Simultaneous Application of High-Intensity Focused Electromagnetic and Synchronized Radiofrequency for Fat Disruption: Histological and Electron Microscopy Porcine Model Study.

BACKGROUND: Radiofrequency (RF) and high-intensity focused electromagnetic (HIFEM) technologies are used for noninvasive body shaping as standalone modalities. OBJECTIVE: To examine the effects of novel synchronized RF and HIFEM on subcutaneous adipose tissue in a porcine animal model. MATERIALS AND METHODS: Seven large white pigs aged 6 months received 3 abdominal treatments of simultaneous application of synchronized RF and HIFEM (30 minutes, once per week). Punch biopsies of treated and control subcutaneous tissue were collected at the baseline, 4 days, 2 weeks, 1 month, and 2 months. Specimens were examined by light and scanning electron microscopy. Adipocyte volume was analyzed. Fat tissue temperature was measured in situ (fiber optic probes) and superficially (thermal imager). RESULTS: Fat layer was heated to temperatures of 42 to 45°C. Signs of fat apoptosis (shape alternations and pyknotic nuclei) appeared at day 4 and peaked between 2 weeks and 1 month. Adipocyte volume decreased significantly (p < .001) by 31.1% at 2 weeks, 1 month (-23.6%), and 2 months (-22.0%). Control samples showed healthy adipocytes. Scanning electron microscopy micrographs corroborated histology findings, showing flattened, volume-depleted and disrupted adipocytes. CONCLUSION: Synchronized RF with HIFEM procedure resulted in a significant and sustained fat reduction with no adverse events.

Morphogenesis of Olfactory Organ of Bushymouth Catfish Ancistrus dolichopterus (Teleostei: Loricariidae) before Switching to Exogenous Feeding.

Olfaction plays an important role in a fish's life. Its value may differ at different developmental stages, depending on the feeding style of the species. The goal of the present study was to investigate the olfactory organ of a species that feeds mainly on algae- the bushymouth catfish, Ancistrus dolichopterus-at developmental stages from olfactory placode to the definitive olfactory chamber. For this study, we used light and electron (scanning) microscopy. The topography of the olfactory placode of A. dolichopterus is typical for teleostei. Formation of olfactory pit takes place at the same time as rostral elevation formation. Rostral elevation participates in the formation of the nasal bridge and anterior tubular nostril. It was found out that the anlage of olfactory rosette in A. dolichopterus arises earlier than in most teleostei. However, the number of lamellae does not increase until switching to exogenous feeding. We suppose that the early development of olfactory organ is necessary for intraspecific communication, not just for finding food.

Changes in the mast cell distribution in the canine ovary and uterus throughout the oestrous cycle.

This is the first study describing the relation between the phase of the oestrous cycle and the number of mast cells (MCs) in the canine ovaries and uterine horns. The total number of MCs was counted in cortex ovarii, medulla ovarii, endometrium and myometrium. The number of MCs was compared in all of these areas among the bitches in the early follicular phase, luteal phase and anoestrus. MCs were the most numerous in the early follicular phase and in the luteal phase; however, they were significantly less numerous during anoestrus. Based on the results, it was concluded that the number of MCs in ovaries and uterine horns fluctuates during the oestrous cycle. The results of this study may contribute to clarifying the role of MCs in the reproductive organs of bitches.

Sodium chloride treatment effects on rainbow trout suffering from proliferative kidney disease caused by Tetracapsuloides bryosalmonae.

The aim of the present study was to evaluate the effect of a long-term sodium chloride bath on rainbow trout Oncorhynchus mykiss naturally infected by Tetracapsuloides bryosalmonae. A total of 106 infected fish were divided into 2 groups. One group was left untreated and the other was treated with sodium chloride in increasing doses up a concentration of 0.8%. After 14 d, treatment was stopped and for a further 7 d the fish response to the sodium chloride bath was observed. Cumulative mortality was significantly lower in the treated group (19.2%) compared to the untreated group (31.5%) after 21 d. This corresponded to the lower but non-significant parasite intensity in kidney and spleen in the treated group after 14 d of treatment. However, lower prevalence of parasites in both tissues was recorded in the untreated group after 21 d of treatment, but a significant difference was observed only in spleen tissue. Furthermore, significant increases in leukocytes, hemoglobin, haematocrit, ferric reducing ability of plasma, and ceruloplasmin, and significant decreases in alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities were noticed in the treated group compared to the untreated group. In contrast, significant decreases in lysozyme concentration in the mucus and phagocyte oxidative burst in the blood were observed in the treated group. Histopathological examination revealed proliferative and reparative changes in parenchymatous tissues in the treated group. The 14- and 21-d salt bath used in rainbow trout with proliferative kidney disease was associated with a reduction in mortality and enhanced the reparative phase in the treated group.

