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This study found S. aureus to be a major pathogen in OIs, although it is not common conjunctival flora. The data caution that prevalence of methicillin resistance in coagulase-negative staphylococci is more than S. aureus in OIs and must be considered in their treatment. Despite methicillin resistance, staphylococci from OIs and NC remain sensitive to vancomycin and cefazolin.
All isolates from ocular infections collected between 2000 and 2009 were prospectively tested against several antibiotics in vitro. S. pneumoniae isolates (n = 93) were tested against 20 and S. aureus (n = 120) and CoNS (n = 214) against 19 antibiotics. To identify changes in susceptibility patterns, we compared results from 2000-2004 with those from 2005-2009. We also compared the antibiotic susceptibilities against aminoglycosides and quinolones between methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) isolates.
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A retrospective analysis of consecutive microbiologically confirmed enteric fever isolates.
Fifty isolates of Escherichia coli serogroup O111 recovered from humans and various animal species over a 24-year period (1976-1999) were examined for typical virulence-associated factors and susceptibilities to antimicrobials of human and veterinary significance. Nine H (flagellar) types were identified including nonmotile (n = 24), 32 (n = 12), negative (n = 5), and 56 (n = 3). Thirty-five (70%) isolates possessed at least one Shiga-toxin-producing E. coli (STEC)-associated virulence determinants (eae, stxl, stx2, hlyA) via PCR analysis. Of these 35 isolates, 20 possessed eae, stxl, and hlyA genes, whereas three isolates possessed eae, stxl, stx2, and hylA genes. Multiple antibiotic resistance was observed in 70% of the 50 E. coli O111 isolates. The majority of isolates displayed resistance to streptomycin, sulfamethoxazole, tetracycline, and kanamycin. Bacterial resistance to ampicillin, gentamicin, chloramphenicol, trimethoprim and apramycin was also observed. Integrons were identified in 23 (46%) of the E. coli isolates assayed, with a 1-kb amplicon being most frequently observed. DNA sequencing of these integrons revealed the presence of the aadA gene, encoding resistance to streptomycin. Two integrons of 1.5 and 2 kb contained the aadA2 and either dfrI or dfrXII genes, encoding resistance to streptomycin and trimethoprim, respectively. Integrons were also identified from isolates dating back to 1982. Isolates were further genetically characterized via ribotyping, which identified 15 distinct ribogroups, with 62% of isolates clustering into four major ribogroups. Certain riboprint patterns from different animal species, including humans, were observed in isolates spanning the 24-year collection period, suggesting the dissemination of specialized pathogenic O111 clones.
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This five-year analysis from southern India shows a high burden and increasing trend of rickettsial infections among children. The occurrence of retinal vasculitis and a high rate of severe complications draw attention to the need for early diagnosis and management of these infections.
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One hundred seventy-six autologous serum containers used in home therapy were studied; 48 of them included an adapted filter (Hyabak; Thea, Clermont-Ferrand, France), and the other 128 were conventional containers. Containers equipped with a filter were tested at 7, 14, 21, and 28 days of use, whereas conventional containers were studied after 7 days of use. In addition, testing for contamination was carried out in 14 conventional containers used during in-patient therapy every week for 4 weeks. In all cases, the preparation of the autologous serum was similar. Blood agar and chocolate agar were used as regular culture media for the microbiologic studies, whereas Sabouraud agar with chloramphenicol was the medium for fungal studies.
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Among 25526 recorded isolates of salmonellae, 5086 isolated from humans and 20440 from animals in 1994 and 1997 in France, the antibiotic resistance phenotype was determined for all human and 5336 animal isolates. In Salmonella enterica serovar typhimurium, one of the two most frequently isolated serovars from humans as well as animals, resistance to ampicillin was observed in 61% of both human and animal isolates in 1994 and in 73% of human and 53% of animal isolates in 1997. During these periods, resistance to co-amoxiclav was between 45% and 66% for both types of isolate. Resistance to ampicillin was associated with resistance to streptomycin, spectinomycin, sulphonamide, tetracycline and chloramphenicol in over 70% of isolates. Resistance to ampicillin as well as co-amoxiclav never exceeded 7% in Salmonella enteritidis. While Salmonella hadar was practically absent among the human isolates in 1994, this serovar was the third most frequent in 1997, and at that time 92% were resistant to nalidixic acid. Among the animal S. hadar isolates, the prevalence of resistance to nalidixic acid increased from 3% in 1994 to 72% in 1997. None of these isolates manifested high-level resistance to ofloxacin. The levels of resistance to aminoglycosides (< or =3%) and trimethoprim-suphamethoxazole (< or =14%) remained practically unchanged in all three serovars. The resistance markers of 463 ampicillin-resistant S. typhimurium isolated in 1997 were determined. Among the 24 phenotypes observed, six multiresistance phenotypes, representing 82% of these isolates (as compared with 80% in 1994), were associated with the PSE-1 gene typically found in the lysotype DT104 of this serovar.
