Effects of Probiotic Use on Bacterial Translocation in Created Rat Models with Biliary Obstructions.

OBJECTIVE: The aim of this study was to investigate the protective effects of the combined use of probiotic strains on the development of bacterial translocation in addition to liver and intestinal tissue damage due to biliary obstruction in rats. MATERIALS AND METHODS: Here, 3 groups each consisting of 10 rats were created:group 1 (sham group), group 2 (obstructive jaundice), and group 3 (obstructive jaundice+probiotic). Groups 1 and 2 were given 1 cc physiological saline solution by oral gavage twice a day; group 3 was given a probiotic solution that included Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium bifidum, Enterococcus faecium, and Bifidobacterium longum microorganisms by oral gavage twice a day. RESULTS: Markers for liver damage were also found to be significantly improved (p<0.05) in the treatment group (group 3). When compared with groups 2 and 3 in terms of liver histology, damage was found to be significantly more severe in group 2 (p<0.01). With regard to ileal villous depth and ileal inflammation, the pathology was found to be significantly more severe in group 2 than that in group 3 (p<0.05). In blood, spleen, and mesenteric lymph node cultures, group 2 showed a microbiological growth rate of 33.8-58.8%, whereas group 3 showed a microbiological growth rate of 14.3-28.6%. This reduction was evaluated to be statistically significant (p<0.05). CONCLUSION: Our study showed that the combined use of a probiotic in bile duct obstructions reduced bacterial translocation and alleviated pathological changes arising in the liver and terminal ileum histology.

Identification of medically important Candida species by polymerase chain reaction-restriction fragment length polymorphism analysis of the rDNA ITS1 and ITS2 regions.

AIM: We aimed to identify the distribution of species in candidal strains isolated from clinical samples and restriction fragment length polymorphism (RFLP) method based on Msp I and Bln I restrictive enzyme cuts of polymerase chain reaction (PCR) products after the amplification of ITS1 and ITS2 regions of rDNA genotypically. MATERIALS AND METHODS: One hundred and fifty candidal strains isolated from various clinical samples were studies/ included. Phenotypic species assessment was performed using automated VITEK-2 system and kit used with the biochemical tests. Common genomic region amplification peculiar to candidal strains was carried out using ITS1 and ITS2 primer pairs. After the amplification, PCR products were cut with Msp I and Bln I restriction enzymes for species identification. RESULTS: The majority of Candida isolates were isolated from urine (78.6%) while other isolates were composed of strains isolated from swab, wound, blood and other samples by 11.3%, 3.3%, 2% and 4.7%, respectively. The result of RFLP analysis carried out with Msp I and Bln I restriction enzymes showed that candidal strains were Candida albicans by 45.3%, Candida glabrata by 19.3%, Candida tropicalis by 14.6%, Candida parapsilosis by 5.3%, Candida krusei by 5.3%, Candida lusitaniae by 0.6% and other candidal strains by 9.3%. CONCLUSION: When the ability to identify Candida to species level of phenotypic and PCR-RFLP methods was assessed, a great difference was found between these two methods. It may be argued that Msp I and Bln I restriction enzyme fragments can be used in the identification of medically important Candida species. Further studies are needed to develop this kind of restriction profile to be used in the identification of candidal strains.

Determination of CTX-M beta-lactamase in Escherichia coli strains isolated from clinical samples.

The aim of this study was to determine the ESBL with phenotypic tests and investigate the bla(CTX-M) genes with the PCR method in Escherichia coli strains. The presence of ESBL in E. coli strains was determined with the Vitek 2 automated system. ESBL-positive 100 and ESBL-negative 50 E. coli strains were included in the study. The ESBL disk diffusion screening test (DDST) and the combined disk confirmation tests (CDCT) were performed on these strains and the results of these tests were compared with each other. bla(CTX-M) genes were investigated with the PCR method. The results of CDCT-CAZ-CZC and CDCT-CTX-CTC were found to be consistent in 90% of strains. Those of the automated system, DDST and CDCT-CAZ-CZC were compatible with each other in 83.3% of strains. Also the results of the automated system and CDCT-CTX-CTC were found to be compatible in 83.3% of strains. Based on PCR, bla(CTX-M) genes were found in 67.3% of 150 strains. According to the order of frequency, 46%, 38.7%, 20%, 7.3% of strains were determined to carry groups I, IV, II and III, respectively.