Effects of therapeutic formalin bath on selected indices in fish.

OBJECTIVES: The aim of this study is to assess the effects of a treatment bath in a formalin solution on fish, focusing on haematological, biochemical and histopathological profiles. METHODS: A total of 96 common carps (Cyprinus carpio) were randomly assigned to eight groups. Four experimental groups were placed in the test solution for 60 minutes. The concentration of the formalin bath was 0.17 ml/l (38% formaldehyde), with the water temperature of 20°C. The effects of the bath were monitored immediately (E0) and 24 hours (E24h), 48 hours (E48h) and 10 days (E10d) after the bath. There was a control group (C0, C24h, C48h, C10d) for each of the experimental groups. RESULTS: Histopathological indices were strongly affected. Extensive changes were found on gill and skin immediately and also after 24 h, 48 h and 10 d of the formalin treatment bath. A plurality of mucinous elements was observed on the skin. Moreover, structural devastation of lamellas and numerous mucinous cells were observed on the gill. Changes were also found in haematological and biochemical indices. CONCLUSIONS: It can be concluded that after applying the treatment bath in the formalin solution, the monitored profiles were affected immediately after the bath and also after 10 days (histopathological changes of skin and gill). Such significant impact of therapeutic formalin bath should be considered in treatment of fish.

The Effects of Salicylic Acid on Juvenile Zebrafish Danio rerio Under Flow-Through Conditions.

The aquatic environment is becoming increasingly contaminated with pharmaceuticals. Salicylic acid (SA), which can be used individually or appear as a degradation product of the widely used acetylsalicylic acid was chosen for testing. Juvenile zebrafish Danio rerio were subjected to OECD test No. 215 (fish, juvenile growth test) with salicylic acid concentrations of 0.004; 0.04; 0.4; 4 and 40 mg/L. Specific growth rate (SGR), histological changes, and parameters of oxidative stress were evaluated. SA had no effects on histological changes, SGR, glutathione reductase, and lipid peroxidation. Increased catalytic activity of GPx was found at 0.04 mg/L compared to control, increased catalytic activity of catalase was found at 0.04 and 4 mg/L compared to control, and increased catalytic activity of glutathione-S-transferase was found at 0.004 and 0.04 mg/L compared to control (P < 0.05). Juvenile zebrafish turned out to be relatively insensitive to both environmentally relevant (0.004 mg/L) and higher concentrations of salicylic acid.

Oxidative stress indices and histopathological effects of the nonsteroidal antiinflammatory drug naproxen in adult zebrafish (Danio rerio).

OBJECTIVES: The aim of this study was to investigate the effects of the nonsteroidal antiinflammatory drug naproxen on adult zebrafish (Danio rerio). METHODS: Three months old zebrafish (Danio rerio) were exposed to naproxen at concentrations of 0.001, 0.1 and 5 mg.L(-1). We focused on the changes in oxidative stress indices during and at the end of the experiment and histopathological examination of tissues after a two week long exposure period. RESULTS: We found that a 3 day long exposure to naproxen causes mild oxidative stress and affects detoxification in zebrafish, which is demonstrated by the increased activity of glutathione peroxidase and glutathione S-transferase at 0.001 and 0.1 mg.L(-1) of naproxen, respectively. After a 7 day long exposure to 0.1 and 5 mg.L(-1), more potent effects on enzymes occur. However, these effects are only short lasting. At the end of the experiment, the activities of the target enzymes recover back to homeostatic baseline levels. Except catalase, which is induced only after a two week long exposure to the environmental concentration of naproxen. Despite the fact that naproxen causes mild oxidative stress in zebrafish, exposure to this drug does not result in lipid peroxidation. Histopathological examination revealed obvious changes to the gills and liver even at exposure to the environmental concentration of naproxen. CONCLUSION: This study demonstrates that the environmental concentration of naproxen can slightly influence both the antioxidant defense system and histopathology of non-target fish.

