Molecular characterization of pseudomonas aeruginosa isolated from clinical bovine mastitis cases

This study aimed to investigate the presence and distribution of virulence determinants and their antimicrobial susceptibilities of 44 Pseudomonas aeruginosa isolates obtained from clinical bovine mastitis cases. In addition, selected 6 P. aeruginosa isolates were further characterized using whole-genome sequencing (WGS). Based on the presence of T3SS-related genes, 25% of the isolates were found to carry exoU and/or exoS genes belonging to invasive (exoU-/exoS+, 18.2%) and cytotoxic (exoU+/exoS-, 6.8%) strains. But, none of the isolates carried both exoU and exoS genes. In terms of other virulence genes examined, various virulence gene profiles were observed among the isolates. The majority of the isolates (72.7%) were susceptible to all tested antimicrobials. Resistance rates to ciprofloxacin and carbapenems (imipenem and meropenem) were determined as 25% and 4.5%, respectively. WGS analysis indicated the presence of different resistome, but fosA genes in all isolates, and different combinations of mutations in gyrA, parC, oprD, efflux pump, and genes playing a role in the regulation of ampC gene expression. Different sequence types (STs) and serotypes were found in representative isolates with the occurrence of the O11-ST235 clone, which is a worldwide multidrug-resistant high-risk clone representing a serious public health threat. The findings of this study provide valuable information on P. aeruginosa isolated from clinical bovine mastitis cases and current antimicrobial resistance levels and virulence determinants

Molecular characterization of trichophyton verrucosum strains isolated from cattle by PCR-RFLP

Dermatofitler, insan ve hayvanlarda keratin içeren dokuları infekte ederek dermatofit infeksiyonuna neden olmaktadırlar. Trichophyton verrucosum sığır dermatofitozis olgularının en yaygın etkenidir. Trichophytosis, bütün dünyada hayvancılık sektöründe önemli ekonomik kayıplara neden olması yanında zoonoz olmasıyla da insan sağlığını tehdit etmektedir. Sığırlardan genellikle T. verrucosum izole edilmektedir. Sığırlar bu etkenin doğal rezervuarıdırlar. Bu çalışmanın amacı, sığırlarda hastalığa neden olan dermatofitlerinin izolasyonu ve izole edilen T. verrucosum suşlarının Internal Transcribed Spacer (ITS) bölgelerinin PCR-RFLP ile moleküler ayrımının yapılmasıdır. Bu amaçla dermatofitozisli sığırlardan 90 adet örnek alınarak kültürleri yapıldı. Bu örneklerin kültürü sonucunda 35 (%38,8) adet T. verrucosum izole ve identifiye edildi. Bu suşların DNA izolasyonu gerçekleştirilerek ITS bölgelerin amplifikasyonu gerçekleştirildi. T. verrucosum suşlarının MvaI ve HinfI enzimleri kullanılarak yapılan Restriction Fragment Lenght Polymorphism (RFLP) analizleri sonucunda bir adet RFLP profiline rastlandı. Sonuç olarak, izole edilen T. verrucosum suşlarının PCR-RFLP sonucunda tek bir profile sahip olduğu, farklı profil örneklerinin saptanması için farklı bölgelerden hatta farklı ülkelerden suşların PCR-RFLP’lerinin yapılması gerektiği kanısına varıldı.Dermatophytes infect tissues containing keratin in humans and animals, causing dermatophytosis infection. Trichophyton verrucosum is the most common agent of bovine dermatophytosis cases. Trichophytosis causes big economic lossess throughout the world and also threatens human health by being a zoonosis. T. verrucosum is usually isolated from cattle. Cattle are the natural reservoirs of this agent. The aim of this study is to isolate disease-causing dermatophytes in cattle and to carry out molecular separation of Internal Transcribed Spacer (ITS) regions of the isolated T. verrucosum strains by PCR- Restriction Fragment Lenght Polymorphism (PCR-RFLP). For this purpose, 90 samples were taken from the cattle with dermatophytosis for cultural examination. As a result of the culture of these samples, 35 (38.8%) T. verrucosum were isolated and identified. DNA isolation of these strains was made and amplification of ITS regions was performed. It was only one RFLP profile was found according to the results of RFLP analysis of T. verrucosum strains using MvaI and HinfI enzymes. At the end of study, it was founded that the isolated T. verrucosum strains showed a single profile by PCR-RFLP analysis and PCR-RFLP was a useful tool for the molecular characterization of the strains. İt was also concluded that PCR-RFLPs of strains from different regions or even from different countries might be necessary in order to detect different profiles of the tested samples

Comparing restrictive versus liberal oxygen strategies for trauma patients-the TRAUMOX2 trial:protocol for a randomised clinical trial

