Effects of long-term administration of various dietary prebiotic supplements on the growth, immune cell activity and digestive tract histology of juvenile vimba (Vimba vimba).

Introduction: The experiment was set to determine the effects of long-term (55-day) use of three commercial prebiotics including Saccharomyces cerevisiae-derived ß-glucans and one including inulin on juvenile vimba (Vimba vimba) reared intensively under controlled conditions. Material and Methods: Six-month-old fish were fed commercial feed (Control group, n = 90), or the same feed supplemented with 0.02% Leiber Beta-S (BS group, n = 90), 0.20% Biolex MB40 (MB group, n = 90), 0.30% CeFi (CE group, n = 90) or 1.00% inulin Orafti GR (IN group, n = 90) for 55 days. Results: In the BS group, the final growth parameters were significantly lower than in the Control group, while the feed conversion ratio was significantly higher. No significant differences were found between any other group and the Control group in the respective parameters. The respiratory burst activity of the head-kidney phagocytes was significantly lower in all fish groups fed the prebiotic-supplemented diets compared to the Control group. The proliferative response of the head-kidney lymphocytes stimulated by concanavalin A was lower in the BS group than in the Control group, while in other groups this response was not affected. No significant differences were found in histopathological analyses of the digestive tract, liver or pancreas. Conclusion: The long-term supplementation of fish diets with prebiotics can negatively influence the growth, feed conversion, nonspecific cellular resistance and proliferative activity of the T lymphocytes of vimba juveniles.

The Effect of Silver Nanoparticles on the Digestive System, Gonad Morphology, and Physiology of Butterfly Splitfin (Ameca splendens).

The aim of this study was to determine the effects of silver nanoparticles (AgNPs) on the morphology and enzymatic activity of butterfly splitfin (Ameca splendens). Individuals of both sexes, aged about five months, were exposed to AgNPs at concentrations of 0 (control group), 0.01, 0.1, and 1.0 mg/dm3 for 42 days. On the last day of the experiment, the fish were euthanized, subjected to standard histological processing (anterior intestine, liver, and gonads), and analysed for digestive enzyme activity in the anterior intestine and oxidative stress markers in the liver. Fish in the AgNP 0.01 and 0.1 groups had the lowest anterior intestinal fold and enterocyte height. However, there were no statistically significant changes in the digestive enzyme activity in the anterior intestine. Analysis of enzymatic activity in the liver showed an increase in superoxide dismutase activity in fish in the AgNP 0.1 group. Histological analyses showed that AgNPs inhibited meiotic divisions at prophase I in a non-linear manner in ovaries and testes. In the AgNP 0.1 and 1.0 groups, the area occupied by spermatocytes was lower compared to the other groups. These results indicate that exposure to AgNPs may lead to disturbances in morphology and enzymatic activity in the liver and intestine and may lead to disruption of reproduction in populations.

Histomorphometric evaluation of melanomacrophage centers (MMCs) and CD3+ T cells of two morphs of brown trout (Salmo trutta) fed diets with immunostimulants.

The brown trout (Salmo trutta) is a salmonid residing in riverine and coastal waters throughout the Northern Hemisphere, whose various populations evolved into distinct ecological morphs, differing in their migratory tendencies and preferred habitats. Unfortunately, due to progressing degradation of natural environment, the conservation of these populations is of growing importance and is undoubtedly a challenging task. Therefore, various means to refine the preparatory protocols for restocking using hatchery-reared fish are being pursued, some of which involve the administration of immunity-boosting substances. The current study assessed the effects of two dietary immunostimulants: Bioimmuno (4% inosine pranobex and 96% ß-glucan) and Focus Plus (commercial preparation by Biomar, Denmark) on two morphs of the brown trout - the river trout (S. trutta morpha fario) and the sea trout (S. trutta morpha trutta). Tissue samples were obtained from ∼75 to 100g fish after 0, 2 and 4 weeks of experimental feeding. Multi-factorial analysis of conducted histological measurements of melanomacrophage centers (MMCs) revealed no changes of their parameters within spleens, but showed a decrease of the occupied tissue area and MMC counts in the livers, progressing with time regardless of the applied diet. Immunohistochemical analysis of CD3+ T cells showed their increased recruitment into mucosal folds of pyloric caeca in the 2-week sampling of trouts fed with the diet with 2% Bioimmuno addition, but this effect was not present in the 4-week sampling. When studying all groups jointly within each morph, there was a significant difference in terms of maintained CD3+ T cells levels, as sea trouts showed significantly higher tissue areas occupied by these cells than river trouts, both in the pyloric caeca and hepatic parenchyma. The study revealed that feeding with a diet enriched with Bioimmuno for 2 weeks may be a favorable enhancement of rearing protocols of brown trout stocks prior to their release, but more studies need to be conducted to test the possibility of an even shorter feeding period.

