Long-term dietary exposure to the non-steroidal anti-inflammatory drugs diclofenac and ibuprofen can affect the physiology of common carp (Cyprinus carpio) on multiple levels, even at "environmentally relevant" concentrations.

The aim of the study was to evaluate the effects of emerging environmental contaminants, the non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBP), on physiological functions in juvenile common carp (Cyprinus carpio). Fish were exposed for 6 weeks, and for the first time, NSAIDs were administered through diet. Either substance was tested at two concentrations, 20 or 2000 µg/kg, resulting in four different treatments (DCF 20, DCF 2000, IBP 20, IBP 2000). The effects on haematological and biochemical profiles, the biomarkers of oxidative stress, and endocrine disruption were studied, and changes in RNA transcription were also monitored to obtain a comprehensive picture of toxicity. Fish exposure to high concentrations of NSAIDs (DCF 2000, IBP 2000) elicited numerous statistically significant changes (p < 0.05) in the endpoints investigated, with DCF being almost always more efficient than IBP. Compared to control fish, a decrease in total leukocyte count attributed to relative lymphopenia was observed. Plasma concentrations of total proteins, ammonia, and thyroxine, and enzyme activities of alanine aminotransferase (ALT), aspartate aminotransferase, and alkaline phosphatase (ALP) were significantly elevated in either group, as were the activities of certain hepatic antioxidant enzymes (superoxide dismutase, glutathione-S-transferase) in the DCF 2000 group. The transcriptomic profile of selected genes in the tissues of exposed fish was affected as well. Significant changes in plasma total proteins, ammonia, ALT, and ALP, as well as in the transcription of genes related to thyroid function and the antioxidant defense of the organism, were found even in fish exposed to the lower DCF concentration (DCF 20). As it was chosen to match DCF concentrations commonly detected in aquatic invertebrates (i.e., the potential feed source of fish), it can be considered "environmentally relevant". Future research is necessary to shed more light on the dietary NSAID toxicity to fish.

Development and evaluation of a fish feed mixture containing the probiotic Lactiplantibacillus plantarum prepared using an innovative pellet coating method.

Introduction: Due to the intensification of fish farming and the associated spread of antimicrobial resistance among animals and humans, it is necessary to discover new alternatives in the therapy and prophylaxis of diseases. Probiotics appear to be promising candidates because of their ability to stimulate immune responses and suppress the growth of pathogens. Methods: The aim of this study was to prepare fish feed mixtures with various compositions and, based on their physical characteristics (sphericity, flow rate, density, hardness, friability, and loss on drying), choose the most suitable one for coating with the selected probiotic strain Lactobacillus plantarum R2 Biocenol™ CCM 8674 (new nom. Lactiplantibacillus plantarum). The probiotic strain was examined through sequence analysis for the presence of plantaricin- related genes. An invented coating technology based on a dry coating with colloidal silica followed by starch hydrogel containing L. plantarum was applied to pellets and tested for the viability of probiotics during an 11-month period at different temperatures (4°C and 22°C). The release kinetics of probiotics in artificial gastric juice and in water (pH = 2 and pH = 7) were also determined. Chemical and nutritional analyses were conducted for comparison of the quality of the control and coated pellets. Results and discussion: The results showed a gradual and sufficient release of probiotics for a 24-hour period, from 104 CFU at 10 mi up to 106 at the end of measurement in both environments. The number of living probiotic bacteria was stable during the whole storage period at 4°C (108), and no significant decrease in living probiotic bacteria was observed. Sanger sequencing revealed the presence of plantaricin A and plantaricin EF. Chemical analysis revealed an increase in multiple nutrients compared to the uncoated cores. These findings disclose that the invented coating method with a selected probiotic strain improved nutrient composition and did not worsen any of the physical characteristics of pellets. Applied probiotics are also gradually released into the environment and have a high survival rate when stored at 4°C for a long period of time. The outputs of this study confirm the potential of prepared and tested probiotic fish mixtures for future use in in vivo experiments and in fish farms for the prevention of infectious diseases.

