A solid-liquid microextraction approach from a spot in an adsorbent layer of a chromatographic plate.

A new approach to extracting substances from a spot on a chromatographic plate for subsequent liquid chromatography-mass spectrometry analysis is described. This method involves extraction in a solid phase (an adsorbent layer of a chromatographic plate) - a liquid system using a simple device. For a single extraction of six selected coccidiostats from the adsorbent layer on the chromatographic plate with silica gel, 50 µL of methanol was used for 5 min. The data from the extraction experiments and liquid chromatography-mass spectrometry measurements demonstrated a good correlation between the ratio of the peak areas of the coccidiostats to the internal standard and the concentration of the substances in the range of two orders of magnitude. The coefficients of determination for the mentioned correlations range from 0.962 to 0.999. Moreover, the repeatability and reproducibility, expressed as the percentage values of relative standard deviation, do not exceed 7.5 % for any of the coccidiostats.

The relationship between fear of COVID-19 infection, fear of COVID-19 vaccination and Big Five personality traits: a mediation model.

BACKGROUND: The primary objective of our correlational, cross-sectional study was to determine the relationship between fear of COVID-19 infection and fear of COVID-19 vaccination and the role of the Big Five personality traits in this relationship. PARTICIPANTS AND PROCEDURE: The study sample consisted of 462 participants, including 286 women and 176 men, aged 18 to 75, recruited from the general population. The Big Five personality traits were diagnosed using the Polish version of the Ten-Item Personality Inventory. We used an 11-point numerical rating scale to measure the intensity of fear of COVID-19 infection as well as fear of COVID-19 vaccination. RESULTS: The results indicate that fear of infection has a weak positive correlation with fear of vaccination. Both types of fear have a weak negative correlation with emotional stability and openness to experience. People diagnosed with COVID-19 have significantly higher levels of fear of vaccination and lower levels of emotional stability compared to those without a COVID-19 diagnosis. Fear of infection proves a good predictor of fear of vaccination and a likely mediator between emotional stability and fear of vaccination against COVID-19. CONCLUSIONS: The results may increase our understanding of the role of personality traits, such as emotional stability, in the fear of infection and COVID-19 vaccination hesitancy.

Misophonia in Children and Adolescents: Age Differences, Risk Factors, Psychiatric and Psychological Correlates. A Pilot Study with Mothers' Involvement.

Misophonia is a type of disorder characterized by decreased sound tolerance. While it typically begins in childhood, research on its characteristics in this population is limited. We assessed 90 children aged 7-18 with and without misophonia, along with their mothers, using interviews, questionnaires, and performance-based tests. Younger children with misophonia were more likely to use aggression in response to triggers than older, while adolescents largely reported self-harm during triggers. Children with misophonia did not differ from their peers in terms of ADHD, ODD, ASD, dyslexia, social and emotional competencies, head injuries, epilepsy, tinnitus, being prematurely born, or delivered via cesarean sections. However, they had significantly higher symptoms of anxiety and depression, more frequent occurrences of OCD, migraines, and psychosomatic complaints. Their mothers self-reported postpartum depression significantly more frequently than mothers in the control group. There is a need for further research on pediatric misophonia, with the involvement and assessment of parents.

Comparison of the Determination of Some Antihypertensive Drugs in Clinical Human Plasma Samples by Solvent Front Position Extraction and Precipitation Modes.

