MacConkey Research Articles

XDR Salmonella- An Emerging Threat in Swabi

Objective: To highlight the threat to effective treatment in more adversely affected areas like Swabi, with XDR Salmonella and stringent implementation of preventive measures to curb the spread of the pathogen.
 Methodology: This was a retrospective study conducted from August, 2022 to July, 2023 at Microbiology Laboratory MTI- GKMC/BKMC, Swabi. A total of 2996 blood cultures from patients were submitted at the Microbiology Department, Swabi. All samples were incubated in Bactec 9120 and processed as per standard guidelines, followed by being subcultured on Blood and Mac Conkey agar. On obtaining the non- lactose fermenting colonies, further confirmation of Salmonella species was carried out by biochemical tests followed by final confirmation with API 10S. Antibiotic sensitivity was done by Kirby Bauer disc diffusion technique. Demographic data of the patients was also recorded.
 Results: A total of 2996 blood cultures were received in the said duration, out of which 208 were found culture positive for Salmonella species. Out of all the confirmed Salmonella typhi 80% of the isolates were XDR.
 Conclusion: Reporting this high incidence of XDR Salmonella species, for the first time from Swabi, KPK is a serious concern, to scrutinize the level of AMR in this region and therefore Pakistan. It is imperative that strict evidence based, tailored as per region, policies need to be implemented strictly to stop the misuse of antibiotics.

Tridax procumbens L. Extracts: Ethnomedicinal Wound Healing Investigation

This research aims to assess the wound healing activity and antimicrobial potential of various extracts (Methanol, Ethanol, n-Hexane, Chloroform, and Aqueous) derived from Tridax procumbens, a plant traditionally used in wound treatment by tribal communities. The study includes evaluating the antimicrobial activity of the extracts using the agar well diffusion method and preparing plant extract-infused gauze and ointment. Different solvents were used to extract the plant compounds using the Soxhlet apparatus. Antimicrobial activity of the extracts was tested against microorganisms isolated from infected wound samples, cultivated on various media (Nutrient agar, Cetrimide agar, Blood agar, MacConkey agar, and MRSA). Minimum inhibitory concentration was determined using the agar well diffusion method, identifying the antimicrobial efficacy of the plant extracts. The ethanolic extract of Tridax procumbens exhibited the highest antimicrobial activity against Staphylococcus aureus and Escherichia coli, among other test organisms (Streptococcus, Pseudomonas, Proteus sp.,) isolated from the infected wound samples. Phytochemical tests were conducted to identify bioactive compounds responsible for wound healing, which were subsequently isolated using column chromatography and purified via thin layer chromatography. FTIR analysis was performed to determine the functional groups present, and HPLC was conducted for further characterization. The presence of flavonoids, tannins and terpenoids in the plant extracts was identified as the active ingredients responsible for wound healing. Furthermore, a gauze infused with plant extracts was prepared to facilitate wound healing and an ointment was formulated using the ethanolic extract of Tridax procumbens and an ointment base.

Epidemiology and Molecular Confirmation of E. coli Isolated from Diseased Fish in Muzaffargarh, Punjab, Pakistan

Background Fish is an important source of protein and vitamins, such as vitamin D and B2 (riboflavin) for human beings. However, they are plagued with a variety of disease-causing pathogens, resulting in significant economic losses. Among these pathogens, Escherichia (E.) coli are prominent, worldwide. This study aimed to conduct epidemiological surveillance and identification of E. coli strains isolated from diseased fish in District Muzaffargarh, Punjab, Pakistan. Methodology A total of fifty (50) diseased fish samples were collected from various fish farms in the district. The isolation process involved enriching the samples in nutrient broth and incubating them at 37°C for 24 hours. After enrichment, the samples were inoculated on MacConkey agar and incubated again at 37°C for 24 hours. Following incubation, Gram staining was performed to identify E. coli and confirm its presence. These isolates were subjected to PCR using the uspA gene for confirmation. Results Among fish diseases, Hemorrhagic septicemia was reported to have the highest prevalence (22%), while 12% of fish samples were infected with abdominal dropsy and fin rot. In total, six (06) E. coli isolates were obtained from five different diseased fish samples and confirmed by PCR-based detection of uspA gene. Conclusion The current study found a link between disease-affected fish and naturally occurring E. coli, with molecular confirmation using the uspA gene. Effective management of soil, stock, water, nutrition, and environment is crucial to control losses caused by E. coli as opportunistic fish pathogens and spoilage agents.

Assessment of Antibiotic Resistance among Clinical Isolates of Enterobacteriaceae in Nepal.

