Abstract
The aim of this study was to investigate multi-drug-resistant (MDR) Escherichia coli in urine of adult male patients with enlarged prostate. Three hundred and sixty-eight samples of urine and blood were collected. Escherichia coli was isolated, purified, and identified and prostate-specific antigen (PSA) was determined. Multi-drug resistance test and specific drug resistance genes were assessed. Prevalence of Escherichia coli was high (38.5%) in patients with PSA of 60–79 ng ml−1 and 60% were MDR. The isolates showed highest resistance to tetracycline (53.3.0%) and least to cephalosporins (5%). They had intL and gyrA genes, which are integron, and quinolone resistance genes and sul1 and sul2 which are sulphonamide resistance–associated genes. Levofloxacin, ertapenem, and Augmentin (100% susceptibilities) were considered choice drugs for treatment of Escherichia coli infection in patients with elevated PSA.
Supplementary Information
The online version contains supplementary material available at 10.1007/s42770-024-01260-x.
Keywords: Prostate-specific antigen, Antibiotic resistance genes, Escherichia coli, MDR
Introduction
Multi-drug-resistant Escherichia coli is an E. coli non-susceptible to at least one agent in three different classes of antimicrobial agents, excluding the broad-spectrum penicillin without a β-lactamase inhibitor [1]. Uropathogenic Escherichia coli (UPEC) is one of the main bacteria causing urinary tract infections (UTIs) [2]. The rates of UPEC with high resistance towards antibiotics and multi-drug-resistant bacteria have increased dramatically in recent years and this could make treatment difficult [3].
Prostatitis and prostate hyperplasia are diseases that commonly occur during men’s lifespan all over the world, and 35–50% of men have prostatitis at some time in their lives [4]. Microbial imbalance is closely associated with the initiation and development of prostate diseases, such as prostate cancer (PCa) and benign prostatic hyperplasia (BPH) [5, 6]. Elderly males, in the peak age group with prostate enlargement, are more likely to experience severe infection [7]. The emergence of drug-resistant microorganisms among UPEC strains increases the serious threat to global health [8].
The present study determined the multi-drug-resistant Escherichia coli in the urine of adult male patients with enlarged prostate attending general hospitals in Benue state Nigeria.
Materials and methods
Study sample
In total, 368 urine and 368 blood samples were collected from 368 patients from 23 general hospitals in Benue state. The samples were collected from adult male patients forty (40) years and above with bladder outlet obstruction secondary to benign prostatic hyperplasia or prostate cancer. The samples were collected between November 2021 and April 2022.
Sample size was determined using the formula of Charan and Biswa [9].
Blood sample
The blood samples were separated and dispensed into “cryo” vials and ice-packed, and plasma was analyzed for prostate-specific antigen (PSA) using Finecare FIA meter (MNCHIP Chemistry Analyzer, Model: Pointcare M4; Serial No: PM2110001Z02 35,287, China).
Quantitative determination of prostate-specific antigen using Finecare FIA meter (ELISA analyzer)
The blood samples were spun using a bucket centrifuge at 3000 rpm for 10 min and using an automated pipette, 75 µl of the plasma samples was pipetted into each of the buffer tubes and gently inverted 10 times. Thereafter, 75 µl of the mixture was transferred into pre-labeled PSA cartridges coated with prostate-specific antibodies. Each of the cartridges was loaded into the Finecare ELISA analyzer and allowed to run for 15 min. The results were displayed on the screen as quantitative values in ng ml−1 with respective labels. Each Finecare™ PSA rapid quantitative test contains internal control that satisfies routine quality control requirements. The internal control was performed each time a patient sample was tested. An invalid result from the internal control caused an error message on the Finecare™ FIA system indicating that the test should be repeated [10]. Additional in-house control was used in the determination of PSA which was a sample previously ran in Ichroma II (Boditech Med. Inc. with Serial number- IR22K057900).
Urine sample
The urine was inoculated on cystine-lactose-electrolyte-deficient (CLED) agar (Oxoid, CM 0398) and incubated at 37 °C for 24 h. Colonies were sub-cultured repeatedly on eosin methylene blue (EMB) agar (Oxoid, CM 0069) to obtain pure cultures. Escherichia coli was isolated, purified, and identified by morphological, biochemical, and molecular characteristics as reported by Iwodi [11]. Escherichia coli isolates were streaked onto nutrient agar slants and incubated at 37 °C for 18 h. They were stored in the refrigerator at 4 °C in agar slants in bijou bottles until used. The isolates were sub-cultured monthly on fresh medium to maintain viability.
