Isolates Of Gram-positive Bacteria Research Articles

The Influence of COVID-19 on Antimicrobial-Resistance in Gram-Positive Bacteria at a Private Saudi Hospital: A Five-Year Evaluation

Due to the emergence of antimicrobial-resistance (AR) as a public health threat, the Saudi National Action Plan (SNAP) was implemented in 2017, incorporating various strategies to combat AR. To evaluate the effectiveness of SNAP and the impact of COVID-19, the study analyzed pre- and post-pandemic rates of AR, methicillin-resistant Staphylococcus aureus (MRSA), and multidrug resistance (MDR) among Gram-positive bacteria at a private medical center in Saudi Arabia. This study reviewed the cases of all patients who had been diagnosed with Gram-positive bacterial infection between January 2017 and December 2021. Bacterial strain identification was conducted using VITEK-2 ID-GP cards, while AR, MRSA, and MDR were defined using AST-GP 67 and AST-ST02 cards, all adhering to the manufacturer’s recommended protocols. The five-year study from 2017 to 2021 yielded 6,271 Gram-positive bacteria isolates from patients in a Saudi private hospital. Though the rate of AR initially declined between 2017 and 2019, it spiked significantly from 2020 to 2021. Similarly, the MRSA rate exhibited a substantial decrease (p < 0.05) from 2017 to 2019, followed by a significant rise (p < 0.05) between 2020 and 2021. Out of the isolated pathogens, 1,031 (16.44%) exhibited MDR, with all isolates showing a marked increase (p < 0.05) in MDR from 2020 to 2021. The study highlights the aggravating impact of the COVID-19 pandemic on AR, underscoring the need for the SNAP to intensify its efforts in combating AR.

Bacterial Co-Infections and Antibiotic Resistance in Urinary Tract Infection of Covid-19 Patients in Erbil City

The COVID-19 pandemic has raised concerns over secondary infections because it has limited treatment options and empiric antimicrobial treatment poses serious risks of aggravating antimicrobial resistance. Many studies have shown that COVID-19 patients are predisposed to developing secondary infections. The prevalence of urinary tract infections (UTI) was evaluated in 248 patients admitted to different hospitals in Erbil city, Iraq. Results showed 138 (55.65%) patients were positive for bacterial growth, of which 72 isolates of Gram-negative and 66 isolates of Gram-positive bacteria were isolated and identified from the urine of COVID-19 patients depending on the 16S rRNA gene using PCR. The predominant obtained bacteria were Staphylococci species, of which isolates belong to 6 species, and the predominant Gram-negative species was Pseudomonas spp. which belongs to 4 species. Also, their susceptibility to 15 antibiotics was tested and it was found that most of the Gram-negative isolates were highly resistant to ampicillin 66(91.66%), amoxicillin 60(83.33%), cephalexin 52(72.22%) and cefixime 50(69.44%). While the most effective antibiotic was imipenem, and with percentage of 26(36.11%), it also showed variable sensitivity to other antibiotics. For Gram-positive bacteria, the highest resistance was against cefixime 40(60.60%), ampicillin 28(42.42%), and 18(27.27%) for both amoxicillin and ciprofloxacin, while the most effective antibiotic was gentamicin, which showed effectiveness against all the Gram-positive isolates. Another part of the study concerned with detection of the existence of ESBLs genes responsible for antibiotic resistance. The results revealed that all the isolates possessed the 16S rRNA gene, whereas blaTEM and blaCTX-M were found to be the most possessed genes in all the isolates, with a percentage of (88.33%).

Diagnoses Bacterial Wound Infection and Detection of Some Immune Parameters in its

This study aims to diagnoses bacterial wound infection and to estimate the serum levels of TLR-3, CXCL8, TGF-β1, IL-8, IL-17 and IL-22 in wound patients. This study had been included 140 samples (blood and swab) obtained from patients found in Baquba-Teaching Hospital from 15/ 6 /2023 to 20 /3 /2024. Swab samples taken from wound were culture on the blood agar and MacConkey agar, and then Bacteria diagnoses was done by using an automatic VITEK 2 system. The serum levels of immune parameters done by ELISA. The results shown that sixty isolates were grow in culture, the results indicated of 38(63.3%) isolates of gram-negative bacteria, represented by: 12 (20%) isolates E. coli, 18 (30%) Pseudomonas aeruginosa, 6 (10%) Protus spp., 2 (3.3%) Acinetobacter spp. the results indicated of 22 (36.6%) isolates of gram-positive bacteria, represented by: 16 (26.6%) isolates Staphylococcus aureus, 4 (6.6 %) another Staphylococcus spp., 2 (3.3%) Enterococcus spp. The study also shown the a increase in the concentration of TLR-3 , CXCL8 , IL-2 , IL-17 and IL-22 in the serum of bacterial patients compared with the healthy, Where the results of the current study show decreases in the concentration of TGF-β1 among compared with healthy. In conclusion, it was discovered that the infection of bacteria in wound infection was lower in females than males. Pseudomonas aeruginosa and S. aureus were found to be the highly isolated bacteria wound infection, with increases levels of TLR-3, CXCL8, IL-17 and IL-22 in serum in patients and decreases level of TGF-β1 in serum of patients.

