Características clínicas y microbiológicas de la infección de vías urinarias bajas en población ambulatoria / Clinical and Microbiological Characteristics of Lower Urinary Tract Infections in an Outpatient Population

Introducción: El tratamiento de las infecciones del tracto urinario es casi siempre empírico, lo que genera una serie de problemas en la consulta diaria. Objetivo: Caracterizar clínica y microbiológicamente las infecciones de vías urinarias bajas no complicadas en pacientes de una clínica de primer nivel. Métodos: Se realizó un estudio transversal descriptivo. La identificación de las bacterias del cultivo de orina se efectuó por métodos establecidos. La prueba de susceptibilidad a los antimicrobianos se realizó por la técnica Kirby-Bauer. Se utilizó el programa estadístico SPSS versión 26, con la prueba de ji al cuadrado y un análisis multivariado discriminante. Se calculó también razón de momios con el programa Epi-Info. Resultados: Se incluyeron 270 pacientes, con frecuencia de 39,3 por ciento de cultivos positivos, y Escherichia coli como la especie predominante. Se identificaron, además, 31,3 por ciento de bacterias Gram positivas. Se presentó significancia estadística entre la infección urinaria y factores como el sexo, y la infección del tracto urinario previa en las mujeres. Se obtuvo 100 por ciento de cepas resistentes a ampicilina. En general, se obtuvieron porcentajes de resistencia altos en los antimicrobianos probados. Conclusiones: Escherichia coli fue la especie más frecuentemente aislada, sin embargo, existe una serie de microorganismos implicados en enfermedades del tracto genital como Gardnerella vaginalis, que parecen estar involucrados en la etiología de las infecciones del tracto urinario. Se identificaron factores de riesgo como el sexo biológico y las infecciones previas en mujeres. Se obtuvieron porcentajes de resistencia altos en los antimicrobianos probados(AU)

Introduction: The management of urinary tract infections is almost always empirical, which generates a series of problems in the daily consultation. Objective: To characterize, clinically and microbiologically, uncomplicated lower urinary tract infections in patients of a primary level clinic. Methods: A descriptive and cross-sectional study was carried out. Bacterial identification in urine culture was performed by established methods. Antimicrobial susceptibility testing was performed using the Kirby-Bauer technique. The statistical software SPSS (version 26) was used, with the chi squared test and multivariate discriminant analysis. Odds ratios were also calculated with the Epi-Info program. Results: A total of 270 patients were included, with a 39.3percent frequency of positive cultures and Escherichia coli as the predominant species. In addition, 31.3percent of Gram-positive bacteria were identified. There was statistical significance between urinary tract infection and factors such as sex or previous urinary tract infection in women. One result was 100percent of ampicillin-resistant strains. In general, high percentages of resistance were obtained for the tested antimicrobials. Conclusions: Escherichia coli was the most frequently isolated species; however, there is a number of microorganisms implicated in genital tract diseases, such as Gardnerella vaginalis, which appear to be involved in the etiology of urinary tract infections. Risk factors such as biological sex and previous infections in women were identified. High percentages of resistance were obtained for the tested antimicrobials(AU)

Multicenter Evaluation of a Gradient Diffusion Method for Antimicrobial Susceptibility Testing of Helicobacter pylori.

Helicobacter pylori is an important human pathogen associated with peptic ulcer disease, dyspepsia, and gastric malignancy. Antimicrobial susceptibility testing (AST) is often requested for patients who fail eradication therapy. The Clinical and Laboratory Standards Institute (CLSI) reference method, agar dilution (AD), is not performed in most laboratories and maintaining organism viability during transit to a reference laboratory is difficult. We assessed the performance of the Etest (bioMérieux) as a method for H. pylori AST in comparison to AD. Etest MICs were determined for 83 H. pylori isolates at ARUP and Cleveland Clinic (CC). Categorical agreement (CA), very major, major, and minor errors (VME, ME, and mE) were determined for Etest using AD performed at Mayo Clinic Laboratories as the reference method. Testing on isolates with errors was repeated to determine final results summarized below. For clarithromycin, 66.3% of isolates were resistant (R) by AD; Etest results at each laboratory showed 1mE (1.2%) and 1 ME (3.8%). For tetracycline, only 2 isolates were R by AD; a single VME occurred at both sites (98.8% CA, 50% VME) with the same isolate. Applying EUCAST levofloxacin breakpoints to interpret ciprofloxacin results, 60.2% of isolates were R by AD; ARUP CA was 97.6% (1 ME (3%), 1 VME (2%)) and CC CA was 96.3% (1 ME (3%), 2 VMEs (4%)). Despite high error rates, the categorical agreement was acceptable (>90%) for all three antibiotics between AD and Etest. In-house susceptibility testing by gradient diffusion can allow for testing of fastidious organisms that may not survive transport to specialized laboratories; however, the method is not without technical challenges. Characterization of resistance mechanisms, increased AD dilutions, and testing from the same inoculum may determine if the observed errors reflect technical issues or breakpoints that need optimization. IMPORTANCE Routine antimicrobial susceptibility testing (AST) of Helicobacter pylori by agar dilution is difficult to perform and not practical in most clinical microbiology laboratories. The Etest gradient diffusion method can be a reliable alternative for H. pylori AST with the advantage of being a less laborious quantitative method. This work reveals that an optimized Etest method can provide acceptable performance for H. pylori AST and describes the challenges associated with this methodology.

