Ruminal fatty acid outflow in dry cows fed different sources of linoleic acid: reticulum and omasum as alternative sampling sites to abomasum

This study was designed to determine the rumen outflow of fatty acids (FA) and biohydrogenation (BH) extent using alternative sampling sites (reticulum and omasum) to abomasum in dry cows fed different sources of FA. Four Holstein non-pregnant dry cows (≥3 parturitions, and 712 ± 125 kg BW), cannulated in the rumen and abomasum, were randomly assigned to a 4 × 4 Latin square design experiment, containing the following treatments: 1) control (CON); 2) soya bean oil (SO), dietary inclusion at 30 g/kg; 3) whole raw soya beans (WS), dietary inclusion at 160 g/kg; and 4) calcium salts of FA (CSFA), dietary inclusion at 32 g/kg. Rumen outflow of nutrients was estimated using the three markers reconstitution system (cobalt-EDTA, ytterbium chloride, and indigestible neutral detergent fibre [NDF]). Diets with FA sources decreased feed intake and increased FA intake. No differences in nutrient intake and digestibility were detected among cows fed diets supplemented with different FA sources. Diets with FA sources reduced the rumen outflow of DM and NDF, hence decreasing their passage rates. In addition, SO diet reduced the ruminal outflow of DM and NDF in comparison with WS and CSFA. Omasal sampling yielded the highest values of rumen outflow of NDF and potentially degradable NDF (pdNDF), whereas the reticular and abomasal samplings yielded intermediate and least values, respectively. The interaction effect between diet and sampling site was observed for rumen outflow of majority FA (except for C16:0, C18:0, and C18:2 trans-10, cis-12) and BH extension of C18:1 cis, C18:2, and C18:3. Calculations derived from abomasal sampling revealed that WS and CSFA diets had lower BH extent of C18:1 cis and C18:2 in comparison with SO, whereas cows fed CSFA had greater BH extent of C18:3 and lower BH extent of C18:1 cis compared to those fed WS. However, the latter results were not similar when calculations were performed based on the reticular and omasal samplings. Thus, there is evidence that neither reticular nor omasal samplings are suitable for estimating rumen outflow of FA in dry cows. In addition, WS and CSFA diets can increase the abomasal flow of polyunsaturated FA in dry cows.     

Distinct sensitivity of fungal freshwater guilds to water quality

Multiple anthropogenic stressors have been shown to impact animal and plant communities in freshwater ecosystems, but the responses of aquatic fungi remain largely unknown. Stressor effects on fungal communities may, however, result in changes of decomposition of plant litter and, thus, impact nutrient cycling, a key process for ecosystem functioning. We tested the impact of increased chloride and sediment levels, as well as reduced water flow velocity, on eukaryotic freshwater communities, with an emphasis on fungi, in a mesocosm experiment. Each of the three stressors was applied individually and in all combinations in a full-factorial design. Litterbags with non-sterilised tree leaves and sterile ceramic tiles were added to the mesocosms, to analyse the responses of communities in decaying plant material and in biofilms. Fungi preferably occurring in biofilms were supposed to represent indigenous aquatic fungi, while litterbag communities should be predominantly composed of fungi known from terrestrial litter. Community composition was assessed by high-throughput sequencing of amplified barcoding regions. Similarity matrices of operational taxonomic unit (OTU) tables calculated by UCLUST and CD-HIT-OTU-Illumina were significantly correlated. Preferred occurrence in biofilm and litter communities, respectively, was used for the grouping of OTUs into three ecological guilds. Stressor sensitivity varied among the guilds. While non-fungal, in particular autotrophic, OTUs responded to several treatments, two of the fungal guilds, i.e. those exclusively colonising litter and those preferably occurring on the ceramic tiles, showed no response to any applied treatment. Only fungi preferably, but not exclusively, colonising litter significantly responded to chloride addition. Their distribution patterns again correlated significantly with those of non-fungal OTUs, indicating possible interdependencies between both groups. The results indicate that eukaryotic freshwater communities are composed of different guilds, with distinctive sensitivity and tolerance to anthropogenic stressors.     

Recurrent gene loss correlates with the evolution of stomach phenotypes in gnathostome history

The stomach, a hallmark of gnathostome evolution, represents a unique anatomical innovation characterized by the presence of acid- and pepsin-secreting glands. However, the occurrence of these glands in gnathostome species is not universal; in the nineteenth century the French zoologist Cuvier first noted that some teleosts lacked a stomach. Strikingly, Holocephali (chimaeras), dipnoids (lungfish) and monotremes (egg-laying mammals) also lack acid secretion and a gastric cellular phenotype. Here, we test the hypothesis that loss of the gastric phenotype is correlated with the loss of key gastric genes. We investigated species from all the main gnathostome lineages and show the specific contribution of gene loss to the widespread distribution of the agastric condition. We establish that the stomach loss correlates with the persistent and complete absence of the gastric function gene kit—H⁺/K⁺-ATPase (Atp4A and Atp4B) and pepsinogens (Pga, Pgc, Cym)—in the analysed species. We also find that in gastric species the pepsinogen gene complement varies significantly (e.g. two to four in teleosts and tens in some mammals) with multiple events of pseudogenization identified in various lineages. We propose that relaxation of purifying selection in pepsinogen genes and possibly proton pump genes in response to dietary changes led to the numerous independent events of stomach loss in gnathostome history. Significantly, the absence of the gastric genes predicts that reinvention of the stomach in agastric lineages would be highly improbable, in line with Dollo''s principle.     

