Tris–Egg Yolk–Glycerol (TEY) Extender Developed for Freezing Dog Semen is a Good Option to Cryopreserve Bovine Epididymal Sperm Cells

Cryopreservation of epididymal spermatozoa is often performed after shipping the excised testis–epididymis complexes, under refrigeration, to a specialized laboratory. However, epididymal spermatozoa can be collected immediately after excision of the epididymis and sent extended and refrigerated to a laboratory for cryopreservation. In this experiment, we evaluated the effect of both methods of cold storage bovine epididymal spermatozoa as well as of two different extenders on spermatozoa characteristics after freeze–thawing. For that, spermatozoa collected from the caudae epididymis of 19 bulls were extended and cryopreserved in either AndroMed^® or a Tris–egg yolk (TEY)‐based extender. Cryopreservation of sperm cells was performed immediately after castration (Group A, n = 9) or after cold storage for 24 h diluted in the two extenders and (Group B, n = 9) and also after cold storage for 24 h within the whole epididymis (Group C, n = 10). Sperm subjective progressive motility (light microscopy), plasma membrane integrity (hypoosmotic swelling test) and sperm viability (eosin–nigrosin) were evaluated. In vitro fertilization and culture (IVF) was performed to assess the blastocyst rate. No differences (p > 0.05) were observed on post‐thaw sperm parameters between samples from Group A, B and C. TEY extended samples presented a higher (p < 0.01) percentage of progressive motile and live sperm, than those extended in AndroMed^®. Blastocyst rate after IVF differed only (p < 0.05) between the reference group (IVF performed with frozen semen with known in vitro fertility) and Group A extended in AndroMed^®. We conclude that when cryopreservation facilities are distant from the collection site, bovine epididymal sperm can be shipped chilled overnight either within the epididymal tail or after dilution without deleterious effect on post‐thaw sperm quality. TEY extender was more suitable for cold storage and freezing bovine epididymal sperm, than the commercial extender AndroMed®.     

Application of image analysis to the prediction of EBC barley kernel weight distribution

It is known that the barley kernel size is an important factor regarding the uniformity of the malting and brewery processes, barley valuation, approval and market value. In order to facilitate the barley purchasing process, a fast field technique for kernel size evaluation, such as the image analysis technique proposed in this work, would be greatly appreciated as a fast and simple procedure for barley selection. In this study a close correlation between the image analysis and the standard EBC was obtained with a correlation factor of 0.999 and a regression coefficient of 0.991 between the two methodologies. The proposed IA methodology was found to accurately predict the Scarlett and Prestige barley varieties weight distribution especially when considering the crucial ‘business transactions selection’ classes.     

Alpha-amylase inhibitors from Brazilian seaweeds and their hypoglycemic potential

The acetone crude extract of Lobophora variegata did not show any alpha-amylase inhibition activity. On the contrary, acetone crude extract of both Spatoglossum schroederi and Caulerpa racemosa inhibited the enzyme activity, with an ED(50) of 0.58 and 0.09 mg/ml.     

Spatial variability of chemical attributes of the soil,plant and yield in a pear orchard

For fruiting trees, fertilization is usually recommended based on the analyses of soil and leaves, adopting a standard value for the whole orchard. This management can lead to undesirable imbalances to the plants and local agroecosystem. Thus, this trial aimed at mapping the spatial variability of properties of soil, plant and yield in a commercial area of pears production and investigating the yield response to a distinguished fertilization. The trial was carried out in Nova Laranjeiras, Paraná, where 36 trees were selected to receive conventional fertilization (CF) and other 36 trees received a site-specific fertilization (SSF). All variables (chemical and physical attributes of soil, leaf tissue, and yield) showed some spatial dependence. For all years, yield variability was considered very high. The yield of SSF system was slightly higher (2,625 kg ha^?1) and took up a larger area (59%) when compared to SF system (2,548 kg ha^?1 at 41% area).     

On the drivers of cost changes in the Norwegian salmon aquaculture sector: a decomposition of a flexible cost function from 2001 to 2014

Abstract

Since 2005, Norwegian salmon farmers have experienced increasing unit costs, contrasting pre-2005 trends characterized by innovations, rapid productivity growth, and diminishing unit costs. This article investigates these cost changes using a panel of salmon producers. The drivers behind cost changes in the industry are identified for the period of 2001–2014 using a flexible cost function. In particular, it is explored how cost changes can be attributable to scale economies, negative productivity shocks, production expansion, and input prices. The results indicate that cost increases in the sector are affected by external factors out of the control of individual firms such as input prices and environmental conditions like sea lice.     