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.

Extraskeletal myxoid chondrosarcoma: A study of 17 cases focusing on the diagnostic utility of INSM1 expression and presenting rare morphological variants associated with non-EWSR1::NR4A3 fusions.

Extraskeletal myxoid chondrosarcoma (EMC) is a rare sarcoma of uncertain lineage. Insulinoma-associated protein 1 (INSM1) has recently been described as a highly specific and sensitive immunohistochemical marker for EMC. The goal of this study was to evaluate the diagnostic significance of INSM1 immunohistochemistry in EMC. Furthermore, correlations between molecular and morphological findings were performed. Sixteen of 17 EMC cases were stained with the INSM1 antibody. Tumors with at least 5% INSM1-positive cells and any staining intensity were considered positive. Molecular testing was successfully performed in 12/17 cases. The immunohistochemical analysis detected 13 INSM1-positive (81%) and 3 INSM1-negative tumors (19%). The extent of the staining was classified as 1+ in 7 cases (44%), 2+ in 2 cases (13%), 3+ in 2 cases (13%) and 4+ in 2 cases (13%). Intensity of immunostaining was weak in 5 cases (31%), moderate in 2 cases (13%) and strong in 6 cases (38%). Molecular assays revealed 8 EWSR1::NR4A3 positive tumors (67%), 2 TAF15::NR4A3 positive tumors (17%), 1 TCF12::NR4A3 positive tumor (8%) and 1 NR4A3 positive tumor (8%) in which no other gene alteration was identified. Two of them, namely TCF12 positive and one TAF15 positive tumors, were highly cellular and partially associated with pseudopapillary architecture. Our study found that moderate/strong expression of INSM1 in more than 25% of tumor cells was present in only 31% of cases. Thus, the diagnostic utility of INSM1 is rather low. Two morphologically unique cases of non-EWSR1 rearranged EMC with an extremely rare pseudopapillary growth pattern are also reported.

Medicine designed to combat diseases of affluence affects the early development of fish. How do plastic microparticles contribute?

The incidence of diseases of affluence, such as diabetes mellitus, cardiovascular diseases, high blood pressure, and high cholesterol has been reported to rise. Consequently, the concentrations of residues of drugs designed to treat these diseases have been rising in water bodies. Moreover, the toxicity of these pharmaceuticals towards fish and other non-target organisms can be even enhanced by microplastic particles that are reportedly present in surface water. Therefore, the aim of this study was to describe the effects of three highly prescribed drugs, in particular metoprolol, enalapril, and metformin on fish early-life stages. Also, it was hypothesized that polystyrene microparticles will increase the toxicity of metoprolol to fish early-life stages. Embryonal acute toxicity tests on Danio rerio and Cyprinus carpio were carried out in order to describe the possible toxic effects of metoprolol, enalapril, and metformin. Also, the acute toxicity of polystyrene microparticles and the combination of metoprolol with polystyrene microparticles were tested on D. rerio embryos. Additionally, a 31-day long embryo-larval subchronic toxicity test was carried out with C. carpio in order to describe the long-term effects of low concentrations of metoprolol. The results of the study show that both metoprolol and enalapril have the potential to disrupt the early development of the heart in the embryonal stages of fish. Also, enalapril and metformin together with polystyrene microparticles seem to possibly disrupt the reproduction cycle and act as endocrine disruptors. Both pure polystyrene microparticles and the combination of them with metoprolol affect inflammatory processes in organisms. Additionally, metformin alters several metabolism pathways in fish early-life stages. The results of the study bring new evidence that even low, environmentally-relevant concentrations of pharmaceuticals have the potential to disrupt the early development of fish, particularly on a molecular level.

Pesticides and Parabens Contaminating Aquatic Environment: Acute and Sub-Chronic Toxicity towards Early-Life Stages of Freshwater Fish and Amphibians.