Identification of the isolates
Colonies of Escherichia coli on CLED were identified by colonial morphology, Gram staining, motility test, oxidase test, citrate test, triple sugar iron agar, and indole test.
Molecular identification of Escherichia coli using polymerase chain reaction (PCR)
Successive washing in nuclease-free water and centrifugation as described by Sambrook and Russell [12] were used for the extraction and purification of bacterial DNA for Escherichia coli identification. Polymerase chain reaction (PCR) was carried out to amplify the 16SrRNA of the Escherichia coli gene using the primer pair F: CGTGAT CAGCGG TGA CTA TGA C and R: CGATTCTGG AAATGG CAAAAG. The PCRreaction was carried out using the Solis BioDyne ready-to-load master mix (5x). Polymerase chain reaction was performed in 25 µl of a reaction mixture. The reaction concentration was brought down from 5 × concentration to 1 × concentration containing 1 × blend master mix buffer (Solis Biodyne). One and a half (1.5 mM MgCl2, 200 µMol) of each deoxynucleoside triphosphates (dNTP) (Solis Biodyne), 25 pMol of each primer (BIOMERS, Germany), 2 units of Hot FIREPol DNApolymerase (Solis Biodyne), proof-reading enzyme, 5 µl of the extracted DNA, and sterile distilled water were used to make up the reaction mixture. Thermal cycling was conducted in a TECHNE 3 Prime thermal cycler for an initial denaturation of 95 °C for 5 min. Thereafter followed by denaturation at 95 °C for 30 s, annealing at 61 °C for 1 min (the annealing temperature is determined by the primer used), and extension at 72 °C for 2 min, and final extension step of 10 min at 72 °C. The total number of cycles was 35 cycles.
Antimicrobial susceptibility testing
Escherichia coli isolates were subjected to antimicrobial susceptibility test using the standard disc diffusion method described by Ochei and Kolhatkar [13] and results were interpreted using the criteria of the Clinical Laboratory Standard Institute [14]. The antibiotic-impregnated discs (MASTDISC UK) used were ceftriaxone (CRO 30 µg), streptomycin (S 30 µg), ampicillin (AP 10 µg), Augmentin (AUG 30 µg), ciprofloxacin (CIP 5 µg), ofloxacin (OFX 5 µg), levofloxacin (LEV 5 µg), tetracycline (T 30 µg), gentamicin (GM10 µg), and ertapenem (ETP 10 µg). Escherichia coli ATCC 25922 was the control bacterium.
Identification of multi-drug-resistant (MDR) Escherichia coli isolate
The Escherichia coli isolates were identified as multi-drug-resistant following the combined guidelines of the European Center for Disease Prevention and Control (ECDC), the Center for Diseases Control and Prevention (CDC), and the Guidelines for Clinical Laboratory Standard Institute [14]. The isolates were defined as multi-drug-resistant (MDR) when they showed resistance to ≥ 1 agent in ≥ 3 antibiotic classes [14].
Extended spectrum β-lactamase (ESBL) production in Escherichia coli isolates
The MDR isolates were separately inoculated on Mueller–Hinton agar plates. Antibiotic discs of ceftazidime (30 µg) and ceftriaxone (30 µg) were placed at 35 mm equidistance to a central disc of amoxicillin-clavunate (30 µg). Inoculated plates were incubated at 37 °C for 24 h. The test isolates were confirmed as ESBL producers if the zone of inhibition around any of the antibiotic discs increased towards the amoxicillin/clavulanic acid disc or if a difference of ≥ 5 mm was observed between the zone diameters of either of the cephalosporins discs and their respective cephalosporin/clavulanic acid discs [15].
Extraction of DNA for the detection of some resistance genes in the isolates
Extraction of DNA for the detection of resistance genes in the E. coli isolates was done according to the method of Anjum et al. [16] as reported below.
Multiplex PCR for the detection of some Escherichia coli resistance genes: determination of some genes responsible for multi-drug resistance
The isolates were subjected to multiplex PCR using specific primers for different enzymes such as ESBL, carbapenemase, aminoglycoside, fluoroquinolones/quinolones resistant-conferring enzymes, methicillinase, oxacillinases, specific porins, and efflux pumps [17]. Multiplex PCR amplification was carried out in a 100-µl volume, comprising approximately 40 ng of template DNA, 10 pmol of each of the primers, a final concentration of 0.4 mM each of deoxyribonucleoside triphosphate, and 5 U of Taq DNA polymerase (Amersham Pharmacia Biotech) in 1 × PCR buffer; the MgCl2 final concentration in the PCR mixture was adjusted to 4 mM. The PCR cycles included initial denaturation at 94 °C for 3 min, followed by 30 cycles of amplification at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s (except for the final cycle, which had an extension step of 4 min). The final PCR products were analyzed on a 1.5% agarose gel electrophoresis [17].