Synthesis, Characterization and Antimicrobial Analysis of Nickel and Cobalt Complexes of Condensates of Salicylaldehyde and 2-Hydroxy Aniline

This study on the synthesis, characterization and antimicrobial analysis of Nickel and Cobalt complexes of condensates of salicylaldehyde and 2-hydroxy aniline constitutes an attempt towards the development of new molecular compounds that could destroy the barrier of resistance of pharmaceutical products. The ligands of 2-phenyliminomethylphenol (PMP) and 2,2-hydroxy benzylidene aminophenol (HAP) were first prepared by separately reacting Aniline with Salicylaldehyde on the one hand, and 2-aminophenol with Salicylaldehyde on the other hand. The unmixed metal complexes were prepared by reacting either of 0.29 mmol 2-phenylimino methyl phenol ligand with the metal salt or 0.29 mmol 2,2-hydroxybenzylideneaminophenol ligand with the metal salt while the mixed metal complexes were prepared by addition of equimolar amount of ligands of 2-phenyliminomethylphenol and 2,2-hydroxybenzylideneaminophenol into the metal salt. The molecular structures of the ligands and their complexes have been determined and characterized using GC-MS, FTIR, UV-Vis, 1H-NMR, and X-ray diffraction techniques. The melting point and solubility of the ligands and synthesized complexes were equally determined. Antimicrobial studies were carried out with ligands PMP and HAP and the metal complexes using clinical isolates of Gram Positive Bacteria (Staphylococcus aureus and Bacillus substilis), Gram Negative Bacteria (Escherichia coli and Pseudomonas aeruginosa) and Fungi (Candida albicans and Aspergillus niger). All complexes and ligands were found to be soluble in dimethylsulfoxide (DMSO). Asymmetric Octahedral geometry is observed for Nickel (II) and Cobalt (II) complexes with PMP, HAP, and the mixed ligands. A bidentate ligand is coordinated to metal ions through oxygen atoms of carbonyl groups and nitrogen atoms of two imine groups for Nickel (II) and Cobalt (II) complexes. The findings from the XRD pattern indicate that the ligand and metal complexes are crystalline with a high crystallite size of 119.23nm to 638.67nm. The FTIR spectra for 2-phenyliminomethylphenol (PMP) showed IR absorption band at 1569.2 cm-1 suggesting the stretching vibration of the C=C bond in the benzene rings, while the bands at 1613.9 cm-1 indicated the =C-N stretching in the benzene ring. The complexes showed electronic spectra which suggest that energy was absorbed in the ultraviolet/visible region, producing changes in the electronic energy of the compound and resulting from the transition of valence electrons in the complexes. The XRD findings reveal that the crystal formed by the ligands and their respective complexes are large. The ligands were found to be larger in size than the complexes and this could be due to the complexation reactions. The X-ray diffractograms of the metal complexes produced good intense peaks that suggest high crystallinity. The ligand of 2-phenyliminomethylphenol (PMP) and 2,2-hydroxy benzylidene aminophenol (HAP) showed partial inhibitory action on the clinical isolates of Gram Postive, Gram Negative and zero inhibitory effect on the fungi while the divalent metal complexes of both mixed and unmixed ligands exhibited either stronger or strongest inhibition. The study concludes that the synthesized ligands and their metal complexes are purely crystalline as revealed by the chemical analysis, exhibited inhibitory actions on Gram Positive, Gram Negative and fungi species.

A STUDY ON IN VITRO ACTIVITY OF FOSFOMYCIN AGAINST ENTEROBACTERIACAE UROPATHOGEN ALONG WITH MICROBIOLOGICAL PROFILE AND ANTIBIOGRAM OF UROPATHOGENS