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In the current study forty eight compounds belonging to anthraquinones, naphthoquinones, benzoquinones, flavonoids (chalcones and polymethoxylated flavones) and diterpenoids (clerodanes and kauranes) were explored for their antimicrobial potential against a panel of sensitive and multi-drug resistant Gram-negative and Gram-positive bacteria. The minimal inhibitory concentration (MIC) determinations on the tested bacteria were conducted using modified rapid INT colorimetric assay. To evaluate the role of efflux pumps in the susceptibility of Gram-negative bacteria to the most active compounds, they were tested in the presence of phenylalanine arginine β-naphthylamide (PAβN) (at 30 µg/mL) against selected multidrug resistance (MDR) bacteria. The anthraquinone, emodin, naphthaquinone, plumbagin and the benzoquinone, rapanone were active against methicillin resistant Staphylococcus aureus (MRSA) strains of bacteria with MIC values ranging from 2 to 128 μg/mL. The structure activity relationships of benzoquinones against the MDR Gram-negative phenotype showed antibacterial activities increasing with increase in side chain length. In the chalcone series the presence of a hydroxyl group at C3' together with a methoxy group and a second hydroxyl group in meta orientation in ring B of the chalcone skeleton appeared to be necessary for minimal activities against MRSA. In most cases, the optimal potential of the active compounds were not attained as they were extruded by bacterial efflux pumps. However, the presence of the PAβN significantly increased the antibacterial activities of emodin against Gram-negative MDR E. coli AG102, 100ATet; K. pneumoniae KP55 and KP63 by >4-64 g/mL. The antibacterial activities were substantially enhanced and were higher than those of the standard drug, chloramphenicol. These data clearly demonstrate that the active compounds, having the necessary pharmacophores for antibacterial activities, including some quinones and chalcones are substrates of bacterial efflux pumps and therefore should be combined to efflux pump inhibitors in the fight against MDR bacterial infections.
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The objective of this study was to determine if the current dosage regimen for chloramphenicol CAP administered to children with severe malaria SM for presumptive treatment of concomitant bacterial meningitis achieves steady state plasma CAP concentrations within the reported therapeutic range of 10-25 mg/l. Fifteen children (11 male, 4 female) with a median age of 45 months (range: 10-108 months) and having SM, were administered multiple intravenous doses (25 mg/kg, 6 hourly for 72 h) of chloramphenicol sodium succinate CAPS for presumptive treatment of concomitant bacterial meningitis. Blood samples were collected over 72 h, and plasma CAPS, CAP and CSF CAP concentrations determined by high performance liquid chromatography. Average steady state CAP concentrations were approximately 17 mg/l, while mean fraction unbound (0.49) and CSF/plasma concentration ratio (0.65) were comparable to previously reported values in Caucasian children. Clearance was variable (mean = 4.3 l/h), and trough plasma concentrations during the first dosing interval were approximately 6 mg/l. Simulations indicated that an initial of loading dose of 40 mg/kg CAPS, followed by a maintenance dose of 25 mg/kg every 6 h would result in trough CAP concentrations of approximately 10 mg/l and peak concentrations <25 mg/l throughout the treatment period. The current dosage regimen for CAP needs to include a loading dose of 40 mg/kg CAPS to rapidly achieve plasma CAP concentrations within the reported therapeutic range.