Preparation and Evaluation of a Dosage Form for Individualized Administration of Lyophilized Probiotics.

Probiotics have been used in human and veterinary medicine to increase resistance to pathogens and provide protection against external impacts for many years. Pathogens are often transmitted to humans through animal product consumption. Therefore, it is assumed that probiotics protecting animals may also protect the humans who consume them. Many tested strains of probiotic bacteria can be used for individualized therapy. The recently isolated Lactobacillus plantarum R2 Biocenol™ has proven to be preferential in aquaculture, and potential benefits in humans are expected. A simple oral dosage form should be developed to test this hypothesis by a suitable preparation method, i.e., lyophilization, allowing the bacteria to survive longer. Lyophilizates were formed from silicates (Neusilin® NS2N; US2), cellulose derivates (Avicel® PH-101), and saccharides (inulin; saccharose; modified starch® 1500). They were evaluated for their physicochemical properties (pH leachate, moisture content, water absorption, wetting time, DSC tests, densities, and flow properties); their bacterial viability was determined in conditions including relevant studies over 6 months at 4 °C and scanned under an electron microscope. Lyophilizate composed of Neusilin® NS2N and saccharose appeared to be the most advantageous in terms of viability without any significant decrease. Its physicochemical properties are also suitable for capsule encapsulation, subsequent clinical evaluation, and individualized therapy.

Does dietary exposure to 17α-ethinylestradiol alter biomarkers related with endocrine disruption and oxidative stress in the adult triploid of Danio rerio?

This study was conducted to investigate a comprehensive effect of 17α-ethinylestradiol (EE2) in zebrafish (Danio rerio) with the emphasis on endocrine disruption, oxidative stress and detoxification processes at different levels. Adult male triploid zebrafish were exposed to EE2 administered in feed at two concentrations - 10 and 1000 µg/kg for six weeks. The estrogenic potential of EE2 was evaluated using an analysis of vitellogenin, gene expression focused on reproductive disorders and gonad histological examination. The alterations in antioxidant and detoxification status were assessed using analyses of enzyme activities and changes in transcriptional levels of selected genes. The most significant changes were observed especially in fish exposed to a high concentration of EE2 (i.e., 1000 µg/kg). Such high concentration caused extensive mortality (25 %) mainly in the second half of the experiment followed by a highly significant decrease in the length and body weight. Similarly, highly significant induction of vitellogenin level and vtg1 mRNA expression (about 43,000-fold compared to the control) as well as a significant downregulation of gonad aromatase expression (cyp19a1a) and histological changes in testicular tissue were confirmed in this group. In the group exposed to environmentally relevant concentration of EE2 (i.e., 10 µg/kg), no significant differences in vitellogenin were observed, although all fish were positive in the detection of vitellogenin compared to control, where only 40 % of individuals were positive. In addition, the high concentration of EE2 resulted in significant alterations in most monitored antioxidant and detoxifying enzymes with the exception of catalase, followed by strongly significant upregulation in mRNA expression of gsr, gpx1a, cat and cyp1a genes. Furthermore, a significant decrease in the glutathione reductase activity was recorded in fish exposed to 10 µg EE2/kg. To our knowledge, this is the first study which reports the effects of subchronic per oral exposure to EE2 in adult triploid zebrafish.

Use of all-male triploid population of zebrafish (Danio rerio) for evaluation of 17α-ethinylestradiol effects.

OBJECTIVES: We tested the toxicity of ethinylestradiol, a semisynthetic estrogen used in oral contraceptives, on all-male triploid zebrafish using commercial feeds and three different doses concentrations. We aimed to determine whether ethinylestradiol peroral administration resulted in vitellogenin production and whether all-male triploid zebrafish could serve as a model species for xenoestrogen testing. METHODS: The actual concentrations of 17α-ethinylestradiol were 0.0035 (low); 0.0315 (medium) and 0.365 (high) µg/g. Positive control represented commercial feeds containing 0.0465 µg/g of ß-estradiol. The experiment lasted 8 weeks. RESULTS: Our results indicate that 17α-ethinylestradiol consumption does induce vitellogenin production in triploid zebrafish. CONCLUSIONS: The simple presence of vitellogenin is a definite symptom indicative of the potential for such changes due to the action of estrogenic substances. As such, this experiment has shown that the use of all-male triploid zebrafish populations, rather than the mixed-sex populations of other species previously used, could serve as a suitable alternative model population for controlled testing of the effects of xenoestrogens on fish.