The determination of the selected antihypertensive drugs in human plasma samples with the novel solvent front position extraction (SFPE) technique is presented. The SFPE procedure combined with LC-MS/MS analysis was used for the first time to prepare a clinical sample containing the drugs mentioned above from different therapeutic groups. The effectiveness of our approach was compared with the precipitation method. The latter technique is usually used to prepare biological samples in routine laboratories. During the experiments, the substances of interest and the internal standard were separated from other matrix components using a prototype horizontal chamber for thin-layer chromatography/high-performance thin-layer chromatography (TLC/HPTLC) with a moving pipette powered by a 3D mechanism, which distributed the solvent on the adsorbent layer. Detection of the six antihypertensive drugs was performed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring (MRM) mode. Results obtained by SFPE were very satisfactory (linearity R2 ≥ 0.981; %RSD ≤ 6%; LOD and LOQ were in the range of 0.06-9.78 ng/mL and 0.17-29.64 ng/mL, respectively). The recovery was in the range of 79.88-120.36%. Intra-day and inter-day precision had a percentage coefficient of variation (CV) in the range of 1.10-9.74%. The procedure is simple and highly effective. It includes the automation of TLC chromatogram development, which significantly reduced the number of manual operations performed, the time of sample preparation and solvent consumption.

Shiga Toxin-Producing Escherichia coli in Faecal Samples from Wild Ruminants.

Wildlife can harbour Shiga toxin-producing Escherichia coli (STEC). In the present study, STEC in faecal samples from red deer (n = 106) and roe deer (n = 95) were characterised. All isolates were non-O157 strains. In red deer, STEC were detected in 17.9% (n = 19) of the isolates, and the eae/stx2b virulence profile was detected in two isolates (10.5%). One STEC strain harboured stx1a (5.3%) and eighteen STEC strains harboured stx2 (94.7%). The most prevalent stx2 subtypes were stx2b (n = 12; 66.7%), stx2a (n = 3; 16.7%), and stx2g (n = 2; 11.1%). One isolate could not be subtyped (NS) with the applied primers (5.6%). The most widely identified serotypes were O146:H28 (n = 4; 21%), O146:HNM (n = 2; 10.5%), O103:H7 (n = 1; 5.3%), O103:H21 (n = 1; 5.3%), and O45:HNM (n = 1; 5.3%). In roe deer, STEC were detected in 16.8% (n = 16) of the isolates, and the eae/stx2b virulence profile was detected in one isolate (6.3%). Two STEC strains harboured stx1a (12.5%), one strain harboured stx1NS/stx2b (6.3%), and thirteen strains harboured stx2 (81.3%). The most common subtypes were stx2b (n = 8; 61.5%), stx2g (n = 2; 15.4%), non-typeable subtypes (NS) (n = 2; 15.4%), and stx2a (n = 1; 7.7%). Serotype O146:H28 (n = 5; 31.3%) was identified. The study demonstrated that the zoonotic potential of STEC strains isolated from wildlife faeces should be monitored in the context of the 'One Health' approach which links human health with animal and environmental health.

Combined Effect of Climate and Anthropopressure on River Water Quality.

This study was a continuation of our investigation of the spatio-temporal variability of the Bzura River's water chemistry. Our research is of particular importance in the context of the recent ecological disaster on the Oder River and concerns the international problem of surface water contamination. The study area was a 120 km section of the Bzura River. We tested more measurement points and with a higher sampling frequency than those used in the national monitoring of river water quality. During two hydrological years, 360 water samples were collected. The selected parameters: electrical conductivity, temperature, dissolved oxygen, dissolved organic carbon, nitrates, phosphates, bicarbonates, chlorides, sodium, potassium, calcium, and magnesium were determined. Numerous results exceeded the Polish threshold limits. Spatio-temporal variability and water quality were assessed using principal component analysis (PCA), cluster analysis (CA), and water quality index (WQI) approaches. Many point sources of pollution related to urbanization, agriculture, and industry were detected. Moreover, due to the changing climatic conditions, a significant difference between temporal variability in both years was observed. Our results indicated that it is necessary to increase the number of measurement stations for surface water monitoring; it will allow for a faster detection of the threat.

Solvent Front Position Extraction and some conventional sample preparation techniques for the determination of coccidiostats in poultry feed by LC-MS/MS.