Clinicians face a global challenge treating infections caused by Enterobacteriaceae because of the high rate of antibiotic resistance. This cross-sectional study from the Nepal Armed Police Force Hospital, Kathmandu, Nepal, characterized resistance patterns in Enterobacteriaceae across different antimicrobial classes and assessed incidences of multidrug-resistant (MDR) and extensively drug-resistant (XDR) infections. Enterobacteriaceae from clinical samples were isolated on blood and MacConkey agar, except for urine samples on cysteine lactose electrolyte-deficient agar. To determine antimicrobial susceptibility patterns, including MDR and XDR, the Kirby-Bauer disc diffusion method was used. Statistics were performed using SPSS, v. 17.0. Members of the family were identified in 14.5% (95% CI: 16.2-12.8%) of the total samples (N = 1,617), primarily in urine (54.7%, 128/234), blood (19.7%, 46/234), and sputum (15.0%, 35/234). Escherichia coli (n = 118, 44.2%) was the most predominant bacteria, followed by Citrobacter freundii (n = 81, 30.3%). As much as 95.6% (392/410) of the isolates were penicillin-resistant, whereas only 36.2% (290/801) were carbapenem-resistant. A total of 96 (36.0%) MDR and 98 (36.7%) XDR Enterobacteriaceae were identified. Proteus mirabilis (44.4%, 8/18) predominated MDR cultures, whereas C. freundii (53.1%, 43/81) predominated XDR cultures. Multidrug resistant (38.4%, 71/154) and XDR Enterobacteriaceae (22.7%, 35/154) were chiefly uropathogens. Fluoroquinolone resistance rates in non-MDR, MDR, and XDR isolates were 19.9%, 63.2%, and 96.2%, respectively, whereas cephalosporin resistance rates were 28.6%, 72.9%, and 95.4% and penicillin resistance rates were 67.0%, 97.4%, and 98.0%. One-seventh of patients visiting the hospital were found to be infected with Enterobacteriaceae, and of these patients, at least one-fourth were infected with MDR strains.

Bacterial Contamination of Drinking Wells in Makurdi Metropolis

This study was carried out to assess the bacterial contamination of drinking wells in Makurdi metropolis. Fifteen (15) untreated water samples, five (5) each were collected from wells in High Level, Wurukum and Wadata areas for physicochemical and bacteriological analysis was done using pour plating and the bacteriological quality was assessed by measuring the total viable, total coliform and fecal coliform counts of bacteria in the samples using Mac Conkey agar and Nutrient agar. The results of the analysis showed that the pH ranged from 6.70 ± 0.16 to 7.40± 0.87, the temperature levels ranged from 28.72 ± 0.04 to 28.87 ±0.05; biological oxygen demand values ranged from 0.02 ±0.04 to 0.02 ± 0.41; the dissolved oxygen values ranged from 1.50 ±0.16 to 170 ± 0.14 while the Electrical conductivity values ranged from 599.20 ±88.98 to 1582.80±199. The results also showed the total viable count, total coliform count and fecal coliform counts ranging from 75.80±16.01cfu/ml to 117.40±34.45cfu/ml; 5.60±5.13cfu/ml to 6.60±6.88cfu/ml and 0.40±0.89cfu/ml to 4.20±3.77cfu/ml respectively. The total viable counts ranged from 75.80±16.01 to 117.40±34.4 (Wadata having the highest and Wurukum having the least), the total coliform counts ranged from 5.60±2.61 to 6.60±6.88 while the fecal coliform count ranged from 0.40±0.89 to 4.20±3.77. The variation in Total Viable Count (TVC) across locations could be as a result of localized factors influencing microbial populations and geographical differences such as land use, anthropogenic activities, and environmental conditions. The bacterial isolates were Bacillus, Escherichia coli, Klebsiella spp, Proteus spp, Enterobacter spp, and Staphylococcus spp. The frequency of occurrence and prevalence percentages of the bacteria isolated from the well water samples show that Klebsiella spp had the highest percentage (23.33%) while Enterobacter spp and Escherichia coli had the least prevalence percentage, representing 10.00% each. Overall, High Level had the highest contamination rate, accounting for 43.33% of the total contamination while Wurukum had the least contamination, representing 23.33% of the total contamination. The distribution patterns of bacterial isolates among the studied locations illuminate discernible trends. The prevalence of pathogenic bacteria in well water sources could be possibly linked to higher urbanization and population density, highlighting the dynamic and site-specific nature of microbial communities. The study highlights the importance of using specific plans to manage water quality. These plans should take into account the special features of each area.

