Characterization of Salmonella phages isolated from poultry coops and its effect with nisin on food bio-control.

Salmonella is a bacterium associated with food contaminated by various animals, primarily poultry. Interest and research on bacteriophages are increasing because they can be used as an alternative against increasing antibiotic resistance. In our study, eight Salmonella-specific lytic bacteriophages were isolated from chicken feces. Two of the isolated phages (AUFM_Sc1 and AUFM_Sc3) were chosen for their characterization due to their broader host range. Based on morphological and genomic analysis, AUFM_Sc1 was identified to be close to similar Enterobacteria spp. CC31 (Myoviridae) and AUFM_Sc3 was identified to be close to Salmonella phage vB_Sen_I1 (Demerecviridae (formerly Siphoviridae)). Although these phages have shown promise for use in phage therapy applications for chickens, further studies are needed on their suitability. When a cocktail of these phages (AUFM_Sc1 + AUFM_Sc3) and nisin combination was applied on chicken breast meat, it was determined that it was effective against Salmonella contamination and while a good inhibitory effect was observed on the food, especially during the first 48 h, the effect decreased later, but the bacterial concentration was still low compared to the control group. Therefore, it is considered that the combination of AUFM_Sc1 + AUFM_Sc3 + nisin can be used as a food preservative against Salmonella.

Characterization of two bacteriophages specific to Acinetobacter baumannii and their effects on catheters biofilm.

Multidrug-resistant strains of Acinetobacter baumannii cause major nosocomial infections. Bacteriophages that are specific to the bacterial species and destroy bacteria can be effectively used for treatment. In this study, we characterized lytic bacteriophages specific to A. baumannii strains. We isolated lytic bacteriophages from environmental water samples and then investigated their morphology, host range, growth characteristics, stability, genome analysis, and biofilm destruction on the catheter surface. Our results showed that the efficacy of the phages varied between 32% and 78%, tested on 78 isolates of A. baumannii; 80 phages were isolated, and two lytic bacteriophages, vB_AbaP_HB01 (henceforth called C2 phage) and vB_AbaM_HB02 (henceforth called K3 phage), were selected for characterization. Electron microscopy scans revealed that the C2 and K3 phages were members of the Podoviridae and Myoviridae families, respectively. Whole-genome sequencing revealed that the sequence of the C2 phage is available in the NCBI database (accession number: OP917929.1), and it was found sequence identity with Acinetobacter phage AB1 18%, the K3 phage DNA sequence is closely related to Acinetobacter phage vB_AbaM_phiAbaA1 (94% similarity). The cocktail of C2 and K3 phages demonstrated a promising decrease in the bacterial cell counts of the biofilm after 4 h. Under a scanning electron microscope, the cocktail treatment destructed the biofilm on the catheter. We propose that the phage cocktail could be a strong alternative to antibiotics to control the A. baumannii biofilm in catheter infections.

Synthesis of some new 2-(substituted-phenyl)imidazo[4,5-c] and [4,5-b]pyridine derivatives and their antimicrobial activities.

The synthesis of 5H-imidazo[4,5-c]pyridines analogues (1a - 1h) and 4H-imidazo[4,5-b]pyridines (3a - 3c) was achieved by reacting 3,4-diaminopyridine or 2,3-diaminopyridine with Na2S2O5 adduct of corresponding benzaldehydes (a1 - a8). Alkylation of compounds (1a - 1h) and (3a - 3c) using 4-chlorobenzyl and /or butyl bromide under basic conditions (K2CO3, DMF) predominantly resulted in the formation of N5 regioisomers (2a - 2l) and N4,3 regioisomers (4a - 4c1,2), respectively. The N5,4,3-regioisomeric structures were confirmed using 2D-NOESY (Nuclear Overhauser Effect Spectroscopy) and HMBC (Heteronuclear Multiple Bond Correlation) spectra. The antibacterial and antifungal activities of the synthesized compounds (2a - 2g, 4a - 5d) were evaluated in vitro against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Methicillin resistant S. aureus, Enterococcus faecalis and Candida albicans, Candida parapsilosis. Among the synthesized compounds, promising activities were observed with compounds 2g, 2h, 4a and 4b with lowest MIC values (4-8 µg/mL). The compounds 2i, 2j, 2k, 2l showed moderate activity. Additionally, a computational approach (ADMETlab 2.0) was used to evaluate the drug likeness properties of the compounds.