Performance of QuantiFERON-TB Gold In-Tube test and Tuberculin Skin Test for diagnosis of latent tuberculosis infection in BCG vaccinated health care workers.

BACKGROUND: Tuberculin skin test (TST) has been used for years as an aid in diagnosing latent tuberculosis infection (LTBI) but it suffers from a number of well-documented performance and logistic problems. Quantiferon-TB Gold In Tube test (QFT-GIT) has been reported to have better sensitivity and specifity than TST. In this study, it was aimed to compare the performance of a commercial IFN-gamma release assay (QFT-GIT) with TST in the diagnosis of HCWs at risk for latent TB infection in BCG vaccinated population. MATERIAL AND METHODS: Hundred healthy volunteer health care workers were enrolled. All were subjected to TST and QFT-GIT. Results were compared among Health Care Workers (HCWs) groups in terms of profession, workplace, working duration. RESULTS: TST is affected by previous BCG vaccinations and number of cases with QFT-GIT positivity is increased in accordance with the TST induration diameter range. QFT-GIT result was negative in 17 of 32 TST positive (≥ 15 mm) cases and positive in 4 of 61 cases whose TST diameters are between 6-14 mm, that is attritutable to previous BCG vaccination(s). It was negative in all cases with TST diameters between 0-5 mm. HCWs with positive QFT-GIT results were significantly older than the ones with negative results. Furthermore duration of work was significantly longer in QFT-GIT positive than in negative HCWs. CONCLUSIONS: There was a moderate concordance between QFT-GIT and TST, when TST result was defined as positive with a ≥ 15 mm diameter of induration. We suggest that QFT-GIT can be used as an alternative to TST for detection of LTBI, especially in groups with high risk of LTBI and in population with routine BCG vaccination program.

Antibiotic resistance genes & susceptibility patterns in staphylococci.

BACKGROUND & OBJECTIVES: This study was carried out to evaluate the association between the antibiotic susceptibility patterns and the antibiotic resistance genes in staphylococcal isolates obtained from various clinical samples of patients attending a teaching hospital in Hatay, Turkey. METHODS: A total of 298 staphylococci clinical isolates were subjected to antimicrobial susceptibility testing. The genes implicated in resistance to oxacillin (mecA), gentamicin (aac(6')/aph(2''), aph(3')-IIIa, ant(4')-Ia), erythromycin (ermA, ermB, ermC, and msrA), tetracyclin (tetK, tetM), and penicillin (blaZ) were amplified using multiplex PCR method. RESULTS: Methicillin resistance rate among 139 Staphlococcus aureus isolates was 16.5 and 25.9 per cent of S. aureus carried mecA gene. Of the 159 CoNS isolates, methicillin resistance rate was 18.9 and 29.6 per cent carried mecA gene. Ninety four isolates identified as gentamicin resistant phenotypically, contained at least one of the gentamicin resistance genes [aac(6')/aph(2''), aph(3')-IIIa, ant(4')-Ia], 17 gentamicin-susceptible isolates were found as positive in terms of one or more resistance genes [aac(6')/aph(2''), aph(3')-IIIa, ant(4')-Ia] by multiplex PCR. A total of 165 isolates were resistant to erythromycin, and contained at least one of the erythromycin resistance genes (ermA, ermB, ermC and msrA). Phenotypically, 106 staphylococcal isolates were resistant to tetracycline, 121 isolates carried either tetK or tetM or both resistance genes. The majority of staphylococci tested possessed the blaZ gene (89.9%). INTERPRETATION & CONCLUSIONS: The present results showed that the phenotypic antibiotic susceptibility patterns were not similar to those obtained by genotyping done by multiplex PCR. Rapid and reliable methods for antibiotic susceptibility are important to determine the appropriate therapy decisions. Multiplex PCR can be used for confirmation of the results obtained by conventional phenotypic methods, when needed.

