The cellular basis of compensatory muscle growth in the teleost Odontesthes bonariensis.

This study evaluates white muscle growth and in vivo cell proliferation during a fasting and refeeding trial, using pejerrey (Odontesthes bonariensis) as animal model, in order to better understand the cellular basis governing catch-up growth. Experiments consisted of two groups of fish, a control group continuously fed ad libitum, and a group fasted for 2 weeks and then fed for another 2 weeks. We examined how the formation of new muscle fibers and their increase in size were related to muscle precursor cell (MPC) proliferation under both experimental conditions. During fasting, the number of 5-ethynyl-2'-deoxyuridine-positive (EdU+) cells decreased along with myogenic regulatory factor (MRF) mRNA levels related to myoblast proliferation and differentiation, and the muscle stem cell marker Pax7 mRNA level increased. Analysis of myomere cross-sectional area, distribution of muscle fiber sizes and number of fibers per myomere showed that muscle hypertrophy but not hyperplasia was inhibited during fasting. Both higher igf2 mRNA level and the persistence of cell proliferation could be supporting new myofiber formation. In contrast, an exacerbated MPC proliferation occurred during catch-up growth, and this increase in cell number could be contributing to the growth of both pre-existing and newly formed small fibers. The findings that some MPCs proliferate during fasting and that muscle growth mechanisms, hyperplasia and hypertrophy are differentially regulated could help to explain why re-fed fish could grow at faster rates, and why they return to the lost growth trajectory.

Quantification and composition analysis of plastic pollution in riverine beaches of the lower Paraná River, Argentina.

Plastic pollution and the numerous consequences it has on aquatic life have become a huge concern in recent years. While many studies have been conducted in marine environments, studies in freshwater ecosystems are scarce and insufficient. The Paraná River is the most important water course in the La Plata River basin and the fifth in the world with a mean annual discharge of 18,000 m3 per second. Currently available studies show the presence of plastic in river shores and fish gut, but more research should be carried out in order to know the extension and origin of plastic contamination. Therefore, the aim of this study was to quantify and characterize macro-, meso-, and microplastics found in the riverine beaches next to Rosario city, the most populated city standing by the lower Paraná River coast in Argentina. The results show that plastic pollution is ubiquitous, but the city shores are significantly more polluted than the wetland shore with a mean of 30,780 and 6375 microplastics per square meter respectively (p = 0.024). The food and beverage industry packaging combined were the most frequent macroplastics found. Also, 3 out of 4 meso- and microplastics were white/transparent, the color that is most likely to be ingested by fish and invertebrates. Finally, all micro- and mesoplastics found were secondary and, in the case of microplastics, they were mainly fibers (93.4%) which highlight its ecological relevance. As a whole, plastic contamination is a serious issue in the Rosario area, specially single-use plastics and short-lived products. The anthropic effect of the cities and how it contributes to plastic pollution are evident.

Improving zebrafish embryo xenotransplantation conditions by increasing incubation temperature and establishing a proliferation index with ZFtool.

BACKGROUND: Zebrafish (Danio rerio) is a model organism that has emerged as a tool for cancer research, cancer being the second most common cause of death after cardiovascular disease for humans in the developed world. Zebrafish is a useful model for xenotransplantation of human cancer cells and toxicity studies of different chemotherapeutic compounds in vivo. Compared to the murine model, the zebrafish model is faster, can be screened using high-throughput methods and has a lower maintenance cost, making it possible and affordable to create personalized therapies. While several methods for cell proliferation determination based on image acquisition and quantification have been developed, some drawbacks still remain. In the xenotransplantation technique, quantification of cellular proliferation in vivo is critical to standardize the process for future preclinical applications of the model. METHODS: This study improved the conditions of the xenotransplantation technique - quantification of cellular proliferation in vivo was performed through image processing with our ZFtool software and optimization of temperature in order to standardize the process for a future preclinical applications. ZFtool was developed to establish a base threshold that eliminates embryo auto-fluorescence and measures the area of marked cells (GFP) and the intensity of those cells to define a 'proliferation index'. RESULTS: The analysis of tumor cell proliferation at different temperatures (34 °C and 36 °C) in comparison to in vitro cell proliferation provides of a better proliferation rate, achieved as expected at 36°, a maintenance temperature not demonstrated up to now. The mortality of the embryos remained between 5% and 15%. 5- Fluorouracil was tested for 2 days, dissolved in the incubation medium, in order to quantify the reduction of the tumor mass injected. In almost all of the embryos incubated at 36 °C and incubated with 5-Fluorouracil, there was a significant tumor cell reduction compared with the control group. This was not the case at 34 °C. CONCLUSIONS: Our results demonstrate that the proliferation of the injected cells is better at 36 °C and that this temperature is the most suitable for testing chemotherapeutic drugs like the 5-Fluorouracil.