Coexistence of ESBL and MBL-mediated resistance in Acinetobacter baumannii in a Tertiary Care Hospital, Bangladesh.

Antimicrobial resistance mediated by extended-spectrum beta-lactamases (ESBL), AmpC beta-lactamase and metallo-beta-lactamase (MBL) producing Acinetobacter species is an emerging problem worldwide. In this cross-sectional study total 341 specimens were collected over a period of one year from January 2017 to January 2018. Specimens were collected from ICU and Surgery unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Specimens were collected from ICU and Surgery Unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Samples were processed for culture by standard conventional methods and susceptibility testing and determined by Kirby Bauer disc diffusion method. Antibiotic discs and their strength were according to the CLSI 2017 guideline. Molecular study was done to detect the species by OXA-51 gene and drug resistance genes (IMP, VIM, NDM, TEM, SHV, CTX, SPM, SIM and GIM). Species identification was done by OXA-51 gene which is intrinsic to Acinetobacter baumannii. Among the 46 isolates, 36(78.26%) were positive for Oxa-51 gene, 16(34.8%) for TEM gene, 9(19.6%) for VIM gene, 3(6.5%) for NDM gene and 1(2.2%) for IMP gene. This study gives an alarming sign towards high prevalence of cephalosporin and carbapenem resistance due to production of extended spectrum beta-lactamases and metallo-betalactamases, respectively. Early detection, proper antibiotic policies, and compliance towards infection control practices are the best defenses against these organisms.

Chemical Composition and In Vitro Antioxidant and Antimicrobial Activities of Marrubium vulgare L.

In this study, the polyphenol content and the antioxidant and antimicrobial activities of hydroethanolic (MVE) and hydroacetonic (MVA) leaf extracts of Marrubium vulgare L. were examined. The results indicated that the total phenolic content was higher in MVA (112.09 ± 4.77 mg GAE/DW) compared to MVE extract (98.77 ± 1.68 mg GAE/DW). The total flavonoid content was also higher in MVA extract (21.08 ± 0.38 mg QE/g DW) compared to MVE (17.65 ± 0.73 mg QE/g DW). Analysis of the chemical composition revealed the presence of 13 compounds with a total of 96.14%, with the major compound being malic acid (22.57%). Both extracts possess a good total antioxidant activity. DPPH and FRAP assays indicated that the MVE extract possesses a better antioxidant activity, with IC50 = 52.04 µg/mL ± 0.2 and EC50 of 4.51 ± 0.5 mg/mL, compared to MVA extract (IC50 = 60.57 ± 0.6 µg/mL and EC50 of 6.43 ± 0.0411 mg/mL). Moreover, both extracts exhibited strong antimicrobial activity against certain nosocomial strains as indicted by the MIC values, which ranged between 0.93 mg/mL and 10 mg/mL. Taken together, these results reveal the importance of M. vulgare as a natural antioxidant with important antimicrobial activity.

GC-MS profiling and in vitro antioxidant, cytotoxic and antimicrobial activities of Trianthema triquetra Rottl. ex Willed.

