Alloprevotella rava isolated from a mixed infection of an elderly patient with chronic mandibular osteomyelitis mimicking oral squamous cell carcinoma.

The anaerobic, Gram-negative bacillus Alloprevotella rava has recently been described in the human oral cavity. To our knowledge, this species has not been isolated from chronic osteomyleitis samples. We present the first case of A. rava infection in a 92-year-old woman with polymicrobial chronic mandibular osteomyelitis, mimicking oral squamous cell carcinoma.

Polymicrobial anaerobic meningitis caused by Bacteroides fragilis, Bacteroides thetaiotaomicron, Fusobacterium necrophorum and Slackia exigua in a patient with mastoiditis following otitis media.

We have reported a case of mastoiditis which progressed to meningitis in a 16-year old male patient. CSF (cerebrospinal fluid) anaerobic culture revealed four species of isolated anaerobic bacteria. This is the first case in the literature in which a patient survived childhood polymicrobial anaerobic meningitis diagnosed by MALDI-TOF MS (Matrix Assisted Laser Desorption Ionization - Time of Flight Mass Spectrometry).

Cost-effectiveness of a new system in ruling out negative urine cultures on the day of administration.

Urine samples account for a significant part of the workload in clinical microbiology laboratories. However, the culture process is time-consuming and a large proportion is reported as negative. To reduce unnecessary culture procedures and speed up the reporting of negative results, a reliable screening method is needed. For this purpose, urine samples submitted to our clinical microbiology laboratory were simultaneously screened by a flow cytometry method (Sysmex UF-1000i, Japan). During screening, the evaluation of various combinations of leucocytes and bacteria cut-offs demonstrated that cut-offs of 30 and 50/µL, respectively, were the best threshold values to reach a 100% negative predictive value (NPV) with a culture reduction rate of 44.8% in adults and 61.9% in children between the ages of 6 and 17 years. With the culture reduction rates mentioned above, the screening method has provided at least 24% savings in expenditures of the routine clinical microbiology laboratory. Since we did not reach such an NPV with any combinations of screening parameters in children younger than 5 years of age, we recommend cultivation of all urine samples in those patients without a screening step. In conclusion, Sysmex UF-1000i as a screening method was capable of improving the efficiency of the routine microbiology laboratory by providing negative results in a few minutes in children greater than 6 years of age and in adults.

Results from the Survey of Antibiotic Resistance (SOAR) 2011-13 in Turkey.

OBJECTIVES: Data are presented from the Survey of Antibiotic Resistance (SOAR) for respiratory tract infection pathogens collected in 2011-13 from Turkey. METHODS: MICs were determined using Etest(®). Susceptibility was assessed using CLSI, EUCAST and pharmacokinetic/pharmacodynamic (PK/PD) interpretive criteria. RESULTS: Rates of antibiotic susceptibility were very low among 333 isolates of Streptococcus pneumoniae tested: penicillin 38% using CLSI (oral) and EUCAST breakpoints; erythromycin 51% using CLSI and EUCAST criteria; and cefuroxime 64.6% using CLSI and PK/PD and 46.9% using EUCAST. Of the isolates, >90% were susceptible to amoxicillin/clavulanic acid, ceftriaxone (except using EUCAST criteria: 76%), levofloxacin and high-dose intravenous penicillin. Among 339 Haemophilus influenzae isolates, 6.8% were ß-lactamase positive while 9.1% were ß-lactamase negative but ampicillin resistant (BLNAR) by CLSI (14.7% by EUCAST) criteria. Amoxicillin/clavulanic acid susceptibility was ∼90% by CLSI (with or without BLNAR adjustment, EUCAST and high-dose PK/PD) but lower, at 82.9%, by EUCAST with BLNAR adjustment. Levofloxacin susceptibility was 96% using all three breakpoints. Dramatic differences in rates of susceptibility, depending on the breakpoints used, were seen for cefaclor [94% by CLSI (86.4% BLNAR adjusted), 23% by PK/PD] and cefuroxime [97% by CLSI (89.1% BLNAR adjusted), 85% by PK/PD, 15% by EUCAST (13.0% BLNAR adjusted)]. Streptococcus pyogenes (n = 222) and Moraxella catarrhalis (n = 40) isolates remained highly susceptible to amoxicillin/clavulanic acid, cephalosporins and levofloxacin, with only erythromycin susceptibility dropping below 95% for S. pyogenes. CONCLUSIONS: Overall, amoxicillin/clavulanic acid and levofloxacin were the most active antibiotics based on all three breakpoints against these pathogens. Although susceptibility was not universally low in Turkey, high resistance rates were found in S. pneumoniae and, when using PK/PD and EUCAST breakpoints, in other respiratory pathogens.

Results from the Survey of Antibiotic Resistance (SOAR) 2002-09 in Turkey.