Vinegar Activity against Clinically Isolated Escherichia Coli

Abstract Background: Antibiotic resistance in Escherichia coli (E. coli) has become a worrying issue been increasingly observed worldwide in recent decades. E. coli intrinsically has a great capacity to accumulate resistance, making them difficult to treat with standard therapies. So, using alternative strategies to treat it is urgent. Material and Methods: The bacteria were isolated from urinary tracts and wounds, and then cultured in three media (blood agar, MacConkey agar, and eosin methylene blue agar). They were incubated for 24 h at a temperature of 37oC to grow. The well diffusion method was used to evaluate the activity of vinegar, and the disk diffusion method was used for antibiotic sensitivity test of gentamicin as a control agent after overnight incubation. Statistical analysis was done by using SPSS 26 software. Results: Out of the 24 participants in the study, 66.6% were women, while 33.4% were men with significant differences (P < 0.05). Isolated E. coli (75%) were sensitive to vinegar, while 70.80% of isolates were sensitive to gentamycin. The mean inhibition zone for vinegar was 13 ± 6.04 mm compared to 15 ± 6.43 mm for gentamicin with no significant differences (P = 0.27). Conclusions: Vinegar shows 75% sensitivity to clinically isolated E. coli, and thus it can be a promising treatment option. Gentamicin with high rate of sensitivity can be used as an empirical treatment for E. coli infections. Vinegar has similar activity to gentamicin (P > 0.05).

Investigating the presence of multidrug-resistant Escherichia coliin raw mutton from open marketplaces

The objective of this research was to identify antibiotic-resistant strains of Escherichia coliand examine the occurrence of E. coli in mutton samples sourced from multiple locations within two districts, given the serious public health risks associated with its presence in food. Six mutton samples were procured from open markets in Kushtia, while another six were sourced from markets in Jhenaidah.Serial dilutions were made by processing mutton after samples were gathered from every region. The samples were plated on nutrient agar and cultured for bacterial growth using the spread plate technique. Following incubation, bacterial contamination was evident in every sample, ranging from 1.2×109 CFU/mL to 4.7×109 CFU/mL. After conducting selective agar tests (MacConkey agar and Eosin-methylene blue) and biochemical tests (Methyl Red, Catalase, Citrate,Oxidase, Indole, and Voges-Proskauer test) to verify the presence of E. coli, it was discovered that 66.67% of the samples of mutton were positive for the E. coli. Then, the E. colistrains were subjected to additional investigation, which showed resistance to three (Amoxicillin,30 μg;Penicillin,10 μg;and Co-trimoxazole, 25 μg)of the eight tested antibiotics and another five (Ciprofloxacin, 5 μg;Azithromycin, 30μg; Streptomycin, 10 μg;Levofloxacin, 5μg;and Erythromycin, 15μg)antibiotics were sensitive. The study underscores the possibility of antibiotic-resistant and multidrug-resistantE. coli in mutton samples, underscoring the need forappropriate handling and cooking techniques to prevent foodborne illnesses from mutton contamination.

The Antibacterial Activity of Lactophenol, Safranin, and Methyl Red Dyes Against Clinically Isolated Escherichia Coli

Abstract Background: Escherichia coli (E. coli) is a type of bacteria commonly found in the intestines of humans and animals that can be pathogenic and develop resistance to various antibiotics through different mechanisms, making them difficult to treat with standard therapies. So using alternative strategies to treat E. coli is urgent, such as utilizing certain dyes that may be a promising branch in microbial therapy. Materials and Methods: The bacteria (E. coli) were isolated from the urinary tract and cultured in three media (blood agar, macConkey agar, and EMB agar), then they were incubated for 24-h at a temperature of 37°C to grow. Well diffusion method used to evaluate the antimicrobial activity of lactophenol, methyl red, and safranin stains. The antibiotic discs (Gentamicin) were used by the disc diffusion method. All the statistical analysis was done by using SPSS 26 software and the Excel app. Results: The participants in this study were 66.67% women, while 33.33% were men, with significant differences (P < 0.05). Gentamicin showed high a rate of sensitivity (88.89%). All isolates of E. coli (100%) were sensitive to lactophenol; in contrast, all isolates (100%) were resistant to methyl red dye. E. coli isolates revealed 55.56% resistance to safranin. Conclusions: Lactophenol inhibition zone was 20.78 ± 5.40 mm, which revealed a great inhibition zone compared to gentamicin’s 17.22 ± 6.55 mm with no significant differences (P = 0.22). That means lactophenol is equal in activity to gentamicin to treat E. coli.