Trianthema triquetra Rottl. ex Willed is being used as a herbal remedy for various diseases in India and Pakistan. Still, no scientific data is available about therapeutic potential and phytochemistry of the plant. The aim of the current investigation is to perform GC-MS analysis, antioxidant (total phenolic and flavonoid content, DPPH assay), antimicrobial (disc diffusion assay) and cytotoxic (XTT and RBC's cellular membrane protection assay) studies. Methanolic extract and its fractions (n-hexane, ethyl-acetate, chloroform, n-butanol and water) were investigated for in vitro studies. Results showed that n-butanol fraction exhibited a significant (p<0.05) antioxidant potential (IC50=63.35±0.13 µg/mL) and also possess highest phenolic content (177±4.36 mg/g GAE). Whereas, n-hexane fraction showed highest flavonoid content (14.67±1.53 mg/g QE). 2, 4-Ditert-butyl-6-nitrophenol (26.79%) and Squalene (25.64%) were detected as major components through GC-MS analysis of chloroform fraction, eluted from column chromatography. Moreover, chloroform fraction also exhibited antimicrobial potential. Significant (p<0.05) dose dependent inhibition response on cell growth against CCRF-CEM cell lines was exhibited by methanolic extract. Furthermore, hemolytic potential of methanolic extract was found to be in safe range (2.23%-6.37%). So, it can be inferred that Trianthema triquetra can be exploited as an alternative remedy for cancer, oxidative stress related disorders and various skin diseases.

Anticancer, Enhanced Antibacterial, and Free Radical Scavenging Potential of Fucoidan- (Fucus vesiculosus Source) Mediated Silver Nanoparticles.

The present research displays the green synthesis of stable silver nanoparticles (AgNPs). The aqueous solution of Fucoidan from Fucus vesiculosus source (brown marine algae) is used as a reducing and capping agent. UV-Vis spectroscopy, XRD, FT-IR, SEM, EDX, and TEM with selected area electron diffraction are used to characterize the synthesized silver nanoparticles (AgNPs). The synthesized AgNPs exhibit a surface plasmon resonance at 430 nm after 24 h. The characterization results showed that AgNPs are crystalline in nature and exhibit mostly spherical shapes with an average diameter of 4-45 nm. Silver nanoparticles showed effective antibacterial activity against representative pathogens of bacteria. The activities of commercial antibiotics were enhanced by impregnation with the synthesized AgNPs. It also shows good fungicidal and anticancer activity against liver and lung cell lines and shows significant antioxidant efficacy (84%) at 10 µg/ml AgNP concentration against DPPH. The utilization of environmentally synthesized AgNPs offers numerous benefits of ecofriendliness and compatibility for biomedical applications.

Photodynamic effect of light emitting diodes on E. coli and human skin cells induced by a graphene-based ternary composite.

In this paper, the photodynamic effect of a ternary nanocomposite (TiO2-Ag/graphene) on Escherichia coli bacteria and two human cell lines: A375 (melanoma) and HaCaT (keratinocyte) after exposure to different wavelength domains (blue, green or red-Light Emitting Diode, LED) was analyzed. The results obtained through bioassays were correlated with the morphological, structural and spectral data obtained through FT-IR, XPS and UV-Vis spectroscopy, powder X-Ray diffractometry (XRD) and STEM/EDX techniques, leading to conclusions that showed different photodynamic activation mechanisms and effects on bacteria and human cells, depending on the wavelength. The nanocomposite proved a therapeutic potential for blue light-activated antibacterial treatment and revealed a keratinocyte cytotoxic effect under blue and green LEDs. The red light-nanocomposite duo gave a metabolic boost to normal keratinocytes and induced stasis to melanoma cells. The light and nanocomposite combination could be a potential therapy for bacterial keratosis or for skin tumors.

Clove Buds Essential Oil: The Impact of Grinding on the Chemical Composition and Its Biological Activities Involved in Consumer's Health Security.

This study is aimed at identifying the chemical composition of the essential oil extracted from the Syzygium aromaticum seeds, as well as investigating its biological activities, insecticide effect, and allelopathic properties. The extraction yield was about 14.3 and 7.14% for grounded and ungrounded seeds, respectively. The GC-MS analysis allowed the identification of 17 heterogeneous compounds, including eugenol (68.7-87.4%), as major compound, cyperene (20.5-7.2%), phenethyl isovalerate (6.4-3.6%), and cis-thujopsene (1.9-0.8%), respectively, for grounded and ungrounded seeds. Concerning the antibacterial activity, the diameter of the inhibition zone reached 35 mm when the essential oil extracted from grounded seeds was applied against Escherichia coli. Regarding the antioxidant activity via the DPPH radical scavenging test, the IC50 varied from 1.2 ± 0.1 to 2.8 ± 0.5 µg/mL. With respect to reducing power, the efficient concentration EC50 ranged from 32 to 50 µg/mL. The essential oil exhibited also an allelopathic effect against seeds of Hyoscyamus niger, as well as an insecticide effect against Sitophilus oryzae with a DL50 value of 252.4 µL/L air. These findings enhance the use of this spice as a natural food preservative and encourage its use in several fields, including pharmaceutical, cosmetics, agriculture, and therapy, that could be a strategic way to guarantee the consumer's health.