Solvent Front Position Extraction is a novel technique developed for effective sample preparation of biological samples containing coccidiostats prior to LC-MS/MS. In this study the technique was used for isolation and determination of seven coccidiostats, from both main groups being: ionophores and chemical coccidiostats. Its effectiveness was evaluated by comparing with other sample preparation procedures, used in European routine laboratories. Results obtained by Solvent Front Position Extraction were very satisfactory (linearity R2 ≥ 0.971, recovery 90.1-111.1%, RSDr: 8.7-16.6%, RSDR: 9.0-17.7%) and fulfilled requirements described in Commission Regulation (EU) 2021/808 of 22 March 2021, which showed great potential of the technique in sample preparation of coccidiostats in poultry feed.

Seroprevalence and Molecular Epidemiology of Aleutian Disease in Various Countries during 1972-2021: A Review and Meta-Analysis.

Aleutian disease (AD) poses a serious threat to both free-ranging and farmed mink around the world. The disease is caused by the Aleutian mink disease virus (AMDV), which also poses a health risk for other members of the family Mustelidae, including wild mink, weasels, badgers and other animal species. This article analyses the seroprevalence of AMDV infections in mink and other species around the world, and reviews recent knowledge relating to the molecular epidemiology of the AMDV. Depending on the applied diagnostic technique and the country, the prevalence of anti-AMDV antibodies or AMDV DNA was established at 21.60-100.00% in farmed American mink, 0.00-93.30% in free-ranging American mink and 0.00-25.00% in European mink. Anti-AMDV antibodies or AMDV DNA were also detected in other free-living fur-bearing animals in Europe and Canada, where their prevalence was determined at 0.00-32.00% and 0.00-70.50%, respectively. This may indicate a potential threat to various animal species. AMDV strains are not clustered into genotypes based on the geographic origin, year of isolation or pathogenicity. The isolates that were identified on mink farms around the world originated from North America because American mink were introduced to Europe and Asia for breeding purposes and to restock natural populations.

Preparation of Antihypertensive Drugs in Biological Matrix with Solvent Front Position Extraction for LC-MS/MS Analysis.

Solvent front position extraction procedure was used to prepare biological samples containing selected antihypertensive drugs (ramipril, lercanidipine, indapamide, valsartan, hydrochlorothiazide, perindopril, and nebivolol). Substances separated from the biological matrix components (bovine serum albumin) were quantified by means of liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Sample preparation process was performed with the use of a prototype horizontal chamber with a moving pipette driven by a 3D printer mechanism enabling a controlled eluent flow velocity. Application of this device was advantageous for simultaneous preparation of several samples for further quantitative analysis, with a synchronized reduction of manual operations and solvent consumption. Quantitative results obtained for the majority of the investigated antihypertensive drugs in a complex biological matrix were satisfactory. The values of the %RSD were around 5% for six of the seven substances (with the exception of indapamide). The method exhibits a suitable accuracy (the relative error percentage was below 10% for most drugs). The values of LOD and LOQ were in the range of 1.19 µg/L-8.53 µg/L and 3.61 µg/L-25.8 µg/L, respectively.

Optimization of Adsorbent Layer Type and Developing Solvent in Coccidiostats Sample Preparation with Procedure of Solvent Front Position Extraction.

Coccidiostats are drugs used against coccidiosis, a common disease among breeding animals. Their widespread application leads to the appearance of their residues in food, which is potentially harmful for human health and life. The European Union has established limits of concentrations of these drugs in premixtures and food. Nowadays, there are many methods for monitoring coccidiostats' presence in market products, but their frequent weakness is sample preparation. Solvent Front Position Extraction is a planar chromatography-based sample preparation method that allows for effective assay of samples with coccidiostats when coupled with LC-MS/MS. The purpose of this research was to find common conditions for the effective isolation of eight coccidiostats from biological sample components with both lower and higher retention than the substances of interest. The acquired results were used for effective isolation of monensin and salinomycin from the premixture samples and allowed for their quantitative determination. The application of a semi-automatic device for the development of chromatograms positively impacted the results, confirming the effectiveness of the method for determining coccidiostats in biological samples.