First detection and characterization of mcr-1 colistin resistant E. coli from wild rat in Bangladesh.

Colistin resistance is a global concern warning for a one health approach to combat the challenge. Colistin resistant E. coli and their resistance determinants are widely distributed in the environment, and rats could be a potential source of these isolates and resistant determinants to a diverse environmental setting. This study was aimed to determine the presence of colistin resistant E. coli (CREC) in wild rats, their antimicrobial resistance (AMR) phenotypes, and genotypic analysis of mcr-1 CREC through whole genome sequencing (WGS). A total of 39 rats were examined and CREC was isolated from their fecal pellets onto MacConkey agar containing colistin sulfate (1 μg/ mL). AMR of the CREC was determined by disc diffusion and broth microdilution was employed to determine MIC to colistin sulfate. CREC were screened for mcr genes (mcr-1 to mcr-8) and phylogenetic grouping by PCR. Finally, WGS of one mcr-1 CREC was performed to explore its genetic characteristics especially resistomes and virulence determinants. 43.59% of the rats carried CREC with one (2.56%) of them carrying CREC with mcr-1 gene among the mcr genes examined. Examination of seventeen (17) isolates from the CREC positive rats (n = 17) revealed that majority of them belonging to the pathogenic phylogroup D (52.94%) and B2 (11.76%). 58.82% of the CREC were MDR on disc diffusion test. Shockingly, the mcr-1 CREC showed phenotypic resistance to 16 antimicrobials of 8 different classes and carried the ARGs in its genome. The mcr-1 gene was located on a 60 kb IncI2 plasmid. On the other hand, ARGs related to aminoglycosides, phenicols, sulfonamides, tetracyclines and trimethoprims were located on a 288 kb mega-plasmid separately. The mcr-1 CREC carried 58 virulence genes including genes related to adhesion, colonization, biofilm formation, hemolysis and immune-evasion. The isolate belonged to ST224 and closely related to E. coli from different sources including UPEC clinical isolates from human based on cgMLST analysis. The current research indicates that rats might be a possible source of CREC, and the presence of mcr-1 and other ARGs on plasmid increases the risk of ARGs spreading and endangering human health and other environmental components through this infamous pest.

Molecular identification and antibiotic resistance profiling of Vibrio spp. in diarrhoeal patients from Dinajpur, Bangladesh

This research aimed to characterize Vibrio species from cholera-suspected patients in three hospitals in Dinajpur. We collected 36 stool samples and identified Vibrio spp. using cultural, morphological, and biochemical tests. The highest prevalence was observed in children (18.18%), compared to adolescents (0%), adults (16.67%), and senior adults (16.67%). Cultural identification revealed yellow colonies on TCBS agar, hemolytic colonies on blood agar, pale non-lactose fermenting colonies on MacConkey agar, and white, opaque colonies on nutrient agar. Gram staining showed gram-negative, curved rod-shaped bacteria. Biochemical tests indicated positive reactions for oxidase, catalase, MR, TSI, citrate utilization, MIU, and indole, while VP was negative. Molecular characterization by PCR and gel electrophoresis using 16S rRNA primers confirmed the presence of Vibrio spp. Antibiotic sensitivity testing showed that isolates were sensitive to cotrimoxazole-25µg (21 mm to 30 mm), neomycin-30µg (22 mm), gentamycin-10µg (15 mm), streptomycin-10µg (20 mm), and tetracycline-30µg (17 mm), but resistant to penicillin, mecillinam, and amoxicillin. These findings suggest streptomycin and cotrimoxazole as effective drugs for controlling cholera.

Emerging multidrug-resistant microbial bugs lurking in the ear canal – The hidden horror