OBJECTIVES: To investigate changes in antibiotic susceptibility of Streptococcus pneumoniae and Haemophilus influenzae from the Survey of Antibiotic Resistance (SOAR) in community-acquired respiratory tract infections (CA-RTIs) between 2002 and 2009 in Turkey. METHODS: Previously published SOAR data were used for this analysis. MICs were determined using Etest(®) gradient strips or disc diffusion. Susceptibility against a range of antimicrobial agents was assessed using CLSI breakpoints. RESULTS: A total of 900 S. pneumoniae isolates were analysed: 2002-03 (n = 75), 2004-05 (n = 301) and 2007-09 (n = 524). Four antibiotics were tested consistently throughout and three showed a statistically significant decrease in susceptibility (P < 0.0001): penicillin (74.7% susceptible in 2002-03; 67.8% in 2004-05; and 47.2% in 2007-09); cefaclor (85.3% in 2002-03; 78.7% in 2004-05; and 53.5% in 2007-09) and clarithromycin (85.3% in 2002-03; 82.7% in 2004-05; and 61.9% in 2007-09). Susceptibility to amoxicillin/clavulanic acid did not significantly change (100% in 2002-03; 98.7% in 2004-05; and 97.7% in 2007-09). A total of 930 H. influenzae isolates were analysed: 2002-03 (n = 133), 2004-05 (n = 379) and 2007-09 (n = 418). Four antibiotics were also consistently tested: ampicillin, amoxicillin/clavulanic acid, clarithromycin and cefaclor. All showed >90% susceptibility, but only cefaclor susceptibility significantly reduced (P < 0.0001) over time (99.2% in 2002-03; 96.3% in 2004-05; and 90.4% in 2007-09). CONCLUSIONS: In S. pneumoniae from Turkey, there has been a clear statistically significant reduction in susceptibility to key antibiotics since 2002, but not to amoxicillin/clavulanic acid (or amoxicillin). However, susceptibility in H. influenzae remained stable. Continued surveillance is required to monitor future changes in antibiotic susceptibility for CA-RTI bacteria.

Molecular characterization and risk factors for carbapenem-resistant Gram-negative bacilli colonization in children: emergence of NDM-producing Acinetobacter baumannii in a newborn intensive care unit in Turkey.

BACKGROUND: Multidrug-resistant Gram-negative bacilli are responsible for more than 50% of healthcare-associated infections. Colonization dynamics, characteristics, and risk factor data for CR-GNB are scarce in children. AIM: To examine the molecular characteristics of, and risk factors for nosocomial colonization with, carbapenem-resistant Gram-negative bacilli (CR-GNB) in hospitalized paediatric patients in a tertiary university hospital's paediatric units in Turkey. METHODS: A prospective case-control study was performed at a university hospital in Istanbul, Turkey. FINDINGS: A total of 1840 rectal swab specimens were collected from all 762 hospitalized children between March 2013 and October 2013. Among them, 176 (23%) patients were colonized with CR-GNB. Of these, 72 (9%) patients were colonized with carbapenem-resistant Enterobacteriaceae, 138 (18%) with CR-non-fermenter Gram-negative bacilli (CR-NF) and 34 (4%) with both. The median CR-GNB colonization time was 10 days (range: 1-116). The median duration of rectal colonization with CR-GNB was 8 days (range: 1-160). NDM (31%) was the second most frequent carbapenemase identified in Acinetobacter baumannii isolates, and has not previously been detected in Turkey. All of the 17 patients colonized with NDM-producing A. baumannii were newborns in the neonatal intensive care unit. Independent risk factors for CR-GNB colonization were: age <1 year, nasogastric tube placement, presence of underlying chronic diseases, ampicillin usage, surgical intervention, and carbapenem use. CONCLUSION: This is the first description of NDM in A. baumannii in newborn units in Turkey. Carbapenem usage is a common independent risk factor for both CRE and CR-NF colonization, which underscores the importance of antibiotic stewardship programmes.

Butyricimonas virosa: the first clinical case of bacteraemia.

The strictly anaerobic Gram-negative bacteria Butyricimonas species have recently been described in human faeces and have to our knowledge not been isolated in infectious clinical materials. We report the first case of Butyricimonas virosa bacteraemia in a 72-year-old man with colon adenocarcinoma, who underwent aortic aneurysm replacement surgery.

Antimicrobial resistance in gram-negative hospital isolates: results of the Turkish HITIT-2 Surveillance Study of 2007.