Background:Urinary tract infection (UTI) is one of the most common infection throughout the world that may affect both the lower and the upper urinary tracts. Aims & Objective:The present study was done to determine various organisms causing Urinary tract infections and to analyse antimicrobial sensitivity patterns of those isolates. Materials and Methods:This prospective study was conducted from April 2020 to March 2021at SIMS& RH, Tumkur. All urine samples were received in Microbiology laboratory for routine microbiology investigations were inoculated on Blood agar and MacConkey agar culture media using calibrated wire loop . Urinary isolates with significant bacteriuria were further processed by conventional biochemical tests and antimicrobial susceptibility testing was done as per standard methods. Results: A total of 697 urine samples were processed in that 324 (46.48%) samples were shown as significant growth of bacteria. Most of the cases seen in (67.28%) female and under age group of 21-30(22.22%).The predominantly Gram-negative bacteria (85.18%) were isolated followed by Gram positive bacteria(10.49%) and Candida spp(4.32%). The Gram negative bacteria showed high sensitivity to Ampicillin sulbactum (88.27%) followed by Amikacin (87.24%). All Gram-positive bacteria isolates is showed sensitivity to Vancomycin (100%).We found that 235( 92.88%) Enterobacteriaceace isolates were is susceptible for Fosfomycin. Conclusion:. Antibiotic resistance has become a major clinical problem worldwide and has increased over the years.Hence antibiotic susceptibility testing should be done to diagnose and treat the drug resistant UTI cases. Fosfomycin may be given empirically in patients with uncomplicated urinary tract infection due to Enterobacteriaceace.

The Pattern of Bacterial Infections Among Chronic Suppurative Otitis Media Cases at a Tertiary Care Centre in North-East India.

Chronic suppurative otitis media (CSOM) is a chronic inflammation of the mucoperiosteal lining of the middle ear cleft, presenting with recurrent ear discharge through a tympanic membrane perforation. The present study aims to assess the spectrum of bacterial infection among CSOM cases and detect the isolated organism's antibiotic sensitivity pattern. The prospective hospital-based observational study was conducted from June 2021 to June 2022 and included 94 CSOM cases. An aural swab of the ear discharge was collected from each patient under aseptic precautions. The swab was utilized for Gram's staining and the aerobic bacterial pathogen culture. The organisms isolated were tested for antibiotic sensitivity using the Kirby-Bauer disc diffusion method. The most affected age group was the second decade of life (27.7%, n=26), with a male-to-female ratio of 1.35:1. The mean duration of ear discharge was 24.0±14.7 months, mostly mucoid ear discharge (39.4%, n=37). Among gram-positive bacteria, methicillin-resistant Staphylococcus aureus was isolated in 16 (17.0%) cases. Pseudomonas aeruginosa was the most isolated gram-negative bacteria strain in 26 (27.7%) cases. Cotrimoxazole (67.7%, n=21) had the highest sensitivity towards gram-positive bacteria isolates. Amongst gram-negative bacteria, amikacin and ciprofloxacin were the most sensitive, with 78.0% (n=39) susceptibility. Evaluating the spectrum of infecting organisms of CSOM and their antibiotic sensitivity may help initiate prompt treatment with the appropriate antibiotic regimen, thereby preventing future complications.

Bacteriological assessment of kunu-zaki sold in selected communities in Port Harcourt, Nigeria

Kunu-zaki is a fermented beverage sold in Nigeria; often prepared under unsafe conditions, predisposing it to pathogen contaminations. The bacteria associated with Kunu-zaki sold in the different area of Port Harcourt and an antibiogram of the pathogens was determined using standard methods involving nutrient agar, Salmonella-Shigella agar, Thiosulphate citrate bile salt sucrose agar, MacConkey agar, Mannitol salt agar, Eosin-Methylene blue agar and Mueller Hinton agar. The counts for kunu-zaki obtained from Choba, Mile 1, Rumuodara and Rumuokoro ranged from 2.87±0.11 to 5.39±0.19, 4.50±0.27 to 5.46±0.38, 4.02±0.54 to 5.96±0.27 and 4.24±1.13 to 5.94±0.22 log10cfu/ml, respectively. The confirmation of the isolates as: Staphylococcus spp. (27%), Lactobacillus spp. (20%), Streptococcus spp. (17%), Bacillus spp. (17%), E. coli (10%), and Shigella at (10%) was based on cultural and biochemical characteristics. The Gram-negative bacteria were 100% susceptible to pefloxacin, gentamycin, ciprofloxacin and tarivid while the Gram-positive bacteria isolates showed varying resistance to the antibiotics with Staphylococcus being the most susceptible. The occurrence of the antibiotic resistance isolates possess danger for potential consumers, hence the need to improve the production process.

In vitro Activity of Ceftaroline Against Isolates of Gram-Positive Bacteria from Patients with Bloodstream Infections Collected as a Part of ATLAS Between 2017 and 2020.