Background: The ear discharge is a common presenting symptom in medical practice affecting all age groups but primarily children. It is a big menace in developing countries like India. Ear discharge can result from varied infective and clinical conditions of the ear. Aims and objectives: This study was done to determine the current infective etiological causes of ear discharge, their antibiograms, and associated risk factors. Materials and Methods: This was a retrospective study of laboratory records of all the ear swabs taken from patients with ear discharge in the Calcutta National Medical College and Hospital, for 15 months. A total number of 266 patients of both genders between 1 year and 80 years visiting the E.N.T department with ear discharge are taken as the study population. All pus swabs from infected ears were inoculated on blood agar, MacConkey agar, and Sabouraud Dextrose agar with chloramphenicol. Furthermore, smears for Gram staining and potassium hydroxide mount were made. Identification of the pathogens was done by standard biochemical tests. Sensitivity tests to antimicrobial agents were done on Mueller–Hinton Agar using the Kirby–Bauer disk diffusion method. Results: Out of 266 ear swabs, 231 (86.84%) had microorganisms isolated. Out of 266 patients, 140 (52.64%) were females and 126 (47.36%) were males. Pseudomonas species was the commonly isolated organism 98 (42.42%) followed by Staphylococcus aureus 92 (39.80%) and Klebsiella pneumoniae 17 (7.35%). Pseudomonas species were mostly sensitive to imipenem/cilastatin followed by meropenem and amikacin. Conclusion: Most cases of ear discharge were found in females and the age group of 11–18 years. The most common bacteriological cause of the ear discharge was Pseudomonas species followed by S. aureus. Carbapenems, cefepime, piperacillin-tazobactam, levofloxacin, linezolid, and voriconazole are effective antimicrobial agents for the empirical treatment of ear infections with discharge.

Molecular mechanism of antimicrobial co-resistance Colistin (mcr-1) and ESBLs genes among Escherichia coli isolates from commercial chickens in Pakistan.

Emergence of plasmid mediated colistin and extended spectrum β-lactamases (ESBL) resistant genes has been impacted the efficacy of colistin and β-lactams drugs like 3rd, 4th generation cephalosporin. Current study was aimed to investigate antimicrobial resistance genes (ARGs) among Escherichia coli isolates from meat producing commercial broilers in Pakistan. Two hundred (n=200) fecal samples were collected during January-2018 to August-2019. For isolation of E. coli, pink colonies on MacConkey agar were transferred to EMB agar. Metallic sheen color colonies were tested biochemically using API-20E kit. The molecular identification of E. coli (n=153) was targeted by amplification of uid gene through polymerase chain reaction (PCR) and different ARGs i.e. gentamicin, streptomycin, tetracycline, colistin, β-lactams drugs, quinolone and ampicillin followed by sequence analysis. Genotypically, followed by phenotypically of resistant ARGs of isolated PCR-confirmed E. coli (153) shoed resistant against gentamicin (aac(3)-IV), streptomycin (aadA1), tetracycline (tetA), colistine (mcr-1), ampicillin (bla-TEM) and bla-CTX-M were 86%, 88%, 86%, 88%, 83% & 77% respectively. 33/38 (86%) of the isolate was positive for quinolone resistance. Colistine (mcr-1), ESBLs (bla-TEM) and (bla-CTX-M) resistance genes were 88%, 83% and 77% respectively. About 33 isolated E. coli harbored the both mcr-1 and ESBLs genes. All of E. coli isolates were found sensitive to ceftriaxone (CTX-30) and imipenem (IMP-10). The Isolated E. coli showed single or multi clade decadency. The E. coli and ARGs sequences showed single or multi clade decadency. This is first comprehensive study from Pakistan that described the molecular evidences of ARGs and their co-existence in single isolates originated from commercial poultry. Commercial chicken (Broilers) can act as melting pot of antibiotic resistance genes for human being. It is alarming situation for surveillance of antibiotic resistance program because of more regulated prescription of antimicrobial agents in Pakistan.

Biofilm-Forming Antibiotic-Resistant Bacteria in Water From Distribution Systems: Occurrence and Public Health Implications.

Biofilm is a structurally-connected microbial community, covered by a self-produced polymeric matrix and adhered to biotic or abiotic surfaces. This study aimed to evaluate the occurrence of biofilm-producing antibiotic-resistant bacteria in water from distribution systems. Water samples were taken from 32 tanks across Ondo City and Akure metropolis, Nigeria. Information regarding the sanitation status of the tanks was gathered by observation and oral interviews. The physicochemical properties were determined using standard methods. Using the pour plate technique. Agars included serially diluted water samples were inoculated onto plate count agar, mannitol salt agar, Salmonella-Shigella agar, MacConkey agar, and cetrimide nutrient agar to assess total viable bacteria, Staphylococcus aureus, Salmonella and Shigella, coliforms, and Pseudomonas aeruginosa, respectively. Eosin-methylene blue agar was used to cultivate Escherichia coli and Enterobacter aerogenes. Pure isolates were characterised using API kits and assessed for antibiotic resistance and biofilm production employing the Kirby-Bauer and tissue culture plate techniques, respectively. The ages of the water tanks ranged from 1 to 25 years old; all tanks had cover-lids; 13 (40.63%) had water guards while 12 (37.5%) underwent water treatment. The physicochemical properties chiefly fell within WHO standards for drinking water. One hundred and eighty-seven isolates were obtained. S. aureus (15.51%) had the highest frequency while Salmonella enterica (3.2%) had the lowest frequency. Thirty-six percent of the isolates were strong biofilm producers, while 20.67% Gram-negative and 18.69% Gram-positive bacterial isolates were antibiotic-resistant. This study revealed a high occurrence of biofilm-forming bacteria and prevalence of antibiotic-resistant bacteria in water distribution systems, emphasizing the urgency of improving water quality for public health protection.