Expression of the myosin light chains 1, 2 and 3 in the muscle of blackspot seabream (<Emphasis Type="Italic">Pagellus bogaraveo</Emphasis>, Brunnich), during development

Previous studies on the histochemistry and immunoreactivity of fibres in lateral muscle of blackspot seabream indicated that there is a developmental transition in the composition of myofibrillar proteins, which presumably reflects changes in contractile function as the fish grows. We hypothesize that the phenomenon underscores age and spatial differences in the expression of myosin light chains (MLC), not studied yet in this species. In this study, we examined selected stages in the post-hatching development of the muscle of blackspot seabream: hatching (0 days), mouth opening (5 days), weaning (40 days) and juveniles (70 days). The spatial expression of embryonic MLC 1 (MLC1), 2 (MLC2) and 3 (MLC3) was studied by in situ hybridization. Overall, MLC expression patterns were overlapping and restricted to the fast muscle. At hatching and mouth opening, all MLC types were highly expressed throughout the musculature in fast muscle. The expression levels in fast muscle remained high until weaning when germinal zones appeared on the dorsal and ventral areas. The germinal zones were characterized by small-diameter fast fibres with high levels of MLC expression. This pattern persisted up to day 70, when the germinal zones disappeared and expression of MLCs was observed only in the smaller cells of the fast muscle mosaic. These results support our hypothesis and, together with previous imuno- and histochemistry results, allow a better understanding of the mechanism of muscle differentiation and growth in fish beyond larval stages, and form- the basis for further comparative and experimental studies with this economically relevant species.     

Putative effector genes detected in Fusarium oxysporum from natural ecosystems of Australia

The Fusarium oxysporum species complex (FOSC) causes disease in plants and animals, but is also widely dispersed in natural ecosystems without evidence of disease. The present study screened a population representing natural ecosystems across the Australian continent for the putative effector genes pisatin demethylase 1 (PDA1), pectate lyase (pelD), secreted gene expression (SGE1) and secreted in xylem (SIX). The genes pelD and SGE1 were prevalent in the natural isolates, PDA1 was present at an intermediate level, whereas SIX genes were detected at low levels. Phylogenies of these putative effector genes were compared to the EF‐1α species phylogeny to determine the likely modes of gene transmission: vertical gene transfer (VGT) and horizontal gene transfer (HGT). There was evidence of both modes of gene transmission within the F. oxysporum isolates. PDA1, pelD and SGE1 were likely to be only vertically inherited, whereas the SIX genes had evidence for both VGT and HGT. The phylogenetic relationships of SIX genes in isolates from natural ecosystems and formae speciales from agro‐ecosystems were also established. These findings have important implications for the evolution of effectors in the FOSC.     

Fish proteins as targets of ferrous-catalyzed oxidation: identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry

Protein oxidation in fish meat is considered to affect negatively the muscle texture. An important source of free radicals taking part in this process is Fenton's reaction dependent on ferrous ions present in the tissue. The aim of this study was to investigate the susceptibility of cod muscle proteins in sarcoplasmic and myofibril fractions to in vitro metal-catalyzed oxidation and to point out protein candidates that might play a major role in the deterioration of fish quality. Extracted control proteins and proteins subjected to free radicals generated by Fe(II)/ascorbate mixture were labeled with fluorescein-5-thiosemicarbazide (FTSC) to tag carbonyl groups and separated by two-dimensional gel electrophoresis. Consecutive visualization of protein carbonyl levels by capturing the FTSC signal and total protein levels by capturing the SyproRuby staining signal allowed us to quantify the relative change in protein carbonyl levels corrected for changes in protein content. Proteins were identified using MALDI-TOF/TOF mass spectrometry and homology-based searches. The results show that freshly extracted cod muscle proteins exhibit a detectable carbonylation background and that the incubation with Fe(II)/ascorbate triggers a further oxidation of both sarcoplasmic and myofibril proteins. Different proteins exhibited various degrees of sensitivity to oxidation processes. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), nucleoside diphosphate kinase B (NDK), triosephosphate isomerase, phosphoglycerate mutase, lactate dehydrogenase, creatine kinase, and enolase were the sarcoplasmic proteins most vulnerable to ferrous-catalyzed oxidation. Moreover, NDK, phosphoglycerate mutase, and GAPDH were identified in several spots differing by their pI, and those forms showed different susceptibilities to metal-catalyzed oxidation, indicating that post-translational modifications may change the resistance of proteins to oxidative damage. The Fe(II)/ascorbate treatment significantly increased carbonylation of important structural proteins in fish muscle, mainly actin and myosin, and degradation products of those proteins were observed, some of them exhibiting increased carbonylation levels.