Commercially Available Enteric Empty Hard Capsules, Production Technology and Application.

Currently, there is a growing need to prepare small batches of enteric capsules for individual therapy or clinical evaluation since many acidic-sensitive substances should be protected from the stomach's acidic environment, including probiotics or fecal material, in the fecal microbiota transplantation (FMT) process. A suitable method seems to be the encapsulation of drugs or lyophilized alternatively frozen biological suspensions in commercial hard enteric capsules prepared by so-called Enteric Capsule Drug Delivery Technology (ECDDT). Manufacturers supply these types of capsules, made from pH-soluble polymers, in products such as AR Caps®, EnTRinsicTM, and Vcaps® Enteric, or capsules made of gelling polymers that release their content as the gel erodes over time when passing through the digestive tract. These include DRcaps®, EMBO CAPS® AP, BioVXR®, or ACGcaps™ HD. Although not all capsules in all formulations meet pharmaceutical requirements for delayed-release dosage forms in disintegration and dissolution tests, they usually find practical application. This literature review presents their composition and properties. Since ECDDT is a new technology, this article is based on a limited number of references.

Development and Comparison of Various Coated Hard Capsules Suitable for Enteric Administration to Small Patient Cohorts.

Pharmaceutical technology offers several options for protecting substances from acidic environments, such as encapsulation in enteric capsules or dosage form with enteric coating. However, commercial enteric capsules do not always meet limits for pharmacopeial delayed release, and the coating process is generally challenging. Preparing small enteric batches suitable for clinical use is, therefore, an unsolved problem. This experiment offers a simple coating process of DRcapsTM capsules based on hypromellose (HPMC) and gellan gum to achieve small intestine administration. In addition, DRcapsTM capsules were compared to hard gelatin capsules to evaluate the suitability of the coating method. Both capsules were immersed in dispersions of Eudragit® S 100, Acryl-EZE®, and Cellacefate at concentrations of 10.0, 15.0, and 20.0% and dried. Coated capsules were evaluated by electron microscopy, disintegration, and dissolution test with a two-step pH change (from 1.2 to 6.8, then to 7.5) to simulate passage through the digestive tract. DRcapsTM capsules coated with Eudragit® S and Cellacefate achieved acid resistance. While samples coated with Eudragit® S released their contents within 360 min at pH 6.8 (small intestine), regardless of polymer concentration, capsules with 15.0 and 20.0% coatings of Cellacefate released content at pH 7.5 (colon) within 435 and 495 min, respectively.

Rational Design of Self-Emulsifying Pellet Formulation of Thymol: Technology Development Guided by Molecular-Level Structure Characterization and Ex Vivo Testing.

The growing need for processing natural lipophilic and often volatile substances such as thymol, a promising candidate for topical treatment of intestinal mucosa, led us to the utilization of solid-state nuclear magnetic resonance (ss-NMR) spectroscopy for the rational design of enteric pellets with a thymol self-emulsifying system (SES). The SES (triacylglycerol, Labrasol®, and propylene glycol) provided a stable o/w emulsion with particle size between 1 and 7 µm. The ex vivo experiment confirmed the SES mucosal permeation and thymol delivery to enterocytes. Pellets W90 (MCC, Neusilin®US2, chitosan) were prepared using distilled water (90 g) by the M1−M3 extrusion/spheronisation methods varying in steps number and/or cumulative time. The pellets (705−740 µm) showed mostly comparable properties­zero friability, low intraparticular porosity (0−0.71%), and relatively high density (1.43−1.45%). They exhibited similar thymol release for 6 h (burst effect in 15th min ca. 60%), but its content increased (30−39.6 mg/g) with a shorter process time. The M3-W90 fluid-bed coated pellets (Eudragit®L) prevented undesirable thymol release in stomach conditions (<10% for 3 h). A detailed, ss-NMR investigation revealed structural differences across samples prepared by M1−M3 methods concerning system stability and internal interactions. The suggested formulation and methodology are promising for other lipophilic volatiles in treating intestinal diseases.