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A plasmid containing the qnrS quinolone resistance determinant and the gene encoding the SHV-2 beta-lactamase has been discovered from a clinical Klebsiella pneumoniae strain isolated in Taiwan. The complete 98-kb sequence of this plasmid, designated pK245, was determined by using a whole-genome shotgun approach. Transfer of pK245 conferred low-level resistance to fluoroquinolones in electroporant Escherichia coli epi300. The sequence of the immediate region surrounding qnrS in pK245 is nearly identical (>99% identity) to those of pAH0376 from Shigella flexneri and pINF5 from Salmonella enterica serovar Infantis, the two other qnrS-carrying plasmids reported to date, indicating a potential common origin. Other genes conferring resistance to aminoglycosides (aacC2, strA, and strB), chloramphenicol (catA2), sulfonamides (sul2), tetracycline (tetD), and trimethoprim (dfrA14) were also detected in pK245. The dfrA14 gene is carried on a class I integron. Several features of this plasmid, including three separate regions containing putative replicons, a partitioning-control system, and a type II restriction modification system, suggest that it may be able to replicate and adapt in a variety of hosts. Although no critical conjugative genes were detected, multiple insertion sequence elements were found scattered throughout pK245, and these may facilitate the dissemination of the antimicrobial resistance determinants. We conclude that pK245 is a chimera which acquired its multiple antimicrobial resistance determinants horizontally from different sources. The identification of pK245 plasmid expands the repertoire of the coexistence of quinolone and extended-spectrum-beta-lactam resistance determinants in plasmids carried by various species of the family Enterobacteriaceae in different countries.
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Preexposure of Bifidobacterium longum NCIMB 702259T to cholate caused increased resistance to cholate, chloramphenicol, and erythromycin. The B. longum ctr gene, encoding a cholate efflux transporter, was transformed into the efflux-negative mutant Escherichia coli KAM3, conferring resistance to bile salts and other antimicrobial compounds and causing the efflux of [14C]cholate.
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This study highlights the increasing resistance of Neisseriae meningitidis to empirically used antimicrobial drugs.
The total of 259 coagulase-negative staphylococci isolates gained from aerosols created during dumping and utilisation of municipal waste were investigated. Species and subspecies identification allowed to determine predominance of novobiocin-resistant species which primarily colonise rodents or other animals. However, about 30% of isolates were species originating from humans. Metabolic properties of the studied group of isolates were examined, as well as their sensitivity to a set of 8 commonly used antibiotics: amoxicillin, erythromycin, tetracycline, clindamycin, chloramphenicol, gentamicin, trimethoprim/sulfamethoxazole and ciprofloxacin. Many isolates resistant to erythromycin (19%), tetracycline (13%) and a considerably smaller group resistant to clindamycin (6.2%), chloramphenicol (2%) and trimethoprim/sulfamethoxazole (1%) were found. It appeared that processes of occurring at the time of dumping and utilisation of waste affect the prevalence of only some features. The examined metabolic features were found to be relatively stable and no changes were observed, which would indicate the tendency of adaptation to existence in the inanimate matter environment. Only isolates with active nitrate reductase were more frequently detected. The increase in frequency of occurrence of isolates resistant to tetracycline and chloramphenicol and simultaneous elimination of isolates resistant to other antibiotics were observed.
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Enteric fever is systemic illness caused by Salmonella Typhi and Salmonella Paratyphi A, B and C. It is believed to be a readily treatable illness by many clinicians in the developing world where it is endemic; however, with the emergence of drug resistance to fluoroquinolones, treatment is becoming increasingly difficult. While drugs such as cefixime, previously believed to be effective, have been proven otherwise, new agents such as gatifloxacin and azithromycin have proven to be promising. Re-emergence of chloramphenicol sensitive strains in previously resistant areas points towards the concept of antibiotic recycling, preserving the use of older antibiotics. Antibiotic recycling has been used successfully in hospital settings. However, its usefulness in community settings, where the main burden of enteric fever resides, is challenging to manage due to logistics and a lack of infrastructure. Nalidixic acid resistance used to be a marker for clinical response to flouroquinolones; however, recent studies highlight the importance of decreased ciprofloxacin susceptibility as a better marker. Enteric fever, as a public health problem, has been tackled by protection of food and water supplies in the industrialised countries of the world. Nonetheless, that goal seems too far-fetched in the developing world where there are hundreds of villages, towns and cities without adequate infrastructures. Perhaps the key to solving this problem is combining point-of-use-purification of water (by chlorination) with the treatment of illness in the community. Treatment of chronic carriers is also necessary in order to halt the cycles of transmission.