To assess the in vitro activity of ceftaroline and a panel of comparator agents against isolates of Gram-positive bacteria, including Staphylococcus aureus, Streptococcus pneumoniae, β-hemolytic streptococci, and coagulase-negative staphylococci (CoNS) from blood collected in Africa and Middle East (AfME), Asia Pacific (APAC), Europe, Latin America (LATAM), and North America from 2017 to 2020 as a part of the Antimicrobial Testing Leadership and Surveillance (ATLAS) program. Susceptibility and minimum inhibitory concentration were determined using broth microdilution for all antimicrobial agents by a central reference laboratory according to the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. Ceftaroline showed good activity (susceptibility ≥89.8%, MIC90 0.008-2 mg/L) against all Gram-positive isolates tested. All isolates of methicillin-susceptible S. aureus, penicillin-susceptible S. pneumoniae, S. agalactiae, S. dysgalactiae, and S. pyogenes were susceptible to ceftaroline (MIC90 0.008-0.25 mg/L). Ceftaroline susceptibility for MRSA isolates was 89.8% globally (MIC90 2 mg/L). Among the comparator agents, all isolates were susceptible to vancomycin, except S. epidermis (susceptibility, 99.9%). Among other agents, daptomycin, linezolid, and tigecycline showed potent activity (susceptibility ≥97.9%, MIC90 0.03-2 mg/L) against all isolates tested. Ceftaroline showed potent in vitro activity against global bloodstream isolates of Gram-positive bacteria collected between 2017 and 2020. Monitoring and surveillance of global as well as regional longitudinal trends of resistance rates among Gram-positive isolates causing bloodstream infections are important to limit the spread of AMR, establish stewardship measures, and manage and appropriately treat infections.

The Relationship of Staphylococcus aureus Virulancity with Serum Cytokine Levels in Polycystic Ovary Syndrome and Polycystic Ovary Disease

The current study was done to isolation and indentification of the Staphlococcus aureus isolates associated with polycystic ovarian syndrome patients and determine the immunological status for these patients. Inflammatory markers have been linked to polycystic ovary syndrome (PCOS). Because ovulation is a quasi-inflammatory process, endothelial adipose tissue can stimulate an inflammatory response and maintain inflammation in adipocytes. By influencing the release of inflammatory cytokines from this downregulated inflammation, polycystic ovary syndrome can arise. The study included 105 clinical specimens, that include age (20-45) years which were collected from November 2022 to June 2023. The specimens included blood specimens and vaginal swabs. They visit Imam Al-Sadiq Hospital and Babylon Teaching Hospital for Maternity and Children in Babylon Governorate. The results of vaginal swabs culture, biochemical tests and vitic2 for isolated bacteria from patients were revealed that Gram-positive bacteria isolates were dominant, amounting to 51 (54.83%), while the percentage of Gram-negative bacteria was 37 (39.78%), while fungi percentage was 5 (5.37%). The results showed that Staphylococcus aureus is the most common bacteria among Gram-positive bacteria, as it constituted 38 (40.86%), followed by Lactobacillus spp. As it accounted for 11 (11.82%) of Staphylococcus epidermidis by 2 (2.15%). The Gram-negative isolates included Escherichiae coli, Klebsiella pneumoniae, and Pseudomonase aerogenosa 23 (24.73%), 9(9.67%) and 5 (5.37%), respectively also the result demonstrates that IL-10, IL-13 level significantly elevation in PCOS, PCOD patients group.

Surveillance of bacterial resistance in children aged 0-14 years from 2018 to 2022

Objective: To understand the distribution and antimicrobial resistance of common bacteria from children aged 0-14 years from China Antimicrobial Resistance Surveillance System. Methods: Bacterial resistance data of 2 575 040 strains from children aged 0-14 years were extracted from the national bacterial resistance surveillance reports from October 2018 to September 2022 and resistance changes were further analyzed by comparing with all data in each year. Results: The total number of bacteria isolated from children in 2018-2022 ranged from 415 306-588 016 strains, accounted for 15.9% (514 193/3 234 372), 16.2% (572 107/3 528 471), 12.8% (415 306/3 249 123), 13.0% (485 418/3 743 027), and 12.2% (588 016/4 828 509), respectively. The proportions of gram-positive bacteria among children were 45.4% (233 456/514 193), 44.5% (254 869/572 107), 44.7% (185 756/415 306), 42.6% (206 903/485 418), and 41.7% (245 044/588 016), respectively. The top five isolates of gram-positive bacteria were Staphylococcus aureus (36.0%-38.8%), Streptococcus pneumoniae (27.1%-31.7%), Staphylococcus epidermidis (7.3%-9.3%), Enterococcus faecium (4.0%-4.8%), and Enterococcus faecium (2.5%-3.6%), and the top five isolates of gram-negative bacteria were Escherichia coli (21.8%-26.2%), Haemophilus influenzae (14.4%-26.4%), Klebsiella pneumoniae (10.1%-14.7%), Moraxella catarrhalis (7.3%-11.9%), and Pseudomonas aeruginosa (5.5%-6.8%). The bacteria from children aged 0-14 years commonly isolated from sputum samples (48.8%-57.0%). The prevalence of methicillin-resistant S. aureus was 28.7%-30.1%. The detection rates of vancomycin-resistant E. faecalis or E. faecium were 0.1%-0.3%. The proportions of non-cerebrospinal fluid-derived penicillin-resistant S. pneumoniae were 0.7%-1.6%. The prevalence of cefotaxime and (or) ceftriaxone-resistant E. coli and K. pneumoniae decreased were 43.7%-50.0% and 31.8%-42.7%, respectively. The resistant rates of E. coli to imipenem and meropenem were 1.2%-1.9% and 1.2%-2.0%, respectively, and the resistant rates of K. pneumoniae to imipenem and meropenem were 7.3%-10.1% and 8.2%-12.2%, respectively. About 6.6%-10.2% and 5.3%-9.6% of the P. aeruginosa isolates showed resistant to imipenem and meropenem, respectively, while 17.2%-24.0% and 19.0%-29.4% of the Acinetobacter baumannii isolates were resistant to imipenem and meropenem, respectively. Conclusions: There is no significant change in the composition of common clinical pathogens in children aged 0-14 years from 2018 to 2022. The prevalence of some resistant bacteria such as methicillin-resistant S. aureus and carbapenem-resistant Enterobacterales is decreasing. However, it is necessary to pay attention to antimicrobial resistance of bacteria from children and long-term monitoring of the prevalence of resistant bacteria should be conducted.

