas P. viridiflava conferred moderate effects. Genome mining revealed the absence of key epiphytic fitness-related genes in P. cerasi and P. viridiflava genomic sequences, which could explain the contrasting field and laboratory bioassay results. P. syringae pv. syringae and P. cerasi isolates harboured the ice nucleation protein, which correlated with the ice nucleation phenotype. Results of sensitivity tests to copper and kasugamycin showed a strong linkage to putative resistance genes. Isolates harbouring the ctpV gene showed resistance to copper up to 600 μg/ml. In contrast, isolates without the ctpV gene could not grow on nutrient agar amended with 200 μg/ml copper, suggesting ctpV can be used to phenotype copper resistance. All isolates were sensitive to kasugamycin at the label-recommended rate of 100μg/ml.

Incidence of urinary infections and pharmacological sensitivity in elective urine samples in a prospective cohort from Luanda, Angola

This prospective cohort study evaluated 107 urine samples from patients suspected of urinary tract infection (UTI) treated at Cajueiros General Hospital in Angola from January to March 2023. Inclusion criteria involved obtaining urine samples for clinical examinations from adult patients with suspected UTI, regardless of gender, age, or previous clinical condition. The samples were transported to the reference laboratory of the National Institute of Health Research for microbiological evaluation and antibiotic sensitivity testing using the VITEK 2 system. Of the 107 patients evaluated, 39.3% (n= 42) showed bacterial growth in their urine. Escherichia coli was the predominant pathogen (23.8%), followed by Sphingomonas paucimobilis, Pseudomonas luteola, and Klebsiella pneumoniae. Antimicrobial resistance was observed, with ampicillin (90.5%) being the most resistant, while imipenem (95.2%) and ertapenem (90.5%) exhibited high sensitivity. Statistical analysis revealed common clinical characteristics, such as fever (61.7%) and dysuria (73.8%). This study provides a comprehensive understanding of microbial ecology in urinary infections, highlighting the prevalence of pathogens and antibiotic resistance patterns in a specific patient population in Angola. Importantly, this research represents the first prospective study of its kind in Angola.

PREVALENCE OF FLUOROQUINOLONES RESISTANCE AMONG ESBL-PRODUCING ESCHERICHIA COLI ISOLATED FROM URINE SAMPLES

BACKGROUND&#x0D; Resistance among uropathogens has emerged as a growing concern necessitating re-evaluation of the efficacy of recommended empiric antimicrobial regimens. Misuse and overuse of various antibacterial agents in hospitals is a key cause of the emergence of anti-microbial resistance.&#x0D; Aims: To identify the prevalence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli from urine specimens and to know the prevalence of Fluoroquinolone resistance among ESBL-producing Escherichia coli (E. coli) isolates.&#x0D; Material and method: The study was conducted on 500 E. coli isolates from urine samples received in the Department of Microbiology, MMIMSR, Mullana, Ambala. The organism isolation and identification were done as per the standard procedures and antibiotic sensitivity testing was done following CLSI guidelines. All the strains were screened out for ESBL production as per CLSI guidelines. Ciprofloxacin, Norfloxacin, and Ofloxacin discs were used for the detection of fluoroquinolone resistance.&#x0D; Results: Out of the total 500 E. Coli isolates from urine samples received in our laboratory, 69% were found to be ESBL producers.&#x0D; Conclusion: We found a high level of quinolone resistance among ESBL-producing Escherichia coli strains isolated from patients with UTI. Because of the widespread use of fluoroquinolones in our country, there is a need for sensitive antibiotic stewardship. Further research is needed to ascertain the gravity of quinolone resistance and to swiftly act against its spread among other nosocomial pathogens.&#x0D;

Overexpression of lasB Gene in Klebsiella pneumoniae and its Effect on Biofilm Formation and Antibiotic Resistance

Background: Klebsiella pneumoniae is the second most frequent pathogenic bacterium in the Enterobacteriaceae family, after E. coli. It is also regarded as a major pathogen responsible for healthcare-associated infections around the world. Objective: To look at how overexpressing the elastase gene (lasB) in K. pneumoniae affects biofilm development and antibiotic resistance. Methods: 25 clinical isolates of K. pneumoniae were received from Baghdad's Chemistry Analysis Center (CAC) and re-identified using the Vitek-2 method. The microtiter plate was used to measure biofilm production with ELISA. The disc diffusion method was used in antibiotic sensitivity tests in accordance with the CLSI 2022 criteria. The PlasB plasmid was transformed into K. pneumoniae via electroporation. Results: Out of 25 isolates, 11 (44%), 11 (44%), and 3 (12%) produced strong, moderate, and weak biofilms, respectively. One strong biofilm producer (KA1) was chosen for further investigation. The lasB plasmid was successfully transformed into KA1, yielding the KA1 (plasB) isolate. KA1 (plasB) formed considerably fewer biofilms than KA1, and it was more susceptible to tetracycline, doxycycline, and amoxicillin-clavulanic acid than KA1. Furthermore, KA1 (plasB) has shown a significant decrease in ampicillin resistance and an increase in ciprofloxacin sensitivity, but no variations in susceptibility to levofloxacin, cefotaxime, piperacillin-tazobactam, amikacin, or erythromycin when compared to KA1. Conclusions: Overexpression of the elastase gene (plasB) has a major impact on biofilm development and antibiotic resistance in K. pneumoniae.

