Targeted elimination of Fusobacterium nucleatum alleviates periodontitis.

Background: Fusobacterium nucleatum, a pathobiont in periodontal disease, contributes to alveolar bone destruction. We assessed the efficacy of a new targeted antimicrobial, FP-100, in eradicating F. nucleatum from the oral microbial community in vitro and in vivo and evaluated its effectiveness in reducing bone loss in a mouse periodontitis model. Methods: A multispecies bacterial community was cultured and treated with two concentrations of FP-100 over two days. Microbial profiles were examined at 24-h intervals using 16S rRNA sequencing. A ligature-induced periodontitis mouse model was employed to test FP-100 in vivo. Results: FP-100 significantly reduced Fusobacterium spp. within the in vitro community (p < 0.05) without altering microbial diversity at a 2 µM concentration. In mice, cultivable F. nucleatum was undetectable in FP-100-treated ligatures but persistent in controls. Beta diversity plots showed distinct microbial structures between treated and control mice. Alveolar bone loss was significantly reduced in the FP-100 group (p = 0.018), with concurrent decreases in gingival IL-1ß and TNF-α expression (p = 0.052 and 0.018, respectively). Conclusion: FP-100 effectively eliminates F. nucleatum from oral microbiota and significantly reduces bone loss in a mouse periodontitis model, demonstrating its potential as a targeted therapeutic agent for periodontal disease.

FP-100 eliminates F. nucleatum from an in vitro multispecies microbial community at low doses without affecting bacterial diversity. FP-100 treatment leads to the in vivo elimination of F. nucleatum, reducing alveolar bone loss and levels of pro-inflammatory cytokines in the gingiva. FP-100 is a new antimicrobial to target F. nucleatum-mediated periodontal disease.

Oral microbial dysbiosis in patients with oral cavity cancers.

OBJECTIVES: The pathogenesis of oral cavity cancers is complex. We tested the hypothesis that oral microbiota dysbiosis is associated with oral cavity cancer. MATERIALS AND METHODS: Patients with primary oral cavity cancer who met the inclusion and exclusion criteria were included in the study. Matching healthy individuals were recruited as controls. Data on socio-demographic and behavioral factors, self-reported periodontal measures and habits, and current dental status were collected using a structured questionnaire and periodontal chartings. In addition to self-reported oral health measures, each participant received a standard and detailed clinical examination. DNA was extracted from saliva samples from patients and healthy controls. Next-generation sequencing was performed by targeting V3-V4 gene regions of the 16 S rRNA with subsequent bioinformatic analyses. RESULTS: Patients with oral cavity cancers had a lower quality of oral health than healthy controls. Proteobacteria, Aggregatibacter, Haemophilus, and Neisseria decreased, while Firmicutes, Bacteroidetes, Actinobacteria, Lactobacillus, Gemella, and Fusobacteria increased in oral cancer patients. At the species level, C. durum, L. umeaens, N. subflava, A. massiliensis, and V. dispar were significantly lower, while G. haemolysans was significantly increased (p < 0.05). Major periodontopathogens associated with periodontal disease (P. gingivalis and F.nucleatum) increased 6.5- and 2.8-fold, respectively. CONCLUSION: These data suggested that patients with oral cancer had worse oral health conditions and a distinct oral microbiome composition that is affected by personal daily habits and may be associated with the pathogenicity of the disease and interspecies interactions. CLINICAL RELEVANCE: This paper demonstrates the link between oral bacteria and oral cancers, identifying mechanistic interactions between species of oral microbiome.

Assessment of Epstein-Barr virus, Candida albicans, and some periodontal pathogens in rheumatoid arthritis patients with periodontitis.

OBJECTIVE: To comparatively investigate the periodontal results and microbial load in subgingival biofilm samples (SBS) in rheumatoid arthritis subjects and healthy volunteers. METHODS: One hundred twenty subjects were classified into different cohorts: healthy (H-C); periodontitis with good systemic health (H-P); rheumatoid arthritis (RA) and good periodontal health (RA-C); and periodontitis with RA (RA-P). The periodontal parameters were recorded, and SBS were collected to determine periodontal pathogens including Epstein-Barr Virus (EBV) and Candida albicans using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Subjects that had greater disease course, determined by moderate or high disease activity scores 28 (DAS28), suffered from worse oral health conditions (higher plaque index, gingival index, bleeding on probing, probing depth, and excessive clinical attachment loss) than those with low DAS28 scores. A higher prevalence of Treponema denticola (T. denticola) was observed in the RA-P group. Cyclic citrullinated peptide was associated with the occurrence of T. denticola and Campylobacter rectus. DAS28 using C-reactive protein (DAS28-CRP) had a significant association with Capnocytophaga gingivalis and EBV. The duration of the RA disease was associated with the presence of T. denticola. CONCLUSION: Subgingival microbial difference could reliably discriminate RA from healthy individuals. Especially, T. denticola and EBV may play a key role in periodontitis associated with RA.