INTRODUCTION: Supplemental oxygen is commonly used in trauma patients, although it may lead to hyperoxaemia that has been associated with pulmonary complications and increased mortality. The primary objective of this trial, TRAUMOX2, is to compare a restrictive versus liberal oxygen strategy the first 8 hours following trauma. METHODS AND ANALYSIS: TRAUMOX2 is an investigator-initiated, international, parallel-grouped, superiority, outcome assessor-blinded and analyst-blinded, randomised, controlled, clinical trial. Adult patients with suspected major trauma are randomised to eight hours of a restrictive or liberal oxygen strategy. The restrictive group receives the lowest dosage of oxygen (>21%) that ensures an SpO(2) of 94%. The liberal group receives 12–15 L O(2)/min or FiO(2)=0.6–1.0. The primary outcome is a composite of 30-day mortality and/or development of major respiratory complications (pneumonia and/or acute respiratory distress syndrome). With 710 participants in each arm, we will be able to detect a 33% risk reduction with a restrictive oxygen strategy if the incidence of our primary outcome is 15% in the liberal group. ETHICS AND DISSEMINATION: TRAUMOX2 is carried out in accordance with the Helsinki II Declaration. It has been approved by the Danish Committee on Health Research Ethics for the Capital Region (H-21018062) and The Danish Medicines Agency, as well as the Dutch Medical Research Ethics Committee Erasmus MS (NL79921.078.21 and MEC-2021-0932). A website (www.traumox2.org) is available for updates and study results will be published in an international peer-reviewed scientific journal. TRIAL REGISTRATION NUMBERS: EudraCT 2021-000556-19; NCT05146700

Difficult tracheal intubation in neonates and infants. NEonate and Children audiT of Anaesthesia pRactice IN Europe (NECTARINE): a prospective European multicentre observational study

BACKGROUND: Neonates and infants are susceptible to hypoxaemia in the perioperative period. The aim of this study was to analyse interventions related to anaesthesia tracheal intubations in this European cohort and identify their clinical consequences. METHODS: We performed a secondary analysis of tracheal intubations of the European multicentre observational trial (NEonate and Children audiT of Anaesthesia pRactice IN Europe [NECTARINE]) in neonates and small infants with difficult tracheal intubation. The primary endpoint was the incidence of difficult intubation and the related complications. The secondary endpoints were the risk factors for severe hypoxaemia attributed to difficult airway management, and 30 and 90 day outcomes. RESULTS: Tracheal intubation was planned in 4683 procedures. Difficult tracheal intubation, defined as two failed attempts of direct laryngoscopy, occurred in 266 children (271 procedures) with an incidence (95% confidence interval [CI]) of 5.8% (95% CI, 5.1–6.5). Bradycardia occurred in 8% of the cases with difficult intubation, whereas a significant decrease in oxygen saturation (SpO2<90% for 60 s) was reported in 40%. No associated risk factors could be identified among co-morbidities, surgical, or anaesthesia management. Using propensity scoring to adjust for confounders, difficult anaesthesia tracheal intubation did not lead to an increase in 30 and 90 day morbidity or mortality. CONCLUSIONS: The results of the present study demonstrate a high incidence of difficult tracheal intubation in children less than 60 weeks post-conceptual age commonly resulting in severe hypoxaemia. Reassuringly, the morbidity and mortality at 30 and 90 days was not increased by the occurrence of a difficult intubation event

Peri-operative red blood cell transfusion in neonates and infants: NEonate and Children audiT of Anaesthesia pRactice IN Europe: A prospective European multicentre observational study

BACKGROUND: Little is known about current clinical practice concerning peri-operative red blood cell transfusion in neonates and small infants. Guidelines suggest transfusions based on haemoglobin thresholds ranging from 8.5 to 12 g dl-1, distinguishing between children from birth to day 7 (week 1), from day 8 to day 14 (week 2) or from day 15 (≥week 3) onwards. OBJECTIVE: To observe peri-operative red blood cell transfusion practice according to guidelines in relation to patient outcome. DESIGN: A multicentre observational study. SETTING: The NEonate-Children sTudy of Anaesthesia pRactice IN Europe (NECTARINE) trial recruited patients up to 60 weeks' postmenstrual age undergoing anaesthesia for surgical or diagnostic procedures from 165 centres in 31 European countries between March 2016 and January 2017. PATIENTS: The data included 5609 patients undergoing 6542 procedures. Inclusion criteria was a peri-operative red blood cell transfusion. MAIN OUTCOME MEASURES: The primary endpoint was the haemoglobin level triggering a transfusion for neonates in week 1, week 2 and week 3. Secondary endpoints were transfusion volumes, 'delta haemoglobin' (preprocedure - transfusion-triggering) and 30-day and 90-day morbidity and mortality. RESULTS: Peri-operative red blood cell transfusions were recorded during 447 procedures (6.9%). The median haemoglobin levels triggering a transfusion were 9.6 [IQR 8.7 to 10.9] g dl-1 for neonates in week 1, 9.6 [7.7 to 10.4] g dl-1 in week 2 and 8.0 [7.3 to 9.0] g dl-1 in week 3. The median transfusion volume was 17.1 [11.1 to 26.4] ml kg-1 with a median delta haemoglobin of 1.8 [0.0 to 3.6] g dl-1. Thirty-day morbidity was 47.8% with an overall mortality of 11.3%. CONCLUSIONS: Results indicate lower transfusion-triggering haemoglobin thresholds in clinical practice than suggested by current guidelines. The high morbidity and mortality of this NECTARINE sub-cohort calls for investigative action and evidence-based guidelines addressing peri-operative red blood cell transfusions strategies. TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT02350348