The effect of phage-antibiotic combination strategy on multidrug-resistant Acinetobacter baumannii biofilms.

Acinetobacter baumannii (A. baumannii) is considered a critical human pathogen due to multi-drug resistance and increased infections. As a result of the resistance of A. baumannii biofilms to antimicrobial agents, it is necessary to develop new biofilm control strategies. In the present study, we evaluated the efficacy of two previously isolated bacteriophage C2 phage, K3 phage and phage cocktail (C2 + K3 phage) as a therapeutic agent in combination with antibiotic (colistin) against biofilm of multidrug-resistant A. baumannii strains (n = 24). The effects of phage and antibiotics on mature biofilm were investigated simultaneously and sequentially in 24 and 48 h. The combination protocol was more effective than antibiotics alone in 54.16% of the strains in 24 h. The sequential application was more effective than the simultaneous protocol compared with the 24 h single applications. When the application of antibiotics and phages alone was compared with their combined administration in 48 h. The sequential and simultaneous applications were more effective than single applications in all strains except two. We observed that combination of phage and antibiotics could increase biofilm eradication and provides new insights into the use of bacteriophages and antibiotics in the treatment of biofilm-associated infections caused by antibiotic-resistant bacteria.

Assessment of synergistic activity of rhamnolipid and linezolid against methicillin-resistant Staphylococcus aureus in-vitro and in-vivo with Galleria mellonella larvae model.

Antibiotic resistance, one of the most crucial public health problems, has increased the interest in synergy studies of antibiotics with existing antibiotics and natural compounds to make current treatment more effective in addition to new drug development. In this study, the effectiveness of rhamnolipid and linezolid on the Galleria mellonella larvae model in-vitro and in-vivo against Methicillin-resistant Staphylococcus aureus isolates, which are problematic in treatment, were investigated. Four S.aureus (One ATCC 29213 strain and three methicillin-resistant strains) were used in the study. Two MRSA isolates were resistant to linezolid, and one was susceptible. Partial synergy was observed in one resistant strain, and although no synergy was observed in the other resistant strain, the minimum inhibitory concentration of the resistant strain decreased from 16 to 4 µg/mL with a four-fold decrease and reached the susceptibility limit. No change was observed in the MIC of linezolid-susceptible strains. The G.mellonella larval model demonstrated that combined therapy was more effective than monotherapy by survival function tests and CFU determination. RML/LNZ combination improved survival compared to monotherapy and decreased the bacterial burden from 108 to 103.

The comparison of lytic activity of isolated phage and commercial Intesti bacteriophage on ESBL producer E. coli and determination of Ec_P6 phage efficacy with in vivo Galleria mellonella larvae model.