Oral Microbiota Signatures in the Pathogenesis of Euthyroid Hashimoto's Thyroiditis.

One of the most prevalent autoimmune illnesses in the world is Hashimoto's thyroiditis, whose pathogenesis is still unknown. The gut-thyroid axis is frequently examined, and although oral health affects thyroid functions, there are limited data on how oral microbiota is linked to Hashimoto's thyroiditis. The study aims to identify the oral microbiota from saliva samples taken from treated (with levothyroxine) and untreated female euthyroid Hashimoto's thyroiditis patients as well as healthy controls who were age- and sex-matched to compare the oral microbiota across the groups and to contribute preliminary data to the literature. This study was designed as a single-center cross-sectional observational study. Sixty (60) female patients with euthyroid Hashimoto's thyroiditis (HT) and eighteen (18) age- and gender-matched healthy controls were included in this study. Unstimulated saliva samples were collected. After DNA isolation, sequencing was performed by targeting the V3-V4 gene regions of the 16S rRNA on the MiSeq instrument. R scripts and SPSS were used for bioinformatic and statistical analysis. No significant differences were found in the diversity indices. However, Patescibacteria phylum showed a significantly higher abundance (3.59 vs. 1.12; p = 0.022) in the oral microbiota of HT patients compared to HC. In the oral microbiota, the euthyroid HT group had approximately 7, 9, and 10-fold higher levels of the Gemella, Enterococcus, and Bacillus genera levels than healthy controls, respectively. In conclusion, the results of our study demonstrated that Hashimoto's thyroiditis causes changes in the oral microbiota, whereas the medicine used to treat the condition had no such effects. Therefore, revealing the core oral microbiota and long-term follow-up of the HT process by conducting extensive and multicenter studies might provide some important data for understanding the pathogenesis of the disease.

A Longitudinal Study in Turkiye of Host Ability to Produce Antibodies following a Third Homologous BNT162b2 Vaccination.

Obesity is a multifaceted, complex condition that has negative impacts on one's health. There are conflicting reports regarding the COVID-19 vaccine's ability to induce antibody formation in obese people. Our study aimed to determine anti-S-RBD IgG and surrogate neutralizing antibody (snAb) levels before and after the third Pfizer-BioNTech (BNT162b2) vaccination (at 15, 60, 90, and 120 days) in normal-weight adults, overweight, and obese individuals without any comorbidity or previous SARS-CoV-2 infection history, but it did not evaluate the response to the first two doses. In this longitudinal prospective study in Istanbul, Turkey, a total of 323 consecutive adult individuals (141 normal weight, 108 overweight, and 74 patients with obesity) were included. Peripheral blood samples were collected. Anti-S-RBD IgG and surrogate neutralizing antibody levels were detected using the ELISA method. After the third dose of BNT162b2 vaccination, obese patients had significantly lower levels of snAb against SARS-CoV-2 compared with normal-weight controls, but the levels otherwise did not differ between the study groups. Across all individuals in our cohort, titers peaked about a month after this third vaccination and then gradually faded. Anti-S-RBD IgG and snAb IH% levels against SARS-CoV-2 were not correlated with IL-6 and TNF-α levels. In conclusion, anti-S-RBD IgG titers and snAb IH% levels against SARS-CoV-2 were determined longitudinally for 120 days after the third homologous BNT162b2 vaccination. Although there were no significant differences in anti-S-RBD IgG, we found significant differences in the snAb IH% levels against SARS-CoV-2 between obese and healthy control subjects.

Could Prior COVID-19 Affect the Neutralizing Antibody after the Third BNT162b2 Booster Dose: A Longitudinal Study.

