Prevalence of House Dust Mites and Presence of Der p 1 and Der f 1 in Ordu, Giresun, Trabzon and Rize Provinces.

OBJECTIVE: This study was carried out to detect house dust mites in houses and to investigate group 1 antigens of Dermatophagoid species in Ordu, Giresun, Trabzon and Rize provinces of the Central and Eastern Black Sea Region. METHODS: Dust samples obtained from the beds were subjected to both microscopic and antigenic examination. Samples prepared by the lactic acid method for microscopic examination were evaluated under a light microscope. Antigenic analysis was performed by investigating Der p 1 and Der f 1 belonging to D. pteronyssinus and D. farinae by ELISA test. RESULTS: 90.3% of the dust samples were evaluated positive by microscopic examination (10x, 40x) and 149 mites were detected. D. pteronyssinus 74%, D. farinae 13%, Dermatophagoides spp. growth forms 5%, Cheyletus spp. 1%, E. maynei 1%, C. arcuatus 1%, T. putrescentiae 1%, L. destructor 1% and unidentified mites were detected at the rate of 3% respectively. Der p 1 antigen was detected in 93% and Der f 1 antigen in 84.7%. The highest amount of antigen detected in one gram of powder was 1,272 µg for Der p 1 and 0,482 µg for Der f 1. CONCLUSION: No difference was observed between mite species and distribution in the provinces where the study was conducted (p<0.05). Dermatophagoides were found in 93% of the population. The low (4%) rate of storage/food mites is related to the fact that samples were not taken from the floors. Antigen accumulation may be important in the beds since the activity of the mites is observed throughout the year in temperate and humid regions. It is thought that this diagnosis method can be used and can be taken into account in terms of the environments in which sensitive people live.

Synthesis, Spectroscopic Characterization, Antibacterial Activity, and Computational Studies of Novel Pyridazinone Derivatives.

In this work, a novel series of pyridazinone derivatives (3-17) were synthesized and characterized by NMR (1H and 13C), FT-IR spectroscopies, and ESI-MS methods. All synthesized compounds were screened for their antibacterial activities against Staphylococcus aureus (Methicillin-resistant), Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, and Acinetobacter baumannii. Among the series, compounds 7 and 13 were found to be active against S. aureus (MRSA), P. aeruginosa, and A. baumannii with the lowest MIC value range of 3.74-8.92 µM. Afterwards, DFT calculations of B3LYP/6-31++G(d,p) level were carried out to investigate geometry structures, frontier molecular orbital, molecular electrostatic potential maps, and gap energies of the synthesized compounds. In addition, the activities of these compounds against various bacterial proteins were compared with molecular-docking calculations. Finally, ADMET studies were performed to investigate the possibility of using of the target compounds as drugs.

Bis-1,2,4-triazol derivatives: Synthesis, characterization, DFT, antileishmanial activity and molecular docking study.

In this study, triazol derivatives, 4,4'-(((1E, 1E')-1,2-phenylenebis (methanylyidene)) bis (azanylidene)) bis (5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one (2), 4,4'-(((1E, 1E')-1,3-phenylenebis (methanylyidene)) bis (azanylidene)) bis (5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one (3) and 4,4'-(((1E, 1E')-1,4-phenylene bis (methanyl yidene)) bis (azanylidene)) bis (5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one (4) were synthesized from the reaction of 4-amino-5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one and phthalaldehyde/isophthalaldehyde/terephthalaldehyde, respectively. Compounds 2-4 were characterized by Fourier transform infrared (FTIR), proton and carbon-13 nuclear magnetic resonance (1H- and 13C- NMR) spectroscopic methods. Theoretical study for compounds 2-4 were carried out by DFT/B3LYP/6-311++G(d,p). Structural and spectroscopic parameters were determined theoreticaly and compared with experimental ones. Also, the molecular electrostatic potential (MEP) maps of compounds were obtained. Leishmanicidal activity of compounds 2-4 against to Leishmania infantum was determined by microdilution broth method containing alamar blue. As a result of the study, compounds 2-4 were found to be effective against the specie of Leishmania. Molecular docking analysis against Trypanothione Reductase (TRe) with compound 2 was carried out to see the necessary interactions responsible for antileishmanial activity. The docking calculations of compound 2 supported the antileishmanial activity exhibiting high inhibition constant.Communicated by Ramaswamy H. Sarma.

