[Legionella Species Isolated in the Water Systems of Ships and Their Molecular Characterization]. / Gemilerin Su Sistemlerinden Izole Edilen Legionella Türleri ve Moleküler Karakterizasyonu.

Legionella species are generally found in nature and in water resources, and they are gram negative bacilli that can cause pneumonia by being transmitted from water systems to humans via aerosol or aspiration. Legionnaires' disease caused by this agent continues to be a public health problem in cruise ships. In this study, it was aimed to determine the prevalence of the colonization of Legionella species by culture method and to determine the molecular characterization of the isolated Legionella in water samples taken from the water systems of the ships docking in Mersin International Port. A total of 158 cold water samples were taken from 18 ferry and/or cargo ships docking in Mersin International Port between December 2014 and June 2015. Fifty-four of the samples were obtained from tanks, 68 from taps and 36 from shower heads. All samples were centrifuged and inoculated from the pellet onto "Buffered Coal Yeast Extract" (BCYE) (Oxoid, CM0655, UK) agar medium supplemented with iron pyrophosphate, L-cysteine and α-ketoglutarate (Oxoid, SR0110, UK). The culture plates were incubated for 10-15 days in microaerophilic environment in a desiccator at 37°C. The suspicious colonies grown in cultures were serogrouped by latex agglutination test (Oxoid, DR0800M, UK) and fluorescent antibody method (m-Tech Monoclonal Technologies, Inc., USA). For the molecular analysis of Legionella species grown in culture, DNA isolation was made from Legionella colonies and then polymerase chain reaction amplification was performed using specific primer sequences targeting the rpoB gene region of the Legionella genome. Direct DNA sequencing of rpoB gene products was performed in the "ABI PRISM 3130XL Genetic Analyzer" (Applied Biosystems, USA). The DNA sequences were typed by BLAST analysis and the determined types, and NCBI (National Center for Biotechnology Information) reference Legionella sequences were phylogenetically compared with the Neighbor-Joining comparison method by using the Mega 7 program. Legionella spp. was isolated in 18 (11.4%) of 158 samples. Of these, four (7.4%, 4/54) were detected from the tank, 11 (16.2%, 11/68) from the tap and three (8.33%, 3/36) from the shower head. After the latex agglutination test performed from the growing bacterial colonies, five (27.8%) were serogrouped as Legionella spp., four (22.2%) as Legionella pneumophila sg 5, two (11.1%, each) as L.pneumophila sg 1,L.pneumophila sg 8 and Legionella bozemanii and one (5.6%) as L.pneumophila sg 3. Two (11.1%) of the isolates grown in culture could not be serogrouped. Molecular characterization of 12 Legionella isolates could be performed. One of them was serologically serogrouped as L.bozemanii, and it was found to be 99% similar to Legionella rubrilucens when compared with NCBI Legionella sequence data in the BLAST program. One isolate that could not be differentiated by serogrouping was identified as Legionella erytra in the BLAST program after DNA sequence analysis. The remaining 10 isolates (55.6%, n= 18) were confirmed as L.pneumophilia after the comparison with reference NCBI sequences. In this study, it was determined that 11.4% of the water samples collected from the water systems of the ships docking in Mersin International Port were contaminated with Legionella species. The detected Legionella species have an important potential source of infection for the captain, ship workers and passengers travelling on the ships. In this respect, this study reveals the necessity of establishing studies to improve the risk management of Legionella in the water systems of ships.

[Detection and Molecular Characterization of Human Adenoviruses from Acute Conjunctivitis Cases]. / Akut Konjunktivitli Olgularda Insan Adenovirüslerinin Belirlenmesi ve Moleküler Karakterizasyonu.

Human adenoviruses (hAdV) can cause a wide range of clinical diseases in children and adults that mainly affect respiratory, eye and gastrointestinal systems. Ocular hAdV infections have various clinical manifestations such as epidemic keratoconjunctivitis, pharyngoconjunctival fever and non-specific follicular conjunctivitis. The hAdV genotypes which can cause conjunctivitis vary according to geographic distribution. In the study, we aimed to determine the frequency of the presence of hAdV by molecular methods and to determine the types with phylogenetic analysis in conjunctival swab samples taken from patients diagnosed clinically as acute conjunctivitis. Conjunctival swab samples (n= 100) were taken from the patients with acute conjunctivitis who have admitted to Mersin University Faculty of Medicine Hospital Ophthalmology Clinic and 50 conjunctival swab samples taken from healthy individuals as a control, between September 2014-July 2017 were included in the study. Following the DNA isolation from swab samples, polimerase chain reaction (PCR) amplification was performed using specific primer sequences targeting the hexon gene region of the hAdV genome. In order to determine hAdV types, direct DNA sequence analysis of hexon gene products was performed in "ABI PRISM 3130XL Genetic Analyzer" (Applied Biosystems, Foster City, CA, USA). The obtained hAdV DNA sequences were typed by BLAST analysis and the identified genotypes were compared phylogenetically with the reference hAdV sequences of the NCBI In the study, 30 (30%, 30/100) of the swab samples of the patients with acute conjunctivitis were found positive for hAdV hexon gene PCR. The hAdV DNA was not found in the conjunctival swab samples belonging to the healthy individuals included as controls. A total 27 samples found as positive of the hexon gene PCR were genotyped by direct DNA sequence analysis. A total of 5 genotypes were identified and the most common genotypes were hAdV-8 (n= 17, 63%) and followed by hAdV-53 (n= 4, 14.8%), hAdV-4 (n= 4, 14.8%), hAdV-7 (n= 1, 3.7%) and hAdV-37 (n= 1, 3.7%). In this study, the prevalence of adenoviral conjunctivitis determined by hexon gene PCR in patients with clinical diagnosis of acute conjunctivitis was similar to the prevalence rate reported in other regions of the world. In our region, more than one type of hAdV type was associated with acute conjunctivitis. The predominant type was determined as hAdV-8 with a 63% ratio. These results will significantly contribute to the molecular epidemiology of hAdV types in conjunctivitis cases.