Caudal Fin Tissue Research Articles

Glyphosate- and aminomethylphosphonic acid (AMPA)-induced mortality and residues in juvenile brown trout (Salmo trutta f. fario) exposed at different temperatures

BackgroundGlyphosate is a broad-spectrum, non-selective systemic herbicide with a commonly assumed low potential for accumulation in biota. Nevertheless, glyphosate has been shown to bioaccumulate in the tissues of several organisms. To understand the bioconcentration dynamics of glyphosate in fish, brown trout (Salmo trutta forma fario) of different age were exposed to different concentrations of glyphosate, the formulation Roundup® LB Plus, and the major transformation product aminomethylphosphonic acid (AMPA) for two, three, or four weeks at different temperatures in the laboratory. Mortality rates were determined, and tissue samples were collected at the end of the experiment to ascertain concentrations of glyphosate and AMPA residues by liquid chromatography coupled to mass spectrometry (LC–MS/MS).ResultsBrown trout mortality during exposure to glyphosate or AMPA was considerably higher at 15 °C than at 7 °C. Also, a significant increase in glyphosate concentrations in samples containing muscle, head, backbone, and caudal fin tissue with increasing exposure concentrations and temperatures was observed. Six-month-old fish contained more glyphosate per kg wet weight after exposure than ten-month-old fish. The bioconcentration factors (BCFs) for glyphosate and AMPA were much higher at 15 °C than at 7 °C, but in both cases decreased with higher glyphosate concentrations. The BCF for glyphosate formulated in Roundup® was higher than the one for the parent compound. Approximately 30–42% of the organ-absorbed glyphosate and AMPA remained in the tissues even when the fish were kept in clean water lacking the test substances for three weeks after termination of exposure.ConclusionOur study demonstrated that there is an interaction between glyphosate and ambient temperature in terms of toxicity. Further it was shown that increasing concentrations of glyphosate and AMPA in the surrounding media lead to significantly increased concentrations of these substances in brown trout tissues, although neither bioconcentration nor bioaccumulation of glyphosate in animal tissues is expected due to the high water solubility of this chemical. As a consequence, the uptake of glyphosate by humans through the consumption of contaminated edible fish is very likely.

Unlocking the genomes of formalin‐fixed freshwater fish specimens: An assessment of factors influencing DNA extraction quantity and quality

AbstractObjectiveRecent technological developments may facilitate the description of evolutionary relationships and population genetic structure as well as other information relevant to fisheries management by using readily available natural history collections. Contemporary sequence capture and short‐read sequencing methods offer opportunities to analyze highly fragmented DNA from formalin‐fixed specimens so long as enough DNA of sufficient quality is recovered.MethodsWe compared two protocols developed to extract DNA from formalin‐fixed tissues using specimens of three freshwater fishes: the Southern Brook Lamprey Ichthyomyzon gagei, Slimy Sculpin Cottus cognatus, and Brown Trout Salmo trutta. Extractions were attempted using hot alkali digestion with and without buffer wash pretreatments to compare the DNA concentration, purity, and fragment length of DNA recovered between extraction protocols, tissue types (muscle and caudal fin tissue for Brown Trout and Slimy Sculpin), and preservation periods (5 or 7 years for Southern Brook Lamprey).ResultLikelihood models generally did not detect DNA quantity differences between extraction protocols nor tissue types; however, 6.0–8.7× more DNA was recovered from Slimy Sculpin caudal fins than from muscle tissue. Extraction protocol had mixed effects on DNA purity; the wash protocol outperformed the no‐wash protocol for Slimy Sculpin and Brown Trout, but the reverse was true for the lamprey. Purer DNA was recovered from the caudal fins; however, fragment lengths were generally greater from muscle tissue for both ray‐finned species.ConclusionOur results suggest that the best tissue for sampling may depend on the quality metric considered most important for a study's objectives and that omitting time‐consuming tissue wash steps can yield DNA of quantity and quality comparable to DNA from more complex methods. Regardless of species, the DNA extracted from most samples using both protocols met quantity and quality thresholds that are likely to result in short‐read sequencing success. These results provide optimism for unlocking the wealth of genetic information in natural history collections for use in fisheries management and conservation genomics.