Efflux pump genes and antimicrobial resistance of Pseudomonas aeruginosa strains isolated from lower respiratory tract infections acquired in an intensive care unit.

The aim of this study was to determine the antimicrobial resistance rates and the resistance genes associated with efflux pumps of Pseudomonas aeruginosa strains isolated from the patients who acquired lower respiratory tract infection (LRTI) in intensive care unit (ICU). Fifty P. aeruginosa strains isolated from the lower respiratory tract specimens of the patients who acquired LRTIs in ICU were included in this study. P. aeruginosa strains were isolated from tracheal aspirate (27), bronchoalveolar lavage (14) and sputum (9). The susceptibilities of the isolates were investigated by the disk diffusion method. Multiplex PCR assay was carried out for the detection of 13 antibiotic-resistance genes. Antimicrobial resistance rates of the isolates were found high and the highest resistance rate of the isolates studied was determined against to mezlocillin (50%) followed by norfloxacin (48%), ciprofloxacin (46%), meropenem (40%). Fourty-three isolates (86%) were determined to carry one and more resistance genes. NfxB gene was most often determined in the genes that were investigated. The significant relation between the resistance to cefepime, piperacilline/tazobactam and the mexC gene, that between the resistance to mezlocillin, piperacilline/tazobactam, ceftazidime, cefepime and ampC genes, and that between the resistance to ciprofloxacin, norfloxacin and oprJ, oprN and nfxB genes was identified. Resistance caused by genes for carbapenemases, aminoglycoside-modifying enzymes and other mechanisms were not identified in this study. Understanding the prevalence and mechanism of antimicrobial resistance in P. aeruginosa may help to select empirical therapy for nosocomial LRTIs due to P. aeruginosa in our ICU.

Evaluation of nosocomial infections and risk factors in critically ill patients.

BACKGROUND: Nosocomial infections are one of the most serious complications in intensive care unit patients because they lead to high morbidity, mortality, length of stay and cost. The aim of this study was to determine the nosocomial infections, risk factors, pathogens and the antimicrobial susceptibilities of them in intensive care unit of a university hospital. MATERIAL/METHODS: The patients were observed prospectively by the unit-directed active surveillance method based on patient and the laboratory. RESULTS: 20.1% of the patients developed a total of 40 intensive care unit-acquired infections for a total of 988 patient-days. The infection sites were the lower respiratory tract, urinary tract, bloodstream, wound, and the central nervous system. The respiratory deficiency, diabetes mellitus, usage of steroid and antibiotics were found as the risk factors. The most common pathogens were Enterobacteriaceae, Staphylococcus aureus, Candida species. No vancomycin resistance was determined in Gram positive bacteria. Imipenem and meropenem were found to be the most effective antibiotics to Enterobacteriaceae. CONCLUSIONS: Hospital infection rate in intensive care unit is not very high. The diabetes mellitus, length of stay, usage of steroids, urinary catheter and central venous catheter were determined as the risk factors by the final logistic regression analysis. These data, which were collected from a newly established intensive care unit of a university hospital, are important in order to predict the infections and the antimicrobial resistance profile that will develop in the future.

GC-MS analysis and antileishmanial activities of two Turkish propolis types.