Antileishmanial Activity of New Synthesized Schiff and Mannich (Morpholine) Base Compounds.

OBJECTIVE: Leishmaniasis is an important parasitic disease in many countries, including ours. A variety of drugs are currently used for its treatment. However, certain side effects of these drugs, such as teratogenicity, hepatotoxicity and nephrotoxicity, have been reported in some patients. The goal of this research is to determine the antileishmanial effects of eight different previously synthesised compounds containing Schiff and Mannich bases (morpholine) against Leishmania infantum (L. infantum) promastigotes by the liquid microdilution method utilising alamarBlue. METHODS: Compounds containing Schiff bases (a-d) and both Schiff bases and morpholine rings (e-h) were tested. Compounds were diluted in the range of 20000-39 µg/mL. L. infantum promastigotes were added to the wells, which were then incubated at 27 °C. The proliferation of Leishmania promastigotes was evaluated after 24, 48 and 72 hours. RESULTS: In this study, compounds b, c and d (MIC values 156 µg/mL, 78 µg/mL and 156 µg/mL) were found to be effective against L. infantum promastigotes, whereas compound f (MIC >20000 µg/mL) was found to be more the most ineffective compound. CONCLUSION: These compounds may be potential drug candidates for the treatment of leishmaniasis. According to the results, there is a need for further studies, such as in vivo experimental animal models and ex vivo Leishmania amastigote macrophage cultures for compounds showing antileishmanial effects.

Bis benzothiophene Schiff bases: synthesis and in silico-guided biological activity studies.

Since benzo [ b ] thiophene scaffold is one of the privileged structures in drug discovery as this core exhibitsactivities for different biological problems, in this study bis (benzo[ b ]thiophene-2-yl) alkyl methanimine derivatives (1-9) were synthesized by reacting benzo[ b ]thiophene-2-carbaldehyde with diamines. All newly compounds were characterized by IR, 1H NMR and 13C NMR spectroscopic methods. Synthesized compounds were investigated using binary QSARbased models on therapeutic activity prediction of synthesized compounds and they showed high predicted activities in following diseases: bacterial, angina, allergy, depression and obesity. Thus, they were then tested for their antimicrobial and antileishmanial activities as a result of this theoretical study. Compound 1(N, N'- (propane-1,3-diyl) bis (1-(benzo [ b ] thiophene-2-yl)) methanimine) was found the most active compound in both diseases. Thus, its molecular docking studies were also carried out.

Role of Adenosine Deaminase in Patients with Erythematotelangiectatic Rosacea and Demodex folliculorum Positivity.

BACKGROUND: Adenosine deaminase (ADA) is an aminohydrolase involved in the catabolism of purine nucleotides and irreversibly deaminizes adenosine and deoxyadenosine to inosine and deoxyinosine. ADA enzyme deficiency results in the loss of functional properties of B and T lymphocytes. Demodex species have been reported to be transmitted between humans through close contact and to play a role in the pathogenesis of rosacea, acne vulgaris, perioral dermatitis, seborrhoeic dermatitis, micropapillary-pruritic dermatitis and blepharitis. The present study aimed to compare serum ADA levels in D. folliculorum positive patients with the healthy control individuals. METHODS: Serum ADA levels were examined for 30 patients diagnosed with erythematotelangiectatic rosacea and 40 healthy individuals in Malatya Inonu University in 2017. Standardized skin surface biopsy (SSSB) method was used to diagnose D. folliculorum. A significant decrease was found in the ADA levels of Demodex-positive rosacea patients when compared to the control group. RESULTS: ADA levels were decreased in the Demodex-positive group. The mean ADA level in patient group was significantly lower than the mean in the control group (P<0.001). There was no significant difference between the patient and control groups in terms of age and gender. CONCLUSION: During and after treatment of Demodex-positive rosacea patients, determination of ADA levels may give more detailed information on the immune mechanisms.