Genome-wide association study (GWAS) analysis of black color trait in the leopard coral grouper (Plectropomus leopardus) using whole genome resequencing

The leopard coral grouper (Plectropomus leopardus) is a coral reef fish species that exhibits rapid and diverse color variation. However, the presence of melanoma and the high proportion of individuals displaying black color in artificial breeding have led to reduced economic and ornamental value. To pinpoint single nucleotide polymorphisms (SNPs) and potential genes linked to the black pigmentation characteristic in this particular species, This study gathered a cohort of 360 specimens from diverse origins and conducted a comprehensive genome-wide association analysis (GWAS) employing whole-genome resequencing. As a result, 57 SNPs related to the black skin trait were identified, and a grand total of 158 genes were annotated within 50 kb of these SNPs. Subsequently, GWAS was applied to three populations (LED, QHH, and QHL), and the corresponding results were compared with the analysis results of the total population. The results of the four GWAS models showed significant enrichment in Rap1 signaling pathway, melanin biosynthesis, metabolic pathways, tyrosine metabolism, cAMP signaling pathway, AMPK signaling pathway, PI3K-Akt signaling pathway, EGFR tyrosine kinase inhibitor resistance, HIF-1 signaling pathway, Ras signaling pathway, MAPK signaling pathway, etc. (p < 0.05), which were mainly associated with eleven genes (POL4, MET, E2F2, COMT, ZBED1, TYRP2, FOXP2, THIKA, LORF2, MYH16 and SOX2). Significant differences (p < 0.05) were observed in the expression of all 11 genes in the dorsal skin tissue, in 10 genes except COMT in the ventral skin tissue, and in all 11 genes in the caudal fin tissue. These findings imply that the control of body color in the P. leopardus is the result of the joint action of multiple genes and signaling pathways. These findings will contribute to a more profound comprehension of the genetic attributes that underlie the development of black skin in the vibrant P. leopardus, thus furnishing a theoretical foundation for genetic enhancement.

Caudal fin as a proxy for dorsal muscle for nutrient enrichment monitoring using stable isotope analysis: the case of Gerres filamentosus and G. oyena from mangrove creeks of Tanzania

The use of fish dorsal muscles in stable isotope studies, which is an invasive method that results in fish deaths, limits their applicability for rare and endangered fish species, as well as when large sample sizes and replicates are required, prompting research into feasible non-lethal sampling methods. The possibility of employing fin clippings (a non invasive approach) was investigated as a proxy for dorsal muscle in nutrient pollution monitoring studies using two common mangrove fish species, namely Gerres filamentosus and G. oyena, which are known to spend their early life stages primarily within mangroves. The dorsal muscles and caudal fin tissues of fish from the mangrove creeks of Kunduchi and Mbegani, Tanzania, were examined for 13C and 15N signatures. Dorsal muscles from Kunduchi (mean SD: 13C = -16.8 ± 2.86, 15N = 9.34±1.15) were more enriched than from Mbegani (mean SD: 13C = -18.60 ± 2.11, 15N = 7.27±1.09), and this enrichment was consistent across the two studied species. Caudal fins indicated similar enrichment trends. Fin tissue stable isotope values explained between 62 % and 87 % of dorsal muscle 13C and between 89 % and 98 % of dorsal muscle 15N variability. These findings support the use of fin-clipping as a non-lethal proxy for stable isotope analysis of the studied species for nutrient enrichment, and additional research into non-lethal sampling methods is recommended.

Regulatory T cells regulate blastemal proliferation during zebrafish caudal fin regeneration.

The role of T cells in appendage regeneration remains unclear. In this study, we revealed an important role for regulatory T cells (Tregs), a subset of T cells that regulate tolerance and tissue repair, in the epimorphic regeneration of zebrafish caudal fin tissue. Upon amputation, fin tissue-resident Tregs infiltrate into the blastema, a population of progenitor cells that produce new fin tissues. Conditional genetic ablation of Tregs attenuates blastemal cell proliferation during fin regeneration. Blastema-infiltrating Tregs upregulate the expression of igf2a and igf2b, and pharmacological activation of IGF signaling restores blastemal proliferation in Treg-ablated zebrafish. These findings further extend our understandings of Treg function in tissue regeneration and repair.

Understanding the complexity of epimorphic regeneration in zebrafish caudal fin tissue: A transcriptomic and proteomic approach.