Vaccination is an essential public health measure for preventing the spread of illness during this continuing COVID-19 epidemic. The immune response developed by the host or the continuation of the immunological response caused by vaccination is crucial since it might alter the epidemic's prognosis. In our study, we aimed to determine the titers of anti-S-RBD antibody and surrogate neutralizing antibody (snAb) formed before and after the third dose of the BNT162b2 vaccination (on the 15th, 60th, and 90th days) in healthy adults who did not have any comorbidity either with or without prior SARS-CoV-2 infection. In this longitudinal prospective study, 300 healthy persons were randomly included between January and February 2022, following two doses of BNT162b2 immunization and before a third dosage. Blood was drawn from the peripheral veins. SARS-CoV-2 NCP IgG and anti-S-RBD IgG levels were detected by the CMIA method, and a surrogate neutralizing antibody was seen by the ELISA method. Our study included 154 (51.3%) female and 146 (48.7%) male (total 300) participants. The participants' median age was 32.5 (IQR:24-38). It was discovered that 208 individuals (69.3%) had never been infected with SARS-CoV-2, whereas 92 participants (30.7%) had SARS-CoV-2 infections in the past. Anti-S-RBD IgG and nAb IH% levels increased 5.94- and 1.26-fold on day 15, 3.63- and 1.22-fold on day 60, and 2.33- and 1.26-fold on day 90 after the third BNT162b2 vaccine dosage compared to pre-vaccination values (Day 0). In addition, the decrease in anti-S-RBD IgG levels on the 60th and 90th days was significantly different in the group without prior SARS-CoV-2 infection compared to the group with past SARS-CoV-2 infection (p < 0.05). In conclusion, it was observed that prior SARS-CoV-2 infection and the third BNT162b2 vaccine dose led to a lower decrease in both nAb and anti-S-RBD IgG levels. To evaluate the vaccine's effectiveness and update immunization programs, however, it is necessary to perform multicenter, longer-term, and comprehensive investigations on healthy individuals without immune response issues, as there are still circulating variants.

Could Long Non-Coding RNA MEG3 and PTENP1 Interact with miR-21 in the Pathogenesis of Non-Alcoholic Fatty Liver Disease?

NAFLD is the most common cause of chronic liver disease worldwide. The miRNAs and lncRNAs are important endogenous ncRNAs families that can regulate molecular mechanisms. The aim of this study was to analyze the miRNA and lncRNA expression profiles in serum samples of NAFLD patients with different types of hepatosteatosis compared to healthy controls by the qPCR method. A total of180 NAFLD patients and 60 healthy controls were included. miRCURY LNA miRNA miRNome PCR human panel I + II kit and LncProfiler qPCR Array Kit were used to detect miRNA and lncRNA expression, respectively. DIANA miRPath and DIANA-lncBase web servers were used for interaction analysis. As a result, 75 miRNA and 24 lncRNA expression changes were determined. For miRNAs and lncRNAs, 30 and 5 were downregulated and 45 and 19 were upregulated, respectively. hsa-miR-21 was upregulated 2-fold whereas miR-197 was downregulated 0.25-fold. Among lncRNAs, NEAT1 was upregulated 2.9-fold while lncRNA MEG3 was downregulated 0.41-fold. A weak correlation was found between hsa-miR-122 and lncRNA MALAT1. As a conclusion, it is clear that lncRNA-miRNA interaction is involved in the molecular mechanisms of the emergence of NAFLD. The lncRNAs MEG3 and PTENP1 interacted with hsa-miR-21. It was thought that this interaction should be investigated as a biomarker for the development of NAFLD.

The link between total antioxidant status, total oxidant status, arylesterase activity, and subgingival microbiota in psoriasis patients.

BACKGROUND: Studies focusing on the relationship between periodontitis and systemic diseases have suggested a possible association between these two chronic and inflammatory disorders. We aimed to comparatively investigate the salivary oxidative status, biomarker levels, clinical findings, and the microbial load on subgingival biofilm samples in psoriasis patients and controls. METHODS: Forty participants were allocated into four groups as follows: (1) systemically and periodontally healthy (C group); (2) systemically healthy with periodontitis (P group); (3) psoriasis (Ps) and periodontally healthy (Ps-C group); and (4) Ps with periodontitis (Ps-P group). Subgingival biofilm samples were obtained to detect the periodontopathogenic agents by Real-time PCR (qPCR). The total antioxidant status (TAS) (mmol/l), total oxidant status (TOS) (µmol/l), and arylesterase (ARE) activity (U/L) were analyzed using saliva samples. RESULTS: The level of TOS and oxidative stress index (OSI) were significantly higher in patients with Ps-P and P compared to controls (P = 0.001, and P ˂ 0.001, respectively). ARE levels were higher in controls compared to Ps and P (P ˂ 0.001). The prevalences of bacteria detected in subgingival biofilm samples were similar between all groups (P > 0.05). CONCLUSIONS: This study reported that psoriasis may amplify TOS and OSI, and the co-existence of psoriasis and periodontitis may aggravate oxidative stress.