The Investigation of the Prevalance and Epidemiology of House Dust Mites in Giresun

Objective: The aim of this study was to determine the species of house dust mites and their prevalence in Giresun. Methods: Dust samples taken from 15 houses which were visited monthly for one year were examined by the lactic acid method. Results: A total of 2251 mites were detected in the study. The distribution of mites was as follows: 81.8% Dermatophagoides pteronyssinus, 0.5% Dermatophagoides farinae, 0.04% Euroglypus maynei (E. maynei), 4.2% Dermatophagoides spp. 0.06% A. siro, 2.4% Glycphagus domesticus, 0.9 % Lepidoglyphus destructor, 1.4% Tyrophagus putrescentiae, 4.5% Campunatus arcuatus, 1.3% Cheyletus spp. Pyroglyphid species were detected in all houses (100%). Dermatophagoides pteronyssinus was found in 100%, D. farinae 5% and E. maynei 4% of the houses. Conclusion: The mites in Giresun were found in all houses throughout the year and were detected in all of the samples. Although they were detected in greater amounts in the spring and summer, only a moderate relationship could be detected with temperature. In August-October period, mite existence was significantly higher than the January-March period (p<0,05). D. pteronyssinus was found in higher numbers on the mezzanine floors between May and August and on the ground floors in September and October (p<0,05). We think that the climate characteristics of Giresun are suitable for the development and proliferation of house dust mites and this can pose a risk for sensitive people.

The Investigation of the Presence of Mites in Some Served Dry Foodstuffs.

OBJECTIVE: Mites are microscopic organisms that lower the quality of life of people who are sensitive to them by causing conditions such as atopic dermatitis, allergic rhinitis, and asthma. These organisms are found in every habitat where humans live. This study was conducted to determine the presence of storage mites in dry food items. METHODS: Various food items were procured 10 times each in 300-gram samples. Mites were extracted with a Berlese funnel apparatus over Erlenmeyer flasks containing 70% alcohol placed at the end of the funnel stems for over 48 h. RESULTS: Of 25 food items examined in the study, only six were contaminated by mites. Species of the mites found were Acarus siro (34.6%), Glycyphagus domesticus (22.8%), Tyrophagus putrescentiae (16.8%), Tyrophagus spp. (7.9%), Rhizoglyphus spp. (1%), Lepidoglyphus destructor (7.9%), Cheylettus malacensis (4%), and Cheylettus spp. (2%). CONCLUSION: Although the results of the study show that the presence of mites in food items sold in open containers at open-air markets or stores was low, we suppose that they can cause important health problems for sensitive people.

[Investigation of the frequency and distribution of beta-lactamase genes in the clinical isolates of Acinetobacter baumannii collected from different regions of Turkey: a multicenter study]. / Türkiye'nin farkli bölgelerinden toplanan klinik Acinetobacter baumannii izolatlarinda beta-laktamaz gen sikligi ve dagiliminin arastirilmasi: Çok merkezli bir çalisma.