Agarose gel electrophoresis for the resistance gene PCR products
The amplification product was separated on a 1.5% agarose gel with Ethidium bromide stain (Carl Roth, Germany) and electrophoresis was carried out at 80 V for 1 h. Agarose (1.5 g) was dissolved in Tris–acetate EDTA (TAE) buffer (100 ml) and heated to boil, to completely dissolve. The gel was allowed to cool to 45 °C and poured into an electrophoresis tray. A casting comb was placed at the end of the tray. The casting comb was removed after the agarose had completely solidified. Electrophoresis was run at 80 V for 1 h. After electrophoresis, DNA bands were visualized and captured under a UV transilluminator (Biobase, China). A 100-bp DNA ladder (Solis Biodyne, Estonia) was used as the DNA molecular weight standard [18].
Primer sequences and PCR conditions for the detection of some resistance genes in Escherichia coli (multiplex PCR)
The primer sequences and PCR conditions for the molecular identification of some of the resistance genes in the Escherichia coli isolates are shown in Supplementary Table 1. They included primers for genes qnrB, qnrS, gyrA, oxqxB, sul1, sul2, cmlA, strA-strB, aac1, tetA, tetB, nsA, nfsB Tolc, Int1, blaTEM, bla VIM, blaOXA, and aac(6´) cr.
Plasmid DNA extraction and purification
Norgen Biotek Plasmid DNA kit (Cat. 13,300) was used and the extraction protocol was based on the manufacturer’s recommendation and as reported [19].
Agarose gel electrophoresis for plasmid
The agarose gel electrophoresis for the plasmid was essentially run as described above for the genomic DNA and was preformed according to Robicsek [20].
Statistical analysis
Statistical analyses were done using SPSS version 17 (2008). Pearson’s chi-square test was used to determine associations between variables at 95% confidence level with p ≤ 0.05 being a statistically significant relationship between two or more variables.
Results
The prevalence of Escherichia coli and their multi-drug-resistant isolates with respect to PSA level is presented in Table 1. The highest Escherichia coli isolation rate was among patients within the PSA range of 60–79 ng ml−1 at 5 (38.5%). Patients with PSA 40–49 ng ml−1 had the least Escherichia coli isolation rate of 2 (6.3%). For multi-drug resistance, the highest MDR Escherichia coli was found in patients with PSA levels of 60–79 ng ml−1. They had an MDR rate of 3 (60.0%), followed by patients with PSA 0–19 ng ml−1 with multi-drug-resistant rate of 8 (47.1%). Patients with PSA 40–59 ng ml−1 had no MDR Escherichia coli.
Table 1.
Isolation rate of Escherichia coli and its multi-drug resistance with respect to prostate-specific antigen (PSA) level
| PSA (ng/ml) | Urine examined | Positive (%) | Negative (%) | Isolate | MDR positive (%) | MDR negative (%) |
|---|---|---|---|---|---|---|
| 0–19 | 182 | 17 (9.3) | 165 (90.3) | 17 | 8 (47.1) | 9 (52.9) |
| 20–39 | 53 | 11 (20.8) | 42 (97.2) | 11 | 2 (18.2) | 9 (81.8) |
| 40–59 | 32 | 2 (6.3) | 30 (93.7) | 2 | 0 | 2(100) |
| 60–79 | 13 | 5 (38.5) | 8 (61.5) | 5 | 3 (60.0) | 2 (40.0) |
| 80–99 | 20 | 7 (35.0) | 13 (65.0) | 7 | 2 (28.6) | 5 (71.4) |
| ≥ 100 | 68 | 18 (26.5) | 50 (73.5) | 18 | 3 (16.7) | 15 (83.3) |
| Total | 368 | 60 (16.3) | 308 (83.7) | 60 | 18 (30.0) | 42 (70.0) |
The antibiotic susceptibility profile of Escherichia coli is presented in Table 2. One hundred percent of the isolates were susceptible to Augmentin, levofloxacin, and ertapenem. Ninety-five percent were susceptible to ceftriaxone and ceftaxidime. Ninety percent were susceptible to streptomycin, whereas 88.3%, 70%, 55%, 51.7%, and 46.7% were susceptible to gentamycin, ofloxacin, ampicillin, ciprofloxacin, and tetracycline respectively.