Synergistic effect of thymoquinone and nystatin in the treatment of oral candidiasis; an in vitro study.

The effectiveness of antifungal agents may be insufficient against resistant strains in some cases of oral candidiasis. The aim of this study was to evaluate the antifungal effect of thymoquinone against Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei strains and the synergistic antifungal activity of these strains in combination with nystatin. To evaluate in vitro antifungal activity and interactions between thymoquinone and nystatin, substances were tested against Candida albicans ATCC 10,231, C. tropicalis ATCC 750, C.krusei ATCC 6258 and C. glabrata ATCC 2001 standard strains both individually and combinationally via microdilution method. MIC and ΣFIC index value were analysed. The Kruskal Wallis test and Bonferroni test were used for statistical evaluations. Statistical significance was set at p < 0.05. A statistically significant difference was observed between the mean ranks of all Candida species and doses of thymoquinone, nystatin, and the combination thymoquinone-nystatin (p < 0.05). MIC values for thymoquinone were determined as 15 µg/mL for C. albicans, C. tropicalis and C. krusei while it was 30 µg/mL for C. glabrata. Moreover, MIC for nystatin was found as 1.875 µg/mL for C. albicans, C. tropicalis and C. krusei, whereas it was 7.5 µg/mL in C. glabrata. Interaction assays and ΣFIC index value revealed that, TQ and nystatin have a synergistic effect against to all strains. Thymoquinone was found to have antifungal activity on Candida species and synergistic effect when combined with nystatin.

Epidemic Klebsiella pneumoniae ST258 incidence in ICU patients admitted to a university hospital in Istanbul.

INTRODUCTION: Klebsiella pneumoniae sequence type 258 (ST258) strains are globally distributed multi-drug resistant pathogens and can spread rapidly throughout the world, causing severe healthcare-associated invasive infections with limited antimicrobial treatment options. The aim of this study was to reveal the incidence of Klebsiella pneumoniae ST258 strains among the intensive care unit patients in a university hospital in Istanbul. METHODOLOGY: Consecutive nonreplicated 83 K. pneumoniae strains were isolated from various clinical samples of intensive care unit patients admitted to a university hospital in Istanbul, between November 2016 to December 2018. Bacterial identifications were performed via VITEK2. Antimicrobial susceptibility tests were conducted with Kirby Bauer's disc diffusion test except for colistin which was performed with broth microdilution. Real-time PCR method was utilized in order to reveal ST258 positivity among the strains. RESULTS: Antimicrobial susceptibility results revealed that 56 (67%) K. pneumoniae strains were carbapenem-resistant. Real-time PCR results demonstrated that 15 out of 83 (18%) K.pneumoniae strain were ST258. According to antimicrobial susceptibility test results of ST258 strains, 8 were found as carbapenem-resistant whereas 7 were found as carbapenem susceptible. 3 out of 8 (37.5%) carbapenem-resistant ST258 strains were found as resistant against all antibiotics tested. CONCLUSIONS: Our study revealed that K. pneumoniae ST258 which caused severe infections worldwide so far has also spread to Istanbul. We believe that rapid molecular methods for monitorization of these clones are useful. our results showed that ST258 is not linked to a multi-resistant strain and suggested that it does not contribute to multi-resistance formation alone.

Investigating antimicrobial features and drug interactions of sedoanalgesics in intensive care unit: an experimental study.