Antibiotic resistance is one of the crucial public health challenges. As a result of rising resistance, as an alternative to antimicrobials, demands for bacteriophage therapy have increased significantly over the years. The objective of this study was to isolate and characterize potentially therapeutic phages active against Escherichia coli (E. coli) and compare the efficacy with commercial Intesti bacteriophage on the extended-spectrum beta-lactamase (ESBL) positive E. coli (ESBL-EC) and performed the effectiveness of bacteriophage using the Galleria mellonella (G. mellonella) larvae model. Intesti bacteriophage is a polyvalent bacteriophage-based drug. The isolated bacteriophages were obtained from the river and clinical isolates of E. coli were used for the enrichment of bacteriophage isolation. The phages were first screened based on plaque morphology and host ranges determined on clinical strains. The susceptibility of phages was determined against 50 clinical isolates of E. coli and eight different laboratory isolates using the spot test technique. E. coli lytic phage Ec_P6 was used to determine the therapeutic and preventive effects on the G. mellonella larvae model. The slides were prepared by G. mellonella hemolymph for cytologic examination, stained with May Grünwald Giemsa (MGG), and evaluated by light microscopy. The results of the activities revealed lytic spectra ranging from 24% to 97%. Overall strains were susceptible to one or more phages from the panel. It was proved that Intesti bacteriophage is very effective in a wide variety of strains of E. coli including test strains, also showed that isolated Ec_P6 phage is as effective as commercial phage. The best MOI of this phage was 0.01, and infectivity decreased above 60 °C. The results suggest that phage is stable at pH values ranging between 5.0 and 9.0. In vivo study was found that in E. coli infection to achieve a survival high rate the infected larvae should be after 2 h treated with 0.01 MOI phage (10 µL, 106 PFU/mL) and colistin doses (10 µL, 2.5 mg/kg). It also prevented infection, increasing the survival of the larvae compared to the untreated control group. Ec_P6 phage was found to have a potential for the treatment of E. coli infections.

Antifungal and Anti-Virulent Activity of Origanum majorana L. Essential Oil on Candida albicans and In Vivo Toxicity in the Galleria mellonella Larval Model.

The aim of this study was to investigate and compare in detail both the antifungal activity in vitro (with planktonic and biofilm-forming cells) and the essential oil composition (EOs) of naturally growing (OMN) and cultivated (OMC) samples of Origanum majorana L. (marjoram). The essential oil composition was analyzed using GC-MS. The major constituent of both EOs was carvacrol: 75.3% and 84%, respectively. Both essential oils showed high antifungal activity against clinically relevant Candida spp. with IC50 and IC90 less than or equal to 0.5 µg mL-1 and inhibition of biofilm with a concentration of 3.5 µg mL-1 or less. Cultivated marjoram oil showed higher anti-biofilm activity against C. albicans. In addition, OMC showed greater inhibition of germ-tube formation (inhibition by 83% in Spider media), the major virulence factor of C. albicans at a concentration of 0.125 µg mL-1. Both EOs modulated cell surface hydrophobicity (CSH), but OMN proved to be more active with a CSH% up to 58.41%. The efficacy of O. majorana EOs was also investigated using Galleria mellonella larvae as a model. It was observed that while the larvae of the control group infected with C. albicans (6.0 × 108 cells) and not receiving treatment died in the controls carried out after 24 h, all larvae in the infected treatment group survived at the end of the 96th hour. When the treatment group and the infected group were evaluated in terms of vital activities, it was found that the difference was statistically significant (p < 0.001). The infection of larvae with C. albicans and the effects of O. majorana EOs on the hemocytes of the model organism and the blastospores of C. albicans were evaluated by light microscopy on slides stained with Giemsa. Cytological examination in the treatment group revealed that C. albicans blastospores were phagocytosed and morphological changes occurred in hemocytes. Our results indicated that the essential oil of both samples showed strong antifungal activities against planktonic and biofilm-forming C. albicans cells and also had an influence on putative virulence factors (germ-tube formation and its length and on CSH).

The evaluation of five commercial bacteriophage cocktails against methicillin-resistant Staphylococcus aureus isolated from nasal swab samples.