Ethical and legal requirements for vaccination against COVID-19.

In the current context, there is an extraordinary interest of states in vaccinating the population to prevent Covid-19. In the Czech Republic, gene mRNA and vector DNA vaccines are approved only with conditional marketing authorization, for which a complete and long-term safety assessment is currently lacking. Vaccines show some potential risks, such as penetration of lipid nanoparticles into surrounding tissues, incorporation of DNA into the host genome, ADE syndrome, development of resistant mutations, myocarditis, pericarditis, and thromboembolic events. Since the level of antibodies after vaccination is soon decreasing, immunity after the disease persists longer, and the disease's fatality rate is very low, especially in adolescents, only voluntary vaccination is ethically acceptable, without any direct or indirect restrictions on the unvaccinated. The conclusion is in line with the principles of medical ethics of nonmaleficence, beneficence, autonomy, and justice.

Surviving of production probiotic strains in a selected application form.

Research in probiotics for aquaculture is at an early stage of development and much work is still needed. Lactiplantibacilli belong to the microorganisms most frequently used to prepare the probiotics. The available information is inconclusive, since few experiments with sufficiently robust design have been conducted to permit critical evaluation. The development of probiotics applicable to commercial use in aquaculture is a multistep and multidisciplinary process requiring both empirical and fundamental research, full-scale trials, and an economic assessment of its use. The aim of the study was to prepare a probiotic aquafeed via excipients and subsequently to observe the survival of probiotic bacterial cells in the feed during the nine months storage period at a refrigerator (4 °C) or room temperature (22 °C). The strain Lactobacillus plantarum R2 Biocenol (CCM 8674) (according to the new taxonomy Lactiplantibacillus plantarum), potentially usable as a probiotic in aquaculture, was administered to prepare the aquafeed. Better survival of probiotic bacterial cells was recorded in a samples of pellets A (Aquatex 41 HMD) compared to the samples of probiotic pellets B (Inicio 918-2). Since oxidation of fatty acids in feed affects the nutritional quality of individual feed components, we assume that higher amounts of oil in feed B negatively affected the survival of probiotic bacterial cells. The highest numbers of viable probiotic bacteria cells were recorded at 4 °C storage of probiotic feed samples. The number of lactiplantibacilli dropped from 7.30 log10CFU . g-1 to 5.57 log10CFU . g-1 after the nine months storage period of feed samples A at 4 °C. Temperature is considered as a critical factor influencing probiotic viability and survival during storage period.

Technology of Processing Plant Extracts Using an Aluminometasilicate Porous Carrier into a Solid Dosage Form.

A method of preparing tablets called liquisolid technique is currently emerging. In these formulations, an important role is played by porous carriers, which are the basic building blocks of liquisolid systems (LSSs). The most common are microcrystalline cellulose (MCC), magnesium aluminometasilicates, silica aerogels, mesoporous silicates, clays, etc. In this study, magnesium aluminometasilicate is used to prepare modified LSS formulations with plant extracts as model drugs dissolved in water (W) or ethanol (E). The modification involves drying tablets in a microwave (MW) and hot air dryer (HA) for a specified period. Powder blends and tablets were evaluated for physical properties, and their antioxidant activity (AA) was measured in a modified dissolution by ferric reducing antioxidant power assay (FRAP). PLS and ANOVA were used to compare tablets properties depending on the composition and technology. The experiment is based on a previous one, in which the plant extracts were processed into tablets using a similar method. Therefore, extending the study to include more plants and the robust statistical evaluation and comparison of the products was a procedure to justify the suitability of the presented method for a wide range of liquid plant extracts. As a result, we obtained tablets with excellent physical properties, including a short disintegration and dissolution, which is problematic in tableted extracts.