Propolis is a honeybee product with a very complex chemical composition and various pharmacological properties. This study was aimed to investigate antileishmanial activities of "Bursa" and "Hatay" propolis samples against Leishmania infantum and Leishmania tropica strains. Propolis samples were analysed with the gas chromatography-mass spectrometry technique. Promastigotes were incubated in Roswell Park Memorial Institute culture medium in the absence and presence of several concentrations (50, 100, 250, 500, 750, and 1,000 µg/mL) of each propolis sample. The viability and cell morphology of promastigotes in each concentration were examined after 24, 48, 72, and 96 h of incubation. The growth of leishmania parasites was significantly suppressed in the presence of 500, 750, and 1,000 µg/mL of Hatay propolis. Bursa propolis was found to be efficient in inhibiting the growth of leishmania promastigotes in culture media at these concentrations, 250, 500, 750, and 1,000 µg/mL. Thus, the in vitro results showed that the Hatay and Bursa propolis samples decreased significantly the proliferation of L. infantum and L. tropica parasites (p < 0.001); however, Bursa propolis was found to be more effective than Hatay propolis against leishmania promastigotes. These two natural products may be useful agents in the prevention of leishmanial infections.

Effect of Thymoquinone on Oxidative Stress in Escherichia coli-Induced Pyelonephritis in Rats.

BACKGROUND: Recurrent urinary tract infections are important in children and adults with diabetes mellitus and/or incontinence due to risk of pyelonephritis (PYN) and renal damage. There is a positive correlation released free radicals during PYN and renal damage. Experimental studies showed that antioxidant agents improve renal damage when used immediately after bacterial inoculation. OBJECTIVE: The aim of the present study was to evaluate whether treatment by thymoquinone (TQ) before or during Escherichia coli inoculation prevents oxidative damage in acute pyelonephritis (PYN) in an ascending obstructive rat model. METHODS: In this study, 42 Wistar rats were grouped as follows: control, PYN (24, 48, and 72 hours), and TQ-PYN (24, 48, and 72 hours). E. coli (1 ×10(9) colony forming units) was inoculated into the bladder via urethral catheterization in both the PYN and TQ groups. TQ injections were performed 24 hours before bacteria inoculation and repeated at 24-hour intervals during the indicated time at a dose of 10 mg/kg body weight intraperitoneally in TQ groups. RESULTS: Superoxide dismutase activity was statistically lower in the TQ-PYN-48 and -72 groups than the PYN-48 and -72 groups (P < 0.001, P = 0.004, respectively). Catalase activity was significantly higher in PYN-24, -48, and -72 groups than the control group (P < 0.001). In addition, there was a significant difference between the TQ-PYN-24, -48, and -72 groups and PYN groups in terms of glutathione peroxidase activity (P < 0.001, P = 0.026, P = 0.046, respectively). When the TQ-PYN-72 group was compared with the PYN-72 group, malondialdehyde levels were significantly lower in the TQ-PYN-72 group than in the PYN-72 group (P = 0.033). A histologic examination also confirmed the protective effect of TQ. In statistical analysis of histopathologic findings, there were significant differences between the PYN-24 and TQ-PYN-24, PYN-48 and TQ-PYN-48, and PYN-72 and TQ-PYN-72 groups (P = 0.008, P < 0.001, P < 0.001, respectively). CONCLUSIONS: The results indicate that TQ administration attenuated the oxidative damage that occurred in PYN and, therefore, could be used as a supportive agent to protect the kidneys from oxidative damage caused by PYN.

Detection of slime and methicillin resistance genes in Staphylococci isolated from nasal samples of patients with orthopaedic implants.

BACKGROUND: The purposes of the present study were (1) to determine the prevalence of mecA and femA genes, (2) to investigate the presence of icaA and icaD genes responsible for slime synthesis, and (3) to search in vitro slime synthesis by staphylococcal strains isolated from the nares of patients with orthopaedic implants using the Congo red agar (CRA) plate test. MATERIAL/METHODS: Staphylococci strains were defined by multiplex polymerase chain reaction (PCR) technique to determine intercellular adhesion genes icaA and icaD. Slime production capability was searched by the CRA plate test, phenotypically. Also, the presence of mecA and femA genes was determined by PCR in all strains. RESULTS: The presence of icaA and icaD was detected in 101 isolates of 134 (75.4%) strains. This ratio was 74.8% (89 of 119) among the Staphylococcus epidermidis and 80% (12 of 15) among the Staphylococcus aureus isolates. A total of 63.4% of all the strains were found to be icaA and icaD positive as well as slime-forming on the CRA plate test. The percentage of icaA- and icaD-negative strains was 36.6%, and all of them were negative on the CRA plate test. Although femA presence was detected in all 15 (11.2%) S. aureus isolates, a total of 5 (3.7%) isolates carried the mecA gene. CONCLUSIONS: The frequency of icaA and icaD genes was determined to be of high prevalence among staphylococcal isolates. The staphylococcal strains that were found in the nasal flora of patients with orthopaedic implants may be important potential sources of infection for these patients.