Infections due to methicillin-resistant Staphylococcus aureus (MRSA) are a growing concern for public health resulting in increase in morbidity, length of hospital stay, and cost of treatment. MRSA nasal swab screening may give clinicians additional information for decision of empiric antimicrobial agents. While increasing antibiotic resistance leads to new treatment approaches, bacteriophages are one of the most promising methods for these alternatives. It was aimed to determine the effectiveness of bacteriophages against MRSA isolates. Nasal swab samples were collected from outpatients without any evidence of infection who applied to Hatay, Mersin and Gaziantep family and immigration health centers. A series (35) were isolated from Turkish patients, and G series (64) were isolated from Syrian immigrants. Methicillin resistance was determined phenotypically and genotypically. Also, antibiotic susceptibilities of all isolates were determined against erythromycin, clindamycin, gentamicin, linezolid, rifampicin, and mupirocin. The total antimicrobial resistance rates of isolates were found to be 11%, 28%, 8%, 5%, 16%, 19%, and 29% respectively. The high susceptibility rate against ciprofloxacin (88.8%) was remarkable. The overall susceptibility of MRSA strains to ENKO, INTESTI, PYO, SES, and staphylococcal bacteriophages was 67.7%, 55.5%, 53.5%, 61.6% and 44.4%, respectively. The antibiotic susceptibility rates (except erythromycin) and efficacy of bacteriophages were higher in group A. Considering that high efficacy rates were not achieved in the study and the sensitivity rates of Turkish isolates to all phages were found to be higher than those of Syrian isolates, searching for phages in the geographic regions where the pathogen is common may be helpful to obtain suitable phages for treatment.

Maldi-TOF MS identification and antibiotic resistance of Escherichia coli isolated from playground.

In this study, it was aimed to determine the antibiotic resistance of Escherichia coli strains isolated from samples taken from various children's parks of Ankara and to confirm the resistance by molecular methods. Five hundred fifty-four samples, including soil samples from 140 different parks and 414 swab samples from slides, swings, ferris wheels, seesaws, and other toys from 176 different parks, were taken. Fourty E. coli strains isolated from these samples were included in the study. Antibiotic susceptibility tests of 40 E. coli isolates were performed by EUCAST recommendations. The resistance rates of E. coli isolates were found as ciprofloxacin 5%, ampicillin 17%, trimethoprim/sulfamethoxazole 15%, streptomycin 12.5%, tobramycin 5%, gentamicin 5%, cefotaxime 2.5%, and ceftazidime 2.5%. Intermediate rates were found as 95%, 90%, and 70% for tobramycin, gentamicin, and streptomycin respectively. blaCTX-M ß-lactamase gene was investigated for an isolate determined to be resistant to both cefotaxime and ceftazidime but blaCTXM gene could not be detected. Aminoglycoside resistance of strains has been investigated because of high intermediate sensitivity rates. For this purpose, aac(6')-Ib, aac(3')-IIa, aph(3')-VI, ant(3')-I, aac(3')-IV, ant(2')-Ia genes scanned, and were detected 97.5% of our isolates ant (3')-I, %25 aac(6')-Ib', 5% aac(3')-IIa, 2.5% ant(2')-Ia. Also, aph(3')-VI, and aac(3')-IV genes could not be detected in any of the isolates. Consequently, it has been revealed that resistant E. coli strains isolated from children's parks can pose a potential risk in public health for transmission of resistant genes.

Microbial exopolysaccharide production of Streptococcus thermophilus and its antiquorum sensing activity.

Interest in the production of exopolysaccharides by microorganisms has increased in the recent years. Using low-cost product is the main step of microbial production to reduce cost and compete with chemical production. In this work, EPS production of Streptococcus thermophilus isolates from yogurt (S2), kefir (S3), and S. thermophilus ATCC 19258 (S1) isolate which was used as control strains were investigated by using different fruit pulps. S. thermophilus isolates were identified by morphological and 16S sequence analysis. The amount of EPS obtained was measured spectrophotometrically using glucose as standard with phenol sulfuric acid method. All three isolates produced higher amounts of EPS on M17 medium than Nutrient medium. When the fruit pulp was added to the medium, EPS production increased in all three isolates. When different nitrogen sources were added together with fruit pulp juice, EPS production increased. The highest amount of EPS produced by ATCC 19258 strain (21.570 mg/L) and S3 isolate (29.131 mg/L) is the medium where mixed fruit pulp juice and nitrogen source is tryptophan. It has been shown that EPS production is increased by adding fruit pulps to the prepared media. It is thought that apricot pulp can be a good alternative in EPS production especially in the evaluation of wastes. Also, antiquorum sensing activity of the highest amount EPS was determined by using Chromobacterium violaceum CV026 strain and found effective on violacein pigment inhibition and C6-AHL production of biosensor strain.