Evaluation of an optimal agar medium for detecting hypervirulent Klebsiella pneumoniae using string test.

The string test is a screening method for detecting hypervirulent Klebsiella pneumoniae (hvKp). Agar media are used for string tests; however, the effect of the type of media on the test results remains unclear. We aimed to determine the optimal agar medium and cutoff value for the string test. We performed the string test on 99 Klebsiella strains using different agar media: sheep blood, chocolate, Drigalski's and MacConkey. The diagnostic accuracy was calculated in concordance with the rmpA, rmpA2 or iucA gene levels. The diagnostic accuracy rates for sheep blood, chocolate, Drigalski's and MacConkey agar were 0.79, 0.75, 0.73 and 0.64, respectively. When the cutoff was changed from 5 to 10 mm, the diagnostic accuracy rate for sheep blood agar decreased from 0.79 to 0.65. Our findings suggest that the type of agar medium impacts the string test results and sheep blood agar with a 5-mm cutoff is the optimal condition for detecting hvKp.

Phylogenetic analysis of Klebsiella pneumoniae isolates of respiratory tract infections in humans and sheep.

Klebsiella pneumoniae is an important opportunistic pathogen, which is capable of colonizing the respiratory system in both humans and animals causing mild to severe infections. This study aims to isolate K. pneumoniae from the nasal discharges of human and sheep as well as identify the antibiotic resistance and molecular phylogeny of local isolates. A total of 100; 50 humans and 50 sheep, positive nasal swab isolates were selected, and confirmed biochemically and by the VITEK-2 system. Molecular testing using the polymerase chain reaction (PCR) and phylogeny was conducted. On MacConkey agar, Klebsiella isolates appeared as large, pinkish, and mucoid colonies; while microscopically, it appeared as Gram-negative rods. Traditional biochemical tests revealed that 62% and 78% of human and sheep isolates were positive Klebsiella isolates, whereas respectively, 54.84% and 71.8% of these isolates were positive by VITEK-2. Antibiotic susceptibility tests showed that the human isolates were sensitive to aztreonam, piperacillin-tazobactam, ciprofloxacin, and cefuroxime. Subsequently, sheep isolates were sensitive to cefuroxime, ciprofloxacin, piperacillin-tazobactam, ampicillin, cefoxitin, and tetracycline. Targeting 16S rRNA gene, a total of 17 human and 28 sheep isolates were molecularly positive K. pneumoniae. Phylogenetic analysis of study human and sheep isolates showed their identity to NCBI Indian (LC747146.1) and Iraqi (LC711141.1) isolates, respectively. Comparative analysis between the local human and sheep isolates revealed a significant identity that ranged from 99.82% to 99.88% with a percentage of mutation ranging from 0.008% to 0.002%. Klebsiella pneumoniae is a highly prevalent bacterium in both human and sheep with an observable resistance to antibiotics. Molecular phylogeny of study isolates demonstrated their close relation, suggesting the possible direct or indirect transmission of the bacterium from sheep to human or vice versa. Moreover, studies are greatly important to estimate the routes of bacterial transmission. Also, extensive hygiene practices could be lowered the spreading of K. pnuemoniae to farm workers.

Prevalence of Extended-Spectrum Beta-Lactamase Producer Gram-Negative Rods and Associated Factors Among Patients With Wound Infection at University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia.