Skin diseases in patients with beta-thalassemia major.

BACKGROUND: Beta-thalassemia major affects multiple organs and is associated with considerable morbidity and mortality. The goal of this analysis was to document the frequency of skin diseases among patients with beta-thalassemia major. METHODS: A sample of 78 patients with beta-thalassemia major was recruited and interviewed at the thalassemia clinic between April and June 2008. A dermatologist completed a skin examination of each patient and recorded any skin disease present. The patients' laboratory results and treatment modalities were recorded from the charts. RESULTS: Sixty-five (83.3%) of the 78 patients examined had a diagnosed skin disease: pruritus (37.2%), xerosis (34.6%), scars (24.4%), ephelides (23.1%), skin irritation/erythema caused by deferoxamine pump (12.8%), idiopathic guttate hypomelanosis (6.4%), pityriasis alba (6.4%), tinea infections (5.1%), verruca vulgaris (5.1%), urticaria (3.8%), hyperhidrosis, contact dermatitis, and acne (2.6%), necrobiosis lipoidica (1.3%), melasma (1.3%), and others (14.1%). Pruritus and xerosis increased in frequency with age. The mean serum ferritin level was higher in patients with xerosis than in those without (5607.7 +/- 2997.5 ng/mL and 4285.2 +/- 4732.4 ng/mL, respectively) (P1/4 0.005). Moreover, xerosis was observed significantly more frequently inpatients using deferoxamine and deferiprone than in those using deferasirox (P1/4 0.047, P1/4 0.027, and P1/4 0.273, respectively). CONCLUSION: Skin diseases, especially pruritus and xerosis, are observed highly frequently inpatients with beta-thalassemia major. Treatment is needed for these patients who already have other significant morbidities.

Cutaneous infection caused by Aspergillus terreus.

Aspergillus species are widely distributed in nature, and more than 30 species have been reported to be involved in human and animal infection. Cutaneous infections due to Aspergillus terreus are particularly rare. In this report, we describe a case of cutaneous infection caused by A. terreus in a paediatric patient who underwent surgical treatment for an open tibial fracture secondary to an agricultural accident.

High prevalence of Staphylococcus aureus cultivation and superantigen production in patients with psoriasis.

The aim of this study was to evaluate the association of Staphylococcal enterotoxins (se) a through e, exfoliative toxin (et) a and b, toxin and toxic shock syndrome toxin (tst) and mecA with psoriasis. We also investigated the distribution of Staphylococcus aureus (S. aureus) in the skin and nares. Fifty consecutive patients with chronic plaque-type psoriasis and 50 sex- and age-matched healthy controls were included in this study. There was a statistical difference in cultivation of S. aureus between lesional (64%) and non-lesional skin (14%) in patients with psoriasis (p = 0.037). S. aureus was cultivated from the nares in 25 (50%) of 50 patients with psoriasis and in 17 (34%) of 50 healthy controls (p > 0.05). In psoriasis patients, 31 (96.8%) out of the 32 strains isolated from the lesional skin and 3 (42.3%) out of the 7 strains isolated from the non-lesional skin were toxigenic (p = 0.01). Isolated strains from the nares were toxigenic in 96% (24/25) for patients with psoriasis and in 41.2% (7/17) for healthy controls, respectively (p = 0.006). Patients with cultivation-positive in lesional skin had a significantly higher PASI score than patients who were cultivation-negative in lesional skin (8.28 +/- 3.97 vs. 5.89 +/- 2.98, p = 0.031). Our results confirm that S. aureus colonization and its toxigenic-strains are associated with psoriasis. According to our findings, non-classical superantigens such as methicillin resistance gene (mecA), see and etb may also be associated with psoriasis.