Penicacids H-J, three new mycophenolic acid derivatives from the marine-derived fungus Rhizopus oryzae.

Chemical investigation of secondary metabolites in crude methanol extract of a solid rice medium of a marine-derived fungus, Rhizopus oryzae, has enriched the metabolic profile of this genus by affording three mycophenolic acid derivatives recognized as new fungal metabolites trivially named as penicacids H-J (1-3), along with two known naphtho-γ-pyrone dimers, asperpyrone A (4) and dianhydroaurasperone C (5). Structure elucidation of isolated compounds was unambiguously determined based on extensive 1D and 2D NMR spectroscopic analyses together with comparing coupling constant and optical rotation values with those reported for related congeners in literature. All isolated compounds were assessed for their antibacterial activity against four different bacterial microorganisms and they revealed moderate to weak activities with minimum inhibitory concentration (MIC) values ranging from 62.5 to 250 µg mL-1.

Synthesis & Anticancer Evaluation of New Substituted 2-(3,4- Dimethoxyphenyl)benzazoles.

BACKGROUND: The benzazole nucleus is found in many promising small molecules such as anticancer and antibacterial agents. Bendamustine (Alkylating agent), Nocodazole (Mitotic inhibitor), Veliparib (PARP inhibitor), and Glasdegib (SMO inhibitor) are being clinically used as anticancer therapeutic which bear benzimidazole moiety. Based on the principle of bioisosterism, in the present work, 23 compounds belonging to 2-(3,4-dimethoxyphenyl)benzazoles and imidazopyridine series were synthesized and evaluated for their anticancer and antimicrobial activities. OBJECTIVE: A series of new 2-(3,4-dimethoxyphenyl)-1H-benz(or pyrido)azoles were synthesized and evaluated for their anticancer and antimicrobial activities. METHOD: N-(5-chloro-2-hdroxyphenyl)-3,4-dimethoxybenzamide 1, was obtained by the amidation of 2-hydroxy-5-chloroaniline with 3,4-dimethoxybenzoic acid by using 1,1'-carbonyldiimidazole. Cyclization of 1 to benzoxazole derivative 2, was achieved by p-toluenesulfonic acid. Other 1H-benz(or pyrido)azoles were prepared by the reaction between 2-aminothiophenol, ophenylenediamine, o-pyridinediamine with sodium metabisulfite adduct of 3,4-dimethoxybenzaldehyde. The NMR assignments of the dimethoxy groups were established by the NOESY spectra. RESULTS: Compound 12, bearing two chlorine atoms at the 5(4) and 7(6) positions of the benzene moiety of benzimidazole was found the most potent analogue against A549 cells with the GI50 value of 1.5 µg/mL. Moreover, 24 showed remarkable cell growth inhibition against MCF-7 and HeLa cells with the GI50 values of 7 and 5.5 µg/mL, respectively. The synthesized compounds have no important antibacterial and antifungal activities. CONCLUSION: It could be concluded that the introduction of di-chloro atoms at the phenyl ring of 2-(3,4-dimethoxyphenyl)-1H-benzimidazoles increases significant cytotoxicity to selected human tumor cell lines in comparison to other all benzazoles synthesized. Unsubstituted 2-(3,4- dimethoxyphenyl)-imidazopyridines also gave good inhibitory profile against A549 and HeLa cells.

High rate of colistin and fosfomycin resistance among carbapenemase-producing Enterobacteriaceae in Turkey.