A single-center study on the distribution and antibiotic resistance of pathogens causing bloodstream infection in patients with hematological malignancies

Objective: To study the incidence of bloodstream infections, pathogen distribution, and antibiotic resistance profile in patients with hematological malignancies. Methods: From January 2018 to December 2021, we retrospectively analyzed the clinical characteristics, pathogen distribution, and antibiotic resistance profiles of patients with malignant hematological diseases and bloodstream infections in the Department of Hematology, Nanfang Hospital, Southern Medical University. Results: A total of 582 incidences of bloodstream infections occurred in 22,717 inpatients. From 2018 to 2021, the incidence rates of bloodstream infections were 2.79%, 2.99%, 2.79%, and 2.02%, respectively. Five hundred ninety-nine types of bacteria were recovered from blood cultures, with 487 (81.3%) gram-negative bacteria, such as Klebsiella pneumonia, Escherichia coli, and Pseudomonas aeruginosa. Eighty-one (13.5%) were gram-positive bacteria, primarily Staphylococcus aureus, Staphylococcus epidermidis, and Enterococcus faecium, whereas the remaining 31 (5.2%) were fungi. Enterobacteriaceae resistance to carbapenems, piperacillin/tazobactam, cefoperazone sodium/sulbactam, and tigecycline were 11.0%, 15.3%, 15.4%, and 3.3%, with a descending trend year on year. Non-fermenters tolerated piperacillin/tazobactam, cefoperazone sodium/sulbactam, and quinolones at 29.6%, 13.3%, and 21.7%, respectively. However, only two gram-positive bacteria isolates were shown to be resistant to glycopeptide antibiotics. Conclusions: Bloodstream pathogens in hematological malignancies were broadly dispersed, most of which were gram-negative bacteria. Antibiotic resistance rates vary greatly between species. Our research serves as a valuable resource for the selection of empirical antibiotics.

Setting of the tentative epidemiological cut-off values of contezolid for Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae and Streptococcus agalactiae.

To set the tentative epidemiological cut-off values (TECOFFs) of contezolid for Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae and Streptococcus agalactiae based on the distributions of inhibition zone diameters and MICs. A total of 1358 non-duplicate clinical isolates of Gram-positive bacteria were collected from the patients across China from 2017 to 2020. The isolates were tested for susceptibility to contezolid and the comparator linezolid by broth microdilution and disc diffusion methods in three microbiology laboratories. The zone diameters and MICs of linezolid WT strains were used to set the WT TECOFFs of contezolid by normalized resistance interpretation calculations. Contezolid showed an aggregate MIC range from 0.03 to 8 mg/L and MIC90 value of 1-2 mg/L against all of the Gram-positive bacterial strains tested. The TECOFF of contezolid based on MIC distributions was 4 mg/L for both S. aureus and Enterococcus species, and 2 mg/L for S. pneumoniae and S. agalactiae. The TECOFF of contezolid based on zone diameter was 24 mm for S. aureus, 18 mm for E. faecalis, 20 mm for E. faecium and S. pneumoniae, and 17 mm for S. agalactiae. The epidemiological cut-off values of contezolid were set tentatively for selected Gram-positive bacteria using the MIC and zone diameter distributions. These data are helpful for clinical microbiologists and clinicians to interpret the antimicrobial susceptibility results of contezolid.