Comparison of the Retention and Separation Selectivity of Aromatic Hydrocarbons with Polar Groups in RP-HPLC Systems with Different Stationary Phases and Eluents.

In this manuscript, the retention of aromatic hydrocarbons with polar groups has been compared for systems with various nonpolar columns of the types from C3 to C18 and different mobile phases composed of methanol, acetonitrile, or tetrahydrofuran as modifiers. The selectivity separation of the solutes in systems with different adsorbents, when one eluent modifier is swapped by another, has been explained, taking into account molecular interactions of the solutes with components of the stationary phase region (i.e., extracted modifier depending on the chain length of the stationary phase).

The Prevalence of Salmonella spp. in Two Arctic Fox (Alopex lagopus) Farms in Poland.

The objective of the study was to determine the occurrence of Salmonella spp. infections in two Arctic fox (Alopex lagopus) farms in Poland, and to analyse the correlations between animals that tested positive for Salmonella spp and breeding results. Faecal samples were taken from 1094 clinically healthy blue foxes from the basic stock of farms A and B. Salmonella spp. were detected in 18.06% (56/310) of the samples collected in farm A and in 15.94% (125/784) of the samples collected in farm B. All isolated strains belonged to S. enterica subsp. enterica serotypes Salmonella Saintpaul (S. Saintpaul), Salmonella Reading (S. Reading), and Salmonella Heidelberg (S. Heidelberg). All three serotypes are typically isolated from commercial poultry flocks. Salmonella spp. infections significantly increased the risk of female infertility, but further research is needed to confirm the results. This is the first report on the prevalence of S. Heidelberg, S. Saintpaul, and S. Reading in faecal samples collected from Arctic fox (Alopex lagopus) farms in Poland.

Comprehensive Evaluation of Metal Pollution in Urban Soils of a Post-Industrial City-A Case of Lódz, Poland.

The pollution of urban soils by metals is a global problem. Prolonged exposure of habitants who are in contact with metals retained in soil poses a health risk. This particularly applies to industrialized cities with developed transport networks. The aim of the study was to determine the content and spatial distribution of mobile metal fractions in soils of the city of Lódz and to identify their load and sources. Multivariate statistical analysis (principal component analysis (PCA), cluster analysis (CA)), combined with GIS, were used to make a comprehensive evaluation of the soil contamination. Hot-spots and differences between urban and suburban areas were also investigated. Metals were determined by atomic absorption spectrometry (AAS) after soil extraction with 1 mol L-1 HCl. In most sites, the metal content changes in the following order: Zn > Pb > Cu > Ni > Cd. About one-third of the samples are considerably (or very highly) contaminated, (contamination factor, CF > 3) with Cu, Pb, or Zn. In almost 40% of the samples, contaminated soils were found (pollution load index, PLI > 1). All metals have a strong influence on the first principal component (PC1), whereas second principal component (PC2) is related to pH. Polluted soils are located in the downtown, in the south and east part of the city. The distribution of contamination coincides with the urban layout, low emission sources and former industrial areas of Lódz.

Solvent front position extraction with semi-automatic device as a powerful sample preparation procedure to quantitatitation of tryptophan in human plasma.

In the paper the results of the tryptophan determination in human plasma samples prepared with the novel Solvent Front Position Extraction (SFPE) technique are presented. The SFPE procedure is used for preparation of real biological sample for the first time. The results obtained using SFPE are compared with those using the classical sample preparation procedure. Under the optimal conditions, tryptophan and its internal standard were separated from other plasma compounds (matrix) as a small common zone/spot on a chromatographic plate using semiautomatic device equipped with moving pipet, which distributed developing solvent on the adsorbent layer. Tryptophan and the internal standard were evenly distributed within the small common zone from that the both substances were extracted and the solution obtained was transferred to quantitation with LC-MS and MS techniques. The determination results are satisfactory, the percentage values of relative error and RSD relative standard deviation do not exceed 5%. The procedure is characterized by simplicity, high analysis throughput, very good sample purification and seems to be easy applicable to other biological samples with these advantages mentioned.