The complex epimorphic regeneration of zebrafish caudal fin tissue is hasty and absolute. This study was executed to understand the role of various genes/proteins involved in the regeneration of zebrafish caudal fin tissue through differential transcriptomics and proteomics analysis. Based on our study 1408 genes and 661 proteins were found differentially regulated in the regenerating caudal fin tissue for having at least 1-log fold change. Interleukin, Solute carrier, Protein arginine methyltransferase, Homeobox, Neurotransmitter and several novel genes were found to be associated with regeneration for its differential regulation during the mechanism. Based on the network and pathway analysis the differentially regulated genes and proteins were found allied with activation of cell proliferation, cell viability, cell survival & cell movement and inactivation of organismal death, morbidity, necrosis, death of embryo & cell death. This study has mapped a detailed insight of the genes/proteins expression associated with the epimorphic regeneration more profoundly.

A DNA methylation age predictor for zebrafish.

Changes in DNA methylation at specific CpG sites have been used to build predictive models to estimate animal age, predominantly in mammals. Little testing for this effect has been conducted in other vertebrate groups, such as bony fish, the largest vertebrate class. The development of most age-predictive models has relied on a genome-wide sequencing method to obtain a DNA methylation level, which makes it costly to deploy as an assay to estimate age in many samples. Here, we have generated a reduced representation bisulfite sequencing data set of caudal fin tissue from a model fish species, zebrafish (Danio rerio), aged from 11.9-60.1 weeks. We identified changes in methylation at specific CpG sites that correlated strongly with increasing age. Using an optimised unique set of 26 CpG sites we developed a multiplex PCR assay that predicts age with an average median absolute error rate of 3.2 weeks in zebrafish between 10.9-78.1 weeks of age. We also demonstrate the use of multiplex PCR as an efficient quantitative approach to measure DNA methylation for the use of age estimation. This study highlights the potential further use of DNA methylation as an age estimation method in non-mammalian vertebrate species.

The effects of Piper sarmentosum aqueous extracts on zebrafish (Danio rerio) embryos and caudal fin tissue regeneration

In Malaysia, Piper sarmentosum or ‘kaduk’ is commonly used in traditional medicines. However, its biological effects including in vivo embryonic toxicity and tissue regenerative properties are relatively unknown. The purpose of this study was to determine zebrafish (Danio rerio) embryo toxicities and caudal fin tissue regeneration in the presence of P. sarmentosum aqueous extracts. The phytochemical components and antioxidant activity of the extract were studied using GC–MS analysis and DPPH assay, respectively. Embryo toxicity tests involving survival, heartbeat, and morphological analyses were conducted to determine P. sarmentosum extract toxicity (0–60 µg/mL); concentrations of 0–400 µg/mL of the extract were used to study tissue regeneration in the zebrafish caudal fin. The extract contained several phytochemicals with antioxidant activity and exhibited DPPH scavenging activity (IC50 = 50.56 mg/mL). Embryo toxicity assays showed that a concentration of 60 μg/mL showed the highest rates of lethality regardless of exposure time. Slower embryogenesis was observed at 40 µg/mL, with non-viable embryos first detected at 50 µg/mL. Extracts showed significant differences (p < 0.01) for tissue regeneration at all concentrations when compared to non-treated samples. In conclusion, Piper sarmentosum extracts accelerated tissue regeneration, and extract concentrations at 60 µg/mL showed the highest toxicity levels for embryo viability.

CARACTERIZACIÓN GENÉTICA DE REPRODUCTORES DE PROCHILODUS MAGDALENAE (PISCES: PROCHILODONTIDAE) USADOS EN PROGRAMAS DE REPOBLAMIENTO EN COLOMBIA

Prochilodus magdalenae is an endemic fish species of Colombia known as an important resource of commercial interest for many communities related to fishing activities as a livelihood activity. However, population deterioration has been observed in natural environments due to factors such as overfishing, fragmentation of ecosystems, among others. This makes it necessary to characterize the genetic diversity of P. magdalenae in the productive systems of some fish farms, which are used to restocking in other basins of Colombia and, thus, to propose technical and scientific criteria that allow the development of management strategies for the conservation of this species. Therefore, in 2013, caudal fin tissue was collected from 1044 individuals in seven fish farms, which were processed in the laboratory. Through the use of seven microsatellites, genetic metrics such as: observed and expected heterozygosity, number of alleles, fixation indexes, F statistics, Bayesian grouping and AMOVA were evaluated. We observed low heterozygosity, correlated with inbreeding processes, which contrast with the high values obtained in the expected heterozygosity index and the number of alleles detected in P. magdalenae productive systems. A moderate genetic differentiation between fish centers was detected and the existence of three genetic groups was observed through the Bayesian analysis. Despite the low diversity reported regarding the others species of the same genus, populations held in Bocachico captivity have the potential to restore the diversity of wild populations. Therefore, it is suggested that each fish station should establish batches of breeders separately, based on their genetic information so that there is congruence between the released individuals and those that inhabit the natural environment. Key words: Bocachico; Reophilic fish; Magdalena Basin; Genetic structure; Fish farms.