Foodborne fluoxetine impacts the immune response in rainbow trout (Oncorhynchus mykkis).

The aim of this study was to reveal the effects of foodborne fluoxetine on morphological and condition profile, hematological profile, biochemical and oxidative stress indices on juvenile rainbow trout. The study was performed according to OECD Guideline No. 215. Fluoxetine was incorporated into Biomar 921 pellets at a dose of 0.047 mg/kg (environmental concentration), 0.577 mg/kg and 6.7 mg/kg. There was statistically significant change in hematological profile, including an increasing trend in neutrophil/lymphocyte ratio and a decreasing trend in the number of lymphocytes. Measurements of oxidative stress indicated decreased activity of the detoxifying enzyme glutathione-S-transferase in the liver and kidney. However, the measurement of GR, GPx, CAT, SOD activity, and TBARS showed no changes. Histopathological examination revealed damage to the proximal tubules of caudal kidney in exposed groups. This study confirms that fluoxetine has a significant effect on immune response.

The effects of dietary exposure to Magnéli phase titanium suboxide and titanium dioxide on rainbow trout (Oncorhynchus mykiss).

Magnéli phase titanium suboxides (Magnéli TiOx) are promising, novel materials with superior properties compared to TiO2, they are substoichiometric titanium oxides with the chemical formula TinO2n-1 (where n ≥ 1). In this study, for the first time, subchronic effects of dietary intake of Magnéli TiOx were evaluated and compared with TiO2 particles of similar size, in concentrations 0.1% and 0.01% of feed. The experiment consisted of 38 d of an exposition period and 14 d of a depuration period. Minor effects on plasma biochemical profile and morphological parameters were recorded. A reduced count of leukocytes was found in the blood of both Magnéli TiOx and TiO2 exposed fish, suggesting immunotoxic effects. Erythrocytosis was specific for Magnéli TiOx. Indices of oxidative stress, namely increased lipid peroxidation in liver, increased activity of superoxide dismutase in liver, kidney and gills and glutathione S-transferase (GST) in gills, as well as decreased activity of ceruloplasmin and GST in liver were found predominantly in fish exposed to TiO2. Histopathological examination revealed increased lipid-like vacuolation in the liver, the presence of hyaline droplets in renal tubules and multiplication of mucous glands in the epidermis in both tested substances and intestine damage in TiO2 groups. Overall, in Magnéli TiOx exposed groups, fewer adverse effects compared to TiO2 expositions were recorded. Their wider practical implementation in place of TiO2 is therefore beneficial.

Structural Changes of Sodium Warfarin in Tablets Affecting the Dissolution Profiles and Potential Safety of Generic Substitution.

At present, the risk of generic substitutions in warfarin tablets is still being discussed. The aim of this study was to assess whether API interactions with commonly used excipients may affect the safety of generic replacement of warfarin sodium tablets. These interactions were observed during an accelerated stability study, and the effect of the warfarin solid phase (crystalline/amorphous form) as well as the API particle size distribution was studied. Commercial tablets and prepared tablets containing crystalline warfarin or amorphous warfarin were used. In addition, binary mixtures of warfarin with various excipients were prepared. The structural changes before and after the stability study were monitored by dissolution test in different media, solid-state NMR spectroscopy and Raman microscopy. During the stability study, the conversion of the sodium in warfarin to its acid form was demonstrated by some excipients (e.g., calcium phosphate). This change in the solid phase of warfarin leads to significant changes in dissolution, especially with the different particle sizes of the APIs in the tablet. Thus, the choice of suitable excipients and particle sizes are critical factors influencing the safety of generic warfarin sodium tablets.