Study Objective: Aim of this study was to evaluate antimicrobial effects and interaction between analgesic combinations of fentanyl citrate, dexmedetomidine hydrochloride and tramadol hydrochloride on Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa and Candida albicans which are some of the most common nosocomial infection related microorganisms. Design: In vitro prospective study. Setting: University Clinical Microbiology Laboratory. Measurements: In order to evaluate in vitro antimicrobial effects and interaction between analgesic combinations, tramadol hydrochloride, fentanyl citrate and dexmedetomidin were used against S. aureus ATCC 29213, K. pneumoniae, E. coli ATCC 25922, P. aeruginosa ATCC 27853 and C. albicans ATCC 10231 standard strains by microdilution method. Main Results: According to microdilution assays tramadol has shown the most efficient antimicrobial activity also it has been observed that 10 µg/ml concentrated dexmedetomidine has antimicrobial effects on S. aureus, K. pneumoniae and P. aeruginosa. Fentanyl has displayed evident inhibitory potency on the pathogens except for Klebsiella pneumoniae, nevertheless our predefined minimum concentration inhibited growth by 9.5 %. Fentanyl and dexmedetomidine together exhibited more antimicrobial effect on P. aeruginosa and E. coli growth. Additionally, when the three drugs examined together, microbial inhibition occurred more than expected on E. coli again and also on C. albicans growth. Conclusions: Our results revealed the antimicrobial properties and synergy with the different combinations of fentanyl, dexmedetomidine and tramadol against the most common nosocomial infection agents in the ICU. This is the first study in the literature looking into the microbial "interactions" of opioids and sedative drugs but more research is needed in order to define clinico-laboratory correlation.

Detection of carbapenem-resistant Klebsiella pneumoniae strains harboring carbapenemase, beta-lactamase and quinolone resistance genes in intensive care unit patients.

Aim: Carbapenem-resistant Klebsiella pneumoniae (CR-Kp) strains are important nosocomial pathogens worldwide. In this study, we aimed to reveal the antibiotic resistance of clinical CR-Kp strains and determine the presence of KPC, OXA-48, VIM and IMP carbapenemase genes. CTX-M-1, TEM-1, SHV-1 extended-spectrum beta-lactamase (ESBL) genes, qnrA, qnrB, qnrS plasmid-mediated quinolone resistance genes and sul1 and sul2 sulfonamide resistance genes provided molecular epidemiological data. Methods: A total of 175 K. pneumoniae strains were isolated from clinical samples of patients hospitalised in an intensive care unit (ICU) betweent April and October 2017. The strains were identified with conventional methods, with VITEK 2 (BioMerieux, France) and MALDI-TOF MS (Bruker, USA). Antimicrobial susceptibilities were tested using the disc-diffusion method and E-test (BioMerieux, France). Antimicrobial resistance genes were investigated via real-time PCR in strains identified as CR-Kp. Results: High frequencies of bla TEM-1 (86.36%), bla SHV-1 (86.36%), and bla CTX-M-1 (95.45%) genes were found in CR-Kp strains. Morever, all three ESBL genes coexisted in 77.3% of all strains. bla KPC was detected in 12 (54.55%) of the strains, and 4 of them which had an MIC> 16 µg/mL to imipenem showed bla OXA-48 positivity as well. The qnrS gene determinant (86.36%) had the highest frequency, and strains carrying qnrA showed higher MICs for ciprofloxacin. Conclusion: CR-Kp strains are able to develop different antimicrobial resistance patterns according to regional changes in antimicrobial therapeutic policies. Thus, it is important to monitor the regional molecular epidemiological data for efficient treatment.

Comparison of microbial adhesion and biofilm formation on orthodontic wax materials; an in vitro study.

BACKGROUND/PURPOSE: Orthodontic wax materials are available on the dental market and are given by orthodontists due to pain, sores and irritation caused by treatment. The aim of the study was to compare biofilm formation and microbial adhesion at different time points on different protective materials used against orthodontic wounds in vitro. MATERIALS AND METHODS: Microbial adhesion and biofilm formation were evaluated against Streptococcus mutans ATCC 25175 and Lactobacillus acidophilus ATCC 4356 standard strains on orthodontic wax materials at the 0, 24th, 48th, 72nd, 96th and 120th hour. The Kruskal Wallis test and Bonferroni test were used for statistical evaluations. Statistical significance was set at p < 0.05. RESULTS: It was observed that S. mutans formed statistically significantly more biofilm on OrthoDots®CLEAR (OrVance) than Ora-Aid (TBM Corporation) at the 48th hour (p < 0.05). Furthermore, L. acidophilus formed statistically significantly more biofilm on OrthoDots®CLEAR (OrVance) than Brace Gard®(Infa-Lab Inc.) at the 72nd, 96th and 120th hours (p < 0.05). CONCLUSION: Significant differences were noted among the different orthodontic wax materials and both S. mutans and L. acidophilus created biofilm on all waxes at different time points in vitro. To prevent biofilm formation, these waxes need to be refreshed and should not be used for more than 24 h. According to our study, biofilm production performances of pathogens on Brace Gard®(Infa-Lab Inc.) are minimal and therefore it may be a better option to use in clinics. However, to our knowledge, this is the first study investigating biofilm formation on waxes and more studies are needed in this field.