A combination of osmotic shock and ultrasound pre-treatments and the use of enzyme for extraction of proteins from Chlorella vulgaris microalgae: Optimization of extraction conditions by RSM

In this study, the effect of a combination of osmotic shock, the use of carbohydrase enzyme and ultrasound waves was investigated on protein content (PC), total phenolic content (TPC) and antioxidant activity (AOA) by CUPRAC and DPPH methods of protein extracts from Chlorella vulgaris green microalgae. The extraction parameters (enzyme/substrate ratio (E/S), ultrasound probe time and extraction time) were optimized by using Response Surface Methodology (RSM) based on Box-Behnken design (BBD). According to experimental results, PC, TPC and AOA by CUPRAC method of the extracts ranged from 95.80 to 263.08 mg protein/g dw, 9.68–33.06 mg GAE/g dw and 17.69–34.52 mg TE/g dw, respectively. Water absorption capacity, oil aborption capacity, emulsion activity, emulsion stability, foaming capacity and foaming stability of the proteins extracted under optimum extraction conditions (E/S of 0.87, ultrasound probe time of 119.30 s and extration time of 2.74 h) from C. vulgaris were determined as 12.19 ± 3.84%, 455.44 ± 5.30%, 208.11 ± 0.22%, 73.10 ± 4.68%, 49.17 ± 5.89% and 26.36 ± 1.29%, respectively. All these suggest that protein extracts obtained from C. vulgaris might be a good candidate to improve foods in terms of techno-functional properties

Investigation of rheological and bioactive properties of dough and bread enriched with protein extracts from Spirulina platensis

Spirulina platensis, simbiyotik, çok hücreli ve ipliksi yapıda mavi-yeşil bir mikroalgdir. Bu tez çalışması kapsamında, Spirulina platensis tozu (SP) ve Spirulina platensis'den ultrases destekli ekstraksiyon ile ekstrakte edilen protein ekstraktları (SPE) ile zenginleştirilen buğday unlarının reolojik ve teknofonksiyonel özellikleri araştırılmıştır. Daha sonra, bu unlar ile üretilen ekmek örneklerinin tekstürel, duyusal ve renk özellikleri ile in vitro mide-bağırsak sindirim sırasında toplam fenolik madde içeriği, antioksidan aktivite ve % protein sindirilebilirliğindeki değişim belirlenmiştir. Çalışmanın sonuçlarına göre, farklı oranlarda SP ve SPE ile zenginleştirilen buğday ununun ekstensograf ve farinograf analizleri sonucunda en uygun SP ve SPE oranları %0,125, 0,25 ve 0,5 olarak belirlenmiştir. Örneklerin in vitro sindirim öncesi toplam fenolik madde içeriği 26,06±1,29 ile 31,69±0,09 mg GAE/100 g arasındadır. Örneklerin in vitro sindirim öncesi antioksidan aktivitesi (CUPRAC) 36,17±2,19 ile 51,78±7,95 mg TE/100 g arasında değişmiştir (p≥ 0,05). SP-%0,125, SP-%0,5 ve SPE-%0,25'in antioksidan aktivitesi, in vitro mide sindirimden sonra artış göstermiştir (p<0,05). Çalışmamızda örneklerin % protein sindirilebilirliği ise %81,86±2,07- %85,02±2,58 arasında değişmiştir (p≥0,05). Ekmek örneklerinin sıkılık değeri en yüksek olan örnek kontrol örneği (919,4±34,7 g, p< 0,05)'dir. Benzer şekilde, SP ve SPE ilaveli ekmeklerin kontrol örneklerine göre daha yüksek hacim değerlerine sahip olduğu tespit edilmiştir (p<0,05). Renk analizi sonuçları incelendiğinde, örneklerin L*, a* ve b* değerlerinin SP ve SPE ilavesi ile farklılaştığı görülmüştür (p<0,05). Duyusal analiz sonuçlarına göre, en tercih edilebilir ekmek örneği SPE-%0,50'dir.Spirulina platensis is a simbiotic, multicellular and filamentous blue-green microalgae. In context of this thesis, enrichment of wheat flour with Spirulina platensis powder (SP) and protein extracts from Spirulina platensis by ultrasound-assisted extraction (SPE) and investigation of rheological nad technofunctional properties of the flours were carried out. Moreover, texturel, sensory and colour properties of the enriched bread samples and changes in the total phenolic content, antioxidant activity and in vitro protein digestibility (%)during in vitro gastro-intestinal digestion were investigated. According to the results of exensograph and farinograph analysis of the enriched flours with SP and SPE, the best ratios for SP and SPE were determined as 0.125%, 0.25% and 0.5%. Total phenolic content and antioxidant activity by CUPRAC method of the samples were 26.06±1.29-31.69±0.09 mg GAE/100 g and 36.17±2.19-51.78±7.95 mg TE/100 g dry weight, before in vitro digestion, respectively (p≥0.05). Antioxidant activity of the samples (SP-%0.125, SP-%0.5 ve SPE-%0.25) increased after in vitro gastric digestion (p<0.05). The protein digestibility (%) of the breads ranged from 81.86±2.07% to 85.02±2.58% (p≥0.05). The firmness value of the control sample was the highest (919.4±34.7 g, p<0.05). The breads with SP and SPE were different from the others in terms of L*, a* and b* values (p<0.05). The L*, values ranged from 52.2 to 60.8 (p<0.05) and it decreased with addition of SP and SPE. According to the results of sensory analysis, the most preferable bread sample was SPE-0.50%

