Immobilization of gelatin onto poly(glycidyl methacrylate)-grafted polycaprolactone substrates for improved cell-material interactions

To enhance the cytocompatibility of polycaprolactone (PCL), cell-adhesive gelatin is covalently immobilized onto the PCL film surface via two surface-modified approaches: a conventional chemical immobilization process and a surface-initiated atom transfer radical polymerization (ATRP) process. Kinetics studies reveal that the polymer chain growth from the PCL film using the ATRP process is formed in a controlled manner, and that the amount of immobilized gelatin increases with an increasing concentration of epoxide groups on the grafted P(GMA) brushes. In vitro cell adhesion and proliferation studies demonstrate that cell affinity and growth are significantly improved by the immobilization of gelatin on PCL film surfaces, and that this improvement is positively correlated to the amount of covalently immobilized gelatin. With the versatility of the ATRP process and tunable grafting efficacy of gelatin, this study offers a suitable methodology for the functionalization of biodegradable polyesters scaffolds to improve cell-material interactions.     

Ribosome profiling shows that miR-430 reduces translation before causing mRNA decay in zebrafish

MicroRNAs regulate gene expression through deadenylation, repression, and messenger RNA (mRNA) decay. However, the contribution of each mechanism in non-steady-state situations remains unclear. We monitored the impact of miR-430 on ribosome occupancy of endogenous mRNAs in wild-type and dicer mutant zebrafish embryos and found that miR-430 reduces the number of ribosomes on target mRNAs before causing mRNA decay. Translational repression occurs before complete deadenylation, and disrupting deadenylation with use of an internal polyadenylate tail did not block target repression. Lastly, we observed that ribosome density along the length of the message remains constant, suggesting that translational repression occurs by reducing the rate of initiation rather than affecting elongation or causing ribosomal drop-off. These results show that miR-430 regulates translation initiation before inducing mRNA decay during zebrafish development.     

Analysis of bacterial community and bacterial nutritional enzyme activity associated with the digestive tract of wild Chilean octopus (Octopus mimus Gould, 1852)

Information on the bacterial community associated with octopus is very scarce, unlike fish and other molluscs. This study revealed the bacterial community associated with digestive tract of wild Chilean octopus Octopus mimus using a culture‐dependent method and 16S rDNA clone library. Moreover, we analysed the bacterial nutritional enzyme activity of culturable bacteria. A culture‐dependent method showed that the composition of the culturable bacterial community was substantially different between female and male octopus. The predominant species in female octopus were Vibrionaceae and Streptococcaceae, whereas only Vibrionaceae was dominated in male octopus. Bacterial nutritional enzyme activities of culturable bacteria from male octopus were much higher than female octopus. The 16S rDNA clone library analysis showed that the bacterial community of male octopus exhibited a higher diversity than that of female octopus. The genus Mycoplasma was the predominant bacteria in the digestive tract of all octopus samples. The results obtained in this study raise the possibility that each octopus has different food consumption due to different bacterial community and nutritional enzyme activity, although Mycoplasma sp. is one of the predominant bacteria in the digestive tract. Moreover, our results are useful for the future of microbiological investigation associated with the octopus and for probiotics in the octopus aquaculture.     

Effect of a size-based selection program on blood metabolites and immune response of Litopenaeus vannamei juveniles fed different dietary carbohydrate levels

Blood metabolites in wild and seventh-generation cultivated shrimp were measured to determine how size-based selection could alter the nutritional and immunological conditions of Litopenaeus vannamei. Wild L. vannamei juveniles and a sample of seventh-generation cultured shrimp were acclimated under identical conditions. During 55 days, shrimp were fed a high (HCHO: 44%) or a low (LCHO: 3%) carbohydrate diet for 55 days. Wild shrimp showed a direct relation between dietary CHO and lactate, protein and hemocyte levels indicating that dietary CHO was used for protein synthesis via transamination pathways. In seventh-generation cultured shrimp these parameters were inversely proportional to dietary CHO level, indicating the capacity to synthesize protein from dietary CHO was repressed in cultured shrimp. Farmed shrimp showed a limited capacity to respond to LCHO diets demonstrating high protein dependence in their metabolism and immune response. These results demonstrate that during size-based breeding programs other metabolic process than CHO catabolism can be selected. The incapacity of shrimp to use dietary CHO could limit protein reduction of diets and limit the efforts of the shrimp industry to be ecologically and environmentally profitable.     

Challenge studies of European stocks of redfin perch, Perca fluviatilis L., and rainbow trout, Oncorhynchus mykiss (Walbaum), with epizootic haematopoietic necrosis virus

A challenge model for comparison of the virulence of epizootic haematopoietic necrosis virus (EHNV) to European stocks of redfin perch, Perca fluviatilis L., and rainbow trout, Oncorhynchus mykiss (Walbaum), was tested. The model investigated intraperitoneal (IP), bath and cohabitation routes at 10, 15 and 20 °C for 5–6 g fish and 15 °C for 20 g perch. In the IP challenges of perch, significant mortality occurred at 15 °C and 20 °C. In challenge trials for rainbow trout, significant mortalities were observed in IP and bath challenges at 20 °C. The mortality observed in IP challenged 20 g perch was not significantly different from that recorded for 6 g fish challenged IP. No significant mortality was observed in any other treatment groups. Re-isolation of ranavirus was confirmed by IFAT and was consistently associated with dead or moribund fish in the trial groups challenged with EHNV. The findings indicate that EHNV does not pose a high risk for wild perch and trout populations in Europe by natural exposure. Mortality appears to be primarily a function of environmental factors, with temperature playing an important role, and not just the presence of the virus in the fish.     