Functional role of annexins in zebrafish caudal fin regeneration – A gene knockdown approach in regenerating tissue

Regeneration is an adaptive phenomenon with wide biological implications spread heterogeneously in almost all the organism including human beings. The ability of regeneration varies from species to species for its complexity. Epimorphic regeneration of zebrafish caudal fin tissue is the most widely studied regeneration mechanism for its discrete and rapid regenerative capability. Several genes and proteins were found to be associated with regenerative mechanisms of zebrafish caudal fin tissue. In this study we have evaluated the functional role of Annexin 2a and 2b genes during zebrafish caudal fin tissue regeneration using inventive electroporation techniques for targeting the gene involving CRISPR-Cas9 technology. The electroporation of the CRISPR was performed on the adult zebrafish caudal fin tissue post amputation. We report retarded growth during the regeneration of caudal fin tissue when Annexin 2a and 2b genes were knocked down, which was validated through gene expression & sequencing analysis and further supported by high-throughput quantitative proteomic analysis of the fin tissue. Annexin family genes such as ANXA13, ANXA1a, ANXA5b were also found to be repressed with their expression. Knocking down of ANXA2a and 2b in regenerating caudal fin tissue compromises regenerating capacity as these genes are involved in cell to cell communication and extracellular matrix growth. This study proves that ANXA2a and 2b plays a significant role in epimorphic regeneration of zebrafish caudal fin tissue.

Bioconcentration of Essential and Nonessential Elements in Black Sea Turbot (Psetta Maxima Maeotica Linnaeus, 1758) in Relation to Fish Gender

This study investigates the influence of gender in the bioconcentration of essential and nonessential elements in different parts of Black Sea turbot (Psetta maxima maeotica) body, from an area considered under high anthropogenic pressure (the Constanta City Black Sea Coastal Area in Romania). A number of 13 elements (Ca, Mg, Na, K, Fe, Zn, Mn, Cu, Ni, Cr, As, Pb and Cd) were measured in various sample types: muscle, stomach, stomach content, intestine, intestine content, gonads, liver, spleen, gills and caudal fin. Turbot adults (4–5 years old) were separated, according to their gender, into two groups (20 males, 20 females, respectively), and a high total number of samples (1200 from both groups) were prepared and analyzed, in triplicate, with Flame Atomic Absorption Spectrometry and High-Resolution Continuum Source Atomic Absorption Spectrometry with Graphite Furnace techniques. The results were statistically analyzed in order to emphasize the bioconcentration of the determined elements in different tissues of wild turbot males vs. females, and also to contribute to an upgraded characterization of the Romanian Black Sea Coast, around Constanta City, in terms of heavy metals pollution. The essential elements Mg and Zn have different roles in the gonads of males and females, as they were the only elements with completely different patterns between the analyzed groups of specimens. The concentrations of studied elements in muscle were not similar with the data provided by literature, suggesting that chemistry of the habitat and food plays a major role in the availability of the metals in the body of analyzed fish species. The gender influenced the bioaccumulation process of all analyzed elements in most tissues since turbot male specimens accumulated higher concentration of metals compared to females. The highest bioaccumulation capacity in terms of Ca, Mg, Na, Ni, As, Zn and Cd was registered in caudal fin, liver and intestine tissues. Also, other elements such as K, Fe, Cu and Mn had the highest bioaccumulation in their muscle, spleen, liver and gills tissues. The concentrations of toxic metals in Black Sea turbot from this study were lower in the muscle samples compared with the studies conducted in Turkey, suggesting that the anthropogenic activity in the studied area did not pose a major impact upon the habitat contamination.