Retardation of some drugs in thin-layer chromatographic systems with impregnated silica gel plates with hen's egg white and bovine serum albumin.

The influence of impregnation the chromatographic plate adsorbent layer, silica, with hen's egg white albumin (OVA) or bovine serum albumin (BSA) on the retention of some popular medicines (paracetamol, aminophenazone, theophylline, caffeine, acetanilide, ciprofloxacin, tramadol, acetylsalicylic acid, acebutolol) is investigated. The effect of composition and buffer pH of the mobile phase on solute separation selectivity is also studied. The chromatographic systems with and without above mentioned albumins and their influence on investigated drug retention are compared. In general, it has been turned out that retention of tested medicines in systems with the sorbent impregnated with albumin significantly increase relative to those with non-impregnated.

Fallow Deer (Dama dama) as a Reservoir of Shiga Toxin-Producing Escherichia coli (STEC).

Shiga toxin-producing Escherichia (E.) coli (STEC) are responsible for the outbreaks of serious diseases in humans. Only a few reports on fallow deer as a reservoir of foodborne pathogens have been published to date. The purpose of this study was to determine the occurrence of STEC strains in the fallow deer population in Poland. In all, 94 fallow deer swabs were tested. Polymerase chain reaction (PCR) was performed to detect the virulence profile of stx1, stx2 and eae or aggR genes, to identify the subtypes of stx1 and stx2 genes and to perform O and H serotyping. STEC and attaching and effacing (AE)-STEC were identified in 13 isolates (13.83%). The most hazardous virulence profile was detected in three strains, namely stx2d serotype O103:HNM, eae/stx1a serotype O26:HNM and eae/stx1a serotype O157:H7. The predominant stx gene was stx2, which was identified in 76.92% of isolates. E. coli O157 was detected in 4/94 (4.26%). Other E. coli serogroups, O26, O103, O111 and O145, were identified in 14/94 fallow deer (14.89%). The present findings suggest that fallow deer are carriers of STEC/AE-STEC that are potentially pathogenic to humans.

The Prevalence of Yersinia enterocolitica and Yersinia pseudotuberculosis in Small Wild Rodents in Poland.

Rodents are a large group of mammals that can be carriers of zoonotic pathogens such as Yersinia strains that cause yersiniosis. The prevalence of Yersinia enterocolitica and Yersinia pseudotuberculosis was determined in 214 small wild rodents from south-eastern Poland. Samples were analyzed by precultivation and PCR. Nine (4.2%) Y. enterocolitica and one (0.5%) Y. pseudotuberculosis isolates were received. Most of them (n = 5) were obtained from the common vole (Microtus arvalis). All Y. enterocolitica strains were classified as biotype (BT) 1A. A PCR analysis of virulence markers revealed that all Y. enterocolitica isolates contained the ystB gene and five isolates harbored a rare genetic combination of ail/ystB. Three of the four ail/ystB-positive isolates belonged to serotype O:5.27. The Y. pseudotuberculosis inv-positive isolate was classified as BT 1. A genetic analysis of Y. enterocolitica harboring the ystB gene revealed 100% similarity between the analyzed sequences and the sequences from diarrhea patients in India and the United Kingdom as well as high similarity with the sequences from different species of wild animals from Poland. The Y. pseudotuberculosis inv sequence was 100% identical to the sequence isolated from fully virulent clinical strain from France and Australia. The results of our study suggest that small wild rodents, especially voles and yellow-necked mice, may act as carriers of Yersinia strains. The high similarity of the tested gene sequences between our isolates and the isolates from other free-living animals indicates that small wild rodents can play a role in the epidemiology of yersiniosis and can shed Yersinia spp. into the environment.

Optimization of the procedure of solvent front position extraction for preparation of multi-component sample for instrumental analysis.