The diversity and distribution of TEM, SHV and CTX-M type of extended-spectrum beta-lactamases (ESBLs) are important for the treatment and control of infections. Determination of ESBL genes in clinical isolates by polymerase chain reaction (PCR) and DNA sequencing can obtain useful data for their molecular epidemiology and risk. The aim of this study was to investigate the frequency of beta-lactamase genes in Acinetobacter baumannii strains isolated from different regions of Turkey. A total of 519 A.baumannii strains collected from hospitals located at 12 different provinces of Turkey (Bolu (n= 67), Tokat (n= 47), Trabzon (n= 25), Ordu (n= 27), Diyarbakir (n= 47), Nigde (n=31), Kayseri (n= 36), Ankara (n= 41), Kirikkale (n= 26), Kahramanmaras (n= 25), Mersin (n= 40), Istanbul (n= 107)] between 2011-2012 period were included in the study. Identification of the isolates were performed by both conventional methods and automated systems, VITEK2 Compact (BioMerieux, France) and API 32GN (BioMerieux, France). Disc diffusion method was used for the detection of antibiotic susceptibilities of the isolates and the results were evaluated according to CLSI (Clinical and Laboratory Standards Institute) criteria. Tigecycline and colistin sensitivities of the isolates were evaluated according to BSAC (British Society for Antimicrobial Chemotherapy) criteria. The presence of beta-lactamase genes, namely blaoxa-51, blaTEM, blaSHV, blaCTX-M1, blaCTX-M2, blaGES and blaVIM were detected by PCR. In our study, the resistance rates against colistin, tigecycline, ampicillin-sulbactam, amoxicillin-clavulanic acid, cefoperazone/sulbactam, tobramycin, ceftriaxone, piperacillin-tazobactam, gentamicin, ampicillin, tetracycline, cefepime, piperacillin, amikacin, trimethoprim-sulfamethoxazole, meropenem, levofloxacin, ciprofloxacin, imipenem and ceftazidime were detected as; 0.6%, 2.7%, 11.9%, 15.2%, 21%, 22.9%, 23.9%, 48.6%, 59.5%, 61.8%, 66.3%, 67.8%, 69.2%, 71.1%, 77.5%, 78.6%, 81.1%, 82.9%, 87.5% and 89.4%, respectively. All of the isolates (100%) were OXA-51 positive, while 443 (85.4%) out of 519 strains harbored other beta-lactamase genes searched in the study. When the distribution of the genes were evaluated, blaTEM-1 was found as the predominant one with a frequency rate of 55.7% (n=289/519), followed by blaCTX-M2 (63/519, 12.1%), blaCTX-M1 (42/519, 8.1%), blaSHV (40/519, 7.7%), blaGES (8/519, 1.5%) and blaVIM (1/519, 0.2%). Cooccurence of ESBL genes was detected in 16.3% (72/443) of the strains, being mostly TEM+CTX-M2 (20/72, 27.8%), TEM+SHV (11/72, 15.3%) and TEM+CTX-M1 (10/72, 13.9%). In addition, it was noted that the distribution of ESBL genes between isolates showed differences according to the provinces. Accordingly, none of the strains isolated from four provinces (Bolu, Nigde, Mersin, Kahramanmaras) and from three provinces (Bolu, Kahramanmaras, Diyarbakir) harbored blaCTX-M1/M2 and blaSHV genes, respectively. The blaTEM gene was detected in isolates collected from all of the provinces, with a highest frequency in Nigde (28/31, 90.3%) and lowest in Trabzon (1/25, 4%). The presence of GES-11 type ESBLs was found only in the isolates sent from Nigde province (8/31; 25.8%). Screening of metallo-beta-lactamase VIM gene also yielded a single positive result amongst only Nigde isolates (1/31; 3.2%), and this gene was identified as VIM-5 type by DNA sequencing. This study which is the first comprehensive national research to characterize ESBLs in A.baumannii isolates by molecular methods, showed that the most prevalent ESBL type is TEM (289/519, 55.7%) amongst A.baumannii strains isolated from different regions of our country. The data of our study is parallel to the results of previous studies carried out from Turkey.

[Antimicrobial susceptibility and molecular characterization of multidrug-resistant Acinetobacter baumannii isolated in an university hospital]. / Bir üniversite hastanesinde izole edilen çok ilaca dirençli Acinetobacter baumannii izolatlarinin antimikrobiyal duyarliligi ve moleküler karakterizasyonu.