Table 2.
Antibiotics profile of Escherichia coli of adult male patient with prostate enlargement attending general hospitals in Benue state
| Antibiotic concentration (µg) | Sensitive (%) | Resistant (%) | Chi-square | p-value |
|---|---|---|---|---|
| Ceftriaxone (30) | 57 (95.0) | 3 (5) | 15.527 | 0.004 |
| Ceftaxidime (30) | 57 (95.0) | 3 (5) | 15.527 | 0.004 |
| Ampicillin (10) | 33 (55) | 27 (45) | 149.574 | 0.001 |
| Augmentin (30) | 60 (100) | 0 | ||
| Ciproxin (5) | 31 (51.7) | 29 (48.3) | 161.602 | 0.001 |
| Ofloxacin (5) | 42 (70) | 17 (28.3) | 97.152 | 0.001 |
| Levofloxacin (5) | 60 (100) | 0 | ||
| Tetracycline (30) | 28 (46.7) | 32 (53.3) | 179.911 | 0.001 |
| Gentamycin (10) | 53 (88.3) | 7 (11.7) | 36.63 | 0.001 |
| Streptomycin (30) | 54 (90) | 6 (10) | 31.31 | 0.01 |
| Eterpenem (10) | 60 (100) | 0 |
Determination of specific MDR genes was carried out. The presence of β-lactam-associated gene (blaTEM) was shown by clear gene bands at 431 bp (Supplementary Plate 1). Isolates 1 (OB13), 3 (BU12), 5 (GJ 15), 6 (AL2), 7 (NK5), 8 (TA3), 9 (TA10), 10 (AD5), 13 (OT14), 14 (OJ 13), 15 (WA6), and 17 (VA11) show band to blaTEM gene at 431 bp. Lane 2 (BU4), 4 (GJ8), 11 (UA3), 12 (MK7), 16 (VA3), and 18 (MK4) are all negative to the ESBL gene as there is no band. Carbapenem-hydrolyzing genes (blaVIM and OXA) showed that gene (blaVIM) was present at 247 bp in the MDR isolate and OXA at 585 bp (Supplementary Plate 2). Sample 3 (BU4) showed band to VIM at 247 bp. The rest showed no band. Sample 9 (TA10) showed a band to blaOXA at 585 bp; the rest showed no band and did not possess those genes. Figure 1 is a representative image of an agarose gel electrophoresis showing the presence of the integron-1 gene. There were bands in 17 lanes. This showed the presence of the integron-1 gene in the isolates at 280 bp.
Fig. 1.
Representative image of an agarose gel electrophoresis showing integron-1 in MDR Escherichia coli isolates. Key: M = DNA ladder, 1–18 E. coli isolates, − ve = control (PCR reaction without DNA)
The presence of nitrofurantoin-resistant genes (nfsA and nfsB) in the MDR Escherichia coli is shown by the gene bands of nfsA at 1036 bp and nfsB at 932 bp (Supplementary Plate 3). Isolate 10 (AD5) possesses nfsA at 1036 bp and nfsB at 932 bp. The rest isolates were negative to nfsA and nfsB. Supplementary Plate 4 is a representative image of an agarose gel electrophoresis showing the presence of quinolone resistance gene B (qnrB) (at gene band 405 bp), quinolone resistance gene S (qnrS) (456 bp), and quinolone efflux pump protein (oqxB). The presence of streptomycin-associated genes (strA and strB) in the isolates was determined (Supplementary Plate 5) with gene bands at 891 bp. Figure 2 is a representative image of an agarose gel electrophoresis showing the presence of sulphonamide resistance–associated genes (sul1 and sul2). Isolates in lanes 2 (BU4), 3 (BU12), 4 (GJ8), 5 (GJ15), 6 (AL2), 7 (NK5), 8 (TA3), 10 (AD5), 11 (UA3), 13 (OT14), 14 (OJ13), 15 (WA6), 16 (VA3), 17 (VA11), and 18 (MK4) harbored sul 1 at 433 bp. Isolates in lanes 3 (BU12), 4 (GJ8), 5 (GJ15), 6 (8A2), 7 (NK5), 8 (TA3), 9 (TA10), 14 (0J13), 15 (WA6), and 16 (VA3) harbored sul 2 at 293 bp and isolates in lanes 3 (BU12), 4 (GJ8), 5 (GJ15), 6 (AL2), 7 (NK5), 8 (TA3), 14 (OJ13), 15 (WA6), and 16 (VA3) harbored both sul 1 and sul 2 at 433 and 293 bp respectively.