Response surface optimization of ultrasound-assisted protein extraction from Spirulina platensis: investigation of the effect of extraction conditions on techno-functional properties of protein concentrates

Yücetepe, Aysun ( Aksaray, Yazar )In the present study, optimum extraction conditions of proteins from Spirulina platensis were determined by using response surface methodology. Box–Behnken design of experiments was performed to investigate the effects of temperature (25, 35, 45 °C), pH (7, 8, 9) and time (60, 90, 120 min) on water absorption capacity (WAC), oil absorption capacity (OAC), emulsifying activity (EA), emulsifying stability (ES), foaming capacity (FC) and foaming stability (FS) of the protein concentrates (PC). WAC of the PC varied with time while OAC changed with temperature and pH. The PC had high values of OAC (10.22 g of oil/g of protein) and WAC (5.12 g of water/g of protein) under extraction conditions of pH of 8 temperature of 45 °C and time of 60 min. On the contrary, there were no significant changes in emulsifying properties of PC under different extraction conditions (p > 0.05), while foaming stability of the PC varied with the interactive effect of temperature and time. The optimum extraction conditions were 43.87 °C for temperature, 7.16 for pH and 60 min for time. Under these conditions, WAC, OAC, FC, FS, EA and ES values were obtained as 4.41 g of water/g, 10.13 g of oil/g, 300%, 83.94%, 42.99% and 92.50%, respectively. The results obtained through this investigation indicated that protein concentrates from Spirulina platensis have higher functional properties than other algae. Therefore, Spirulina platensis proteins can be thought as an inexpensive source of protein for food industry due to its extraordinary functionality...

Antioxidant activity and techno-functional properties of protein extracts from Caulerpa prolifera: optimization of enzyme-assisted extraction by response surface methodology

In this study, ultrasound-assisted enzymatic extraction was performed to extract proteins from Caulerpa prolifera and the techno-functional properties of the protein extracts were investigated for the first time. The effect of extraction temperature (30–40 °C), extraction time (60–120 min) and substrate/enzyme (S/E) ratio (5–15) on protein content (PC), total phenolics content (TPC) and antioxidant activity (AOA) was investigated using response surface metho-dology (RSM). According to the results, optimum conditions were determined as extraction temperature of 30 °C, extraction time of 60.03 min and S/E of 5. The values of PC, TPC, AOACUPRAC (AOA determined by cupric reducing antioxidant capacity assay) and AOADPPH (AOA determined by 2,2-diphenyl-1-picrylhydrazyl assay) of the extracts obtained under optimum conditions were 51.57 g·kg-1 dry weight (dw), 31.12 g·kg-1 dw (expressed as gallic acid equiva-lents), 4.45 g·kg-1 dw and 0.83 g·kg-1 dw (expressed as Trolox equivalents), respectively. Water and oil absorption capac-ity, emulsion activity-stability and foaming capacity-stability of the proteins extracted under optimum conditions were determined as 54.0 ± 5.0 %, 210.0 ± 15.0 %, 41.4 ± 2.3 %, 41.7 ± 0.1 %, 26.5 ± 1.5 % and 26.7 ± 2.4 %, respectively. These research findings indicate that C. prolifera proteins, as an alternative protein source, offer promising opportuni-ties for functional food applications