Effect of pearl guinea fowl eggshell ultrastructure and microstructure on keets hatchability.

Variability in shell structure is an evolutionary mechanism in birds that enables them to adapt to specific environmental conditions. This variability may also occur within the same species under the influence of individual indicators, such as the age or health status of females. While interspecies variation is quite obvious and easy to interpret, the reasons for intraspecies variation remain unclear. In this study, we examined the ultra- and microstructure of guinea fowl eggshells to identify the association between variations in shell structure and hatchability outcomes. We analyzed the visual differences between shells with low (L), intermediate (I), and high (H) external porosity using scale invariant feature transform analysis with NaturePatternMatch software. We found that the external pore image was closely related to the overall porosity of the shell before incubation. The total pore area, total porosity, and diffusion index (GH2O) were highest in group H shells (P < 0.001). Posthatching shells were characterized by an increased diameter and total surface area, decreased pore number (P < 0.001), as well as shortened mammillary layer (P < 0.001) and decreased total consumption of mammillary knobs (P < 0.001). The porosity indices of posthatching H shells had intermediate values between L and I. Although the effect of shell structure parameters on hatching was not confirmed, we assumed that all categories (L, I, and H) of shells were ideal for incubation. This suggests that the shell structure adapts to the metabolic rate of developing embryos; however, differences in shell structure affect the duration of incubation and synchronization of hatching. Both L and H shells showed delayed and prolonged hatching. Therefore, we recommended that guinea fowl eggs with different external porosity parameters should be incubated separately for better hatching synchronization. Differences in GH2O between L, I, and H eggs suggest that the shell porosity characteristics of guinea fowl eggs may be a key determinant of the rate of water loss during storage before incubation.

The Effects of Seven-Day Exposure to Silver Nanoparticles on Fertility and Homeostasis of Zebrafish (Danio rerio).

Silver nanoparticles (AgNPs) are found in open waters, but the effect of their low concentrations on an organism's homeostasis is not fully understood. The aim of the study was to determine the short-term exposure effects of AgNPs coated by PvP (polyvinylpyrrolidone) on the homeostasis of livers and gonads in zebrafish. Sexually mature zebrafish were exposed for seven days to silver ions (0.01 mg/dm3) or AgNPs (0.01; 0.05; 0.1; 0.5; 1.0 mg/dm3). On the last day, the liver, testes, and ovaries were subjected to a histology analysis. In the liver, we analyzed the expression of the cat, gpx1a, gsr, sod1, and cyp1a genes. On the last day of the experiment, the lowest survival rate was found in the AgNPs 0.05 mg/dm3 group. The histological analysis showed that AgNPs and silver ions cause an increase in the area of hepatocytes. The highest proliferation index of hepatocytes was found in the AgNP 0.05 mg/dm3 group. Furthermore, AgNPs were found to interfere with spermatogenesis and oogonesis as well as reduce the expression levels of the cat, gpx1a, and sod1 genes in the liver compared with the control group. Based on the results, it can be concluded that exposure to AgNPs causes cytotoxic changes in zebrafish, activates the immune system, negatively affects the process of meiosis in the gonads, and generates oxidative stress.

Tollip-deficient zebrafish display no abnormalities in development, organ morphology or gene expression in response to lipopolysaccharide.