Evaluation of antioxidant, antimicrobial and anticancer activities of compounds reported Saussurea hypoleuca Spreng. roots.

Current study was intended to isolate bioactive compounds from ethyl acetate fraction of Saussurea hypoleuca root extract and evaluation of their antioxidant, antimicrobial and anti-cancerous activities which might be helpful for their chemo preventive potential against selected bacterial strains. Column chromatography was done for isolation of compounds which were characterized on the basis of extensive spectroscopic analysis; Infra-red (IR), Electron Ionization (EI-Positive), Proton Nuclear Magnetic Resonance (1H-NMR) and Carbon Nuclear Magnetic Resonance (13C-NMR). Two compounds were identified, as sesquiterpenes (40mg) and linoleic acid (33mg) from 10 grams of ethyl acetate fraction. Both compounds have shown in vitro antioxidant activity which in regard; 2, 2- diphenyl-1-picrylhydrazyl (DPPH) scavenging potential was high in sesquiterpenes (261.81) as compared to linoleic acid (90.89). The minimum inhibitory concentrations (MIC) of both compounds were evaluated in various bacterial and fungal strains against respective controls. However, in human hepatocellular carcinoma (Hep G2 cell lines) sesquiterpenes exhibited strong anticancer potential than linolenic acid which might be its potential free radical inactivator in MTT assay. This paper directs the ethano medicinal worth of plant root as it possesses bioactive compounds which in our best knowledge these compounds isolated and reported first time from this plant root specie.

A mechanistic study on the inhibition of bacterial growth and inflammation by Nerium oleander extract with comprehensive in vivo safety profile.

BACKGROUND: Nerium oleander (L.) is well known traditionally used medicinal plant with several pharmacological activities. However, the anti-bacterial, anti-inflammatory activity and in vivo toxicity potential of floral parts of this plant are not reported. Therefore the present study was designed to investigate these activities of Nerium oleander ethanolic flower extract (NOEE) in different animal models. METHODS: Antimicrobial activity of plant extract was compared with five different antibiotics using the disk diffusion method. The time-killing kinetic assay and bacterial killing mechanism of NOEE were also performed. Anti-inflammatory activity was assessed using granuloma induced by cotton-pellet, rat paw edema induced by carrageenan and levels of different inflammatory biomarkers on healthy Wistar rats. The protein and mRNA expressions of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) were also measured. Acute (14 days) and sub-acute (28 days) oral toxicity studies were also performed on healthy Sprague Dawley rats. RESULTS: NOEE produced highly significant (P < 0.005) and significant (P < 0.05) zones of inhibition at 30 mg/mL and 20 mg/mL respectively against most of the tested bacterial strains. NOEE produced a more drop in viable counts of Gram-negative isolates within 20 min. After 12 h exposure with NOEE, the SEM images of MRSA showed the destruction of cell membrane. NOEE showed highly significant (P < 0.005) anti-inflammatory activity in cotton-pellet and carrageenan inflammatory models. In addition, treatment with NOEE also decreased the production of NO, PGE2, TNF-α and IL-1ß in the rat paw after treated with carrageenan. Similarly, NOEE also suppressed the inducible nitric oxide synthase (iNOS), TNF-α, IL-1ß, and cyclooxygenase-2 (COX-2) mRNA expressions. It is also showed highly significant reduction in total leukocyte count (73.09%) and C-reactive protein levels (54.60%). NOEE also inhibited COX-1, COX-2, 5-LO and 12-LO in a highly significant manner. Moreover, acute and sub-acute toxicity studies of NOEE in rats confirm the toxicity with hepatotoxicity at higher doses (2000 mg/kg) i.e. four times greater than the therapeutic dose. CONCLUSION: It is concluded that crude flower extract of N. oleander is a potent antimicrobial and anti-inflammatory agent with no toxicity potential at therapeutic doses.