"A Zebra in the Water": Inspiring Science in Spain.

The zebrafish model can play a role in education because of its suitability for manipulation and attractiveness to students compared to traditional lecture-based instruction. Furthermore, zebrafish offer advantages over other model species. Seeing as fewer and fewer students are entering science degree programs, this project has been developed to encourage scientific vocations among secondary school students. To do so, an aquarium was given to 114 schools so that they could look after adult zebrafish, mate them, and visualize embryo development. For training on more sophisticated techniques, a virtual tool was developed to simulate a real genetics laboratory on a personal computer. Results based on teachers' feedback indicate that the students were fully dedicated to the project and achieved better understanding of genetic concepts and techniques. These results demonstrate the potential of alternative teaching methods for engaging students in science learning.

Integrative Transcriptome, Genome and Quantitative Trait Loci Resources Identify Single Nucleotide Polymorphisms in Candidate Genes for Growth Traits in Turbot.

Growth traits represent a main goal in aquaculture breeding programs and may be related to adaptive variation in wild fisheries. Integrating quantitative trait loci (QTL) mapping and next generation sequencing can greatly help to identify variation in candidate genes, which can result in marker-assisted selection and better genetic structure information. Turbot is a commercially important flatfish in Europe and China, with available genomic information on QTLs and genome mapping. Muscle and liver RNA-seq from 18 individuals was carried out to obtain gene sequences and markers functionally related to growth, resulting in a total of 20,447 genes and 85,344 single nucleotide polymorphisms (SNPs). Many growth-related genes and SNPs were identified and placed in the turbot genome and genetic map to explore their co-localization with growth-QTL markers. Forty-five SNPs on growth-related genes were selected based on QTL co-localization and relevant function for growth traits. Forty-three SNPs were technically feasible and validated in a wild Atlantic population, where 91% were polymorphic. The integration of functional and structural genomic resources in turbot provides a practical approach for QTL mining in this species. Validated SNPs represent a useful set of growth-related gene markers for future association, functional and population studies in this flatfish species.

Insulin-like growth factor-I of pejerrey, Odontesthes bonariensis: cDNA characterization, tissue distribution and expression profiles after growth hormone administration.

The liver production of the insulin-like growth factor-I (IGF-I) is a key factor in the endocrine control of body growth by a growth hormone. As pejerrey Odontesthes bonariensis has been reported as a fish with low growth rates in captivity, basic research on this respect is needed in order to understand it. In this context, the pejerrey IGF-I cDNA was cloned and its hepatic expression was examined in fish after recombinant pejerrey growth hormone (pjGHr) administration. The full length of IGF-I transcript showed a high sequence similarity to other teleost sequences. The tissue distribution analysis by reverse transcriptase polymerase chain reaction in adult fish revealed that IGF-I expressed ubiquitously with the highest mRNA levels in the liver, posterior intestine and brain. No alternative IGF-I mRNA was found in the liver, as it was reported for other teleosts. IGF-I transcript was measured in the liver after pjGHr in vivo stimulation by means of quantitative real-time polymerase chain reaction assays. A dose-dependent response of IGF-I mRNA was observed after pjGHr administration, and reached a six-fold IGF-I maximum increase over control group when 2.5 microg pjGH/g-body weight (bw) was injected. Temporal analysis of hepatic IGF-I mRNA level showed that administration of a single dose of pjGHr into juvenile pejerrey resulted in a significant increase (P <0.02) 9 hours post-injection (hpi). These results add novel information on the nucleotide sequence of IGF-I in Atheriniformes and demonstrate that pjGHr could promote a dramatic response in liver, increasing the IGF-I mRNA level.