Detection of O25b-ST131 clone, CTX-M-1 and CTX-M-15 genes via real-time PCR in Escherichia coli strains in patients with UTIs obtained from a university hospital in Istanbul.

BACKGROUND: Escherichia coli sequence type 131 is an important multidrug resistant clone responsible from more than half of ESBL-producing E.coli isolates. Aim of this study was to investigate the presence of O25b-ST131 clone, CTX-M-15 and CTX-M-1 genes in the E. coli strains isolated from both hospital and community acquired UTIs by real-time PCR and to reveal molecular epidemiological data. METHODS: Non-duplicate E. coli (n = 101) strains isolated from UTI patients were included. Bacterial identifications were performed with VITEK Compact. Antimicrobial susceptibility tests, phenotypic ESBL and E-tests were performed conventionally. Real-time PCR was utilized to detect presence of O25b-ST131 clone, blaCTX-M-15 and blaCTX-M-1. RESULTS: O25b-ST131 clone, CTX-M-1 and CTX-M-15 were detected in 22%, 73%, 37% in UTIs, respectively. Presence of O25b-ST131 clones and CTX-M-1 genes among E. coli strains isolated from inpatients were found statistically higher than outpatients. The most effective choice was found to be fosfomycin and nitrofurantoin in outpatients and inpatients, respectively. The MIC90 values of Amikacin, Cefotaxime, Cefepime and Ciprofloxacin were higher in inpatients than in oupatients, whereas Cefotaxime and Ciprofloxacin MIC50 values were found to be higher in inpatients than in outpatients. The highest increase of MIC90 values was observed in O25b-ST131, CTX-M-1 and CTX-M-15 coexistence. CONCLUSION: The presence of O25b-ST131 clone, CTX-M-1 and CTX-M-15 genes in E. coli strains in patients with UTI has been revealed. In the presence of the O25b-ST131 clone, a significant increase was observed in the ciprofloxacin MIC values indicating the importance of monitorization of the clone using molecular epidemiology.

[Detection of HEV RNA amounts and genotypes in raw milks obtained from different animals]. / Farkli hayvanlardan elde edilen çig sütlerde HEV RNA miktarinin ve genotiplerinin tespiti.

Hepatitis E virus (HEV) is one of the major foodborne viral pathogens transmitted through the fecal-oral route. Four genotypes of HEV are known to infect humans and it is reported that different types of HEV are active in zoonotic transitions. It is known that the HEV genotype 1 and HEV genotype 2 infections are generally acute and the HEV genotype 3 infections are chronic. Therefore, in the studies related to HEV infections, it is important to determine the genotypes to monitortreatment regimens. Although raw milk is often used in communities due to its low cost, there are limited data on the rates and the genotypes of HEV in our country and in the world. In light of this information, we aimed to investigate epidemiologically the quantity and genotypes of HEV RNA in 231 raw milk (48 cow milk, 65 goat milk, 65 sheep milk, and 53 donkey milk) samples. Viral RNAs were isolated from raw milk samples and the ORF2 region of HEV was investigated by the qRt-PCR method to determine quantitatively the presence of HEV RNA. In addition, among HEV RNA positive samples, the ORF2 region of HEV was amplified by nested PCR and the amplicons were sequenced. HEV RNA was detected in 47 (20.34%) raw milk samples, Positivity was detected in 14 (29.16%) of cow milk, 12 (18.46%) of goat milk, 8 of sheep milk (12.3) and 13 of donkey milk (24.5%). The amount of HEV RNA in cow milk found as the highest in both proportion and quantity. When the distribution of the HEV genotypes in the 47 positive samples was examined, 27 (57.44%) HEV genotype 1a, 10 (21.27%) HEV genotype 1b, 4 (8.5%) HEV genotype 4c, 2 (4.2%) HEV genotype 3a, (2.13) HEV genotype 1c, 1 (2.13%) HEV genotype 3e, 1 (2.13%) HEV genotype 3f and 1 (2.13%) HEV genotype 3g were determined. Although genotype 1a is more frequent, it has been revealed that different genotypes encountered in our country. In conclusion, it has been determined that HEV, one of the major foodborne viral agents, may be encountered in raw milk, and the genotypes that can cause infections in human are found especially in raw milk from animal sources. For the prevention of foodborne outbreaks, the presence of HEV in raw milk should not be ignored.