The influence of diet composition and tissue type on the stable isotope incorporation patterns of a small‐bodied southern African minnow Enteromius anoplus (Cypriniformes, Cyprinidae)

In trophic ecology, the use of stable isotope data relies on the general understanding of isotope turnover rates and diet-to-tissue discrimination factors (DTDFs). Recent studies on the application of stable isotope data have shown that isotope turnover rates and DTDFs can be influenced by many factors, including diet composition and tissue type. This study investigated the influence of diet composition and tissue type on stable isotope incorporation patterns in a small-bodied African minnow, the chubbyhead barb Enteromius anoplus. The isotopic incorporation patterns of carbon (δ13 C values) and nitrogen (δ15 N values) into white muscle and caudal fin tissues of the chubbyhead barb were examined using two isotopically different diets. Controlled-diet stable isotope feeding trials using a fishmeal-based diet (diet 1) and a soya-based diet (diet 2) were conducted over a 180-day period for the chubbyhead barb. The two diets had contrasting isotopic incorporation patterns: diet 1 was associated with progressively high δ13 C and δ15 N values, whereas diet 2 was associated with progressively low δ13 C and δ15 N values over time for both muscle and fin tissues. The δ13 C turnover rates were similar for both tissues (56 and 61 days), whereas the δ15 N turnover rates differed between fin and muscle tissue in both diets (diet 1 = 4 and 130 days, and diet 2 = 72 and 300 days, respectively). The DTDFs were similar for both tissues in diet 1 (Δ13 C: -3.96 to -2.62‰, Δ15 N: 1.98 to 2.61‰) and diet 2 (Δ13 C: 4.05 to 5.24‰, Δ15 N: 8.45 to 9.69‰). These results suggest that fin tissue can potentially be used as an alternative for muscle tissue in food web studies with a reasonable level of error. The isotopic turnover rate and DTDFs estimates for E. anoplus, however, require consideration of diet composition because different diets may differ in their isotopic incorporation patterns.

Zebrafish fin-derived fibroblast cell line: A model for in vitro wound healing.

The goal of this study was to develop and characterize a cell line from the caudal fin tissue of zebrafish and also its application as an in vitro model to study the effect of H2 O2 in wound healing. Fibroblastic cell line was developed using explant culture method from caudal fin tissue of zebrafish and characterized. This cell line was named as DrF cell line. The DrF cells treated with 0-10µM/ml H2 O2 were tested for viability, proliferation and motility by MTT assay, trypan blue assay and chemotaxis assay, respectively. Among the different concentrations of H2 O2 , 4µM was found to be nontoxic to study cell migration in in vitro scratch wound assay. Furthermore, the expression of proliferating cell nuclear antigen (PCNA) and chemokine receptor (CXCR4) genes was carried by qPCR. The cell survival, proliferation and migration were extremely enriched at 4µM level of H2 O2 . We observed accelerated wound closure in DrF cells treated with H2 O2. The qPCR results indicated that H2 O2 markedly up-regulated mRNA expression of PCNA and CXCR4. The findings from our study suggest that H2 O2 at low levels promotes cell survival, proliferation, migration and wound healing in DrF cells.

A novel cell line established from caudal fin tissue of Carassius auratus gibelio is susceptible to cyprinid herpesvirus 2 infection with the induction of apoptosis

Herpesviral haematopoietic necrosis induced by cyprinid herpesvirus 2 (CyHV-2) in Carassius auratus gibelio has caused huge economic losses in aquaculture. To better characterize this pathogen at molecular and cellular levels, it is necessary to study its pathogenesis in a sustainable cell line. In this study, a new cell line (GiCF) from the caudal fin of Carassius auratus gibelio (C. a. gibelio) was established and characterised. Typical cytopathic effect represented by characteristic morphological changes was induced in the cells at the 7th day post infection with CyHV-2, which was isolated from diseased C. a. gibeli. The susceptibility of GiCF to CyHV-2 was further demonstrated by the presence of viral assembling factory in the cytoplasm unveiled by transmission electron microscopy, and viral protein synthesis through immunofluorescence and Western blot assays. Sustainable propagation of CyHV-2 in GiCF had been demonstrated by virus infection and titration assays in a consecutive manner. The virus titer reached 104.9±0.22 TCID50/ml, and infectious CyHV-2 was produced from the GiCF cells over 30 subcultures from the established cell lines. In addition, apoptosis was observed in the GiCF cells upon CyHV-2 infection by DAPI staining, TUNEL and Annexin V staining, and quantitative expression patterns of caspase genes suggested that caspase activation might involve in CyHV-2 induced apoptosis. In summary, a new cell line, GiCF, susceptible to CyHV-2 infection was established from caudal fin tissue of C. a. gibelio, and it should serve as an ideal infection platform to characterize CyHV-2 and its pathogenic effect including apoptosis.