When the problem with carbapenem-resistant Enterobacteriaceae (CRE) increases, the older antimicrobial agents such as colistin and fosfomycin are used for the treatment of these infections. In this study, the broth microdilution method for colistin and the agar dilution method for fosfomycin were used for a total of 147 multidrug-resistant (MDR) or extensively drug-resistant (XDR) strains of CRE. The study included Klebsiella pneumoniae (91.16%), Escherichia coli (7.48%), Enterobacter cloacae (0.68%), and Serratia marcescens (0.68%). All these strains produce various types of carbapenemase, including OXA-48, NDM, and KPC. Some of these strains also have three different carbapenemase mechanisms, including OXA-48 (78.23%), NDM (2.04%), and KPC (0.68%) or OXA-48 and NDM (10.88%), or OXA-48 and KPC (0.68%). About 76.19% of the strains and 67.35% of the strains were resistant for colistin and fosfomycin, respectively. A total of 21 out of 35 colistin-susceptible strains were found to be susceptible to fosfomycin. This study showed that the resistance rates of colistin and fosfomycin are high. The MDR and XDR strains of CRE are spreading in our region and thus a monitoring system for CRE should be followed. Moreover, the applicability of antimicrobial stewardship programs should be increased in all inpatient and outpatient settings.

Antifungal activity of some Sternbergia taxa: effects on germ tube and biofilm formation

Natural products are rapidly becoming the primary sources of novel antimicrobial agents, as resistance to existing antimicrobial agents is increasing. Apart from determining the antimicrobial activity of natural products, it is also important to understand their effects on the virulence factors of microorganisms. This study aimed to determine the antimicrobial activity of Sternbergia species prevalent in Turkey and investigate their role in the inhibition of germination tube and biofilm formation, both of which are known to be important virulence factors of Candida albicans. The antimicrobial activities of the plant extracts were evaluated using bore-plate and broth microdilution method. The extracts' capacity to inhibit the formation of the germ-tube was also evaluated. The findings of our study revealed that Sternbergia lutea, Sternbergia vernalis possessed antimicrobial activities, with MIC values ranging between 0.048 mg/mL and 0.39 mg/mL. The highest antimicrobial activity was observed against Candida dubliniensis (0.048 mg/mL). While evaluating the inhibition of fungal germination activities, S. vernalis extract (at a concentration of 0.09 mg/mL) was found to be the most effective against C. albicans ATCC 90028 strain. The results also indicated that S. vernalis extracts at sub-MIC levels inhibited germ tube formation and modulated the tail-length of germinated cells, both of which are important virulence factors of C. albicans. Furthermore, the inhibition of biofilm-formation was also investigated, and it was found that two Sternbergia spp. extracts at or below MIC levels inhibited biofilm formation.

RS sample: Can be guide for empirical treatment of haematological malignancy patients?

Neutropenia due to intensive chemotherapy in haematological malignancy patients leaves the host vulnerable and makes them susceptible to infections. Infections are the most important cause of morbidity and mortality especially in haematological malignancy and chemotherapy patients. In addition, the use of multiple or inappropriate antibiotics leads to the development of resistant microorganisms. Therefore, the choice of empirical treatment is of vital important in these patient groups. Escherichia coli, Acinetobacter baumannii and Klebsiella pneumoniae are among the most frequently isolated Gram negative bacteria in neutropenic patients. Rectal swab (RS) samples were obtained from haematological malignancy patients not yet on chemotherapy or have no infection on chemotherapy period, E. coli was isolated from these samples, and A. baumannii and K. pneumoniae colonization were investigated. Susceptibilities of bacteria against antibiotics used in empirical treatment and prophylaxis were determined by using Gradient test strips according to the EUCAST recommendation. All isolates were sensitive against colistin. The resistant rates of antibiotics were detected as 39.1%, 9.4%, 6.8%, 35.1%, 31%, 39.1% for ciprofloxacin, meropenem, imipenem, piperacillin-tazobactam, cefepime, ceftazidime respectively The clonal relationship between Gram negative bacteria of intestinal flora and infection agents of same patient was investigated by Pulsed-Field gel electrophoresis. Twenty-three of the 30 patients (76.6%) were found to have a clonal relationship between the bacterial isolates before and after infection. It was determined that it can be able to predict with RS samples about possible agents of infection and their antibiotic susceptibility patterns.

Chemical composition and antimicrobial activity of the commercial Origanum onites L. oil against nosocomial carbapenem resistant extended spectrum beta lactamase producer Escherichia coli isolates.