Magnificines A and B, Antimicrobial Marine Alkaloids Featuring a Tetrahydrooxazolo[3,2-a]azepine-2,5(3H,6H)-dione Backbone from the Red Sea Sponge Negombata magnifica.

Investigation of the Red Sea sponge Negombata magnifica gave two novel alkaloids, magnificines A and B (1 and 2) and a new ß-ionone derivative, (±)-negombaionone (3), together with the known latrunculin B (4) and 16-epi-latrunculin B (5). The analysis of the NMR and HRESIMS spectra supported the planar structures and the relative configurations of the compounds. The absolute configurations of magnificines A and B were determined by the analysis of the predicted and experimental ECD spectra. Magnificines A and B possess a previously unreported tetrahydrooxazolo[3,2-a]azepine-2,5(3H,6H)-dione backbone and represent the first natural compounds in this class. (±)-Negombaionone is the first ß-ionone of a sponge origin. Compounds 1-3 displayed selective activity against Escherichia coli in a disk diffusion assay with inhibition zones up to 22 mm at a concentration of 50 µg/disc and with MIC values down to 8.0 µM. Latrunculin B and 16-epi-latrunculin B inhibited the growth of HeLa cells with IC50 values down to 1.4 µM.

Smaller Copper Oxide Nanoparticles have More Biological Effects Versus Breast Cancer and Nosocomial Infections Bacteria.

BACKGROUND AND OBJECTIVES: Despite promising successes in developing new drugs and pharmaceutical biotechnology, infectious diseases and cancer are still the principal causes of mortality and morbidity globally. Therefore, finding effective ways to deal with these pathogens and cancers is critical. Metal nanoparticles are one of the new strategies to combat bacteria and cancers. METHODS: We examined the antimicrobial activity of 30 and 60 nm copper oxide nanoparticles (CuO-NPs) against Acinetobacter baumannii and Staphylococcus epidermidis bacteria responsible for nosocomial infections in standard and clinical strains and anti-cancer activity against 4T1 cell line as malignancy breast cancer cells. Synthesis of CuO-NPs was performed by a one-step reduction method and confirmed by DLS and TEM microscopy at 30 and 60 nm sizes. The antibacterial and anti-cancer activities of the nanoparticles were then investigated against the aforementioned bacteria and breast cancer. RESULTS: Using disk, well, MIC, MBC methods, and viability/bacterial growth assay, 30 nm CuO NPs were found to have more antibacterial activity on standard and clinical strains than 60 nm CuO NPs. On the other hand, using MTT, apoptosis, and gene expression method, 30 nm nanoparticles were found to have more anti-cancer potential than 60 nm CuO NPs. CONCLUSIONS: Our findings implicate CuO-NPs to possess antimicrobial and anti-cancer effects and more significant potential in smaller sizes, suggesting their pharmaceutical and biomedical capacity.
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Frequency and expression of genes involved in adhesion and biofilm formation in Staphylococcus aureus strains isolated from periodontal lesions.

BACKGROUND/PURPOSE: The aim of this study was to characterize the Staphylococcus aureus strains isolated from periodontal lesions of patients, to determine the expression of genes involved in cell adhesion upon their infection of human epithelial cells using an in vitro model, its biofilm formation, and its resistance to antibiotics. METHODS: S. aureus was analysed by PCR, Kirby-Bauer, and pulsed-field gel electrophoresis (PFGE), measuring gene expression by real-time PCR after infection of human cells in vitro. RESULTS: S. aureus was identified in 18.6% (50/268) of the samples. All strains (n = 50) possessed the virulence genes spa (Staphylococcal protein A), coa (coagulase), and icaAB (intercellular adhesin); 96% (n = 48) possessed clfB (clumping factor B), and 88% (n = 44) possessed ebps (elastin-binding protein) and sdrD (serine aspartate repeat protein D). All strains were resistant to methicillin, ampicillin, dicloxacillin, cefotaxime, and penicillin, and were multidrug resistant to 6-12 antibiotics. PFGE analysis showed 37 different pulsed-field types and most strains (60.4%) had a unique pulsed-field type. Twenty-four distinct combinations of virulence genes and antibiotic-resistant phenotypes were identified. CONCLUSION: Although S. aureus has been considered a transient member of the oral microbiota, our results indicate a high-level expression of virulence genes and multidrug resistance in the strains isolated from periodontal lesions. These strains might complicate the successful treatment of the disease.