Monitoring changing cellular characteristics during the development of a fin cell line from Cyprinus carpio.

The establishment and in-depth characterization of a novel continuous cell line derived from fin tissue of common carp (Cyprinus carpio), CCApin, is reported. The cells of the cell line could be propagated in Leibovitz's L-15 medium containing 15% foetal calf serum and 0.5% carp serum for >150 passages during the last 24 months, with a stable fast growth. Furthermore, antibody staining indicated that cell types obtained in primary cultures, containing the epithelial stem-cell marker tumorprotein 63, were different from cells in long-term cell cultures, containing tight junction protein zona occludens 1 and cytokeratin 7. These observations suggest a switch of dominant cell types. Molecular analysis of gene expression profiles of caudal fin tissue and CCApin cells showed that genes relevant in epithelial cells but also in mesenchymal cells were expressed. However, during cultivation of CCApin a set of very steadily expressed, primarily mesenchymal genes like collagen 1 alpha 1, fibronectin or cadherin 2 was found. In summary, the long-term cell culture could be described as a stably growing epithelial population with some mesenchymal features. There are several application possibilities, especially for virus susceptibility studies, e.g. cyprinid herpesvirus-3 (CyHV-3). The study leads to a better understanding of molecular and physiological mechanisms of in vitro fish cell cultures.

De novo transcriptome analysis of Bagarius yarrelli (Siluriformes: Sisoridae) and the search for potential SSR markers using RNA-Seq.

BackgroundThe yellow sisorid catfish (Bagarius yarrelli) is a carnivorous freshwater fish that inhabits the Honghe River, Lanchangjiang River and Nujiang River of southern China and other Southeast Asian countries. However, the publicly available genomic data for B. yarrelli are limited.Methodology and principal findingsIllumina Solexa paired-end technology produced 1,706,456 raw reads from muscle, liver and caudal fin tissues of B. yarrelli. Nearly 5 Gb of data were acquired, and de novo assembly generated 14,607 unigenes, with an N50 of 2006 bp. A total of 9093 unigenes showed significant similarities to known proteins in public databases: 4477 and 6391 of B. yarrelli unigenes were mapped to the Gene Ontology (GO) and Clusters of Orthologous Groups (COG) databases, respectively. Moreover, 9635 unigenes were assigned to 242 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. In addition, 8568 microsatellites (simple sequence repeats, SSRs) were detected, and 31 pairs of polymorphic primers were characterized using wild populations of B. yarrelli from the Nujiang River, Yunnan Province, China.Conclusion/SignificanceThese sequences enrich the genomic resources for B. yarrelli and will benefit future investigations into the evolutionary and biological processes of this and related Bagarius species. The SSR markers developed in this study will facilitate construction of genetic maps, investigations of genetic structures and germplasm polymorphism assessments in B. yarrelli.

Validating fin tissue as a non-lethal proxy to liver and muscle tissue for stable isotope analysis of yellow perch (Perca flavescens)

ABSTRACTStable isotope ecology typically involves sacrificing the animal to obtain tissues. However, with threatened species or in long-term longitudinal studies, non-lethal sampling techniques should be used. The objectives of this study were to (1) determine if caudal fin tissue could be used as a non-lethal proxy to liver and muscle for stable isotope analysis, and (2) assess the effects of ethanol preservation on δ15N and δ13C in fin tissue of juvenile yellow perch Perca flavescens. The δ13C of caudal fin was not significantly different from liver (t23 = −0.58; p = 0.57), and was more correlated with δ15N in liver (r2 = 0.78) than muscle (r2 = 0.56). Ethanol preservation enriched 15N and 13C for caudal fins, but by using our developed regression models, these changes in δ15N and δ13C can now be corrected. Overall, caudal fin tissue is a more reliable proxy to liver than muscle for δ15N and δ13C in yellow perch.