Fig. 2.
Representative image of an agarose gel electrophoresis showing sulphonamide resistance–associated genes in MDR Escherichia coli isolates. Key: M = DNA ladder, 1–18 E. coli isolates, − ve = control (PCR reaction without DNA), Sul 1 and sul 2 = sulfonamide resistance–associated genes
The presence of tetracycline resistance–associated genes (tetA and tetB) was determined in the isolates (Supplementary Plate 6). Isolates in lanes 1 (OB13), 3 to 9 (BU12, GJ8, GJ15, AL2, NK5, TA3 to TA10), 11 to 14 (UA3, MK7, OT14, OJ13), 16 (VA3), and 18 (MK4) harbored tetA resistance gene at 210 bp. Lanes 10 (AD5), 15 (WA6), and 17 (VA11) harbored resistance gene tetB at 659 bp. Supplementary Plate 7 is a representative image of an agarose gel electrophoresis showing the presence of a band in lane 3 (GJ8) which indicated that it harbored the aac1 resistance gene at 873 bp (gentamicin resistance–associated gene). Isolates in lanes 5 (GJ15), 9 (TA10), and 14 (OJ13) harbored aac(6)cr resistance gene (gentamicin resistance gene) at 482 bp. The absence of a band in the remaining lanes indicated the absence of aac1 and aac(6)cr genes. Quinolone resistance–associated gene was determined in the multi-drug-resistant Escherichia coli (Fig. 3). Bands were observed in isolates 2 (OB13 and BU4), 4 to 14 (GJ8, GJ15, AL2, NK5, TA3, TA10, AD5, UA3, MK7, OT14, OJ13), and 16 to 18 (VA3, VA11, MK4) which indicated possession of gene gyrA at 311 bp.
Fig. 3.
Representative image of an agarose gel electrophoresis showing quinolone resistance–associated gene (gyrA) in MDR Escherichia coli isolates. Key: M = DNA ladder, 1–18 E. coli isolates, − ve = control (PCR reaction without DNA), gyrA = quinolone resistance–associated gene
Supplementary Plate 8 is a representative image of an agarose gel electrophoresis showing efflux pump genes (tolC, arcB, and cmLA) in the isolates. Areas of intense bands observed in lanes 3 (BU12), 5 (GJ15), 7 (NK5), 11 (UA3), 12 (MK7), 15 (WA6), and 17 (VA11) indicated resistance gene to arcB at 761 bp. The band observed in lane 11 (UA3) showed the presence of the resistance gene tolC at 1170 bp while lane 5 (GJ15) had the resistance cmlA gene at 212 bp. Lane 5 (GJ15) had both arcB at 761 bp and cmlA gene at 212 bp. Bands observed in lanes 1–18 (OB13, BU4, BU12, GJ8, GJ15, AL2, NK5, TA3, TA10, AD5, UA3, MK7, OT 14, OJ13, WA6, VA3, VA11, and MK4) indicate the presence of plasmid in the MDR Escherichia coli isolates at ˃2322 to 23,190 bp (Supplementary Plate 9).
Discussion
In this study, from the 368 urine samples analyzed, 16.3% had Escherichia coli. This slightly differs from the report of Menyfa et al. [21] in the emergency department of KAMC in Riyadh, Saudi Arabia. They reported a higher percentage of Escherichia coli (60.24%).
Sixty percent of the patients with PSA 60–79 ng ml−1 had multi-drug-resistant Escherichia coli. This was the highest and the least being patients with PSA level of ≥ 100 ng ml−1 with an MDR of 16.7%. There was a significant difference in the multi-drug-resistance rate (p < 0.05) among other patients with varying degrees of PSA.
The isolates showed 53.3% resistance to tetracycline which is in line with the findings of Ibrahim et al. [22]. They recorded a 77.1% resistance rate of Escherichia coli to tetracycline in Escherichia coli isolates from a hospital in Khartoum State, Sudan. The implication of this is that patients who will use this antibiotic will not respond well to treatment of infections caused by Escherichia coli even when other antibiotics from the same class (doxycycline) are used. Sapadin and Fleishihmajer [23] stated that resistance to tetracycline by E. coli is worrisome as it is a naturally occurring drug that is used to treat a wide range of illnesses.