Corn starch films as a long-term drug delivery system for chlorhexidine gluconate.

The present study describes the development of a chlorhexidine long-term drug delivery system using starch as a biodegradable polymer base. Three batches of thermoplastic starch films, containing starch particles/nanoparticles and chlorhexidine (CHX), were manufactured by casting. Morphological characterization showed an irregular surface with particles incorporated with chlorhexidine agglomerated in a starch matrix. Nanoindentation showed that the control film (without chlorhexidine) presented a more plastic and rigid behavior in relation to the films containing CHX. CHX was partially bounded to starch and prevented starch crystallization. Starch nanoparticles formed by precipitation were observed through transmission electron microscopy. By incorporating CHX into the solution, the nanoparticles presented different morphology, suggesting absorption of the drug. In vitro drug release was observed for 21 days by UV-vis spectrophotometry and released CHX amounted up to 19 mg/100 ml. Films presented microbiological potential for inhibiting Staphylococcus aureus growth as evaluated by the disk diffusion test in agar. It has been concluded that the developed film met the main requirements for a drug delivery system and that it is possible to be produced from a simple, cheap and reproduceable process.

Comparación de los perfiles de resistencia antimicrobiana de Escherichia coli uropatógena e incidencia de la producción de betalactamasas de espectro extendido en tres establecimientos privados de salud de Perú / Comparison of Escherichia coli antibiotic-resistance profiles and incidence of beta- lactamase phenotypes in three private health facilities in Perú

Introducción. La aparición de enterobacterias multirresistentes y productoras de betalactamasas de espectro extendido en pacientes ambulatorios con infecciones urinarias representa un problema de salud pública en Perú. Objetivo. Comparar los perfiles de resistencia de Escherichia coli uropatógenas e identificar los fenotipos de cepas productoras de betalactamasas de espectro extendido en tres establecimientos privados de salud localizados en las regiones de la costa, la sierra y la selva de Perú. Materiales y métodos. Se llevó a cabo durante el 2016 un estudio descriptivo de 98 muestras de orina de pacientes con infección urinaria, 35 procedentes de Lima (costa), 38 de Juliaca (sierra) y 25 de Iquitos (selva), en el que se determinó la sensibilidad antimicrobiana utilizando ocho discos antibióticos. Asimismo, se evaluó la producción de betalactamasas de espectro extendido con discos de cefotaxima, de ceftazidima o de su combinación, con ácido clavulánico en agar Mueller-Hinton. Resultados. Se identificaron 18 perfiles de resistencia que incluían desde los sensibles a todos los antibióticos hasta los resistentes simultáneamente a siete antibióticos, con el 18,4 % de aislamientos resistentes a un antibiótico y el 54,0 % de multirresistentes. Se detectó producción de betalactamasas en el 28,6 % de las cepas procedentes de la región de Puno. También, se observó un mayor número de casos en el rango de edad de 31 a 45 años con resistencia a ceftazidima, ceftriaxona, gentamicina y trimetoprim-sulfametoxazol en el establecimiento de salud de Puno. Conclusión. Los perfiles de resistencia variaron según la localización geográfica del establecimiento de salud, observándose mayor resistencia a los antibióticos en la región de la sierra de Perú, con el 28,6 % de cepas productoras de betalactamasas de espectro extendido.

Introduction: The appearance of multidrug-resistant and beta-lactamase producing enterobacteria in outpatient care facilities represent a public health problem in Perú. Objective: To compare the resistance profiles of uropathogenic Escherichia coli and to identify extended-spectrum beta-lactamase-producing phenotypes in three private health facilities located in the Peruvian coast, Andean and jungle regions. Materials and methods: We conducted a descriptive study on 98 urine samples from Lima (coast), Juliaca (Andean region) and Iquitos (jungle region) during 2016. We determined the antimicrobial susceptibility in 35 samples from Lima, 38 from Juliaca and 25 from Iquitos using eight antibiotic disks in samples from patients diagnosed with urinary infection. We also evaluated the production of extended-spectrum beta-lactamases with cefotaxime and ceftazidime disks and a combination of both with clavulanic acid on Mueller-Hinton agar. Results: We identified 18 resistance profiles ranging from those sensitive to others simultaneously resistant to seven antibiotics: 18.4% resistant to one and 54.0% to multiple antibiotics. We detected beta-lactamase production in 28.6% of the strains from the Puno region. Likewise, we observed a greater number of cases with resistance to ceftazidime, ceftriaxone, gentamicin, and trimethoprim-sulfamethoxazole in Puno's health facility in patients within the 31 to 45 year age range. Conclusion: Resistance profiles varied according to the geographical location of the health facilities under study. Resistance to antibiotics was higher in the Andean region with 28.6% of strains producing extended-spectrum beta-lactamases.