In recent years rapidly growing antibiotic resistance has increased interest toward natural products, especially essential oils because of their various effects. The aim of this study was to identify the chemical composition of the commercial Origanum onites essential oil (EO) and to investigate the antimicrobial activity by disc diffusion and dilution methods, against ten different ATCC strains, including eight bacteria, two yeasts and seventy-nine clinical nosocomial Escherichia coli isolates that produce extended spectrum beta lactamase (ESBL). The chemical composition of EO was analyzed by GC and GC-MS. The major compounds of the EO were determined as carvacrol (51.4%) followed by linalool (11.2%), p-cymene (8.9%) and γ-terpinene (6.7%). O. onites EO had antimicrobial activity against all standard strains and inhibited microbial growth of ESBL positive E. coli isolates. According to our results, O. onites EO may be an alternative to synthetic drug, used in combination with other antibiotics for treatment of infection caused by multidrug resistant bacteria after testing toxic effects and irritation at preferred doses on human.

Performance of CarbaNP and CIM tests in OXA-48 carbapenemase-producing Enterobacteriaceae.

This study applied two phenotypic tests, namely "Carbapenemase Nordmann-Poirel" (CarbaNP) test and "Carbapenem Inactivation Method" (CIM), against the isolates carrying the carbapenem resistance genes. The study included 83 carbapenem-resistant Enterobacteriaceae isolates producing oxacillinase-48 (OXA-48) and 30 carbapenem-sensitive Enterobacteriaceae isolates. Out of the total isolates studied, 77 isolates (92.77%) were identified as Klebsiella pneumoniae and six isolates (7.23%) were identified as Escherichia coli by Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry. Polymerase chain reaction (PCR) method used to detect resistance genes found that 74 isolates (89.16%) produced OXA-48 carbapenemase, whereas nine isolates (10.84%) produced both OXA-48 and New Delhi metallo-beta-lactamase-1 (NDM-1). The isolates producing both OXA-48 and NDM-1 were found to be positive by both phenotypic tests. Among isolates carrying only blaOXA-48 gene alone, nine isolates (13.04%) for CarbaNP test and two isolates for CIM test (2.90%) displayed false negative results, respectively. The sensitivity of CarbaNP and CIM tests was found to be 89.16% and 97.59%, respectively, whereas the specificity was determined to be 100% for both tests. These findings suggest that CarbaNP and CIM tests are useful tools to identify the carbapenemase producers. Molecular methods like PCR are recommended to verify false negative tests predicted to have OXA-48 activity.

[Comparison of MALDI-TOF and 16S rRNA methods in identification of viridans group streptococci]. / Viridans grup streptokok tanimlamasinda MALDI-TOF ve 16S rRNA yöntemlerinin karsilastirilmasi.