This study was conducted to determine frequency and pattern of antimicrobial susceptibility of Shigella species isolated from stool specimens collected from patients presenting with bloody diarrhoea in Mwanza City, Tanzania. The study was carried out from October 2004 to October 2005 and involved patients attending Sekou Toure Regional Hospital and Butimba Health Centre. Bacteriological cultures were done at the National Institute for Medical Research laboratory. A total of 489 patients (median age = 20 years) participated in the study and were able to provide stool specimens. Shigella species were isolated from 14% (69/489) of the stool specimens collected. Of the sixty nine strains of Shigella spp isolated, 62 (90%) were S. flexneri and 7 (10%) were S. dysenteriae. All Shigella strains isolated showed high resistance to ampicillin, tetracycline, trimethoprim-sulphamethoxazole and chloramphenicol, drugs commonly used for management of shigellosis in Tanzania. However all isolates were fully susceptible to ciprofloxacin, nalidixic acid, erythromycin, cefuroxime and gentamycin. S. flexneri showed resistance to amoxy-clavulanic_acid and azithromycin in 5% and 2% of isolates, respectively. None of the S. dysenteriae isolates were resistant to these two drugs. Entamoeba histolytica, Giardia lamblia and Schistosoma mansoni were microscopically detected in 16.5%, 4.4% and 5.3% of patients, respectively. These findings suggest that there is a need to carry out extensive susceptibility studies in different parts of the country with view of re-appraising the current guidelines for management of bloody diarrhoea in Tanzania.
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BACKGROUND. Increasing resistance to some antimicrobial agents among anaerobic bacteria has made susceptibility patterns less predictable. METHOD. This was a prospective study of the susceptibility data of anaerobic organisms isolated from clinical specimens from patients with suspected anaerobic infections from June 2005 until February 2007. Specimens were submitted to the microbiology laboratory at Charlotte Maxeke Johannesburg Academic Hospital, where microscopy, culture and susceptibility testing were performed the using E test® strip minimum inhibitory concentration method. Results were interpreted with reference to Clinical and Laboratory Standards Institute guidelines for amoxicillin-clavulanate, clindamycin, metronidazole, penicillin, ertapenem, cefoxitin, ceftriaxone, chloramphenicol and piperacillin-tazobactam. RESULTS. One hundred and eighty anaerobic isolates were submitted from 165 patients. The most active antimicrobial agents were chloramphenicol (100% susceptible), ertapenem (97.2%), piperacillin-tazobactam (99.4%) and amoxicillin-clavulanic acid (96.7%). Less active were metronidazole (89.4%), cefoxitin (85%), clindamycin (81.7%), ceftriaxone (68.3%) and penicillin (33.3%). CONCLUSION. Susceptibility testing should be performed periodically to identify emerging trends in resistance and to modify empirical treatment of anaerobic infections.
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Human leptospirosis is generally treated with penicillin or doxycycline. We studied the susceptibilities of 11 serovars (seven species) of Leptospira to 14 antibiotics. With the exception of chloramphenicol, all tested agents were at least as potent as penicillin and doxycycline, with the macrolide and ketolide drugs producing the lowest MICs (and minimal bactericidal concentrations).
Polymorphisms in rpoS are common in Escherichia coli. rpoS status influences a trade-off between nutrition and stress resistance and hence fitness across different environments. To analyze the selective pressures acting on rpoS, measurement of glucose transport rates in rpoS+ and rpoS bacteria was used to estimate the role of F(nc), the fitness gain due to improved nutrient uptake, in the emergence of rpoS mutations in nutrient-limited chemostat cultures. Chemostats with set atmospheres, temperatures, pH's, antibiotics, and levels of osmotic stress were followed. F(nc) was reduced under anaerobiosis, high osmolarity, and with chloramphenicol, consistent with a reduced rate of rpoS enrichment in these conditions. F(nc) remained high, however, with alkaline pH and low temperature but rpoS sweeps were diminished. Under these conditions, F(sp), the fitness reduction due to lowered stress protection, became significant. We also estimated whether the fitness need for the gene was related to its regulation. No consistent pattern emerged between the level of RpoS and the loss of rpoS function in particular environments. This dissection allows an unprecedented view of the genotype-by-environment interactions controlling a mutational sweep and shows that both F(nc) and F(sp) are influenced by individual stresses and that additional factors contribute to selection pressure in some environments.
