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.

An insight into cancer palaeobiology: does the Mesozoic neoplasm support tissue organization field theory of tumorigenesis?

BACKGROUND: Neoplasms are common across the animal kingdom and seem to be a feature plesiomorphic for metazoans, related with an increase in somatic complexity. The fossil record of cancer complements our knowledge of the origin of neoplasms and vulnerability of various vertebrate taxa. Here, we document the first undoubted record of primary malignant bone tumour in a Mesozoic non-amniote. The diagnosed osteosarcoma developed in the vertebral intercentrum of a temnospondyl amphibian, Metoposaurus krasiejowensis from the Krasiejów locality, southern Poland. RESULTS: A wide array of data collected from gross anatomy, histology, and microstructure of the affected intercentrum reveals the tumour growth dynamics and pathophysiological aspects of the neoplasm formation on the histological level. The pathological process almost exclusively pertains to the periosteal part of the bone composed from a highly vascularised tissue with lamellar matrix. The unorganised arrangement of osteocyte lacunae observed in the tissue is characteristic for bone tissue types connected with static osteogenesis, and not for lamellar bone. The neoplastic bone mimics on the structural level the fast growing fibrolamellar bone, but on the histological level develops through a novel ossification type. The physiological process of bone remodelling inside the endochondral domain continued uninterrupted across the pathology of the periosteal part. CONCLUSIONS: Based on the results, we discuss our case study's consistence with the Tissue Organization Field Theory of tumorigenesis, which locates the causes of neoplastic transformations in disorders of tissue architecture.

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.

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.

Long-term consumption of fish oil partially protects brain tissue from age-related neurodegeneration.

The purpose of this study was to determine the effect of fish oil supplementation on aging in the cerebral cortex, hippocampus and cerebellum of rats. Results of biochemical and histological analyses of brain tissue collected from young rats (age: 2 months) prior to the experiment were compared with the results obtained from the 14-month-old animals assigned to the control and supplemented group. Total polyunsaturated fatty acid (PUFA) composition and thiobarbituric acid reactive substance (TBARS) levels were assessed in the examined brain regions. Furthermore, the presence of lipofuscin and amyloid-ß, as well as the number of apoptotic and proliferative cells, was determined in the brain tissue. The analyses revealed that the number of proliferative neurons was significantly higher in the fish oil treated group in the cerebral cortex, hippocampus proper and dentate gyrus. Furthermore, in animals fed the fish oil-supplemented diet, amyloid-ß plaques were not observed in the examined brain regions. These results suggest that fish oil supplementation has a neuroprotective effect on the process of proliferation and may protect against spontaneous amyloidosis in the brain.

Influence of nanoparticles of platinum on chicken embryo development and brain morphology.

Platinum nanoparticles (NP-Pt) are noble metal nanoparticles with unique physiochemical properties that have recently elicited much interest in medical research. However, we still know little about their toxicity and influence on general health. We investigated effects of NP-Pt on the growth and development of the chicken embryo model with emphasis on brain tissue micro- and ultrastructure. The embryos were administered solutions of NP-Pt injected in ovo at concentrations from 1 to 20 µg/ml. The results demonstrate that NP-Pt did not affect the growth and development of the embryos; however, they induced apoptosis and decreased the number of proliferating cells in the brain tissue. These preliminary results indicate that properties of NP-Pt might be utilized in brain cancer therapy, but potential toxic side effects must be elucidated in extensive follow-up research.

Histopathological, histomorphometrical, and immunohistochemical biomarkers in flounder (Platichthys flesus) from the southern Baltic Sea.

Flounder (Platichthys flesus), collected in late fall of 2009 from four coastal sites in the southern Baltic Sea including the Gulf of Gdansk (GoG), were investigated for a suite of biomarkers of contaminant effects. The biomarkers included liver histopathologies, which were diagnosed and assessed using commonly applied lesion categories, the size and density of melano-macrophage aggregates (MMAs), expression of proliferating cell nuclear antigen (PCNA) and of cytochrome P450 1A (CYP1A) in liver as well as the size and density of MMAs and density of Perls'-positive cells in the spleen. The prevalence of liver lesions differed among the sites. Most frequently occurring were non-specific and early toxicopathic non-neoplastic lesions. Mean MMA size was in the range of 264-519 µm(2) and 717-2137 µm(2) in liver and spleen, respectively, and density was in the range of 6-13 and 15-26 MMAmm(-2), respectively. Mean density of PCNA-positive hepatocytes was in the range of 300-1281 cellsmm(-2). These histomorphometrical biomarkers correlated positively with the muscle Hg, Σ(7)PCB, and ΣDDT residues and negatively with the indices of general liver condition. They showed significant differences between the sites, which were in line with the spatial prevalence of liver lesions and CYP1A induction. Overall, the biomarker responses were more pronounced in the GoG sites in comparison to those outside the Gulf, which confirms some earlier results and broadens the knowledge of contaminant effects in the Polish coastal zone of the Baltic Sea.

The effect of peptide absorption on PepT1 gene expression and digestive system hormones in rainbow trout (Oncorhynchus mykiss).

The present study evaluates the effect of protein source (dipeptides, free amino acids, and intact protein) on development and growth of Salmonid fish alevin. Specifically, we follow the expression of oligopeptide transporter protein PepT1 in the intestine of rainbow trout (Oncorhynchus mykiss). Fish were fed exogenously one of four diets: three formulated (lysyl-glycine dipeptide supplemented diet - PP, free lysine and glycine supplemented diet - AA, control diet with no lysine - CON) or commercial starter (Aller Futura - AF). Fish increased mean body weight 8 fold with PP- and AA-supplemented diets resulting in significantly higher weight gain than fish fed CON. Statistical analysis revealed a significant increase in relative PepT1 expression of fish fed experimental diets. Immunohistochemical staining with PepT1 antibody showed the presence of the transporter protein in the brush border membrane of the proximal intestinal enterocytes of fish from all experimental groups. Leptin immunoreactivity occurred not only in the gastric glands but also in proximal intestine and pyloric caeca of fish fed PP, AA and AF diets. Leptin immunoreactivity was also observed in hepatocyte cytoplasm and pancreatic acinar cells. Gastrin/CCK immunoreactive cells were present in the proximal intestine and pyloric caeca.