A novel assembly pipeline and functional annotations for targeted sequencing: A case study on the globally threatened Margaritiferidae (Bivalvia: Unionida)

The proliferation of genomic sequencing approaches has significantly impacted the field of phylogenetics. Target capture approaches provide a cost-effective, fast and easily applied strategy for phylogenetic inference of non-model organisms. However, several existing target capture processing pipelines are incapable of incorporating whole genome sequencing (WGS). Here, we develop a new pipeline for capture and de novo assembly of the targeted regions using whole genome re-sequencing reads. This new pipeline captured targeted loci accurately, and given its unbiased nature, can be used with any target capture probe set. Moreover, due to its low computational demand, this new pipeline may be ideal for users with limited resources and when high-coverage sequencing outputs are required. We demonstrate the utility of our approach by incorporating WGS data into the first comprehensive phylogenomic reconstruction of the freshwater mussel family Margaritiferidae. We also provide a catalogue of well-curated functional annotations of these previously uncharacterized freshwater mussel-specific target regions, representing a complementary tool for scrutinizing phylogenetic inferences while expanding future applications of the probe set.     

Session-to-session variations in external load measures during small-sided games in professional soccer players

The aims of this study were 1) to analyse session-to-session variations in different external load measures and 2) to examine differences in within-session intervals across different small-sided game (SSG) formats in professional players. Twenty professional soccer players (mean ± SD; age 28.1 ± 4.6 years, height 176.7 ± 4.9 cm, body mass 72.0 ± 7.8 kg, and body fat 10.3 ± 3.8%) participated in 3v3, 4v4, and 6v6 SSGs under different conditions (i.e., touch limitations and presence of goalkeepers vs. free touch and ball possession drill) over three sessions. Selected external load measures—including total distance (TD), high-intensity running (HIR, distance covered > 14.4 km^.h^-1), high-speed running (HSR, distance covered > 19.8 km^.h^-1), and mechanical work (MW, accelerations and deceleration > 2.2 m^.s²)—were recorded using GPS technology during all SSG sessions. Small to large standardized typical errors were observed in session-to-session variations of selected measures across SSGs. TD^.min^-1 showed less variability, having a coefficient of variation (CV) of 2.2 to 4.6%, while all other selected external load measures had CV values ranging from 7.2% to 29.4%. Trivial differences were observed between intervals in TD^.min^-1 and HIR^.min^-1 for all SSGs, as well as in HSR^.min^-1 and MW^.min^-1 for most SSG formats. No reductions or incremental trends in session-to-session variations were observed when employing touch limitations or adding goalkeepers. The increased noise observed in higher speed zones (e.g., high-speed running) suggests a need for more controlled, running-based conditional drills if the aim is greater consistency in these measures.     

A systematic review of collective tactical behaviour in futsal using positional data

Although many studies on collective tactical behaviour have been published in the last decade, no study has revised and summarized the findings provided for futsal. The main aim of this systematic review was to identify and discuss the geometrical centre (GC), distance and area tactical variables used to assess team behaviour in futsal. In addition, it summarizes the findings on the tactical response during futsal competition and training. A systematic review of the relevant articles provided on futsal was carried out using seven electronic databases (SPORTDiscus, ProQuest, Cochrane Plus, Scopus, Google Scholar, PubMed and Web of Science) until September 25, 2019. From a total of 1,209 studies initially found, 12 were included in the qualitative synthesis. There were some trends in the analysis of positional data in futsal with the most relevant situations analysed being 1 vs 1 and 5 vs 4+Goalkeeper. The distances and angles between two points were the most assessed tactical variables. Five types of distance variables were used to assess collective tactical behaviour in futsal: GC-GC, GC-player, player-player, player-ball and player-space. Pressure (GC-GC) was greater in shots on goal than in tackles during professional futsal matches. Area variables were reduced to occupied space, exploration space and dominant area. Occupied space was measured only during competition while the dominant area was measured only during training sessions. The surface area and dominant regions were greater when players were attacking in comparison to when they were defending. In addition, two non-linear techniques (i.e. relative phase and entropy) were applied to analyse synchronisation and complexity and regularity or predictability. Defenders were highly synchronous, while attackers tried to break this coordination to achieve possibilities for action. Task constraints are suitable to induce different regularity patterns. This review is an opportunity to develop studies aimed at bridging the gap in collective tactical behaviour in futsal.