Acinetobacter baumannii, an aerobic, non-motile, gram-negative bacterium is an important nosocomial pathogen which shows resistance to the most antibiotics. Carbapenems are the most commonly used antibiotics for the treatment of infections caused by this pathogen. However the emergence of resistance against carbapenems in an increasing rate generates serious problems for antimicrobial therapy. The aims of this study were to detect the antibiotic susceptibility, and the presence of blaOXA resistance genes of clinical A.baumannii isolates and to determine the clonal relationship between these isolates. A total of 79 A.baumannii strains isolated from various clinical specimens (37 respiratory tract samples, 11 wound, 10 blood, 8 catheters, 6 tissue, 5 urine, 2 abscess) of the patients admitted to Mersin University Medical School Hospital between May 2012-January 2013, were included in the study. The isolates were identified by conventional methods and Vitek®2 Compact automated system. Antibiotic susceptibilities of the isolates were determined by Kirby-Bauer disk diffusion method and evaluated according to CLSI criteria. The presence of blaOXA-51, blaOXA-23, blaOXA-24, blaOXA-48 and blaOXA-58 genes were detected by an in-house polymerase chain reaction (PCR), and the clonal relationship between the isolates were identified by pulsed-field gel electroforesis (PFGE) using the ApaI restriction enzyme. In our study, all of the isolates were susceptible to colistin, while the resistance rates against piperacillin-tazobactam, ciprofloxacin, imipenem, meropenem, cefoperazone/sulbactam, trimethoprim-sulfamethoxazole, ceftazidime, levofloxacin, gentamicin, tetracycline, ampicillin-sulbactam, amikacin, netilmicin and tigecycline were 97.5%, 96.2%, 94.9%, 94.9%, 93.6%, 91.1%, 88.6%, 86%, 83.6%, 77.2%, 69.6%, 55.7%, 27.8% and 3.8%, respectively. All the isolates were identified as A.baumannii with the OXA-specific PCR and OXA16S rDNA sequence analysis. All of the isolates (100%) harboured blaOXA-51 and 71 (89.9%) harboured blaOXA-23 gene, however they were all negative for blaOXA-24, blaOXA-48 and blaOXA-58 genes. According to PFGE results 10 pulsotypes were identified, of these eight pulsotypes formed 77 (97.5%) similar strains with indistinguishable PFGE profiles ranging between 3-30 [A (n= 30), B (n= 20), C (n= 9), D (n= 5), E (n= 4), F (n= 3), G (n= 3), H (n= 3)]. When compared with the other clones, clones A and B were dominant among the samples and they have exhibited high level of antibiotic resistance. The rest two pulsotypes [I (n= 1), J (n= 1)] were in close relation with the main cluster. No common outbreak isolate was detected, but the relationship between the majority of the strains pointed out that there was a cross contamination problem in our hospital. In conclusion blaOXA-51 and blaOXA-23 were detected as predominant genes responsible from carbapenem resistance in our clinical A.baumannii strains, and it was considered that the high prevalence of clones A and B may constitute a threat in terms of hospitalized patients.

The new Schiff base 4-[(4-Hydroxy-3-fluoro-5-methoxy-benzylidene)amino]-1,5-dimethyl-2-phenyl-1,2-dihydro-pyrazol-3-one: experimental, DFT calculational studies and in vitro antimicrobial activity.

The synthesized Schiff base, 4-[(4-Hydroxy-3-fluoro-5-methoxy-benzylidene)amino]-1,5-dimethyl-2-phenyl-1,2-dihydro-pyrazol-3-one (I), has been characterized by (13)C NMR, (1)H NMR, 2D NMR ((1)H-(1)H COSY and (13)C APT), FT-IR, UV-vis and X-ray single-crystal techniques. Molecular geometry of the compound I in the ground state, vibrational frequencies and chemical shift values have been calculated by using the density functional method (DFT) with 6-311++G(d,p) basis set. The obtained results indicate that optimized geometry can well reflect the crystal structural parameters. The differences between experimental and calculated results of FT-IR and NMR have supported the existence of intermolecular (O-H⋯O type) and intramolecular (C-H⋯O type) hydrogen bonds in the crystal structure. Molecular electrostatic potential (MEP), frontier molecular orbital analysis (HOMO-LUMO) and electronic absorption spectra were carried out at B3LYP/6-311G++(d,p). HOMO-LUMO electronic transition of 3.92eV is due to contribution of the bands the n→π∗. The antimicrobial activity of the compound I was determined against the selected 11 bacteria and 8 fungi by microdilution broth assay with Alamar Blue. In vitro studies showed that the compound I has no antifungal effect for selected fungal isolates. However, the compound I shows remarkable antibacterial effect for the bacteria; Streptococcus pneumoniae, Haemophilus influenzae and Enterococcus faecalis.

[Ascaris lumbricoides in the nasogastric tube after operation on a patient with the diagnosis of acute mesenteric ischemia: case report]. / Akut Mezenterik Iskemi Tanili Hastanin Operasyon Sonrasi Nazogastrik Sondasindan Çikan Ascaris lumbricoides: Olgu Sunumu.

Ascaris lumbricoides is a comman intestinal helminths in humans. It is a parasite which commonly affects society with a low socioeconomic status, especially in tropical and rural areas. Ascaris lumbricoides infestation can lead to serious complications because of the mobility of the worms. The parasite can cause a variety of complications like intestinal obstruction, perforation, biliary obstruction, pancreatitis, peritonitis, liver abscess, cholangiohepatitis, volvulus, and gangrene, etc. A 59-year-old female patient hospitalized with the diagnosis of mesenteric ischemia was operated on for jejunal resection. On the 6th postoperative day, a worm was noticed emerging through the nasogastric tube. Ascaris lumbricoides was determined as a result of the examination microbiology laboratory. The patient was treated successfully with one dose of albendazole 200 mg 1x2. Our case describes a clinical situation of ascariasis observed after jejunal resection and emphasizes the importance of remaining aware of this rare complication of ascariasis.