Bacterial etiology of ocular and periocular infections, antimicrobial susceptibility profile and associated factors among patients attending eye unit of Shashemene comprehensive specialized hospital, Shashemene, Ethiopia.

BACKGROUND: Eye infection is a public health problem in developing countries including Ethiopia. Bacteria are major causative agents of eye infections that can lead to loss of vision. The objective of this study was to determine bacterial etiology of ocular and periocular infections, antimicrobial susceptibility profile and associated factors among patients who visited the eye unit of Shashamane Comprehensive Specialized Hospital (SCSH). METHOD: A hospital-based cross-sectional study was conducted at SCSH from September 1, 2018, to March 30, 2019. Specimens from the ocular and periocular areas were collected from a total of 332 patients who visited the eye unit. Specimens were inoculated on blood agar, chocolate agar, MacConkey agar, and mannitol salt agar. Isolated bacteria were identified by a series of biochemical tests using the standard bacteriological method. Antimicrobial susceptibility test was performed according to the Clinical and Laboratory Standard Institute by disk diffusion method. Factors that could be associated with ocular and periocular infection were collected by using structured questionnaire. Data analysis was done using SPSS version 22.0 software package. A P value less than 0.05 was considered statistically significant. RESULT: Out of the total 332 study participants with ocular and periocular infections, 198(60%) were culture positive. The proportion of Gram-positive and Gram-negative bacteria were 135(68.2%) and 63(31.8%) respectively. Among Gram-positive bacteria, Staphylococcus aureus were predominant. Among Gram-negative bacteria, Escherichia coli were predominant. Most S. aureus were resistant to penicillin. CONCLUSION: Majority of ocular and periocular infections in this study were caused by bacteria; Gram-positive bacteria were responsible for most cases.

Effect of Amoxicillin in combination with Imipenem-Relebactam against Mycobacterium abscessus.

Infections caused by Mycobacterium abscessus are increasing in prevalence in cystic fibrosis patients. This opportunistic pathogen's intrinsic resistance to most antibiotics has perpetuated an urgent demand for new, more effective therapeutic interventions. Here we report a prospective advance in the treatment of M. abscessus infection; increasing the susceptibility of the organism to amoxicillin, by repurposing the ß-lactamase inhibitor, relebactam, in combination with the front line M. abscessus drug imipenem. We establish by multiple in vitro methods that this combination works synergistically to inhibit M. abscessus. We also show the direct competitive inhibition of the M. abscessus ß-lactamase, BlaMab, using a novel assay, which is validated kinetically using the nitrocefin reporter assay and in silico binding studies. Furthermore, we reverse the susceptibility by overexpressing BlaMab in M. abscessus, demonstrating relebactam-BlaMab target engagement. Finally, we highlight the in vitro efficacy of this combination against a panel of M. abscessus clinical isolates, revealing the therapeutic potential of the amoxicillin-imipenem-relebactam combination.

Antimicrobial activities of silver nanoparticles of extra virgin olive oil and sunflower oil against human pathogenic microbes.

Silver nanoparticles were synthesized using extra virgin olive oil (Olea europaea L.) and sunflower oil (Helianthus annuus L.) and characterized by UV-vis spectroscopy, X-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The brown color solution of olive oil nanoparticles (EVOO-NPs) and sunflower oil nanoparticles (SFO-NPs) showed typical absorption at 418 nm and 434 nm respectively. The morphology of extra virgin olive oil was found to be in semi cubic shapes with particle size of 23.45 nm (XRD) and 42.30 nm (SEM) while particle size of (SFO-NPs) had 42.30 nm (XRD) and 46.80 nm (SEM). Antimicrobial activities of crude extra virgin olive oil (EVOO), crude sunflower oil (SFO), synthesized nanoparticle from (EVOO-NPs) and (SFO-NPs) against human pathogenic strains were investigated. Synthesized nanoparticle from each oil showed a potent antimicrobial activity against all tested micro-organisms than crude oil which increased by (81.14% to 174.65 %) and by (111.65% to 192.31 %) than (EVOO) and (SFO) respectively. Both (EVOO-NPs) and (EVOO) had more antimicrobial activities than (SFO-NPs) and (SFO). EVOO (NPs) and SFO (NPs) showed maximum antibacterial activities against K. pneumoniae. Therefore (EVOO-NPs) and (SFO-NPs) could be used as safe natural product against multidrug resistant microbes.