Tollip is a multifunctional adaptor protein implicated in innate immunity, lysosomal trafficking/autophagy of protein aggregates and various signaling processes in mammalian models. To verify evolutionary conservation of these functions, we used CRISPR/Cas9 editing to construct a zebrafish line bearing a stable tollip knockout. In contrast to previously reported tollip morphants, Tollip-deficient fish display normal development until adulthood, are fertile, and have no apparent physiological defects. When challenged with lipopolysaccharide (LPS), inflammatory gene expression is unaffected. Moreover, Tollip deficiency does not aggravate swimming deficiency resulting from lysosomal dysfunction and proteotoxicity in a fish model of Gaucher disease. Thus, individual functions of Tollip may be organism-specific or manifest only upon certain conditions/challenges or disease backgrounds.

Domestication affected stress and immune response markers in Perca fluviatilis in the early larval stage.

It is already known that domestication modifies stress and immune responses in juveniles and adults of several fish species. However, there is a lack of information on whether these modulations result from adaptability along the life cycle or if they are pre-determined in very early developmental stages. To shed light on mechanisms that help to explain the process of domestication, a study was conducted to analyze comparatively Eurasian perch larval performance, stress, and immune status between wild and domesticated specimens. Eurasian perch larvae obtained from wild and domesticated (generation F5 reared in recirculating aquaculture systems) spawners were reared in the same conditions during the main rearing trial (MRT) and also subjected to a thermal challenge (TC). During the study, larval performance (including survival, growth performance, swim bladder inflation effectiveness, deformity rate), the expression of genes involved in immune and stress response, and the specific activity of oxidative stress enzymes (during MRT only) were analyzed. No significant differences in hatching rate, deformity rate, or swim bladder inflation effectiveness between wild and domesticated larvae were found, whereas specific growth rate, final total length, and wet body weight were significantly lower in wild larvae. Higher mortality was also observed in wild larvae during both MRT and TC. The data obtained in this study clearly indicated that during domestication, significant modifications in stress and immune response, such as complement component c3, were noted as early as just after hatching. Generally, domesticated fish were characterized by a lower stress response and improved immune response in comparison to the wild fish. This probably resulted from the domesticated larvae being better adapted to the conditions of artificial aquaculture. The data obtained provided information on how domestication affects fish in aquaculture, and they contribute to the development of efficient selective breeding programs of Eurasian perch and other freshwater teleosts.

The Role of Transcription Factors in Gonad Development and Sex Differentiation of a Teleost Model Fish-Guppy (Poecilia reticulata).

The guppy (Poecilia reticulata) is one of the world's most popular ornamental fish. Due to lecithotrophic viviparous, it is commonly used in toxicological studies and environmental monitoring. This study aimed to investigate the molecular mechanisms of gonad development and differentiation during guppy ontogenesis. The study mainly focused on the role and localization of potential specific sex markers and transcription factors: Sox9, Dmrt1, Erß. For histological analysis, guppies of both sexes were collected at 1, 60, and 360 dph (day post-hatching). The gonads morphology and immunohistochemistry detection of mentioned markers localization were performed. The expression of Sox9 protein was compared between sexes. Histological analysis revelated all types of male germinal cells in 60 dph guppy's testes. Maturated oocytes were visible in the ovaries of 360 dph fish. The Sox9 expression varied in spermatocytes and spermatids, from cellular to nuclear localization, and was higher in ovaries. Dmrt1 was detected in all testes groups and 360 dph ovaries. The Erß was observed in both sexes at 60 and 360 dph. For the first time, the localization of transcription factors in guppy during ontogenesis was traced. The Sox9 designation as a factor regulating the development of germinal cells in adult guppies may facilitate the analysis of xenobiotics' influence on fish's reproductive system.

Effect of in ovo application of hydroxyapatite nanoparticles on chicken embryo development, oxidative status and bone characteristics.