Comparison of Flow and Compression Properties of Four Lactose-Based Co-Processed Excipients: Cellactose® 80, CombiLac®, MicroceLac® 100, and StarLac®.

The utilization of co-processed excipients (CPEs) represents a novel approach to the preparation of orally disintegrating tablets by direct compression. Flow, consolidation, and compression properties of four lactose-based CPEs-Cellactose® 80, CombiLac®, MicroceLac® 100, and StarLac®-were investigated using different methods, including granulometry, powder rheometry, and tablet compaction under three pressures. Due to the similar composition and the same preparation technique (spray drying), the properties of CPEs and their compacts were generally comparable. The most pronounced differences were observed in flowability, undissolved fraction after 3 min and 24 h, energy of plastic deformation (E2), ejection force, consolidation behavior, and compact friability. Cellactose® 80 exhibited the most pronounced consolidation behavior, the lowest values of ejection force, and high friability of compacts. CombiLac® showed excellent flow properties but insufficient friability, except for compacts prepared at the highest compression pressure (182 MPa). MicroceLac® 100 displayed the poorest flow properties, lower ejection forces, and the best mechanical resistance of compacts. StarLac® showed excellent flow properties, the lowest amounts of undissolved fraction, the highest ejection force values, and the worst compact mechanical resistance. The obtained results revealed that higher compression pressures need to be used or further excipients have to be added to all tested materials in order to improve the friability and tensile strength of formed tablets, except for MicroceLac® 100.

The biological activity of the organic UV filter ethylhexyl methoxycinnamate in rainbow trout (Oncorhynchus mykiss).

UV filters are able to enter the aquatic environment and negatively affect non-target organisms. The aim of this study was to evaluate subchronic exposure to ethylhexyl methoxycinnamate (EHMC) in rainbow trout. The tested EHMC concentrations of 6.9 (low), 96.0 (medium) and 395.6 µg/kg (high) were used. The lowest concentration was based on environmentally relevant concentrations. The higher concentrations were chosen as a multiple of the lowest one to determine the dose-response relationship. EHMC was incorporated into feed pellets. The experiment was conducted for six weeks in a semi-static system. Haematological, biochemical and oxidative stress indices were determined at the end of the experiment and supplemented by histological examination. Significant changes were proven at medium and high concentrations of EHMC. Specifically, increases of glucose, lactate and decrease of albumin and total protein in plasma indicate a stress ethology. Moreover, a decrease of plasma cholesterol, triacylglycerols and ammonia were observed even in the experimental group exposed to the lowest concentration of EHMC, perceived as an environmentally relevant concentration. The ferric reducing ability of plasma was decreased in all tested concentrations. Exposure to the highest concentration of EHMC resulted in a decrease in leukocyte counts. Increased activity of glutathione peroxidase in liver was recorded for the medium and the highest concentration of EHMC. The level of the thiobarbituric acid reactive substances in kidney was elevated for the highest concentration. Decrease of the activity of glutathione-S-transferase in gills for medium concentration of EHMC was registered. Histopathological examination revealed massive destruction of hepatic parenchyma at the highest concentration of EHMC. All these results support the finding of a stress load on the fish organism. In summary, although subchronic exposure to EHMC had no effect on behaviour, mortality or feed intake, this exposure resulted in the alteration of saccharide, lipid and protein metabolism and weakened antioxidant capacity.

Multivariate analysis in the development of bioequivalent tablets containing bicalutamide.

The pharmaceutical industry has to tackle the explosion of high amounts of poorly soluble APIs. This phenomenon leads to numerous sophisticated solutions. These include the use of multifactorial data analysis identifying correlations between the components and dosage form properties, laboratory and production process parameters with respect to the API liberation Example of such API is bicalutamide. Improved liberation is achieved by particle size reduction. Laboratory batches, with different PSD of API, were filled into gelatinous capsules and consequently granulated for tablet compression. Comparative dissolution profiles with Casodex 150 mg (Astra Zeneca) were performed. The component analysis was used for the statistical evaluation of f1 and f2 factors and D(v,0.9) and D[4,3] parameters of PSD to identify optimal PSD values. Suitable PSD limits for API were statistically confirmed in laboratory and in commercial scale with respect to optimized tablet properties. The tablets were bioequivalent with originator (n = 20; 90% CI for ln AUC0-120: 99.8-111.9%; 90% CI for ln cmax: 101.1-112.9%). In conclusion, the micronisation of the API is still an efficient and inexpensive method improving the bioavailability, although there are more complicated and expensive methods available. Statistical multifactorial methods improved the safety and reproducibility of production.

