Phylogenetic Analysis of Brucella melitensis Strains Isolated from Humans Using 16S rRNA Sequencing and Multiple Locus Variable Number of Tandem Repeats Analysis-16.

Background: Brucellosis is the most important public health problem worldwide, and the annual incidence of the disease in humans is 2.1 million. The Brucella genome is highly conserved, with over 90% similarity among species. The aim of this study was to perform species-level identification of Brucella spp. strains isolated from humans diagnosed with brucellosis and to further investigate the phylogenetic relationships using multiple locus variable number of tandem repeats analysis (MLVA)-16 and 16S rRNA sequencing analysis. Materials and Methods: Brucella spp. was isolated from the blood cultures of 54 patients who tested positive for brucellosis through serological examinations. Real-time PCR was used to identify the isolates in species, and the genus level of Brucella was confirmed with 16S rRNA. All isolates were subjected to phylogenetic analysis using variable number of tandem repeat analysis with multiple loci. Results: Subsequent analysis via real-time PCR confirmed these isolates to be of the Brucella melitensis species. The 16S rRNA sequence analysis showed 100% homogeneity among the isolates. MLVA revealed the formation of five different genotypic groups. While two groups were formed based on the 16S rRNA sequence analysis, five groups were formed in the MLVA. Conclusions: The study concluded that 16S rRNA sequence analysis alone did not provide sufficient discrimination for phylogenetic analysis but served as a supportive method for identification. MLVA exhibited higher phylogenetic power. The widespread isolation of B. melitensis from human brucellosis cases highlights the importance of controlling brucellosis in small ruminants to prevent human infections.

Genotyping of Brucella isolates from animals and humans by Multiple-Locus Variable-number Tandem Repeat Analysis (MLVA).

This study investigates country-wide genotype variations through the genotyping of Brucella strains isolated from domestic ruminants and humans. The Brucella spp. isolated from samples taken from animals and humans were first identified as B. abortus and B. melitensis by real-time PCR, and the MLVA-16 approach was then used for the genotyping of the identified isolates. For the study, 416 Brucella spp. were isolated from aborted fetus samples examined between 2018 and 2021, and 74 Brucella spp. from infected humans. Of the 74 human isolates analyzed, 1.3% were identified as B. abortus and 98.7% (73/74) as B. melitensis. The MLVA-16 typing method revealed 30 clonal groups for B. abortus and 37 clonal groups for B. melitensis from which the dominant genotypes and similarities with human isolates in Türkiye were determined.

Growth hormone level in COVID-19 patients.

OBJECTIVE: Coronavirus disease-2019 (COVID-19) disease can cause asymptomatic and mild flu-like symptoms as well as severe symptoms ranging from respiratory failure and death. Growth hormone (GH) is produced in the anterior pituitary and plays an important role in the immune system. COVID-19 is severe in the elderly, men, obese, diabetics, and people with immune deficiency. The probability of GH deficiency is high in these patient groups. In this study, we aimed to investigate the relationship between the severity of COVID-19 infection and GH level. METHODS: A total of 456 patients, between 45 and 80-years-old, who were hospitalized with the diagnosis of COVID-19 disease were evaluated in the study. Our study was a retrospective study. Demographic data of the patients, GH, insulin-like growth factor-I (IGF-1), and biochemical parameters and thorax tomography results were evaluated. Patients with chronic diseases that would affect GH levels and those in need of intensive care were excluded from the study. RESULTS: 456 patients were included in the study, 168 female, 288 male, mean age 67.57±12.60 years. Patients were divided into two groups according to thorax tomography findings, patients with lung involvement in Group-1:352 (77%) and those without pulmonary involvement in Group-2:104 (23%). While the GH of Group-1 was 0.125 ng/ml, the GH of Group-2 was 0.238 ng/ml, the difference between them was statistically significant (p=0.000). IGF-1 in Group-1 was: 55.05 ng/ml, while IGF-1 in Group-2 was: 104.08 ng/ml, the difference between them was statistically significant (p=0.000). In multivariate regression analysis, low IGF-1 (p=<0,01, OR:1,06 [1028-1093]) level was found to be significantly effective in lung involvement in COVID-19 disease. CONCLUSION: In our study, we found GH and IGF-1 deficiency in COVID-19 cases with lung involvement, regardless of age and gender. We can say that COVID-19 infection progresses worse in GH and IGF-1 deficiency.