Management of seed borne ralstonia solanacearum in brinjal (Solanum melongena L.) and tomato (Lycopersicon esculentum Mill.)

Total of 165 bacterial isolates of Ralstonia solanacearum were collected from different seed samples of tomato and brinjal in various seed sources and 22 isolates were tested to find out the management tools among different management approaches. Hundred percent reduction of bacterial incidence was found with Gentamicin (0.1, 0.05%), Erythromycin (0.1, 0.05%) and Doxycycline (0.1%) over control in both brinjal and tomato treated seeds in nutrient agar medium. Seed treatment with BAU-Biofungicide (3%) exhibited 83.61% decreases of bacterial incidence over control in tomato and 90.28% reduction in brinjal seeds. Garlic also showed better performance in controlling R. solanacearum among the plant extracts. Antibiotic sensitivity test revealed that maximum number of isolates of R. solanacearum was found to be sensitive to Gentamicin and Erythromycin, and the highest number of isolate of R. solanacearum of tomato and brinjal seeds marked resistant to Doxycycline. Higher germination 40.38% was achieved over control in tomato seed and 26.67% germination was in brinjal seed when the seeds were treated with BAU-Biofungicide (3%). Among all the treatments, BAU-Biofungicide (3%) also resulted in the highest increase of vigor index (77.36%) in tomato and brinjal seed with an increase of 69.91% over untreated control. Bangladesh J. Bot. 53(1): 173-183, 2024 (March)

Unveiling the High Prevalence of Antibiotic Resistance and Quorum Sensing Genes in Uropathogenic &lt;i&gt;Escherichia coli&lt;/i&gt;

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.

Detection of Uropathogenic Specific Protein Gene (usp) and Multidrug Resistant Bacteria (MDR) of Pathogenic Escherichia coli Isolated from Baghdad City

Escherichia coli is gram negative bacteria and represents a typical resident of the digestive systems of both humans and animals. The stability and equilibrium of the luminal microbial flora are significantly influenced by E. coli. The Escherichia coli uropathogenic-specific protein (Usp) represents type of genotoxins produced by uropathogenic E. coli rather than fecal E. coli isolates. In the current study E. coli was isolated from urine and stool and usp gene was detected in it. Sensitivity test was evaluated by using different types of antibiotics and the usp gene was detected by PCR in all bacterial isolates. Antibiotics sensitivity test showed variable degrees of sensitivity and resistance. High percentage of sensitivity was achieved against amikacin (86%) and that for ciprofloxacin, ceftazidime, gentamicin and tobramycin it was 80%. Whereas trimethoprim and aztreonam showed 64% and 60% sensitivity respectively. In this study, most isolates were resistant to amoxicillin 92%; while they showed different degrees of resistance against other types of antibiotics (from tetracycline 62% to amikacin 4%). The frequency of multi drug resistant (MDR) bacteria was about 64 % (32 isolates), 30 from urine and 2 from the stool. The results showed that usp gene was found in 26 bacterial isolates (52%), whereas other 24 (48%) isolates didn’t have this gene. It was concluded that source of bacterial isolates carry usp gene was urine, with the exception of one isolate from f stool, and E. coli recorded as multidrug resistant bacteria (MDR).