Intensive selection in modern lines of fast-growing chickens has caused several skeletal disorders. Therefore, current research is focused on methods to improve the bones of birds. A new potential solution is in ovo technology using nanoparticles with a high specificity for the bone tissue. Thus, the objective of the present study was to evaluate the effect of in ovo application of hydroxyapatite nanoparticles (HA-NP) in different concentrations (50, 100 and 500 µg/ml colloids) on chicken embryo development, with a particular focus on the oxidative status and bone characteristics of the embryo. The results showed that in ovo treatment with HA-NP did not negatively affect hatchability and body weight. However, bone weight was reduced in 500 µg/ml group. The concentrations of calcium, phosphorus and crude ash were not affected. The modulatory effect of HA-NP was observed on the basis of antioxidative markers - superoxide dismutase, total antioxidant status, malondialdehyde in serum and selected tissues. Glutathione concentration in serum suggested higher metabolic stress. Among bone turnover markers, the concentration of osteocalcin was found to be significantly affected by HA-NP injection. Thus, the in ovo application of HA-NP could modify the molecular responses at the stage of embryogenesis but these changes were not reflected in embryo growth and even slowed down bone development. Nevertheless, the question for the follow-up research is whether in ovo administration of HA-NP would affect the antioxidative status and bone turnover resulting in improved bone conditions and body gain in post hatch chickens.

Author Correction: Domestication process modifies digestion ability in larvae of Eurasian perch (Perca fluviatilis), a freshwater Teleostei.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Domestication process modifies digestion ability in larvae of Eurasian perch (Perca fluviatilis), a freshwater Teleostei.

To date, a comparative analysis of larval performance and digestion abilities between wild and domesticated Eurasian perch has not yet been performed. Eurasian perch larvae from wild and domesticated spawners were reared in the same conditions and at different development stages, growth performance variables, the expression of genes encoding digestive enzymes and specific enzymatic activity were analysed. No significant differences in hatching rate, deformity rate or swim bladder inflation effectiveness between wild and domesticated larvae were found. Specific growth rate, final total length and wet body weight were significantly lower in wild larvae, whereas higher mortality in wild larvae was observed compared to domesticated larvae. The data obtained in this study clearly indicate that during domestication, significant modification of digestion ability occurs at the very beginning of ontogeny, where domesticated fish are characterised by lower enzymatic activity and lower expression of genes encoding digestive enzymes. This probably results from the low diversity of the food offered in culture conditions, which significantly modified digestion capability. The obtained data provide an understanding of how domestication affects fish in aquaculture and may improve the planning of selective breeding programs of Eurasian perch and other freshwater Teleosts.

Acute exposure of zebrafish (Danio rerio) larvae to environmental concentrations of selected antidepressants: Bioaccumulation, physiological and histological changes.

Antidepressants have been detected in surface waters worldwide at ng-µg/L concentration. These compounds can exert adverse effects on fish even at low levels. But, all previous analyses have concentrated on adult fish. The aim of the study was to assess the effect of environmental concentrations of sertraline, paroxetine, fluoxetine and mianserin, and their mixtures on such unusual endpoints as physiological and histological changes of zebrafish (Danio rerio) larvae. We also determined the bioconcentration of the pharmaceuticals. Fish Embryo Toxicity test was used to analyze the influence on developmental progression. Histological sections were stained with hematoxylin and eosin. Proliferating cells in liver were determined immunohistochemically by detection of Proliferating Cell Nuclear Antigens. The bioconcentration factor was measured by liquid chromatography coupled to mass spectrometry. Pharmaceuticals were used at low, medium and high concentrations in mixtures and at medium concentration as single compound. Exposure to the analyzed pharmaceuticals increased the rate of abnormal embryo and larvae development, accelerated the hatching time and affected the total hatching rate. Three-times lower proliferation of hepatocytes was observed in larvae exposed to paroxetine, mianserin, sertraline and the mixture of the pharmaceuticals at the highest concentrations. The highest bioaccumulation factor (BCF) was obtained for sertraline. The BCF of the analyzed compounds was higher if the organisms were exposed to the mixtures than to single pharmaceuticals. To conclude, the exposure of zebrafish larvae to selected antidepressants and their mixtures may cause disturbances in the organogenesis of fish even at environmental concentrations.