Multi-drug and biocide-resistant Gram-positive bacteria prevalent on surfaces of veterinary clinics in Benue State, Nigeria

Objective: In this study, the common aerobic Gram-positive bacteria (GPB) contaminating veterinary clinic surfaces were isolated, and their susceptibility to antimicrobial agents (antibiotics and biocides) was determined. Materials and Methods: Standard cultural and biochemical procedures were used to process 62 swab samples collected from different surfaces in 4 veterinary clinics for the isolation and identification of GPB. The agar dilution technique was used to determine the sensitivity of the isolates to disinfectants, while the disc diffusion technique was used to determine the sensitivity to antibiotics. Results: Isolation rates in veterinary clinics I, II, III, and IV were 39.4%, 40.4%, 7.4%, and 12.8%, respectively. GPB were isolated from all the surfaces sampled. A total of 94 GPB isolates were recovered, comprising Bacillus species (51.1%), Staphylococcus species (38.3%), Corynebacterium species (6.4%), and Enterococcus species (4.3%). Only 13.8% of the GPB were resistant to at least one of the biocides tested. Multiple drug resistance to at least three classes of antimicrobial agents was exhibited by 95% of the isolates tested. Conclusion: The results of this study suggest that veterinary clinic environments and surfaces in Makurdi are reservoirs of wide varieties of antimicrobial-resistant GPB of veterinary and public health importance.

Isolation of Lactic Acid Bacteria from Cocoa Bean Fermentation as Potential Antibacterial Agent against ESKAPE Pathogens

This study aimed to evaluate the antibacterial activity of microorganisms isolated before and during cocoa fermentation against ESKAPE pathogens. Microorganisms from cocoa fermentation process were isolated on the selective media and were tested against ESKAPE pathogens. Total titratable acidity, minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of cell-free supernatant from lactic acid bacteria (LAB) were determined. The presence of bacteriocin and antibacterial activity were determined. LAB was identified through molecular and biochemical tests. Ninety-five and 134 isolates were obtained from the fermentation of Sungai Balung 25 (BR25) clone and mixed clones, respectively. Screening of antibacterial activity showed that 26 isolates of LAB from the cocoa fermentation process had antibacterial activity against ESKAPE pathogens (zone of inhibition ≥11 mm). Cell-free supernatant from some LAB demonstrated potent antibacterial activity against some ESKAPE pathogens through MIC and MBC and evaluation. Furthermore, one isolate (mix48M01) exhibited antibacterial activity against Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterobacter spp., methicillin-resistant Staphylococcus aureus, and Enterococcus raffinousus. An increase in the diameter of inhibition zone was observed as the lactic acid production increased. Gram-positive bacteria isolates were more resistant to LAB than Gram-negative bacteria. Bacteriocin with good antibacterial activity was also detected from the LAB examined. Molecular analysis showed that 13 LAB shared >99% sequence similarity to Lactobacillus fermentum CECT 562 while one isolate shared sequence similarity <98%, indicating that it might be a novel Lactobacillus species. Isolated LAB from this study exhibited high antibacterial activity against ESKAPE pathogens and could be investigated further as potential probiotics and antibacterial agents in the future.

Antibacterial Assessment of Oils from Four Plants Against Selected Gentamicin-Resistant Gram-Positive Bacteria

Background: The continuous rise of the menace of antibiotic resistance in microorganisms remains a global health problem, and this places a significant burden on the effective management of infections caused by multiple-resistant species. Phytocompounds are being constantly assessed for bioactive components to discover new products. This study was designed to investigate the antibacterial activity of oils from four plants – sweet almond (SAO), avocado (AO), black seed (BSO), and coconut (CO) against gentamicin-resistant Gram-positive bacteria from environmental sources. Methods: Fifty-seven water and soil samples were collected, and gentamicin-resistant Gram-positive bacteria isolates were recovered and identified with the ABIS online Microbiology software. Antibiotic susceptibility testing was done using the Kirby-Bauer disc diffusion method and multiple antibiotic resistance indices (MARI) of isolates evaluated. The antibacterial activity of plant oils was evaluated with the agar well diffusion technique, and analyses of bioactive compounds in the oil samples using a gas chromatograph-mass spectrometer (GC-MS). Results: Altogether, 60 Gram-positive Gentamicin-resistant bacterial isolates were recovered. The isolates spanned 34 species belonging to 11 genera, namely Staphylococcus spp. (27), Bacillus spp. (18), Enterococcus spp. (2), Listeria spp. (2), Macrococcus spp. (3), Corynebacterium sp. (1), Lactobacillus sp. (1), Paenibacillus sp. (1), Rothia sp. (1), Salinicoccus sp. (1) and Streptococcus spp. (3). A high proportion of the isolates were resistant to erythromycin and oxacillin at 96.7%, and ampicillin at 86.7%. Meropenem was observed to be the most effective in-vitro. All isolates (100.0%) exhibited multidrug resistance and had MARI above 0.4. In undiluted forms, plant oils exhibited very low inhibitory activity against isolates but improved upon dilution of the plant oils in the order BSO > SAO > CO > AO. Conclusion: The extremely high levels of multidrug resistance suggest the rapid dissemination of antibiotic resistance traits in the community and are quite bothersome. The plant oils exhibited low antimicrobial activity, emphasising the need for a continuous search for bioactive compounds against multidrug-resistant pathogens. This study, therefore, recommends the in-vivo investigation of the plant oils and the possibility of a synergistic relation-ship of these plant oils with conventional antibiotics.