Markers of inflammation as determinants of mortality in intensive care unit patients.

In this study, the investigators explored the relationship between mortality rate and serum levels of C-reactive protein (CRP), erythrocyte sedimentation ratio (ESR), albumin, and hemoglobin, leukocyte, and platelet counts of patients at the time of first admission to the intensive care unit (ICU). A total of 123 patients were admitted to 2 different ICUs. In the emergency departments, serum levels of CRP, ESR, and albumin and hematologic parameters of 81 patients who died and 42 patients who survived were compared. A Student t test and the chi2 test were used for statistical analyses. Mean CRP and ESR levels and leukocyte counts were higher in nonsurvivor than in survivor groups (P<.001 for all). Additionally, serum CRP and ESR elevations and leukocyte counts were determined to be individually related to mortality (P<.001, P<.05, and P<.05, respectively). The investigators concluded that initial serum levels of CRP and ESR and leukocyte counts can be used as determinants of mortality in ICU patients.

First case report of empyema caused by Beauveria bassiana.

Infections with Beauveria bassiana are extremely rare in humans. A 51-year-old man was admitted to hospital with the diagnosis of lung adenocarcinoma. Thoracic wall resection with lobectomy was applied and empyema has developed after prolonged air leakage. B. bassiana was isolated in pleural fluid. The patient improved without antifungal therapy after thoracotomy with securing of air leakage.

The antimicrobial susceptibility of Stenotrophomonas maltophilia isolates using three different methods and their genetic relatedness.

BACKGROUND: Stenotrophomonas maltophilia is inherently resistant to many antimicrobials. So far, antimicrobial susceptibility tests for S. maltophilia have not been fully standardized. The purpose of the study was to compare the susceptibility of S. maltophilia isolates against seven different antimicrobials using three different methods and to investigate their genetic relatedness. RESULTS: Although trimethoprim/sulfamethoxazole (SXT) and ciprofloxacin have the lowest MIC values, SXT (98.1%) and ticarcillin/clavulanate (TLc) (73.1%) were found to be the most effective antimicrobials by agar dilution method, which was in accordance with the breakpoints established by NCCLS. Disc diffusion and E-test was in agreement with agar dilution method for SXT. When the isolation dates, clinics, antibiotyping, and AP-PCR data were investigated, two small outbreaks consisting of five and three cases were determined. CONCLUSION: By using the NCCLS criteria, disc diffusion and E-test were unreliable alternative methods for S. maltophilia, except for SXT. However, the significance of these data should be confirmed by further experimental and clinical studies.

An outbreak of tularemia in Western Black Sea region of Turkey.

The aim of this study was to investigate the source and the size of a tularemia outbreak in a village located in a non-endemic area. Five patients from the same village were admitted to hospital with the same complaints all within one week of September 2001. Tularemia was suspected and a diagnosis was made after physical and anamnesis examinations. The village was visited the same week that the patients were admitted to the hospital, in the January and April 2002. The villagers were examined and screened serologically by microagglutination method and the water sources were investigated bacteriologically. A total of 14 people were found to be infected from the outbreak and the oropharyngeal form was the only clinical presentation. Antibody titers ranged between 1 : 80 and 1 : 640. The patients responded well to the aminoglycoside plus tetracycline therapy. Examination of the pipewater and three springs revealed that all the water sources were contaminated by coliforms, however, Francisella tularensis could not be isolated in glucose-cystine medium. Antibody levels stayed stable or decreased seven months after. Tularemia had not been reported in this area before, so the first patients were misdiagnosed. In conclusion tularemia should be considered in differential diagnosis of patients with fever, sore throat and cervical lymphadenopaties.