In vitro ANALYSIS OF BIOFILM FORMATION AND ANTIBIOTIC RESISTANCE OF UROPATHOGENIC Escherichia coli

Urinary tract infections are an exceedingly common worldwide problem, caused mostly by Gram-negative bacteria, especially Escherichia coli. Microbial biofilms are considered a serious public health problem. The potential of uropathogenic E. coli (UPEC) to produce biofilm was explored in the present study. The Congo red agar, tube- and tissue culture plate methods were used to evaluate the formation of biofilm by E. coli isolates. Of the 155 isolates, 101, 106 and 90 isolates were positive for these three methods, respectively. Subsequently, the sensitivity of E. coli isolates to antimicrobial agents (amikacin, cefepime, cefixime, cefotaxime, cefpodoxime, ceftazidime, ceftriaxone, ciprofloxacin, nitrofurantoin, gentamycin, nalidixic acid, ofloxacin and pefloxacin) was tested. There was no difference in the rate of biofilm detection between Congo red agar method and tube method. The antibiotic sensitivity test revealed that the biofilm producing isolates were multi-drug resistant. The study emphasized the necessity for developing alternative therapeutic approaches to overcome multi drug resistance arising from biofilm formation of UPEC.

Modulating the antibacterial effect of the existing antibiotics along with repurposing drug metformin.

Owing to the extensive prevalence of resistant bacteria to numerous antibiotic classes, antimicrobial resistance (AMR) poses a well-known hazard to world health. As an alternate approach in the field of antimicrobial drug discovery, repurposing the available medications which are also called antibiotic resistance breakers has been pursued for the treatment of infections with antimicrobial resistance pathogens. In this study, we used Haloperidol, Metformin and Hydroxychloroquine as repurposing drugs in in vitro (Antibacterial Antibiotic Sensitivity Test and Minimum Inhibitory Concentration-MIC) and in vivo (Shigellosis in Swiss albino mice) tests in combination with traditional antibiotics (Oxytetracycline, Erythromycin, Doxycycline, Gentamicin, Ampicillin, Chloramphenicol, and Penicillin) against a group of AMR resistance bacteria (Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Shigella boydii). After observing the results of the conducted in vitro experiments we studied the effects of the above non antibiotic drugs in combination with the said antibiotics. As an repurposing adjuvant antibiotic drug, Metformin exhibited noteworthy activity in almost all in vitro, in vivo and in silico tests (Zone of inhibition for 30 to 43mm for E.coli in combination with Doxycycline; MIC value decreased 50 µM to 0.781 µM with Doxycycline on S. boydii).In rodents Doxycycline and Metformin showed prominent against Shigellosis in White blood cell count (6.47 ± 0.152 thousand/mm3) and Erythrocyte sedimentation rate (10.5 ± 1.73mm/hr). Our findings indicated that Metformin and Doxycycline combination has a crucial impact on Shigellosis. The molecular docking study was performed targeting the Acriflavine resistance protein B (AcrB) (PDB ID: 4CDI) and MexA protein (PDB ID: 6IOK) protein with Metformin (met8) drug which showed the highest binding energy with - 6.4kcal/mol and - 5.5kcal/mol respectively. Further, molecular dynamics simulation revealed that the docked complexes were relatively stable during the 100 ns simulation period. This study suggest Metformin and other experimented drugs can be used as adjuvants boost up antibiosis but further study is needed to find out the safety and efficacy of this non-antibiotic drug as potent antibiotic adjuvant.

Molecular identification and characterization of endophytic bacteria from root of chamaecrista rotundifolia growing on the nigerian air force shooting range, kaduna.

Chamaecrista rotundifolia is one of the few plant species growing on heavy metal-contaminated Nigerian Air Force shooting range, Kaduna. This microbially unexplored plant species can have microbial association with endophytes. This study was aimed at isolating, identifying and characterizing endophytic bacteria associated with the roots of Chamaecrista rotundifolia growing on the Nigerian Air Force shooting range, Kaduna. The identification of the endophytic bacteria was done using standard biochemical methods and 16S rRNA sequencing analysis. The isolates were then screened for antibiotic resistance, plant growth promoting properties and heavy metal tolerance using standard techniques. Varied concentrations of heavy metal salts (Pb2+, Cr3+, Ni2+ and Mn2+) were used to test the isolates for their heavy metal tolerance. Four endophytic bacteria isolated from the roots of C. rotundifolia were Aerococcus viridans, Peribacillus simplex, Staphylococcus simplex and Pseudomonas aeruginosa. Results on antibiotic sensitivity test revealed that the four isolates were resistant to most of the antibiotics. The isolates possessed plant growth promoting properties by showing positive results for Indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylic (ACC) deaminase activity and phosphate solubilizing. Results also revealed that the isolates showed some degree of tolerance to the heavy metals. The tolerance of both A. viridans and P. simplex to chromium and manganese showed significant difference (p&lt;0.05) across all the concentration levels of the metal salts. It is therefore concluded that Chamaecrista rotundifolia growing on Nigerian Air Force shooting range, Kaduna, is associated with endophytic bacteria which could be the reason the plant species thrive in heavy metal-contaminated site.&#x0D; &#x0D;