A combined treatment using ethylmethane sulfonate and ultraviolet light to compare amylase production by three Bacillus sp. isolates.

In this study, three Bacillus sp.-producing amylase enzymes were isolated from soil samples and identified using 16S rDNA sequence analysis. Amylase production and total protein productions were spectrophotometrically measured. The following media were tested to increase enzyme production: LB medium and molasses. Three Bacillus sp. were identified as follows: Bacillus subtilis subtilis, Bacillus thuringiensis, and Bacillus cereus. Amylase production levels were in the range of 10 U/mL, whereas total protein production levels were at 15 mg/mL. Higher amylase activity was found in the Bacillus subtilis isolate. Ethylmethane sulfonate (EMS) and ultraviolet (UV) mutagenesis in combination were applied to compare amylase production. Amylase activity was increased to around 58% in the treatment with 0.03 mL of EMS and UV when compared to the control group. A pilot scale bioreactor with a total working volume of 10 liters was used to produce amylase by B. subtilis subtilis. In conclusion, B. subtilis subtilis can be used to produce amylase enzyme for various industrial purposes, and, for the first time, the amylase activities of B. subtilis can be enhanced with EMS and UV treatment.

Analysis of Virulence Factors and Antimicrobial Resistance in Salmonella Using Molecular Techniques and Identification of Clonal Relationships Among the Strains.

A total of 50 Salmonella enterica strains were isolated from clinical samples from 2009 to 2012 and analyzed for the presence of virulence genes found in SPI-1, SPI-2, and plasmids. The distribution and frequency of the antimicrobial resistance genes and plasmids were revealed, and pulsed-field gel electrophoresis (PFGE) patterns were investigated. Five genes were identified from the seven strains with resistance or intermediate resistance to ampicillin: blaSHV-1 (present in six strains), qnrS1 (present in five strains), blaTEM-1 (present in three strains), blaCTX-M-1 (present in one strain), and qnrB1 (present in one strain). One trimethoprim-sulfamethoxazole-resistant strain was positive for sulI but negative for sulII. In addition, we detected TEM-1 and qnrS1 in one strain; SHV-1 and qnrS1 in two strains; TEM-1, SHV-1, CTX-M-1, and qnrS1 in one strain; TEM-1, SHV-1, and qnrB1 in one strain; and SHV-1 and sulI genes in one strain together. Plasmid-based replicon typing assay revealed that all 50 strains carried FIIS, 13 carried I1, 1 carried I2, 4 carried P, 1 carried A/C, and 4 carried X1 replicon. PFGE was used to type 46 of the 50 strains and classify them into 22 major groups, 33 pulsotypes, and 8 major clusters. All strains carried all the virulence genes of interest on both Salmonella Pathogenicity Islands 1 and 2 and plasmids suggested high potential for pathogenicity. All antimicrobial-resistant strains contained at least one of the resistance genes of interest, confirming a phenotype-genotype association in antimicrobial resistance.

Adult onset tubulointerstitial nephritis and uveitis syndrome.

A Turkish woman aged 44 years who presented with a 1 month history of abdominal pain, fatigue and weight loss of 10 kg was diagnosed as having acute tubulointerstitial nephritis. Opthalmological evaluation revealed unilateral uveitis and contralateral chorioretinal scarring. X-ray films of the pelvis revealed unilateral sacroileitis. An elevated erythrocyte sedimentation rate, C-reactive protein, tubular proteinuria and renal glucosuria returned to normal 2 weeks after treatment was started. It is important to be aware of tubulointerstitial nephritis and uveitis syndrome in order to achieve a quick diagnosis in patients with renal impairment and tubular dysfunction with minor symptoms so that appropriate management can be started early.