The use of Matrix-assisted laser desorption ionization-time of flight mass spectrometry in the identification of Francisella tularensis

Francisella tularensis is the cause of the zoonotic disease tularemia and is classified among highly pathogenic bacteria (HPB) due to its low infection dose and potential for airborne transmission. In the case of HBP, there is a pressing need for rapid, accurate and reliable identification. Phenotypic identification of Francisella species is inappropriate for clinical microbiology laboratories because it is time-consuming, hazardous and subject to variable interpretation. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was recently evaluated as a useful tool for the rapid identification of a variety of microorganisms. In this study, we evaluated the use of MALDI-TOF MS for the rapid identification of Francisella tularensis and differentiation of its subspecies. Using national collection of Francisella isolates from the National Tularemia Reference Laboratory (Public Health Institute of Turkey, Ankara), a total of 75 clinical isolates were investigated by species and subspecies-specific polymerase chain reaction (PCR) test and MALDI-TOF MS. All isolates were originally identified as F. tularensis subsp. holarctica due to RD1 subspecies-specific PCR result. For all isolates MALDI-TOF MS provided results in concordance with subspecies-specific PCR analysis. Although PCR-based methods are effective in identifying Francisella species, they are labor-intensive and take longer periods of time to obtain the results when compared with MALDI-TOF MS. MALDI-TOF MS appeared to be a rapid, reliable and cost-effective identification technique for Francisella spp. Shorter analysis time and low cost make this an appealing new option in microbiology laboratories

Brucella Melitensis Misidentified As Roseomonas Gilardii By Maldi-Tof Ms: Experience Of A Clinical Microbiology Laboratory

Misidentification of a blood culture isolate of Brucella melitensis as Roseomonas gilardii by the VITEK (R) MS (bioMerieux, Marcy l'Etoile, France) matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) microbial identification system is herein drawn attention. Blood and serum samples of a 42-year-old male with pre-diagnosed neurobrucellosis were evaluated in bacteriology and serology laboratories. Gram-negative coccobacilli from positive blood culture bottle were identified as R. gilardii by VITEK (R) MS, but it was not identified by VITEK (R) 2 Compact (bioMerieux, Marcy l'Etoile, France) MALDI-TOF MS microbial identification system. Serological tests were positive for brucellosis, so it was an important clue for the possibility of a misidentification. The isolated organism was confirmed as B. melitensis by polymerase chain reaction and biotyping in the National Reference Laboratory. Microbiology laboratories in countries where brucellosis is endemic must be careful while using commercial bacterial identification systems for identification of Brucella spp.WoSScopu

First Isolation of Coxiella burnetii in Turkey from a Patient with Endocarditis; Antigen Production and Phase Change Study

Coxiella burnetii is the causative agent of Q fever, a zoonotic infection. The bacteria is a gram-negative, pleomorphic, coccobacilli and capable to survive and proliferate within the host cell's phagolysosome. There are two morphological cell types of C.burnetii including small and large cell variants. C.burnetii is divided into phase I and phase II serologically variants according to LPS structure in the cell wall. Phase I is the natural phase found in infected animals or humans and is highly infectious. Phase II is not very infectious and could be obtained only in laboratories after serial passages in cell cultures or embryonated egg cultures. Q fever can be asymptomatic (in 50% of the cases), acute or chronic. Major presentations of acute Q fever are flu-like illness, pneumonia, and hepatitis, whereas the chronic form presents mainly as infective endocarditis. The aim of this study was to obtain C.burnetii phase II variant from C.burnetii phase I variant by a phase change study. In this study, C.burnetii was isolated by cell culture method from the heart valve tissue of a Q fever endocarditis case. C.burnetii phase I antigen for the indirect fluorescent antibody test (IFAT) was prepared from the isolated strain. For the isolation and identification of C.burnetii, heart valve tissue of the patient was homogenized and DNA was extracted by tissue extraction kit. C.burnetii DNA in the valve tissue was determined by real-time PCR (Rt-PCR). This C.burnetii DNA positive specimen was inoculated into Vero cells by shell vial centrifugation method. The scraped Vero cells were fixed on the slides after one week of incubation and IFAT was performed using C.burnetii phase I IgG positive sera, bacteria that were grown in and surrounding the Vero cells stained apple green were determined microscopically. Infected cells were disrupted by freeze and thaw method to obtain bacterial suspension. The DNA obtained from the bacterial suspension was again found to be positive for C.burnetii by Rt-PCR. Isolation sample was found to be positive in PCR at an earlier cycle compared to heart tissue sample, thus the bacterial growth was also confirmed with PCR. 16S ribosomal RNA gene of our isolate was amplified by PCR using 27F and 1492 primers and then sequenced. The DNA sequences were compared with reference DNA sequences of GeneBank; and the nucleotide sequence of the 16S ribosomal RNA gene of our isolate was found to be 99% similar to C.burnetii strain ATCC VR-615 an accession number NR104916. Serial cell culture passages of the isolated strain were performed to obtain C.burnetii phase II variant from C.burnetii phase I variant. After each passage, presence of phase change was investigated by IFAT using C.burnetii phase I and phase II IgG positive sera. At the end of 17 cell culture passages, phase change could not be observed. C.burnetii phase I IFAT antigen was prepared from the obtained bacterial suspension. In this study, we presented the isolation and identification of C.burnetii by cell culture, molecular and serological methods from the heart valve of a patient with endocarditis for the first time in our country

Human anthrax in Turkey: A ten years' experience (2009-2018).