Background: Multidrug-resistant (MDR) bacteria have significantly affected the management and treatment of wound infections globally. Data on the prevalence of MDR bacterial profiles that cause wound infections in Ethiopia are scarce. Therefore, this study aimed to determine MDR as well as extended-spectrum beta-lactamase production profiles of Gram-negative rods that are difficult to treat with conventional antibiotics and that cause wound infections. Objective: The aim of the study was to determine the prevalence of extended-spectrum beta-lactamase producer Gram-negative rods and associated factors among patients with wound infection at University of Gondar Comprehensive Specialized Hospital, northwest Ethiopia. Materials and Methods: This hospital-based cross-sectional study was conducted at University of Gondar Comprehensive Specialized Hospital between May and July 2022. Convenience sampling was used to recruit 228 participants. Swabs from different wound types were inoculated onto the MacConkey agar and blood agar plates and incubated overnight at 37°C for 24 h. Biochemical tests were performed on isolated colonies for the identification of bacterial species based on their biochemical reaction. Antimicrobial susceptibility tests were performed using the disk diffusion technique as per the standard Kirby-Bauer method by using Muller-Hinton agar, and the zone of inhibition was interpreted as resistant, intermediate, and sensitive as per the recommendation of Clinical Laboratory Standard Institute. Isolates were tested against ceftriaxone, cefotaxime, and ceftazidime for extended-spectrum beta-lactamase screening using the Kirby-Bauer disk diffusion method, and combined disk tests were applied for phenotypic confirmatory test of extended-spectrum beta-lactamase producing isolates. Result: Of 228 study participants, 162 (71.1%) were culture-positive. Among culture-positive patients, 165 Gram-negative bacteria were recovered. The most common Gram-negative isolates were Pseudomonas aeruginosa (47; 28.5%), followed by Escherichia coli (43; 26.1%) and Klebsiella pneumoniae (24; 14.5%). The susceptibility rates of the isolate for imipenem and tobramycin were 97.0% and 73.2%, respectively, and the overall multidrug resistance rate was 80.5%. Extended-spectrum beta-lactamase producer bacteria were also isolated. Besides, all (100%) of extended-spectrum beta-lactamase producer bacteria were MDR. Living in rural areas (AOR 5.8; 95% CI [2.01-16.7]), hospital admission (AOR 3.95; 95% CI [1.13-13.83]), antibiotic use (AOR 2.83; 95% CI [1.03-7.72]), and comorbidity (AOR 0.25; 95% CI [0.07-0.59]) were significantly associated with wound infection. Conclusions and Recommendations: There was a high prevalence of Gram-negative bacterial isolates in this study. Pseudomonas aeruginosa (28.5%) was the predominant isolate. In addition, high rates of multidrug resistance were observed. The high level of multidrug resistance in this study implies that definitive therapy should be based on culture and susceptibility analysis to promote the rational use of antibiotics and to reduce the emergence of bacterial resistance to antimicrobials.

The Possible Synergism Effect of Amoxyclav Combined with Gentamicin and Paracetamol against Blood Stream Escherichia coli

Abstract Background: Some bacteria have developed resistance to antibiotics that were once commonly used to treat them. Escherichia coli is one of the most common causes of bloodstream bacterial infections. This research was performed to study the effect of some antibiotics on the growth of E. coli by combination with amoxyclav. Materials and Methods: E. coli were isolated from the blood stream of 20 patients and cultured in three media (blood agar, MacConkey agar, and eosin methylene blue agar). The cultures were incubated for 24 h at a temperature of 37°C. The well diffusion method used to evaluate the activity of antimicrobial agents. The antibiotics (paracetamol, gentamicin, and amoxyclav) were added separately in one petri dish and in combination with amoxyclav in another petri dish to evaluate the effect of antibiotics on bacterial growth before and after the addition of amoxyclav. Results: The study showed that males were more infected (65%) than females (35%), without significant differences (P = 0.18). The addition of amoxyclav increased the effect of gentamicin from 28.9 ± 2.7 to 33.7 ± 7.47 mm, showing significant differences (P = 0.013). The addition of paracetamol to amoxyclav showed antagonism, decreasing the effect from 31.75 + 9.41 to 29.95 ± 8.66 mm), without significant differences (P = 0.53). Conclusions: The combination of gentamicin with amoxyclav has a synergistic effect against E. coli clinically isolated from blood streams (P = 0.013). The addition of paracetamol to amoxyclav showed an antagonism effect, but it was still insignificant (P = 0.53).