[Identification of filamentous fungi isolated from clinical samples by two different methods and their susceptibility results]. / Klinik Örneklerden Izole Edilen Filamentöz Mantarlarin Iki Farkli Yöntemle Tanimlanmasi ve Duyarlilik Sonuçlari

Molds are widely distributed in nature. Aspergillus spp. represent the most frequently observed causative agents, however less frequent pathogens Fusarium, Scedosporium and Zygomycetes have also been considered the most important causes of morbidity and mortality in profoundly immunosuppressed hosts. The aims of this study were to identify filamentous fungi isolated from clinical specimens by conventional and molecular methods, and to detect their antifungal susceptibilities. A total of 6742 clinical specimens obtained from hospitalized patients at critical units of Mersin University Medical Faculty Hospital and sent to our laboratory between April 2008-January 2010 were included in the study. The isolates were identified by classical mycological methods and polymerase chain reaction-based DNA sequencing. Susceptibilities to fluconazole and voriconazole were tested by disk diffusion method and to fluconazole, voriconazole, amfoterisin B, caspofungin and posaconazole by E-test. Filamentous fungi were isolated from 71 (1.05%) samples (13 sputum, 4 wound, 4 peritoneal fluid, 3 extrenal ear discharge, 3 abscess and one of each cerebrospinal fluid, blood, tissue biopsy, nasal swab and conjunctival swab) which belonged to 32 patients (13 female, 19 male; age range 7 months-77 years, mean age: 46.6 years). Of the patients 62.3% presented one or more risk factors such as chronic renal failure (n= 8), chronic obstructive lung disease (n= 6), malignancy (n= 6), diabetes mellitus (n= 5) and peripheral vascular disease (n= 5). Of the isolates six were identified as Aspergillus niger, six as Aspergillus flavus, five as Aspergillus fumigatus, four as Aspergillus terreus, five as Fusarium spp., two as Bipolaris spp., and one of each as Acremonium spp., Aurebasidium spp., Mucor spp., and Scedosporium spp. By conventional methods. Three isolates exhibited different identities by DNA sequencing. All Aspergillus isolates were correctly identified at species level by both methods, Other fungi were identified at genus level by conventional methods and at species level by DNA sequencing. Fluconazole minimum inhibitory concentration (MIC) values were determined as > 256 mg/L in all strains, except Scedosporium; voriconazole MIC values were < 0.38 mg/L in all Aspergillus spp. Caspofungin MIC values were > 32 mg/L for Fusarium, Scedosporium, Rhizopus and Bipolaris strains and ≤ 0.006-0.125 mg/L in all Aspergillus isolates, In three strains (Fusarium equiseti, Cylindrocarpon lichenicola and Rhizopus oryzae) posaconazole minimum inhibitory concentration (MIC) values were > 32 mg/L, however it was < 1.5 mg/L, for the other strains. Amphotericin B MIC values were > 32 mg/L for Fusarium, Scedosporium, Rhizopus and all A.terreus strains and < 2 mg/L for the others. E-test and disk diffusion test results were compatible with each other for all the antifungal agents tested. In conclusion, the identification of filamentous fungi such as Aspergillus and Fusarium spp. is easily and reliably achieved by conventional methods. Since the rate of invasive fungal infections is increasing currently, filamentous molds should be searched especially in the clinical specimens of immunocompromised patients for accurate and prompt diagnosis of such infections and to decrease the related mortality risk.