Vaccines from the perspective of a pharmacist.

The field of development, production and safety of vaccines has recently come under the public eye. The aim of this review article is not to provide comprehensive information on the development and production of vaccines, which would not be possible even in a limited space. Its purpose is to outline a brief overview of the principles, development, and production of basic types of vaccines and point out the benefits and risks from the perspective of a pharmacist. He can participate not only in basic research, but his role is mainly in the formulation of a dosage form, registration of a vaccine, its distribution, and educational activities towards the lay and professional public.

The effect of foodborne sertraline on rainbow trout (Oncorhynchus mykiss).

The worldwide consumption of antidepressants is raising as well as their concentrations in the aquatic environment. This increases the risk of food chain contamination and bioaccumulation in aquatic biota. The aim of this study was to describe a potential risk of sertraline as a pollutant from water environment, wherein rainbow trout (Oncorhynchus mykiss) has been chosen as the test organism, because predatory fish are on the top of the food chain in the aquatic environment. The effects of foodborne sertraline were tested on rainbow trout during a 28-day toxicity test according to OECD 215 method. Sertraline was incorporated in commercial feed at a dose of 4.4 µg/kg (environmental concentration), 42 µg/kg and 400 µg/kg. The results confirmed that sertraline has a significant effect on fish behaviour, resulting in suppression of the escape reflex and increased resistance to stress. Moreover, increased Fultons condition factor was found in fish fed with the highest concentration of sertraline. Haematological analysis revealed a statistically significant increase in the number of neutrophilic bands and neutrophil/lymphocyte ratio, and decreased number of lymphocytes. The results of biochemical examination showed a statistically significant decrease in ammonia and lactate concentrations and histological examination revealed changes in gills and caudal kidney. Although sertraline reduces stress in fish, the decline in nonspecific immunity is a risk to fish population stability.

Unique coated neusilin pellets with a more distinct and fast visual detection of nerve agents and other cholinesterase inhibitors.

Pellets with an immobilized enzyme (acetyl- or butyrylcholinesterase) are the up-to-date type of carriers used for the detection of nerve agents (soman, sarin, tabun, VX, Novichok) and other cholinesterase inhibitors such as organophosphate and carbamate insecticides (parathion, malathion). They are used in the glass detection tubes as a layer containing the enzyme together with the second layer, which contains a colorimetric reagent and substrate. The detection method is based on the visually or spectrophotometrically observable Ellman's reaction, which develops a yellow color in the absence of the cholinesterase inhibitor; otherwise, the detector preserves its original color (preferably white). This reaction occurs very fast and has a high sensitivity to nerve agents but it suffers from an indistinctive color transition from white to yellow. In the presented study, a new approach with the use of the synergic effect of magnesium aluminometasilicate with a high surface area marketed as Neusilin®US2 and a protective semipermeable Eudragit® RL layer was utilized. The prepared pellets have been evaluated for their properties such as the activity of the enzyme, intensity of the developed yellow color, sensitivity to cholinesterase inhibitor physostigmine, which acts as a nerve agent simulant, and physical parameters such as hardness, pycnometric density and sphericity. After the initial evaluation, all samples underwent a stability test under three different storage conditions for 24 months during which they were evaluated at given time points (0, 3, 6, 12 and 24 months). It was found that the prepared samples achieved a much higher intensity of developed yellow color than in the published studies while maintaining similar or better sensitivity, speed of detection and suitable physico-chemical properties.