Accurate identification of viridans group streptococci (VGS) frequently encountered as a causative agent of infective endocarditis is always a challenge for the clinical microbiology laboratory. Clinical microbiology laboratories generally use semi automatic/full automatic systems, molecular methods and also conventional methods for the identification of these bacteria. There are recent published studies that have used MALDI-TOF (Matrix Assisted Laser Ionization Mass Spectrometry-Time of Flight) systems in the identification of VGS. The aim of the study was to compare the performance of the conventional methods, semi automatic and MALDI-TOF MS system used in identification of VGS in oral microbiota of persons under the risk of infective endocarditis, with the gold standard method 16S rRNA sequence analysis and to create a diagnosis algorithm for the identification of VGS in clinical microbiology laboratories according to the obtained data.The study was conducted with 51 VGS strains isolated from oral microbiota of the patients with rheumatologic cardiac, valve and/or prosthetic valve diseases, under the risk of development of infective endocarditis, who have admitted to Ankara Numune Training and Research Hospital, Department of Cardiology, between February-June 2015. Standard microbiology procedures, optochin susceptibility and bile solubility tests were done for the isolation of bacteria. Bacteria were also identified with APISTREP (bioMérieux, France) and MALDI-TOF MS Bruker Microflex (Bruker Biotyper; Bruker Daltonics, Bremen, Germany) methods. BSF-8 (5´-AGAGTTTGATCCTGGCTCAG-3´) and BSR-534(5´-ATTACCGCGGCTGCTGGC-3´) primers were used in the 16S rRNA sequence analysis of bacteria. ABI PRISM 3100 Avan t Genetic Analyzer (Applied Biossytems, Foster City, CA, USA) were used for the sequence analysis. Electropherograms were analyzed in SeqScape Software (Applied Biosystems, Foster City, CA, USA) and compared with the reference sequences in GenBank with BLASTN (NCBI). According to the result of optochin and bile solubility tests, with API STREP system, 16 (31,37%) of the isolates were identified as Mitis group, 15 (29.41%) as Anginosus group, 9 (17.5%) as Salivarius group, 7 (13,73%) as Sanguinis group and 4 (7.84%) as Bovis group among optochin and bile resistant alpha hemolytic streptococci. Moreover, of the same isolates 20 (39.22%) were identified as Mitis group, 14 (27.45%) as Anginosus group, 13 (25.49%) as Salivarius group and 4 (7.84%) as Sanguinis group with MALDI-TOF system. In the identification with 16S rRNA, 25 (49.02%) of the isolates were identified as Mitis group, 13 (25.49%) as Anginosus group, 12 (23.53%) as Salivarius group and 1 (1.96%) as Sanguinis group. According to the results, it was determined that 33 (64.70%) of the isolates identified in MALDI-TOF MS system and 31 (60.78%) of the isolates identified in API STREP system were compatible with 16S rRNA sequence analysis method. For Mitis group, API STREP test sensitivity was 48.00% and specificity was 84.62% and MALDI-TOF system sensitivity was 80.00% and specificity was 100%. As VGS identification is a complicated process, we believe a single method will be insufficient for the identification of these isolates in clinical microbiology laboratories. We suggest that MALDI-TOF system can be used for VGS diagnosis, however, optochin test and/or molecular methods should also be included in the diagnosis algorithm when necessary.

Antimicrobial susceptibility against penicillin, ampicillin and vancomycin of viridans group Streptococcus in oral microbiota of patients at risk of infective endocarditis.

The viridans group Streptococci (VGS) are most abundant in the mouth; in some instances they might emerge as pathogens particularly in infective endocarditis (IE). In this study, we aimed to define and determine the susceptibility against antibiotics of VGS that are members of the oral microbiota of patients exhibiting a risk of developing IE. Forty-nine patients at risk of infective endocarditis were included in the study. Identification of the bacteria was performed using API STREP (bioMérieux, France). Gradient test strips (E-Test, France) were used to determine MIC of the bacteria against penicillin, ampicillin, and vancomycin. The distribution of the isolated VGS groups was determined as follows: Streptococcus mitis 32.6% and anginosus group - 32.6%, S. sanguinis group - 16.3%, S. mutans group - 12.2%, and S. salivarius group - 6.1%. The rates of resistance and reduced sensitivity of the isolates for penicillin and ampicillin were determined at 61.2% and 55.1%, respectively. However, all isolates were found to be susceptible to vancomycin. We conclude that the antimicrobial resistance of VGS should be determined on a regular basis locally, and decisions on therapeutic and prophylactic interventions should be given taking this resistance into consideration.

Rhamnolipid Biosurfactants Produced by Pseudomonas Species

ABSTRACT: Surfactants are chemical products widely used in our daily life in toothpaste and other personal hygiene and cosmetic products, and in several industries. Biosurfactants are surfactants of biological origin that can be produced by microorganisms and have many advantages, such as low toxicity and high biodegradability, compared to synthetic counterparts. Unfortunately, high production costs limit the use of biosurfactants. Low-cost production is the most important factor for biosurfactants to be able to compete in the global market place. This review presents general information on rhamnolipid biosurfactant produced by Pseudomonas species, as well as on their production and applications. In addition, industrial products and their wastes used for rhamnolipid production are reviewed in detail based on recent studies.