[Investigation of the presence of Cryptosporidium spp. in different water sources in Mersin province, Turkey]. / Mersin Ilinde Farkli Su Kaynaklarinda Cryptosporidium spp. Varliginin Arastirilmasi

Cryptosporidium is an intracellular protozoon that causes enteritis in human and animals. Contaminated water and food are the major sources for the transmission of oocysts via oral-fecal route. It is reported that the prevalence of cryptosporidiosis is higher in developing countries than developed countries because of inefficient sanitation and disinfection facilities for drinking water. The most frequently detected species is Cryptosporidium parvum leading to high morbidity in healthy subjects and also fatal infections in immunocompromised patients. The acid-fast staining method is widely used in the diagnosis of cryptosporidiosis. Nowadays, Cryptosporidium could easily be detected in water supplies and asymptomatic carriers by molecular techniques to obtain epidemiological data. In this study it was aimed to detect and identify Cryptosporidium oocysts in different water sources in Mersin province, Turkey. A total of 135 water samples (70 taps, 50 wells and 15 sewage) collected from city center (n= 25) and from Tarsus (n= 32), Mezitli (n= 33) and Karaduvar (n= 45) counties between March 2007 and May 2009 were included in the study. Water samples in 10 liter volumes, were filtered by 0.45 µm pore-sized membrane filter vacuum/ pressure pumping technique. Cryptosporidium oocysts in filtrates were detected by modified cold Kinyoun acid-fast stain (MCK) technique and also identified and typed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. MCK yielded three and PCR yielded seven positive results. All the strains were identified as C.parvum by PCR-RFLP method. All of the three MCK-positive samples were also found positive with PCR, however four PCR positive samples were MCK-negative. Thus, the prevalence of C.parvum was estimated as 5.2% (7/135) in our region. Of seven positive samples, one was a sewage water sample collected from the city center, while the remaining (two tap water, two well water and two sewage water samples) belonged to the samples collected from Karaduvar county, interestingly. It was thought that deficient infrastructure and use of well water as drinking water supply in Karaduvar region might be the cause of high rate of Cryptosporidium (6/45; 13.3%). Further studies which will determine the genotypes and investigate the phylogenetic relationship between these Cryptosporidium spp., might aid to the epidemiology of cryptosporidiosis in our region.

[Identification and isolation of non-tuberculous mycobacteria from environmental samples]. / Çevre örneklerinden tüberküloz disi mikobakterilerin izolasyonu ve tanimlanmasi

Non-tuberculous mycobacteria (NTM) found frequently in tap water and environment cause important opportunistic infections in immunocompromised patients. The aim of this study was to isolate and identify non-tuberculous mycobacteria in soil, raw milk and water distribution system samples in Mersin (a province located at Mediterranean region of Turkey). A total of 101 water, 124 soil and 40 milk samples collected from the central part and suburban parts of Mersin during November 2003-May 2004 period were included in the study. Water samples were collected from 29 different water distribution systems; soil samples from different parks and gardens and milk samples from raw milks sold at different districts. After the samples were processed by homogenization and decontamination, acid-fast staining and culture into Löwenstein-Jensen medium were performed. Acid-fast bacilli isolated from culture medium were identified by using conventional methods, polymerase chain reaction (PCR)-RFLP (Restriction Fragment Length Polymorphism) and INNO-LIPA Mycobacteria methods. NTM were identified from 4.9% (5/101) of water samples and 0.8% (1/124) of soil samples by culture and PCR. No NTM were detected in the raw milk samples. Three of the NTM strains isolated from water samples were defined as Mycobacterium chelonae type III and two as Mycobacterium kansasii type II. One NTM strain isolated from soil was defined as Mycobacterium fortuitum. It was of note that two of the five NTM positive water samples were tap water samples collected from hospitals. It was concluded that NTM colonization/contamination of water and environment in the hospitals was a potential risk factor in terms of nosocomial infections. Thus surveillance cultures of the water systems and the medical devices in the hospital are necessary to fix the source of NTM, to identify and type the strains and to establish effective control measures such as sterilization, disinfection, maintenance and modernization of water systems.

[Isolation and identification of Mycobacterium tuberculosis from the urine samples by conventional and molecular methods]. / Konvansiyonel ve moleküler yöntemler ile idrar örneklerinden Mycobacterium tuberculosis izolasyonu ve tanimlanmasi.