Ampicillin resistance rate was 45%. This result differs from the work done by Sabina and Aida [24], University of Sarajevo, who observed high resistance to Escherichia coli for ampicillin (82.79%); also, Habibi and Khameneie [25] gave a similar report that uropathogenic Escherichia coli isolates showed resistance to ampicillin at the rate of 96.42%, tetracycline 85.71%, amikacin 71.42%, ciprofloxacin 67.85%, and gentamycin 58.71% and have been found in pregnant women with history of recurrent urinary tract infections. In this study, all the Escherichia coli isolates 60 (100%) were susceptible to levofloxacin a fluoroquinolone and ertapenem a carbapenem and Augmentin a β-lactam. This agrees with the finding of Idil et al. [26] who obtained a total of 1123 Escherichia coli strains from urine and reported that among other antibiotics, imipenem represents the best efficient antibiotic against all uropathogenic Escherichia coli strains (100%), followed by ertapenem (99.98%), amikacin (99.94%), and nitrofurantoin (99.91%). This makes them the drugs of choice for the treatment of Escherichia coli in patients with enlarged prostate in an area where facility for isolation, identification, and determination of susceptibility pattern is difficult especially in low-resource settings.
Results of the gene amplification by PCR revealed that 94.4% of the isolates harbored intL and gyrA genes, which were integron and quinolone resistance genes. This explains the resistance of the isolates to fluoroquinolones. Other genes that the isolates had at a high rate were sul1and sul2 at 83.3%. This was higher than the report of David et al. [27] who reported a lower percentage of 45.5%.
The multi-drug-resistant Escherichia coli had 77.8% of tetA gene, which codes for tetracycline resistance–associated gene. This result is similar to the work of Margareta et al. [28] who reported that over half the isolates encoding a single determinant were positive for tetA 26% and tetB 32% as well as tetC, tetD, and tetM each with 4%. A 66.7% of the E. coli isolated harbored blaTEM which codes for β-lactam resistance–associated gene. This is in line with the report of Wassan et al. [29], who reported that Escherichia coli and Klebsiella pneumoniae (25%) harbored extended-spectrum-β-lactam producers’ gene and that the prevalence of blaTEM, blaSHV, and blacTXM genotypes was identified as 55, 35, and 45% respectively.
The presence of qnrS (38.9%), qrcB (38.9%), and oqxB at 33.3% in the E. coli calls for concerted effort by all stakeholders. On qnrS, 38.9% was reported in this study as against 2.9% reported by Maryam et al. [30]. This means that the rate of qnrS gene is increasing. Tarchouna et al. [31] reported a frequency of 32% for the qnr genes: 12.5% for qnrB, 5.3% for qnrA, 3.5% qnrS, and 2.6% for both qnrS and qnrB and 3.5% for both qnrA and qnrB genes. The implication of the presence of various resistance genes is that even newer drugs may become resistant shortly as most of these pathogenic organisms can share multi-drug resistance genes through horizontal or vertical gene transfer.
Conclusions
E. coli prevalence was high (38.5%) in patients with PSA of 60–79 ng ml−1 and 60% were MDR. They showed the highest resistance to tetracycline (53.3.0%) and harbored intL, gyrA, sul1, and sul2 genes. They were 100% susceptible to levofloxacin, ertapenem, and Augmentin and were considered choice drugs for the treatment of Escherichia coli infection in patients with elevated PSA.
Supplementary Information
Below is the link to the electronic supplementary material.
Author contribution
In a round table meeting, IC, GMG, IOO, and EOA conceptualized the framework for the study. IC performed the experiment and wrote a sketch of the work. IOO and GMG performed the initial proof-reading of the manuscript and all other things were mutually carried out by IC, GMG, IOO, and EOA including the final approval of the manuscript and decision on submission.
Data Availability
All data relating to this publication are freely available with either the paper or with the corresponding author.
Declarations
Ethics approval and consent to participate
Ethical approval was obtained from the ethical committee of the Benue State Hospitals Management Board (BSHMB) (HMB/OFF/215/VOL.II/453). Ethical approval certificate is available with the corresponding author and ready to be presented anytime upon request.
Consent for publication
Not applicable.
Conflict of interest
The authors declare no competing interests.
Footnotes
Responsible Editor: Beatriz Ernestina Cabilio Guth
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