Thermococcus nautili, strain 30-1T (formerly reported as Thermococcus nautilus), was isolated from a hydrothermal chimney sample collected from the East Pacific Rise at a depth of 2633 m on the 'La chainette PP57' area. Cells were motile, irregular cocci with a polar tuft of flagella (0.8-1.5 µm) and divided by constriction. The micro-organism grew optimally at 87.5 °C (range 55-95 °C), at pH 7 (range pH 4-9) and with 2% NaCl (range 1-4%). Doubling time was 64 min in Zillig's broth medium under optimal conditions. Growth was strictly anaerobic. It grew preferentially in the presence of elemental sulfur or cystine, which are reduced to H2S, on complex organic substrates such as yeast extract, tryptone, peptone, Casamino acids and casein. Slow growth was observed on starch and pyruvate. Strain 30-1T was resistant to chloramphenicol and tetracyclin (at 100 µg ml(-1)) but sensitive to kanamycin and rifampicin. The G+C content of the genomic DNA was 54 mol%. Strain 30-1T harboured three plasmids named pTN1, pTN2 and pTN3 and produced membrane vesicles that incorporate pTN1 and pTN3. As determined by 16S rRNA gene sequence analysis, strain 30-1T is related most closely to Thermococcus sp. AM4 (99.3% similarity) and Thermococcus gammatolerans DSM 15229T (99.2%). DNA-DNA hybridization values (in silico) with these two closest relatives were below the threshold value of 70% (33% with Thermococcus sp. AM4 and 32% with T. gammatolerans DSM 15229T) and confirmed that strain 30-1 represents a novel species. On the basis of the data presented, strain 30-1T is considered to represent a novel species of the genus Thermococcus, for which the name Thermococcus nautili sp. nov. is proposed. The type strain is 30-1T (=CNCM 4275=JCM 19601).
This study was conducted to investigate antimicrobial resistance profiles of Escherichia coli isolates from broiler chickens in Alberta, Canada. Cecal contents of broiler chickens from 24 flocks were collected at slaughter between January and March 2005 for culture and antimicrobial susceptibility testing against a panel of 15 antimicrobials using a broth microdilution technique. Of 600 E. coli isolates tested, 475 (79.2%) were resistant to one or more antimicrobials, 326 (54.3%) were resistant to three or more antimicrobials, 65 (10.8%) were resistant to five or more antimicrobials, and 15 (2.5%) were resistant to seven or more antimicrobials. The most common resistance was to tetracycline (69.2%), followed by streptomycin (48.2%), kanamycin (40.3%), and sulfisoxazole (38.0%). None of the E. coli isolates were resistant to amikacin, ceftriaxone, or ciprofloxacin. Of the isolates that were resistant to two or more antimicrobials, the most common multidrug resistance patterns were streptomycinte-tracycline (44.0%), streptomycin-sulfisoxazole-tetracycline (30.7%), and kanamycin-streptomycin-sulfisoxazole-tetracycline (23.5%). Resistance to tetracycline and kanamycin (odds ratio = 46.7, P = 0.0001) was highly associated, followed by resistance to streptomycin and sulfisoxazole (odds ratio = 12.0, P = 0.0001), and streptomycin and tetracycline (odds ratio = 10.3, P = 0.0001). The flock level prevalence of resistance varied from 16.7% for chloramphenicol to 100.0% for ampicillin, streptomycin, sulfisoxazole, and tetracycline. The results of this study provided baseline information on antimicrobial susceptibility of E. coli isolates of broiler chickens at slaughter in Alberta, which can serve as a bench mark for future research.
The modified U1 snRNA gene can suppress expression of a target transgene. In the present study, its potential utility to inhibit a dominant negative/gain of function mutation is explored. Using a green fluorescent protein (GFP) target gene, inhibition was achieved in all cells transduced with U1antiGFP directed at multiple sites within GFP. Using a chloramphenicol acetyltransferase (CAT) target gene, inhibition was not increased by increasing the hybridization domain from 10 to 16 bp or when a site in an upstream exon or intron was targeted. To determine if a U1 anti-target design could discriminate between two transcripts that differ by a 1-2 bp mismatch, GFPtpz and GFPsaph were chosen as targets because they share sequence homology except for three regions where a 1, 2 or 3 bp mismatch exists. The results demonstrated that U1antiGFP correctly reduced its cognate GFP expression by >90% and therefore U1 anti-target constructs are able to discriminate a 1 or 2 bp mismatch in their target mRNA. Thus, these U1 anti-target constructs may be effective in a strategy of somatic gene therapy for a dominant negative/gain of function mutation due to the discreteness of its discrimination. It may complement other anti-target strategies to reduce the cellular load of a mutant transcript.
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The resistance to ampicillin, erythromycin, sulfadiazine, novobiocin and chloramphenicol is prevalent among the bacteria isolated from EUS-affected fish, and resistant determinants of some of these antibiotics have been transferred to the bacteria of other origin.