Antibacterial Activity of Magnesium Oxide Nanoparticles against MDR Pseudomonas aeruginosa Isolated from Different Clinical Infections

In this study, 180 isolates of Pseudomonas aeruginosa were isolated from patients suffering from various infections, including urinary tract infections, burns, ear infections, tonsillitis, and pneumonia. Specimens were taken from patients hospitalized in Al-Hakim General Hospital, Al-Sadr Medical City, and the Burn Center in Najaf Al-Ashraf, and they were transferred to the Microbiology Laboratory in the Biology Department in the College of Science. P. aeruginosa have been isolated from Specimens taken from patients with these various infections. Isolation methods were different, including culture on MacConkey medium, blood agar, as well as various biochemical and Vitek tests. Susceptibility testing was also performed on these bacteria for six families according to CLSI. These families are penicillins, aminoglycosides, carbopenems, cephalosporins, fluoroquinins, and lipopeptides. It was noted that forty isolates were multi-resistant to these antibiotics. It was also revealed that resistant isolates formed biofilms using a flat microliter. The effectiveness of domestic and imported magnesium oxide nanoparticles on resistant isolates was conducted. These two types were taken at concentrations of 100, 150, and 200 μg/ml. Imported MgO nanoparticles were more effective than domestic ones. It was also observed that the effect of magnesium oxide nanoparticles on resistant bacteria increased with increasing concentration. Antibiotics resistant to Pseudomonas aeruginosa bacteria were also taken and mixed with imported and local magnesium oxide nanoparticles with an optimal concentration of (200) μg/ml. The effect of magnesium oxide nanoparticles combined with antibiotics was greater than if the nanoparticles were alone.

<b>An assessment of bacterial transmission via rebound tonometry: An <i>in vitro</i> pilot study</b>

<b>Background:</b> Bacterial contamination of medical equipment is a significant risk in both human and veterinary medicine, leading to potential cross-contamination between clinicians, technicians, patients, and healthcare professionals. Pathogens can persist on surfaces, resulting in healthcare-associated infections. Tonometry used to measure intraocular pressure (IOP) for diagnosing conditions like glaucoma and uveitis, is crucial in both human and veterinary care. However, there is limited information on bacterial transmission through reused tonometer probes in veterinary patients. This pilot study aimed to explore the potential for cross-contamination of bacteria via a rebound tonometer probe using bacterial isolates linked to bacterial ulcerative keratitis in dogs. <b>Aim:</b> To evaluate the potential for bacterial transmission via rebound tonometry using the ICare® TONOVET Plus in a controlled laboratory environment. The study focused on two bacterial isolates commonly implicated in canine bacterial ulcerative keratitis: Pseudomonas aeruginosa and beta-haemolytic Streptococcus. <b>Methods:</b> Bacterial cultures were inoculated on blood agar and MacConkey agar plates. Tonometer probes were used to perform multiple corneal rebounds (CR) readings on known bacterial samples, followed by multiple CR readings on agar growth plates, incubation and re-examination of the agar plates for evidence of bacterial transfer. Controls included plates where the probes were only used on sterile agar. <b>Results:</b> Bacterial transmission was detected with distinct bacterial colonies corresponding to individual CR readings on the plates inoculated with Pseudomonas and beta-haemolytic Streptococcus. A control plate exhibited a single bacterial colony, likely due to inadvertent contamination. <b>Conclusion:</b> The results indicate that bacterial transmission via rebound tonometer probes is possible. Although this was an in vitro study, the findings raise concerns about cross-contamination in clinical practice. Changing tonometer probes between patients, particularly after those with bacterial ulcerative keratitis is advised until further investigation into the in vivo bacterial transmission is investigated.

Genotypic And Phenotypic Surveillance Of Multidrug Resistance Of Hypermucoid K. Pneumoniae Among Clinical Isolates In AL Najaf City /Iraq

Hypervirulent Klebsiella pneumoniae strains are the major cause of liver abscesses ,diabetes foot ulcers and other infections throughout Iraq, and these strains are usually antibiotics susceptible. Recently, hypervirulent K. pneumoniae isolates have emerged due to acquiring antimicrobial resistance determinants or the transfer of a virulence plasmid into a classic isolates ,.Klebsiella pneumoniae is responsible for hospital- and community-acquired infections. This study aimed to determine the prevalence of hyper irulence K. pneumoniae and investigate the antibiotics resistance profile among clinical specimens at Al Najaf Hospitals in/ Iraq, and detect the genes for molecular identification of K. pneumoniae in comparison with phenotypic and biochemical methods. In total, 100 clinical specimens were collected from patients infected with K. pneumoniae, between January 2023 and May 2023 from both sexes males (38) and Females (62) with age ranged between (20-70) years. The Hypervirulent Klebsiella pneumoniae isolates were cultured on general and selective media as (Blood agar and MacConkey agar and CHROM agar), and confirmed by using Vitek-2 system. The isolates were reported with high resistance towards various types of antibiotics, especially penicillins and cephalosporins. In contrast, K. pneumoniae showed very low resistance to imipenem and amikacin. According to the results the range of multidrug-resistant K. pneumoniae isolates in this study was estimated at 100%.