Resistance rates to amikacin, ciprofloxacin, ceftazidime, cefepime, imipenem, cefoperazone/sulbactam and piperacillin/tazobactam in Escherichia coli (n= 438), Klebsiella pneumoniae (n= 444), Pseudomonas aeruginosa (n= 210) and Acinetobacter baumanni (n=200) were determined with e-test in a multicenter surveillance study (Hitit-2) in 2007. ESBL production in Escherichia coli and K. pneumoniae was investigated following the CLSI guidelines. Overall 42.0% of E.coli and 41.4% of K. pneumoniae were ESBL producers. In E. coli , resistance to imipenem was not observed, resistance to ciprofloxacin and amikacin was 58.0% and 5.5% respectively. In K. pneumoniae resistance to imipenem, ciprofloxacin and amikacin was 3.1%, 17.8% 12.4% respectively. In P. aeruginosa the lowest rate of resistance was observed with piperacillin/tazobactam (18.1%). A. baumanni isolates were highly resistant to all the antimicrobial agents, the lowest level of resistance was observed against cefoperazone/sulbactam (52.0%) followed by imipenem (55.5%). this study showed that resistance rates to antimicrobials are high in nosocomial isolates and show variations among the centers.

A possible role of Bacteroides fragilis enterotoxin in the aetiology of colorectal cancer.

The prevalence of enterotoxigenic Bacteroides fragilis (ETBF) was investigated in stool specimens from 73 patients with colorectal cancer and from 59 control patients. Stool specimens were cultured on Bacteroides Bile Esculin agar and B. fragilis was identified by conventional methods. After DNA extraction, the enterotoxin gene (bft) was detected by PCR in 38% of the isolates from colorectal cancer patients, compared with 12% of the isolates from the control group (p 0.009). This is the first study demonstrating an increased prevalence of ETBF in colorectal cancer patients.

Microbiology of sinusitis and the predictive value of throat culture for the aetiology of sinusitis.

A prospective study of throat cultures and maxillary sinus aspirates from children with chronic sinusitis (n = 21), acute sinusitis (n = 28) or a clinical diagnosis of chronic adenoiditis (n = 41) was performed. Seventy-two bacterial pathogens were isolated from sinus aspirates from 52% of the study population. Haemophilus influenzae was most common pathogen, followed by Moraxella catarrhalis, Streptococcus pneumoniae, Staphylococcus aureus, and group A streptococci. Quantitative throat cultures had positive predictive values of 41%, 53% and 75% for H. influenzae, Strep. pneumoniae and M. catarrhalis, respectively, while negative predictive values were 93-98%, indicating that these three pathogens do not cause sinusitis when absent from the throat.

Typing of Pseudomonas aeruginosa strains in Turkish cystic fibrosis patients.

The majority of cystic fibrosis (CF) patients suffer from chronic respiratory infection with the opportunistic bacterial pathogen Pseudomonas aeruginosa. The virulence of P. aeruginosa is associated with the presence of various extracellular factors, like alginate, elastase, alkaline protease which contribute tissue destruction and assist bacterial invasion. Virulence factor production of P. aeruginosa strains isolated from 46 CF patients followed in two cities in Turkey was detected. Strains were compared genotypically by arbitrarily primed PCR. Antimicrobial susceptibilities to 12 antibiotics were determined by broth microdilution method. Evaluation of virulence factor results revealed that 95.8% of the strains were alginate, 71.7% elastase and 52.1% alkaline protease producers. AP-PCR analysis revealed 35 genotypes indicated almost a complete discrepancy among the strains. The most effective drugs were penems and quinolones. Among aminoglycosides amikacin was the most effective one and a high level resistance to beta lactams was observed. Alginate is the most important virulence factor in the chronic colonisation of CF patients with P. aeruginosa. No evidence for cross infection between patients and for relationship between phenotypes and genotypes of the strains was found.

Elastase and alkaline protease production by Pseudomonas aeruginosa strains: comparison of two procedures.

Pseudomonas aeruginosa is an opportunistic pathogen that can cause fatal infections in immunocompromised hosts. The virulence of P. aeruginosa is associated with the presence of various extracellular factors like elastase and alkaline protease. These enzymes are suggested to contribute to tissue destruction and assist bacterial invasion during infection. Therefore it seems likely that determination of these virulence factors will be an important prognostic marker in the near future especially for follow up of cystic fibrosis patients, to start antimicrobial agents that are directly or indirectly inhibit microbial growth or virulence factor production. Herein, we suggest a simple test procedure to be used in routine laboratories for estimation of elastase and alkaline protease levels and compare them with quantitative methods in the literature. We detected the amount of elastase and alkaline protease in 49 clinical P. aeruginosa isolates by comparing agar plate method and colorimetric assay. The resulting values were in the range reported in the literature and differed from one strain to another(elastase: 0-1390 mg/ml, alkaline protease: 0- 770 mg/ml). Linear relationships were found when assays compared in pairs and significant correlation coefficients were obtained(r>0.788 for alkaline protease, p<0.0001- r>0.926 for elastase, p<0.0001). Our method can be applied in laboratories regardless of the availability of technical equipment.

Immunologic influences of hyperthermia in a rat model of obstructive jaundice.