Solvent Front Position Extraction (SFPE) procedure has been recently introduced as a novel concept for multi-component sample preparation. According to the procedure, thin-layer chromatography (TLC) is used to separate the compounds of interest from matrix components, and to focus them into a common zone from which the compounds are extracted and transferred to apparatus for instrumental analysis. In the paper, we investigate different adsorbent types of the chromatographic plates and various mobile phases, including pH of their buffers, in respect of optimization conditions of the SFPE procedure. The research was carried out using a test sample containing 9 compounds characterised by different chemical properties, hence the conclusions from the obtained results can be applied to other multi-component samples. Under the optimal conditions, all target compounds are separated from other compounds (matrix), and evenly distributed along a narrow strip, which is advantageous for their quantitation. The determination results are good, the percentage values of relative error and relative standard deviation do not exceed 6%.

Shiga toxin-producing Escherichia coli isolates from red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) in Poland.

Shiga toxin-producing Escherichia (E.) coli (STEC) pathogens are responsible for the outbreaks of serious diseases in humans, including haemolytic uraemic syndrome (HUS), bloody diarrhoea (BD) and diarrhoea (D), and they pose a significant public health concern. Wild ruminants are an important environmental reservoir of foodborne pathogens that can cause serious illnesses in humans and contaminate fresh products. There is a general scarcity of published data about wildlife as a reservoir of foodborne pathogens in Poland, which is why the potential epidemiological risk associated with red deer, roe deer and fallow deer as reservoirs of STEC/AE-STEC strains was evaluated in this study. The aim of the study was to investigate the prevalence of STEC strains in red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) populations in north-eastern Poland, and to evaluate the potential health risk associated with wild ruminants carrying STEC/AE-STEC strains. We examined 252 rectal swabs obtained from 134 roe deer (Capreolus capreolus), 97 red deer (Cervus elaphus) and 21 fallow deer (Dama dama) in north-eastern Poland. The samples were enriched in modified buffered peptone water. Polymerase chain reaction (PCR) assays were conducted to determine the virulence profile of stx1, stx2 and eae or aggR genes, to identify the subtypes of stx1 and stx2 genes, and to perform O and H serotyping. E. coli O157:H7 isolates were detected in the rectal swabs collected from 1/134 roe deer (0.75%) and 4/97 red deer (4.1%), and they were not detected in fallow deer (Dama dama). The remaining E. coli serogroups, namely O26, O103, O111 and O145 that belong to the "top five" non-O157 serogroups, were detected in 15/134 roe deer (11.19%), 18/97 red deer (18.56%) and 2/21 fallow deer (9.52%). STEC/AE-STEC strains were detected in 33 roe deer isolates (24.63%), 21 red deer isolates (21.65%) and 2 fallow deer isolates (9.52%). According to the most recent FAO/WHO report, stx2a and eae genes are the primary virulence traits associated with HUS, and these genes were identified in one roe deer isolate and one red deer isolate. Stx2 was the predominant stx gene, and it was detected in 78.79% of roe deer and in 71.43% of red deer isolates. The results of this study confirmed that red deer and roe deer in north-eastern Poland are carriers of STEC/AE-STEC strains that are potentially pathogenic for humans. This is the first report documenting the virulence of STEC/AE-STEC strains from wild ruminants in Poland.

Solvent Front Position Extraction with Semi-Automatic Device as a Powerful Sample Preparation Procedure Prior to Quantitative Instrumental Analysis.

The new prototype device is applied to the Solvent Front Position Extraction (SFPE) sample preparation procedure. The mobile phase is deposited onto the chromatographic plate adsorbent layer by the pipette, which is moved, according to programmed movement path, by a 3D printer mechanism. The application of the prototype device to SFPE procedure leads to the increased repeatability of the results and significant reduction of the analysis time in comparison to the classical procedure of chromatogram development. Additionally, the new equipment allows use procedures that are not possible to run using the classic chromatogram development. In this paper, the results of manual and semi-automatic sample preparation with SFPE are compared and the possible application of this prototype device is discussed.