Enhancing the bioconversion of winery and olive mill waste mixtures into lignocellulolytic enzymes and animal feed by Aspergillus uvarum using a packed-bed bioreactor

Wineries and olive oil industries are dominant agro-industrial activities in southern European regions. Olive pomace, exhausted grape marc and vine shoot trimmings are lignocellulosic residues generated by these industries, which could be valued biotechnologically. In the present work these residues were used as substrate to produce cellulases and xylanases through solid-state fermentation using Aspergillus uvarum. For that, two factorial designs (32) were first planned to optimize substrate composition, temperature and initial moisture level. Subsequently, the kinectics of cellulolytic enzymes production, fungal growth and fermented solid were characterized, Finally, the process was performed in a packed-bed bioreactor. The results showed that cellulase activity improved with the optimization processes, reaching 33.56 U/g, and with the packed-bed bioreactor aeration of 0.2 L/min, reaching 38.51 U/g. The composition of fermented solids indicated their potential use for animal feed because cellulose, hemicellulose, lignin and phenolic compounds were partially degraded 28.08, 10.78, 13.3 and 28.32% respectively, crude protein was increased from 8.47% to 17.08%, and, the mineral contents meet the requirements of main livestock.J.M.S. was supported by Grant SFRH/BPD/84440/2012 from Fundacao para a Ciencia e Tecnologia (FCT), Portugal. L.A. was supported by Grant Incentivo/EQB/LA0023/2014 from O Novo Norte (ON.2). We thank FCT Strategic Project PEst-OE/EQB/LA0023/2013 and the Project "BioInd - Biotechnology and Bioengineering for improved Industrial and Agro-Food processes, Rer. NORTE-07-0124-FEDER-000028" co-funded by the Programa Operacional Regional do Norte (ON.2), QREN, FEDE

Improved canine exome designs, featuring ncRNAs and increased coverage of protein coding genes OPEN

By limiting sequencing to those sequences transcribed as mRNA, whole exome sequencing is a costefficient technique often used in disease-association studies. We developed two target enrichment designs based on the recently released annotation of the canine genome: the exome-plus design and the exome-CDS design. The exome-plus design combines the exons of the CanFam 3.1 Ensembl annotation, more recently discovered protein-coding exons and a variety of non-coding RNA regions (microRNAs, long non-coding RNAs and antisense transcripts), leading to a total size of ≈152 Mb. The exome-CDS was designed as a subset of the exome-plus by omitting all 3' and 5' untranslated regions. This reduced the size of the exome-CDS to ≈71 Mb. To test the capturing performance, four exome-plus captures were sequenced on a NextSeq 500 with each capture containing four precapture pooled, barcoded samples. At an average sequencing depth of 68.3x, 80% of the regions and well over 90% of the targeted base pairs were completely covered at least 5 times with high reproducibility. Based on the performance of the exome-plus, we estimated the performance of the exome-CDS. Overall, these designs provide flexible solutions for a variety of research questions and are likely to be reliable tools in disease studies. In 2014, the first report detailing the design and performance of a whole exome sequencing (WES) enrichment assay for the dog was published by our group 1 . Aiming to selectively sequence all the regions that are transcribed to mRNA, WES is a reliable tool used to identify disease-causing or predisposing mutations at a fraction of the price of whole genome sequencing (WGS) studies. A limitation of WES is that it is based on our current knowledge of the annotation of the genome and that many disease causing mutations are likely to fall outside protein-coding regions. With new information becoming available, updates and extensions are required. Recently, an improved annotation for the dog genome has been published and new data on non-protein coding genes has been obtained 2 . Based on this data, two new target enrichment designs for dogs, called the exome-plus and the exome-CDS, were developed. The exome-plus offers the most comprehensive design. The exome-CDS is a subset of the exome-plus, focusing on the coding DNA sequences (CDS) by excluding the 3′ and 5′ untranslated regions (UTRs). Thes

A comparison of diatrizoate and ioxaglate for positive contrast shoulder arthrography in dogs

In six experimental dogs, arthrographic quality and synovial inflammatory response with shoulder arthrography comparing meglumine-sodium diatrizoate (Urovison) and the monoacidic dimer, meglumine-sodium ioxaglate (Hexabrix) was evaluated. In our study initial films were of equally high diagnostic quality for both contrast media, but delayed films significantly favored ioxaglate for diagnostic quality. The rise in white blood cells in synovial fluid samples collected 24 hours after the arthrographic procedure was significantly lower after the use of ioxaglate. Histologic examination performed 14 days after the intra-articular injection revealed no drug related lesions

Total and toxic arsenic levels in North Sea fish

Levels of arsenic contamination in muscle and liver tissue of 25 sea fish and 4 shellfish species from the North Sea were determined. Analyses were done by both ICP-MS and HG-AFS to distinguish between nontoxic and toxic fractions of As. Highest total As concentrations were found in lemon sole, dogfish, ray, and witch. Average total As concentrations in these fish species were higher than 20 mg kg-1 WW. The same species as well as the other flatfishes contained the highest amounts of toxic As (> 0.1 mg kg-1 WW). Toxic fractions (AsTox/AsT%) above 2% were found in the following six species: seabass, ling, john dory, pouting, dab, and brill. No preferential concentration in the liver compared to the muscle was observed. In a worst-case scenario (when fish has been dried or smoked and the toxic As level is high; for example 0.5 mg kg-1 WW), the As content of North Sea marine food may reach harmful levels. A normalization reflecting the toxic potential of sea fish was made. Shark and ray and most flatfish species have positive (high) normalized AsTox values. By ANOVA testing we compared the individual AsT concentrations of samples of the same species (intraspecies variability). Significant differences for some fish species were observed; significantly higher AsT concentrations were found in dogfish from the French coast versus the western North Sea and in common sole from the Bay of the Seine, in the north of France, versus the western North Sea