In this study, the effect of hyperthermia on immune response and bacterial translocation from the gut in jaundiced rats was assessed. In hyperthermic (HP; N = 8) and normothermic (NP; N = 8) preconditioning groups, rats were preconditioned by hyperthermia for 15 min at 42 degrees C or 38 degrees C, respectively. After 8 hr, the common bile duct (CBD) of each animal was ligated. In thermal (TT; N = 8) and normothermic treatment groups (NT; N = 8) the CBD of the animals was ligated, and after seven days rats were treated by hyperthermia for 15 min at 42 degrees C and 38 degrees C, respectively. The rats in the preconditioning groups (HP and NP) were killed at day 7 and rats in the treatment groups (TT and NT) were killed 8 hr after they were put in a water bath. Determination of the immunophenotypes of lymphocytes and serum levels of bilirubin was done in serum samples taken just after death. The quantity and identify of translocated bacteria were determined in tissue samples of mesenteric lymph nodes, spleen, and liver. NK cell expression as well as CD4+/CD8+ ratio were elevated in HP group when compared to NP group. CD8+ expression was found to be low in HP group when compared to NP group. CD4+, CD11b+, and B cell expressions were not found to be different between HP and NP groups. All immunologic parameters were similar when TT and NT groups were compared to each other. In the TT group, half of the rats revealed bacterial translocation, whereas in all other groups, we determined translocation in only 1/8 rats. The application of hyperthermia as preconditioning rather than applying it after the establishment of jaundice seemed to be beneficial. Hyperthermic preconditioning led an improvement in immune responses whereas the latter resulted an increase in bacterial translocation with no favorable influence on immune system. Bacterial translocation was unrelated with the immune status.

Asymtomatic carriage of Neisseria meningitidis and Neisseria lactamica in relation to Streptococcus pneumoniae and Haemophilus influenzae colonization in healthy children: apropos of 1400 children sampled.

Meningococcal disease is one of the most important causes of morbidity and mortality among children in many parts of the world. Main reservoir of carriage and site of meningococcal dissemination appears to be the upper respiratory tract. Colonization of Neisseria meningitidis and lactamica and factors affecting this carriage were determined in a group of healthy children aged 0-10 years. Meningococcus and N. lactamica carriage were detected in 17 (1.23%) and 245 (17.7%) of 1382 subjects, respectively. Number (%) of serogroups for meningococci was 1 (6), 5 (29), 0 (0), 1 (6), 1 (6), and 9 (53) for A, B, C, D, W135, and Y, respectively. Having more than three household members, elementary school attendance, pharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae were associated with carriage of meningococci, whereas age less than 24-month was associated with carriage of N. lactamica. There was a reverse carriage rate between N. meningitidis and N. lactamica by age which may suggest a possible protective role of N. lactamica against meningococcal colonization among pre-school children.

Streptococcus pneumoniae penicillin resistance in Turkey.

Resistance of Streptococcus pneumoniae (750) to penicillin, erythromycin, chloramphenicol and trimethoprim/sulfamethoxazole isolated in 4 Turkish hospitals between 1996 and 1999 was evaluated according to year of isolation, patients' age groups and specimen. Penicillin susceptibility was determined by E-test strips and the other antibiotics were tested by disk diffusion test following the NCCLS guidelines in each center. Overall high and intermediate resistance to penicillin was 3% and 29%, respectively. There was a significant difference (p<0.001) between the centers with regard to penicillin resistance. However, there was no significant increase in resistance by year. Penicillin resistance varied significantly among children and adults (36% versus 25%) and according to the specimen. Highest rate of penicillin resistance was observed in respiratory specimens (36%) followed by ear exudates (33.5%). In blood isolates, resistance to penicillin was 28.6%. Overall resistance to erythromycin was 8%, to chloramphenicol 5% and to trimethoprim-sulfamethoxazole 47%. Although overall penicillin resistance in these Turkish S. pneumoniae isolates is high, resistance rates vary in each center and have not increased from 1996 to 1999.

The functional assessment of autotransplanted splenic tissue by its capacity to remove oxidatively modified erythrocytes.

Free radicals and reactive oxygen species have been implicated in the pathogenesis of a variety of hematologic diseases and erythrocyte aging. Aged erythrocytes are removed from the circulation primarily by the spleen. In this study, we aimed to determine the functional effectiveness of autotransplanted splenic tissue by its capacity to remove oxidatively modified erythrocytes from the circulation. Our experimental model in rats includes splenectomy with autotransplantation of 80% of the excised splenic tissue into the omental pouch. In this model, free radical damage was estimated by different parameters of lipid peroxidation such as carbonyl content and thiobarbituric acid reactive substances (TBARS), together with Heinz body formation. Our results have shown that splenic autotransplantation was effective in removing oxidatively modified, aged erythrocytes from the circulation.