Molecular epidemiology of Coxiella burnetii detected in humans and domestic ruminants in Turkey.

Q fever is a zoonotic disease that is known to be widespread throughout the world by many researches since its discovery in 1935 and it is important in terms of animal and public health. Coxiella burnetii, which is the etiological agent of the disease, is an obligate intracellular pathogen. While the disease generally manifests itself with abortion in animals, disease manifests as atypical pneumonia or granulomatous hepatitis in the acute form and as endocarditis in the chronic form in humans. Its presence in Turkey has been shown with a large number of studies. The aim of this study was to show the genotypic relationship with MLVA analysis of C. burnetii samples found in cattle, sheep and goat samples in Erzurum and Samsun Veterinary Control Institutes and blood samples collected from humans with atypical pneumonia findings. In the study, MLVA analyses of 100 positive samples from 50 cows, 41 sheep and 9 goats from Northeast Anatolia and Black Sea regions and C. burnetii positive samples found in 6 individuals with atypical pneumonia were performed. As a result of the study, it was found that 106 C. burnetii samples had belong to 16 genotype groups. It was found that genotype XVI was the most prevalent among these groups and it was seen in both regions. In addition to this, genotype IX profile was the second largest group with 83.3% (5/6) of human samples. In this study, the genotypes common in the regions were determined and a data source was created for possible outbreaks.

High-Frequency Chest Wall Oscillation in Patients with COVID-19: A Pilot Feasibility Study.

OBJECTIVE: Coronavirus 2019 disease presents in a spectrum that can range from mild viral infection to pneu- monia. Common symptoms of coronavirus disease 2019 pneumonia include cough, sputum, and shortness of breath. High-frequency chest wall oscillation is a pulmonary rehabilitation method used for the recovery of pulmonary functions and removal of secretions in the lungs. The aim of the study was to evaluate the efficacy of high-frequency chest wall oscillation on patients with coronavirus disease 2019 pneumonia. MATERIALS AND METHODS: In this study, 100 patients, between 18 and 70 years old, with a positive polymerase chain reaction result for coronavirus disease 2019, were included. Standard medical treatment was applied to all patients. In group rehabilitation, high-frequency chest wall oscillation treatment was applied twice a day for 20 minutes for 5 days. No additional intervention was made to the control group. Pulmonary function tests and oxygenation were evaluated on the first and fifth days. Patients' high-flow oxygen, non-invasive mechani- cal ventilation, and invasive mechanical ventilation needs were evaluated and recorded. RESULTS: Compared with the control group, the forced expiratory volume in 1 second, forced vital capacity, and peak expiratory flow rates were statistically higher in the rehabilitation group on the fifth day (P < .05). On evaluating the oxygenation of patients, the fifth day to first-day oxygen saturation difference was signifi- cantly higher in rehabilitation group than in control group (P < .05). Furthermore, the number of patients who needed non-invasive mechanical ventilation was lower in the rehabilitation group (P < .05). CONCLUSION: This study demonstrated that pulmonary rehabilitation applied with the high-frequency chest wall oscillation device in patients with coronavirus disease 2019 in the early period contributed to the improvement of oxygenation by providing significant improvement as observed in the pulmonary function tests of the patients.

Evaluation of apically extruded bacteria associated with different nickel-titanium systems.

INTRODUCTION: The aim of this in vitro study was to evaluate the amount of apically extruded bacteria associated with several root canal preparation systems. METHODS: Forty-four extracted human mandibular premolar root canals were contaminated with an Enterococcus faecalis suspension. After incubation at 37°C for 24 hours, the root canals were instrumented using the Twisted File (SybronEndo, Orange, CA), OneShape (Micro Mega, Besançon, France), and ProTaper Next (Dentsply Maillefer, Ballaigues, Switzerland). During instrumentation, apically extruded bacteria were collected into vials containing 0.9% NaCl. The microbiological samples were taken from the vials and incubated in brain-heart agar medium for 24 hours. The numbers of colony-forming units were determined. The data obtained were analyzed using Welch analysis of variance followed by post hoc Games-Howell tests. RESULTS: ProTaper Next extruded the highest amount of bacteria, whereas OneShape extruded the least compared with all the other instruments (P = .004). There was a significant difference between OneShape and ProTaper Next in the number of colony-forming units (P = .007) but not between OneShape and Twisted File (P > .05). CONCLUSIONS: All instrumentation systems extruded bacteria beyond the foramen. The OneShape system extruded less bacteria compared with the Twisted File and ProTaper Next systems.