Resistance profile of genetically distinct clinical Pseudomonas aeruginosa isolates from public hospitals in central Pakistan.

INTRODUCTION: Pseudomonas aeruginosa (member of ESKAPE group) is predominantly responsible for emerging nosocomial infections and poses serious health concern due to ever-increasing drug resistance trends. The current study investigates the prevalence of such highly resistant P. aeruginosa in major hospital settings and further characterizes and compares them for genetic heterogeneity. MATERIALS AND METHODS: Samples of patients (n=108) with wound infections, bacteremia and burn injuries from major hospitals of Rawalpindi and Islamabad during 2017 to 2018 were collected for the present study. The samples were processed in the COMSATS Microbiology and Public Health lab and screened for the P. aeruginosa by routinely used biochemical tests, drug susceptibility tests and rapid molecular approaches. RESULTS: The results suggested that most of the isolates (88/108) are indeed P. aeruginosa (81.4%) underpinning the need of its active surveillance in hospital settings. Further analysis suggested that 32 of these 88 microbes are multi-drug resistance (36.3%), 16 (18.1%) are extensively drug resistance and 4 (4.5%) are pan-drug resistance. Moreover, double disc synergistic test suggested that 16 (18.1%) are positive for metallo-ß-lactamase production. Molecular screening confirmed that 2 (12.5%) and 3 (18.75%) of these 16 isolates are positive for VIM and NDM gene respectively while all the studied isolates were positive for AmpC ß-lactamase. PAP17 isolate harbors both VIM and NDM genes. ERIC PCR profiling showed that majority of MDR bacteria fall in cluster II and III similarly XDR bacteria also fall in cluster II and III while PDR bacteria fall in cluster IV. CONCLUSION: This study revealed that majority of the isolates are multi drug resistant MDR and extensively drug resistant (XDR). However, the presence of some pan drug resistant (PDR) isolates among such small sample size screened is of utmost concern. Molecular typing of extremely resistant P. aeruginosa revealed high genetic diversity. Therefore, we suggest that regular monitoring and surveillance of such highly resistant P. aeruginosa in hospital settings will help to control their transmission and hence reduce the disease burden.

Ionic liquid functionalized biogenic synthesis of AgAu bimetal doped CeO2 nanoparticles from Justicia adhatoda for pharmaceutical applications: Antibacterial and anti-cancer activities.

The noble metal such as Ag and Au doped CeO2 nanoparticles was successfully prepared by ionic liquid assisted hydrothermal method in the presence of Justicia adhatoda leaves extract and were used as an antibacterial and anticancer agent. The FT-IR and Raman spectrum exhibit the peaks at 460 and 464 cm-1 assigned to CeO stretching vibrations of NPs. The electron microscopic micrographs confirmed the spherical shaped morphology of prepared NPs. The insertion of Ag and Au ions into the CeO2 surface creates lattice defects the leads to reduce the band gap energy of Ag-Au/CeO2 at 3.15 eV. The XRD results suggested the average crystalline size of the silver­gold loaded CeO2 was 28 nm. From the elemental mapping images, we have visualized that existence and uniform distribution of Ag, Au, Ce, and O in the prepared nanomaterials. The antibacterial activity of unloaded and bimetal loaded CeO2 NPs was evaluated with Gram-positive and Gram-negative bacteria using disc diffusion assay. The AgAu loaded CeO2 NPs exhibited the highest zone of inhibition against E. coli and S. aureus strains when compared with pristine CeO2, Ag loaded CeO2, and Au loaded CeO2 NPs. In addition, the 100 µg mL-1 of CeO2, Ag/CeO2, Au/CeO2 and Ag-Au/CeO2 NPs exposed the 50, 51, 52 and 56% of anticancer activity against the HeLa cells respectively. Overall, this study concludes that the ionic liquid functionalized green synthesized bimetal loaded cerium oxide NPs showed potent antibacterial and anticancer activities.