Quantifying Elements in Arctic Grayling and Bull Trout in the South Nahanni River Watershed, Northwest Territories, Using Nonlethal Tissue Samples

AbstractMonitoring of contaminants in fish generally involves lethal sampling, but public scrutiny and increased pressure on fisheries have driven the need to develop nonlethal sampling methods. We examined the ability of adipose, anal, and caudal fin tissues to serve as nonlethal surrogates for lethal muscle tissue samples in the analysis of metals (elements). First, we evaluated the use of biopsies by examining relationships between concentrations of 39 elements in low‐volume dorsal muscle biopsies and high‐volume muscle samples from Arctic Grayling Thymallus arcticus and Bull Trout Salvelinus confluentus collected in the South Nahanni River watershed, Northwest Territories, Canada. Low‐volume dorsal biopsy samples in this study served to most closely model the concentrations of elements found in high‐volume dorsal samples; caudal and anal fins were more representative of high‐volume dorsal samples than were adipose fins. Regressions between high‐ and low‐volume dorsal muscle samples were significant for 12 elements/species, with Cs, Rb, and Tl having the strongest relationships in both species. Regression analyses comparing low‐volume muscle samples and fin samples revealed variation between Arctic Grayling and Bull Trout, but Co, Hg, and Tl concentrations among samples were strongly related for both species. Addition of fish length or age as a covariate did not greatly improve the predictive power of calculated regressions. For future monitoring, selection of a nonlethal sampling strategy (e.g., use of dorsal biopsy or adipose fin samples) will require consideration of the element of interest, the primary route of exposure, interaction with other elements, and the basic biology and ecology of the fish species. Ideally, nonlethal sampling tools can be further developed for the two species to promote inclusion of community partners; these tools offer sustainable, long‐term approaches for monitoring sensitive fish populations in northern Canadian habitats.Received February 29, 2016; accepted September 9, 2016Published online December 20, 2016

Integration of zebrafish fin regeneration genes with expression data of human tumors in silico uncovers potential novel melanoma markers.

Tissue regeneration requires expression of a large, unknown number of genes to initiate and maintain cellular processes such as proliferation, extracellular matrix synthesis, differentiation and migration. A unique model to simulate this process in a controlled manner is the re-growth of the caudal fin of zebrafish after amputation. Within this tissue stem cells differentiate into fibroblasts, epithelial and endothelial cells as well as melanocytes. Many genes implicated in the regeneration process are deregulated in cancer. We therefore undertook a systematic gene expression study to identify genes upregulated during the re-growth of caudal fin tissue. By applying a high stringency cut-off value of 4-fold change, we identified 54 annotated genes significantly overexpressed in regenerating blastema. Further bioinformatics data mining studies showed that 22 out of the 54 regeneration genes where overexpressed in melanoma compared to normal skin or other cancers. Whereas the role of TNC (tenascin C) and FN1 (fibronectin 1) in melanoma development is well documented, implication of MARCKS, RCN3, BAMBI, PEA3/ETV4 and the FK506 family members FKBP7, FKBP10 and FKBP11 in melanoma progression is unclear. Corresponding proteins were detected in melanoma tissue but not in normal skin. High expression of FKBP7, DPYSL5 and MDK was significantly associated with poor survival. We discuss a potential role of these novel melanoma genes, which have promising potential as new therapeutic targets or diagnostic markers.

Genetic Differentiation Studies among Natural Populations of &lt;i&gt;Tilapia zillii&lt;/i&gt;

The population structure of Tilapia zillii (Gervais 1848) from three reservoirs in Nigeria, Osun State (Opa, Osu and Igun) was determined by employing morphological and molecular (Random Amplified Polymorphic DNA) methods. For morphological studies, 25 morphometric measurements and six meristic counts were recorded on 40 individuals within each population. Principal Component Analysis (PCA) was performed on the morphometric and meristic data using the PAST software. For RAPD studies, genomic DNA was extracted from caudal fin tissue using CTAB method and five primers were used to initiate PCR amplifications. All the clusters produced by the Principal components analysis (PCA) of the morphometric and meristic parameters overlapped indicating a low level of genetic differentiation between the three populations of T. zillii studied. The UPGMA cluster diagram from RAPD analysis identified two major genotypic groups with inter and intra group relationships. All individuals in the first cluster were from the Osu reservoir, while individuals from Opa and Igun reservoirs constituted the second cluster. Nei’s unbiased measure of genetic distances was 0.8532, 0.7321 and 0.7111 for Osu, Igun and Opa populations respectively. This revealed that Opa and Igun populations were genetically closer, while Osu populations is distant from them. The results suggest that the RAPD technique could be used to differentiate populations of T. zillii. However, additional methods such as microsatellite and sequence analysis can be used to maximize the efficiency of genetic differentiation studies.