Genitourinary tuberculosis presents a challenge in diagnosis and treatment due to variations in clinical and radiological signs, insufficient patient history and difficulty in the isolation of the bacilli. The aim of this study was to isolate and identify Mycobacterium tuberculosis from the urine samples obtained from patients with suspected urinary tuberculosis admitted to our hospital by using Ehrlich-Ziehl-Neelsen (EZN), culture and polymerase chain reaction-restriction analysis (PCR-RFLP) methods. A total of 1004 urine samples collected from 437 patients who were admitted to our hospital between January 2004-July 2006, were inoculated on Löwenstein-Jensen (LJ) and/or BACTEC 12B (Becton Dickinson, USA) after decontamination and, direct preparations stained with EZN method were evaluated microscopically. M. tuberculosis complex (MTC) and mycobacteria other than tuberculosis (MOTT) were differentiated by nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test and the susceptibility testing for the MTC strains to primary antituberculosis drugs were performed by BACTEC 460 TB (Becton Dickinson, USA) system. PCR-RFLP method was performed for the identification of Mycobacterium spp. Twenty-two (5%) patients have yielded positive results by at least one of the conventional methods (EZN, LJ and/or BACTEC). Fifteen samples were positive for acido-resistant bacilli (ARB) by EZN method, and 17 samples were positive for mycobacterial growth in the cultures. Ten of 22 patients were found positive by both of the methods, while seven were culture positive but ARB negative and five were culture negative but ARB positive. These five patients received BCG treatment because of the presence of bladder tumor. Twelve (70.5%) of 17 strains isolated from culture were identified as MTC, while five (29.4%) were identified as M. fortuitum. Of 12 MTC isolates, eight (66.7%) were found susceptible to all of the antituberculosis agents, while one was found resistant to isoniazide (INH) and ethambutole (ETB), one was resistant to INH and rifampicin (RIF), and two were resistant to only INH. It is concluded that, in order to identify mycobacteria and to perform antituberculous susceptibility tests, cultivation of mycobacteria is a prerequisite.

Consistence of miniexon polymerase chain reaction-restriction fragment length polymorphism and single-copy gene sequence analyses in discriminating Leishmania genotypes.

Recently, a new polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP)-based assay had been developed using the miniexon sequences for genotyping Leishmania isolates. We had used this method for rapid diagnosis and genotyping of visceral and cutaneous leishmaniasis with the combination of microcapillary cultivation. In this study, we have evaluated this approach by examining genomic DNAs from 47 independent isolates, which were grouped into 19 genotypes of Leishmania subgenus complexes by sequence polymorphism of single-copy genes. Results obtained provide miniexon RFLP configurations specific to Leishmania enriettii, Leishmania tarentolae, and Leishmania gerbilli for the first time. Altogether, 92% of the results from miniexon PCR-RFLP are in agreement with those based on the sequence database of single-copy genes from the same isolates. The miniexon PCR-RFLP method is simple, sensitive, and specific method useful for routine diagnosis of different Leishmania.

Does cerumen have a risk for transmission of hepatitis B?

OBJECTIVES/HYPOTHESIS: Chronic hepatitis B virus infection is a significant worldwide health problem. It affects 350 to 400 million people. The patients with chronic hepatitis B virus infection have a significant risk for the development of cirrhosis or hepatocellular carcinoma. Full awareness of the mechanisms of transmission can allows susceptible individuals to refrain from this infection. Cerumen has never been studied as a route for hepatitis B transmission. The aim of the study was evaluate the importance of cerumen in transmission of hepatitis B virus infection. STUDY DESIGN: This study was performed on forty patients with confirmed hepatitis B virus infection. METHODS: Forty cerumen specimens collected from the patients with hepatitis B virus DNA in their sera were prospectively analyzed for the presence of hepatitis B virus DNA by real-time polymerase chain reaction. RESULTS: Eleven of 40 cerumen specimens (27.5%) were positive for hepatitis B virus DNA, with counts ranging from 4.2 x 10 to 4.7 x 10 copies per sample. There was positive correlation between hepatitis B virus DNA concentrations of serum and cerumen. Half of hepatitis B e antigen (HBeAg)-positive patients had detectable hepatitis B virus DNA levels (5.7 x 10 to 4.7 x 10 copies) in cerumen specimens, whereas 12.5% of cerumen specimens from anti-HBe-positive patients had hepatitis B virus DNA levels (4.2 x 10 to 7.0 x 10 copies). CONCLUSION: Cerumen can be a potential source of transmission. Therefore, this route should be investigated in further studies for horizontal, nosocomial, and occupational transmission of hepatitis B.