Antimicrobial and structural effects of different irrigation solutions on gutta-percha cones.

PURPOSE: To evaluate the alterations on the surface of gutta-percha cones (GPCs) on exposure to the different irrigation solutions and their possible antibacterial effect against Enterococcus faecalis. (E. faecalis). MATERIALS AND METHODS: Disinfection ability of different solutions (5.25% sodium hypochlorite, 2% chlorhexidine, 1% peracetic acid, and QMix) were tested with 96 GPCs and the time of exposure to each solution was 5 and 10 minutes, respectively. GPCs used in this study were contaminated with E.faecalis. After disinfection, GPCs were placed in tubes containing the medium and incubated at 37˚C for 7 days. All tubes were visually checked for turbidity at 24-hour intervals. About 92 new GPCs were analyzed by means of SEM/EDS to assess the topography and chemical elements present on their surface. The data generated was analyzed using Pearson chi-square test, p<0.05. RESULTS: There were no significant statistical differences in disinfection quality between the irrigation solutions used on GPCs contaminated with E. faecalis (p>0.05). SEM/EDS analyses showed no alteration in the superficial features of GPCs after treating with various irrigation solutions. CONCLUSION: QMix was found to be an effective agent for rapid disinfection of GPCs as well-known irrigation solutions. Irrigation solutions were found to have sterilized the GPCs after both 5 and 10 minutes of exposure.

[Evaluation of Xpert MTB/RIF results for the detection of Mycobacterium tuberculosis in clinical samples]. / Klinik Örneklerde Mycobacterium tuberculosis Varliginin Gösterilmesinde Xpert MTB/RIF Sonuçlarinin Degerlendirilmesi.

Tuberculosis is still a major global health problem. Nowadays nucleic acid amplification tests which are recommended by the World Health Organization (WHO) become popular methods for the rapid detection of Mycobacterium tuberculosis complex (MTC). Recently introduced commercial Xpert MTB/RIF (Cepheid, USA) system is also a molecular method based on real-time polymerase chain reaction for simultaneous detection of both MTC and rifampicin resistance in the clinical sample. The sample processing, nucleic acid extraction, amplification and detection of known mutations related to rifampicin resistance are performed in a single cartridge in this integrated system and the results are obtained in two hours. The aim of this study was to evaluate the performance of Xpert MTB/RIF system for the detection of M.tuberculosis in pre-processed clinical samples by comparing the results obtained by Bactec 460TB 12B (BD Diagnostic, USA), Löwenstein-Jensen (LJ) culture and direct microscopy of smears stained with Ziehl- Neelsen (ZN). A total of 85 clinical specimens (50 sputum, 25 bronchoalveolar lavage, five thorasynthesis fluid and five urine samples) obtained from tuberculosis-suspected patients were included to the study. All specimens were decontaminated and this decontaminated suspension was used in the diagnostic methods, except for Xpert MTB/RIF process. Twenty-five (29%) of the samples yielded positive result with Bactec 460TB, 25 (29%) were found positive with Xpert MTB/RIF, 15 (18%) were found positive with LJ and 11 (13%) were found positive with ZN staining method. High consistency was detected between the results of Bactec 460TB and Xpert MTB/RIF when Bactec 460TB was considered as the gold standard method (r= 0.943; p= 0.000). One specimen yielded false positive result with Xpert MTB/RIF when compared to the reference method. The sensitivity, specificity, positive and negative predictive values of Xpert MTB/RIF test were then estimated as 96%, 98%, 96% and 98%, respectively. No resistance were detected for the tested isolates. This study suggested that the sensitivity of Xpert MTB/RIF system in direct detection of M.tuberculosis in smear positive and smear negative samples was consistent with the reference methods. Moreover, the MTB/RIF test provided sensitive detection of tuberculosis in less than two hours.