Anthrax is a notifiable disease in Turkey. In order to control the human disease, animal foci are being monitored and prevention and control activities are being implemented by the Ministry of Health in coordination with the Ministry of Agriculture and Forestry. The objective of our study was to evaluate the national surveillance data and control activities in the last decade. A total of 1174 anthrax cases and 9 deaths have been reported. Anthrax was frequent in eastern provinces and in big cities where large animal movements were significant. The incidence rate was 1.5 times higher in males than in females. The disease was more common in the 30-64 age group. The number of cases increased in the summer and autumn seasons. Human anthrax is still being reported though in decreasing numbers in Turkey. A collaborative control programme continues to be needed

Genomic Characterization of the Novel Bartonella refiksaydamii sp. Isolated from the Blood of a Crocidura suaveolens (Pallas, 1811)

International audienc

Seroprevalence of Coxiella burnetii in Stray Cats in Central Anatolia

7th Veterinary Microbiology Congress -- SEP 22-28, 2006-2008 -- Antalya, TURKEYWOS: 000262864300013Coxiella burnetii causes Q fever (Coxiellosis) in humans and animals worldwide. The present study was carried out to determine the seroprevalence of Q fever among stray cats in 3 providences (Ankara, Nigde, and Kayseri) in Central Anatolia, Turkey. A total of 143 sera from stray cats were examined for the presence of IgG against C. burnetii phase II antigen by indirect fluorescent antibody test (IFAT). Seven out of the 143 (4.9%) stray cats were seropositive for Q fever, with titers of 1:64 to 1:256. Seroprevalences in Ankara, Nigde, and Kayseri provinces were 1.6%, 7.4%, and 8.3%, respectively. This is the first report of the presence of C. burnetii in cats in Turkey

Bartonella species in wild small mammals in Western Black Sea Region of Turkey

WOS: 000357000800004The species within the genus Bartonella are intracellular bacteria causing long-lasting bacteremia in humans and animals. In this study, Bartonella spp. in 173 small mammals, which were Apodemus flavicollis, A. witherbyi, A. uralensis, A. mystacinus, Myodes glareolus, Crocidura suaveolens, Rattus rattus and Rattus norvegims species captured from Western Black Sea Region of Turkey, were investigated by blood culture and molecular methods. The positivity of Bartonella was 63.6% (110/173) by blood culture of small mammalian. The gltA gene regions for the isolated strains were identified by DNA sequencing analysis. Isolates were identified as Bartonella taylorii, B. birtlesii, B. coopersplainsensis and a zoonotic B. grahamii

A Serological Investigation of Bartonella henselae Infection in Cats in Turkey

Bartonella hencelae is the causative agent of cat scratch disease (CSD) in humans. Cats are the main reservoir of this bacterium and may infect humans through scratches and bites. The purpose of this study was to determine the B. henselae seroprevalence in cats in Turkey. A total of 298 cats blood samples were collected from six different provinces of Turkey. Sera were tested for the presence of anti-B. henselae IgG antibodies by indirect fluorescent antibody test (IFA). The seroprevalence of B. henselae was 27.9% (83/298) for the cats examined in this study. The seroprevalence of cats by province was significantly higher in Bursa (41.3%), Adana (33.9%), Aydin (27.5%) and Burdur (32.3%) than in Kayseri (17.9%) and Istanbul (12.5%). Statistically significant differences were not observed between cat sexes and living conditions of cats. The results revealed that B. henselae is an important zoonotic pathogen in Turkey

Seroprevalance and co-existence of Brucellosis, Listeriosis and Toxoplasmosis in dairy cattle in Kirikkale province

Celebi, Bekir/0000-0002-4545-5573WOS: 000259238600014This study was conducted on a total of 100 dairy cows of Holstein and Holstein crossbred cows of 3-7 years of age between December 2005 and April 2006. The aim of the Study was to determine seroprevalence of brucellosis, listeriosis and toxoplasmosis, and the frequency of their co-existence in dairy cows. Clinically, the mean pulsus rate, breathing rate and body temperature were determined as 72 +/- 12/min., 24 +/- 6/min and 38.2 +/- 0.6 (degrees) under barC, respectively. Antibodies against Brucella abortus were determined by means of Microplate Agglutination Test (MAT). The titres above 1/40 were considered seropositive. The 'O' antibodies, generated against listeria monocytogenes, were determined by Osebold method. At least 2 (+) results at the titres above 1/100 were considered seropositive. Toxoplasma gondii antibodies were analyzed by Sabin-Feldman Dye Test (SFBT). The titres above 1/16 were considered seropositive. As a result, in dairy cows of Kirikkale region the seroprevalence of brucellosis, listeriosis and toxoplasmosis were 19%, 37% and 53%, respectively. The frequency of co-existence all three together was 5%. The frequency of co-existence of brucella with listeria was 4%, of brucella with toxoplasma was 7%, and of listeria with toxoplasma was 13%