BACTERIAL PATTERN AMONG SEPSIS PATIENTS IN INTERNAL MEDICINE INPATIENT WARD DR. SOETOMO GENERAL ACADEMIC HOSPITAL, SURABAYA, INDONESIA IN 2017-2019

Highlights:1. Bacteria remain a major cause of bacterial sepsis.2. The most common causal agent in bacterial septicaemia was the gram-positive bacterium. Abstract: Background: Bacteria remain the primary cause of bacterial sepsis. Gram-negative bacteria are the most commonly isolated from sepsis patients. However, gram-positive bacterial infections have also increased recently. Objective: To identify the pattern of bacterial infection in sepsis patients in Internal Medicine inpatient ward Dr. Soetomo General Academic Hospital, Surabaya, Indonesia. Material and Method: This retrospective study reviewed the medical records of all sepsis patients in Internal Medicine Ward Dr. Soetomo General Academic Hospital, Surabaya, Indonesia from January 1 – December 31, 2016. All patients were divided according to bacterial species into two groups: patients with gram-positive and gram-negative infection. The collected data were statistically analyzed using SPSS ver. 16.0 to find out the frequency. Result: From 179 eligible data reviewed, there were 103 (57.5%) patients with gram-positive bacterial infection and 76 (43.5%) patients with a gram-negative bacterial infection. The major isolates of gram-positive bacteria were Staphylococcus hominins (30 isolates) and gram-negative bacteria was Escherichia coli (30 isolates), 43 isolates showed multi-drug resistant organisms; Escherichia coli ESBL 23 isolates, Klebsiella pneumoniae ESBL 3 isolates, Klebsiella oxytoca ESBL 2 isolates and Methilcillin Resistance Staphylococcus aureus 5 isolates. Conclusion: The most common causative agent in bacterial sepsis was gram-positive bacteria. The major isolated gram-positive bacteria are Staphylococcus hominis and gram-negative bacteria were Escherichia coli. The species of multi-drug resistant organisms found are Methilcillin-Resistant Staphylococcus aureus (MRSA), Escherichia coli ESBL, Klebsiella pneumonia ESBL and Klebsiella oxytoca ESBL. Among the patients with multi-drug resistant organism infection, Escherichia coli ESBL were the most prevalent one.

Characterisation of Staphylococci species from neonatal blood cultures in low- and middle-income countries

BackgroundIn low- and middle-income countries (LMIC) Staphylococcus aureus is regarded as one of the leading bacterial causes of neonatal sepsis, however there is limited knowledge on the species diversity and antimicrobial resistance caused by Gram-positive bacteria (GPB).MethodsWe characterised GPB isolates from neonatal blood cultures from LMICs in Africa (Ethiopia, Nigeria, Rwanda, and South Africa) and South-Asia (Bangladesh and Pakistan) between 2015–2017. We determined minimum inhibitory concentrations and performed whole genome sequencing (WGS) on Staphylococci isolates recovered and clinical data collected related to the onset of sepsis and the outcome of the neonate up to 60 days of age.ResultsFrom the isolates recovered from blood cultures, Staphylococci species were most frequently identified. Out of 100 S. aureus isolates sequenced, 18 different sequence types (ST) were found which unveiled two small epidemiological clusters caused by methicillin resistant S. aureus (MRSA) in Pakistan (ST8) and South Africa (ST5), both with high mortality (n = 6/17). One-third of S. aureus was MRSA, with methicillin resistance also detected in Staphylococcus epidermidis, Staphylococcus haemolyticus and Mammaliicoccus sciuri. Through additional WGS analysis we report a cluster of M. sciuri in Pakistan identified between July-November 2017.ConclusionsIn total we identified 14 different GPB bacterial species, however Staphylococci was dominant. These findings highlight the need of a prospective genomic epidemiology study to comprehensively assess the true burden of GPB neonatal sepsis focusing specifically on mechanisms of resistance and virulence across species and in relation to neonatal outcome.