Pesticides and personal care products are two very important groups of contaminants posing a threat to the aquatic environment and the organisms living in it.. Therefore, this study aimed to describe the effects of widely used pesticides and parabens on aquatic non-target biota such as fish (using model organisms Danio rerio and Cyprinus carpio) and amphibians (using model organism Xenopus laevis) using a wide range of endpoints. The first part of the experiment was focused on the embryonal toxicity of three widely used pesticides (metazachlor, prochloraz, and 4-chloro-2-methyl phenoxy acetic acid) and three parabens (methylparaben, propylparaben, and butylparaben) with D. rerio, C. carpio, and X. laevis embryos. An emphasis was placed on using mostly sub-lethal concentrations that are partially relevant to the environmental concentrations of the substances studied. In the second part of the study, an embryo-larval toxicity test with C. carpio was carried out with prochloraz using concentrations 0.1, 1, 10, 100, and 1000 µg/L. The results of both parts of the study show that even the low, environmentally relevant concentrations of the chemicals tested are often able to affect the expression of genes that play either a prominent role in detoxification and sex hormone production or indicate cell stress or, in case of prochloraz, to induce genotoxicity.

Unique expression patterns of the embryonal stem cell marker SOX2 and hormone receptors suggest the existence of a subpopulation of epithelial stem/progenitor cells in porcine and bovine endometrium.

BACKGROUND: There are currently insufficient data on the population of endometrial epithelial stem/progenitor cells in farm animals. OBJECTIVES: With the aim of identifying a potential population of epithelial stem/progenitor cells in the porcine and bovine endometrium, this study immunohistochemically examined the expression patterns of the oestrogen and progesterone receptors, as well as that of the embryonal stem cell marker SOX2. METHODS: A total of 24 endometrial tissue samples obtained from cycling pigs (n = 12) and cows (n = 12) were included in our study. Each endometrium was divided into basal, middle and luminal portions. The percentage of marker-positive cells and the intensity of the immunoreaction in each portion of the endometrium were determined. RESULTS: Inverse expression patterns of SOX2 and progesterone receptors were found in both animal species throughout the oestrous cycle. Strong diffuse SOX2 expression was detected in the basal portions of the glands, while a significant decrease in positivity and a weak immunoreaction were found in the luminal two thirds of the glandular epithelium. Strong progesterone receptor expression was observed in at least 90% of glandular cells in the middle and luminal portions, whereas weak staining and significant decrease in positivity were detected in the basal portions of the glands. One oestrogen receptor expression pattern resembled that of progesterone receptors. CONCLUSION: The inverse expression patterns of SOX2 and hormone (especially progesterone) receptors suggest that endometrial epithelial stem/progenitor cells represent a subset of cells that reside in the basal portions of the endometrial glands in both the bovine and porcine endometrium.

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.

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.

Effect of GnRH agonist deslorelin implant on spermatogenesis and testosterone concentration in Guinea pigs (Cavia aperea porcellus).

Guinea pigs are social animals that are often kept in groups regardless of their gender. Due to reproduction control and male aggressiveness prevention, surgical castration is commonly required. In the present study, we evaluated the effect of GnRH agonist implant (4.7 mg deslorelinum) on the serum testosterone concentration (T) and spermatogenesis in male guinea pigs. Twenty-four animals were divided into two groups. All animals in the first group were neutered (Group 1), animals in the second group (Group 2) were administered the implant subcutaneously and then neutered in one-month intervals. A histological examination was performed when cross sections of seminiferous tubules were assessed. Subsequently, these tubules were divided based on the most developed germ cell observed: spermatogonia, spermatocytes, round spermatids, elongating spermatids and elongated spermatids. The anticipated decrease in testosterone concentration and cessation of spermatogenesis was not achieved. Thus, the results obtained proved the inefficacy of the deslorelin implant in male guinea pigs so the alternative methods of contraception remain the methods of choice.

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.

Light and scanning electron microscopy of the tongue of a degu (Octodon degus).

The tongue of an adult degu was examined by light and scanning electron microscopy. It consists of an apex, corpus, and radix and contains a lingual prominence. The aim of this study was to describe the course of muscle fascicles of the proper lingual muscle, the presence and nature of the lingual salivary glands, and particularly the appearance and distribution of the lingual papillae. Three major types of papillae have been observed: filiform, conical, and vallate. The dorsal surface of the lingual apex extends in caudally bent filiform papillae with two spines. The lingual corpus bears long filiform papillae with a single tip. The lingual radix contains crown-like papillae in the region of the prominence and conical papillae in the remaining areas. Two oval vallate papillae were discovered caudally on the lingual radix. This first description of the lingual structures in a degu could be used for comparative studies or as basic data for differentiation of lingual morphology in this species.