Nutritional Contribution of Biofloc within the Diet of Growout and Broodstock of Litopenaeus vannamei,Determined by Stable Isotopes and Fatty Acids

The relative contributions of feed sources were determined through the isotopic signal (δ¹³C and δ¹⁵N) and fatty acid profile of feed items, shrimp muscle, and eggs of Litopenaeus vannamei reared in a biofloc system. In the growout phase, the isotope analysis showed the biofloc particle size class ≥250 μm contributed 55–100%; size ≥50 < 250 μm contributed 0–22%; and artificial feed contributed 0–45%. Principal component analysis applied to fatty acid profiles showed that biofloc ≥250 μm and artificial feed were the most important items in shrimp growout. For the egg production, isotope analysis suggested that the most important feed sources according to their relative contributions were polychaetes (0–100%), followed by artemia biomass (0–86%) and semi‐moist feed (0–66%), with lower contributions from squid, mussel, and the muscle of L. vannamei broodstock that had been cultured in biofloc. In terms of fatty acids, the most important items were artemia, polychaetes, and semi‐moist feeds. This work clarified the importance of feed sources for shrimp during culture in biofloc systems and during reproduction. Analysis of stable isotopes and fatty acids can be successfully used to trace the assimilation of nutrients during the nutrition of shrimp.     

Characteristics of monoclonal antibodies against Piscirickettsia salmonis

A panel of 28 monoclonal antibodies against Piscirickettsia salmonis was produced using a purified fraction of the bacterium. To determine their specificity to the pathogen, the antibodies were assayed by ELISA and indirect immunofluorescence microscopy. Six monoclonal antibodies were selected based on their strong reaction against P. salmonis and absence of cross-reactivity with other common fish pathogens. Western blot analysis showed that the antibodies reacted to several antigens of P. salmonis . Immunofluorescence assays revealed that these antibodies reacted with the same specificity to different isolates of P. salmonis obtained from the south of Chile. This panel of monoclonal antibodies represents an important tool to develop simple, rapid, sensitive and highly specific methods for the detection of the pathogen and diagnosis of the disease.     

Generation of spatially coherent light at extreme ultraviolet wavelengths

We present spatial coherence measurements of extreme ultraviolet (EUV) light generated through the process of high-harmonic up-conversion of a femtosecond laser. With a phase-matched hollow-fiber geometry, the generated beam was found to exhibit essentially full spatial coherence. The coherence of this laser-like EUV source was shown by recording Gabor holograms of small objects. This work demonstrates the capability to perform EUV holography with a tabletop experimental setup. Such an EUV source, with low divergence and high spatial coherence, can be used for experiments involving high-precision metrology, inspection of optical components for EUV lithography, and microscopy and holography with nanometer resolution. Furthermore, the short time duration of the EUV radiation (a few femtoseconds) will enable EUV microscopy and holography to be performed with ultrahigh time resolution.     

A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis

Dynamin 1 is a neuron-specific guanosine triphosphatase thought to be critically required for the fission reaction of synaptic vesicle endocytosis. Unexpectedly, mice lacking dynamin 1 were able to form functional synapses, even though their postnatal viability was limited. However, during spontaneous network activity, branched, tubular plasma membrane invaginations accumulated, capped by clathrin-coated pits, in synapses of dynamin 1-knockout mice. Synaptic vesicle endocytosis was severely impaired during strong exogenous stimulation but resumed efficiently when the stimulus was terminated. Thus, dynamin 1-independent mechanisms can support limited synaptic vesicle endocytosis, but dynamin 1 is needed during high levels of neuronal activity.     

Thermal preference and tolerance of green abalone Haliotis fulgens (Philippi, 1845) and pink abalone Haliotis corrugata (Gray, 1828)

The thermoregulatory behaviour of green abalone Haliotis fulgens and pink abalone H. corrugata was investigated. Haliotis fulgens juveniles ranging in wet weight from 3.0 to 3.3 g and from 28.7 to 30.5 mm shell length and of H. corrugata 2.0 g and 25.7 mm in shell length were exposed to 19°C for 30 days in a flow‐through water system. Temperature preference was determined in a horizontal thermal gradient and was found to be 25.4°C for green abalone and 25.0°C for pink abalone. Displacement velocity was 4.3 cm h⁻¹ for H. fulgens and 12.8 cm h⁻¹ for H. corrugata. The optimum temperature for growth calculated for both abalone species was 24.6 and 24.5°C respectively. The critical thermal maxima (CTMax) of H. fulgens and H. corrugata were determined as a measure of thermal tolerance. Abalones were subjected to increasing water temperatures at a rate of 1°C on 30 min until they detached from the substrate. The CTMax at 50% were 33.6 and 32.0°C for green and pink abalone respectively. The results are discussed in relation to site selection and commercial rearing.