Antimicrobial Resistance of Secondary Bacterial Infections in Patients Hospitalized with Covid-19

Coronavirus disease 2019 (COVID-19) is an emerging serious global health problem. It has been recognised for a considerable time-period, that viral respiratory infections predispose patients to bacterial infections, and that these co-infections have a worse outcome than either infection on its own. This study was carried out on 100 samples of sputum from COVID-19 patients. During the laboratory diagnosis, 156 bacterial isolates were obtained from the positive samples . The Gram-positive bacteria isolates included Strptococcus pneumonia 64(40%) Streptococcus pyogenes, 7 (4%), Streptococcus mitis 1 (1%) Streptococcus mutus 1(1%) , Streptococcus parasanguinis 1(1%) Staphylococcus eidermidis 10(6%), staphylococcus aureus 4 (2%), Micrococcus lutus,1 (1%). Whereas, Gram-negtive bacteria included Pseudomonas aeruginosa 9 (6%), E coli 10 (6%), Serratia marcescens, 3 (2%), Klebsiella pneumonia 31 (19%), H.influenzae 10 (6%) Acinetobacter baumannii , 4 (2%) .The isolates varied in their response against the antibiotics ; and Gram positive bacteria were significantly (p<0.05) more sensitive to the antibiotic then Gram negative. To detect the Streptococcus pneumonia isolates , the house keeping Eno genes was screened. Results showed that all the isolates, had Eno gene (100%). Furthermore, This study was carried out in order to detect tet-L and ermB gene in 10 S.pneumonia isolates . genes were Results showed that all the isolates, numbering 10, had both tetL and ermB genes(100%). Keywords : Bacteria, Co-infections, COVID-19, Eno gene, tet-L gene , ermB gene

The Antimicrobial Potential of Hexane Oils and Polyphenols-Rich Extracts from Pistacia vera L.

Pistachio (Pistacia vera L.) nuts contain nutrients and phytochemicals which have been linked to several positive outcomes. The aim of this research was to examine the antimicrobial effect of natural raw and roasted unsalted polyphenols-rich pistachio extracts (NRRE and RURE) and hexane oil fractions. American Type Culture Collection (ATCC), food and clinical isolates of Gram-positive bacteria (Listeria monocytogenes and Staphylococcus aureus), Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli and Enterococcus faecium) and yeasts (Candida albicans) were used. In addition, the influence of the extraction method was evaluated. Generally, NRRE extracts were richer in polyphenolic compounds compared with RURE extracts. NRRE extracted with n-hexane was the most effective on Listeria monocytogenes food isolates strains (MIC values between 0.25 and 2.0 mg mL−1). All extracts, except for RURE extracted with n-hexane, were active against Listeria monocytogenes ATCC 13932. Both hexane oil fractions were active against Listeria monocytogenes ATCC 13932 and Enterococcus faecium DSZM 17050. The oil obtained from natural pistachio was active against three food isolates of Listeria monocytogenes. In conclusion, the present study indicates an inhibitory effect of pistachio polyphenols against Listeria monocytogenes, one of the most serious pathogens causing foodborne disease.

Genotypic Detection of qnrA and qnrC Genes in Citrobacter koseri Isolated from Patients with Urinary Tract Infection.

The urinary tract infection (UTI) is a prevalent infection that affects people of all ages. Bacterial agents are the most common causes of UTIs. Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and other staphylococcal species, Citrobacter freundii and Citrobacter koseri (C. koseri) account for a smaller number of infections. These pathogens are transported into the urinary tract from the colonic biotope into dysbacteriosis. Urine samples were randomly collected from 249 outpatients who were suspected of having UTIs. After genital cleaning, 10 mL of urine specimens were collected in a sterilized bowel. Then, the specimens were centrifuged at 2,000 rpm for 5 min and the residue was aerobically incubated with the broth infusion of brain flasks at 37°C for 24 h and then applied with a sterile ring onto blood agar plates and MacConkey agar (OxoidTM). Out of 249 urine samples, the results proved that there were 176 (70.7%) and 51(20.5%) gram-negative and gram-positive bacteria isolates, respectively. However, the results demonstrated that there were 22 (8.8%) urine samples with no growth. In addition, the results showed that eight various antimicrobials are used to treat C. koseri. In the current study, C. koseri was treated with eight different antimicrobial agents. The antimicrobial resistance rate for 7 isolates against Cefotaxime, Ceftriaxone, Ciprofloxacin, and Levofloxacin was high for 6 (85.71%) isolates. The results indicated that 6 and 5 isolates had 85.71% and 71.42% antimicrobial resistance against Ceftazidime and Levofloxacin, respectively. Whereas Gentamicin showed a moderate rate of resistance (4 isolates, 57.14%), and Amikacin resistance was found in 5 isolates, accounting for 28.57%. The bacterial isolates had a high susceptibility rate to Imipenem. The qnrA gene was found in 6 (85.71%) isolates. However, the recorded data demonstrated that there is no isolate carrying the qnrC gene. Among all pathogenic bacteria, C. koseri was the lowest causative agent of UTI in this study and was highly resistant to most antimicrobials except Imipenem, which was a good antibiotic with 100% sensitivity.