Toxicity of naproxen sodium and its mixture with tramadol hydrochloride on fish early life stages.

Pharmaceuticals occur in water bodies as a consequence of their incomplete removal during waste water treatment processes. The occurence of pharmaceuticals in surface waters as well as their possible impact on aquatic vertebrates have received considerable attention in recent years. However, there is still a lack of informations on the chronic effects of widely used drugs as well as their possible mixture toxicity on non-target aquatic vertebrates as well as their possible mixture toxicity. The aim of this study was to assess the effects of naproxen sodium on early life stages of fish and evaluate its mixture toxicity with tramadol hydrochloride, which was assessed in our earlier study as a single substance. Two embryo-larval toxicity tests with common carp (Cyprinus carpio) were performed according to the OECD guideline 210 (Fish, Early-life Stage Toxicity Test) in order to assess the subchronic toxicity of naproxen sodium and tramadol hydrochlorid-naproxen sodium mixture at the concentrations of 10; 50; 100 and 200 µg/L. These experiments were conducted for 32 days. The subchronic exposure to naproxen sodium and naproxen sodium and tramadol hydrochloride mixture had a strong effect on the early life stages of common carp. Hatching, developmental rate, morphology, histopathology and, in the case of the naproxen sodium and tramadol hydrochloride mixture, mortality were influenced. The bioindicators of oxidative stress were also influenced. The LOEC was determined at 10 µg/L for both naproxen sodium and naproxen sodium and tramadol hydrochloride mixture.

Effects of selected tricyclic antidepressants on early-life stages of common carp (Cyprinus carpio).

Despite the fact that pharmaceuticals occur in surface water in low concentrations, they can still have adverse effect on aquatic biota, because these substances are designed to have a specific mode of action even at low concentrations. To our knowledge, only little is known about the long-term effect of tricyclic antidepressant residues on non-target animals, especially fish. Hence, the aim of this study was to assess the effect of subchronic exposure to selected tricyclic antidepressants - namely, amitriptyline, nortriptyline, and clomipramine and their mixture - on early-life stages of common carp (Cyprinus carpio). Two embryo-larval toxicity tests with common carp were performed, each taking 30 days. For the experiment, three tricyclic antidepressants as well as their mixtures were tested at three different concentrations - 10, 100 and 500 µg/L. Hatching and mortality were recorded twice a day. In addition, samples were taken at regular intervals in order to record developmental stage, morphometric and condition characteristics and morphological anomalies. At the end of the test, additional samples were taken for histopathological examination and also for the determination of antioxidant and biotransformation enzyme activity and lipid peroxidation and protein carbonylation. Long-term exposure resulted in a significant increase in mortality, developmental retardation, morphological anomalies, and pathological changes in brain, heart, and cranial and caudal kidney. In addition, changes in antioxidant enzyme activity as well as increased lipid peroxidation were observed, even at the lowest tested concentrations. Hence, environmentally relevant concentrations of TCAs have the potential to cause harmful effects on early-life stages of fish.

Development of olfactory epithelium and associated structures in the green iguana, Iguana iguana-light and scanning electron microscopic study.

The ontogenesis of the nasal cavity has been described in many mammalian species. The situation is different with reptiles, despite the fact that they have become relatively common as pets. In this study we focused on the ontogenesis of the olfactory epithelium, as well as other types of epithelia in the nasal cavity of pre-hatched green iguanas (Iguana iguana). Collection of samples began from day 67 of incubation and continued every four days until hatching. Microscopic examination revealed that significant morphological changes in the nasal cavity began approximately at day 91 of ontogenesis. Approximately at this same stage, the nasal cavity epithelium began to differentiate. The cavity was divided into two compartments by a cartilaginous disc. The ventral compartment bulged rostrally and eventually opened up into the external environment. Three clearly demarcated areas of epithelium in the nasal cavity were visible at day 107.