Antarctic Thraustochytrids as Sources of Carotenoids and High-Value Fatty Acids

Eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and carotenoids are needed as human dietary supplements and are essential components in commercial feeds for the production of aquacultured seafood. Microorganisms such as thraustochytrids are potential natural sources of these compounds. This research reports on the lipid and carotenoid production capacity of thraustochytrids that were isolated from coastal waters of Antarctica. Of the 22 isolates, 21 produced lipids containing EPA+DHA, and the amount of these fatty acids exceeded 20% of the total fatty acids in 12 isolates. Ten isolates were shown to produce carotenoids (27.4–63.9 μg/g dry biomass). The isolate RT2316-16, identified as Thraustochytrium sp., was the best producer of biomass (7.2 g/L in five days) rich in carotenoids (63.9 μg/g) and, therefore, became the focus of this investigation. The main carotenoids in RT2316-16 were β-carotene and canthaxanthin. The content of EPA+DHA in the total lipids (34 ± 3% w/w in dry biomass) depended on the stage of growth of RT2316-16. Lipid and carotenoid content of the biomass and its concentration could be enhanced by modifying the composition of the culture medium. The estimated genome size of RT2316-16 was 44 Mb. Of the 5656 genes predicted from the genome, 4559 were annotated. These included genes of most of the enzymes in the elongation and desaturation pathway of synthesis of ω-3 polyunsaturated fatty acids. Carotenoid precursors in RT2316-16 were synthesized through the mevalonate pathway. A β-carotene synthase gene, with a different domain organization compared to the gene in other thraustochytrids, explained the carotenoid profile of RT2316-16.     

Salmon cells SHK‐1 internalize infectious pancreatic necrosis virus by macropinocytosis

We have previously shown that infectious pancreatic necrosis virus (IPNV) enters the embryo cell line CHSE‐214 by macropinocytosis. In this study, we have extended our investigation into SHK‐1 cells, a macrophage‐like cell line derived from the head kidney of Atlantic salmon, the most economically important host of IPNV. We show that IPNV infection stimulated fluid uptake in SHK‐1 cells above the constitutive macropinocytosis level. In addition, upon infection of SHK‐1 cells, IPNV produced several changes in actin dynamics, such as protrusions and ruffles, which are important features of macropinocytosis. We also observed that the Na+/H+ pump inhibitor EIPA blocked IPNV infection. On the other hand, IPNV entry was independent of clathrin, a possibility that could not be ruled out in CHSE 214 cells. In order to determine the possible role of accessory factors on the macropinocytic process, we tested several inhibitors that affect components of transduction pathways. While pharmacological intervention of PKI3, PAK‐1 and Rac1 did not affect IPNV infection, inhibition of Ras and Rho GTPases as well as Cdc42 resulted in a partial decrease in IPNV infection. Further studies will be required to determine the signalling pathway involved in the macropinocytosis‐mediated entry of IPNV into its target cells.     

Characterization of Lay Phase in Three Genetic Types of Heavy Broilers Breeder in Tropical Conditions

The objective of the present work is characterizing some parameters in the lay phase in the lines of heavy broilers breeder Cobb Avian 48, Arbor Acres Plus and Ross 208, reared under the same commercial conditions in the tropics. Monitored three batches of approximately 18,000 females each （one for each genetic type）, from weeks 24 to 64, according to the conditions of handling and power described by the generic lines distribution companies. Although no differences were found for feed intake, Cobb Avian was presented a lower rate of feed conversion, being also that presented the highest percentage of implementation and production of incubables eggs, which meant that it was in line with the highest cumulative production of chickens by bird and week for tropical conditions studied. To adjust the lay data to a Gorssman model, in order to predict the parameters that characterize the lay modeling, estimated the best behavior for Cobb Avian 48 females, because while it takes more time to reach the level of putting constant （5.30 weeks） and maintain it for less time （2.4 weeks）, they manage to put constant high （79.11%） level, considering that the largest number of eggs per bird （171.19 vs. 164.54 and 160.11 in Arbor Acres and Ross 208, respectively）.     

Synthesis, analytical features, and biological relevance of 5-(3',4'-dihydroxyphenyl)-γ-valerolactone, a microbial metabolite derived from the catabolism of dietary flavan-3-ols

The physiological significance of 5-(3',4'-dihydroxyphenyl)-γ-valerolactone, an important metabolite derived from the catabolism of flavan-3-ols by gut microbiota, has been often overlooked due to the lack of the commercial standard. In the present work, this metabolite has been chemically synthesized, and its analytical parameters and antioxidant capacity have been determined in comparison to other chemical analogues [isomer 3-(3',4'-dihydroxyphenyl)-δ-valerolactone and γ-valerolactone] and other structurally related compounds [(+)-catechin, (-)-epicatechin, and 3-(3,4-dihydroxyphenyl)-propionic acid]. The synthesized compound was also used to perform a targeted analysis in samples collected during the in vitro fermentation of a grape seed flavan-3-ol extract with human fecal microbiota from three healthy volunteers. The time-course formation of 5-(3',4'-dihydroxyphenyl)-γ-valerolactone revealed large interindividual differences among volunteers, with concentrations ranging from 3.31 to 77.54 μM at 10 h of fermentation. These results are further discussed in view of the scarce reports quantifying 5-(3',4'-dihydroxyphenyl)-γ-valerolactone in in vitro fermentation studies, and pharmacokinetic and intervention studies.     

Growth and lay modelling in a population of Castellana Negra native Spanish hens

1. This study shows the results of (a) fitting growth data to a Gompertz-Laird model and (b) fitting egg production per housed bird to a Grossman model, in a population of Castellana Negra hens. 2. The maximum growth point for males and females, respectively, occurred at 10.6 and 9.3 weeks and the estimated weights at maturity were 2852 and 1878 g. Growth curve parameters were lower than those of commercial lines and similar to reports of other unselected breeds. 3. The start of lay was at 23 weeks. Total egg production reached 163 at 52 weeks, with a persistency (length of constant production) of 3.4 weeks and a production maximum of 69%. The maximum rate of change segment of the laying curve started at 24.3 weeks of age, and the production maximum started at 5.8 weeks into lay. The rate of decline was 1.4% weekly. 4. The Castellana Negra breed belongs to the class of slow-growing light lines and egg production was 163 eggs per calendar year.     

Temperature Differentially Affects Gene Expression in Antarctic Thraustochytrid Oblongichytrium sp. RT2316-13

Oblongichytrium RT2316-13 synthesizes lipids rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The content of these fatty acids in the total lipids depended on growth temperature. Sequencing technology was used in this work to examine the thraustochytrid’s response to a decrease in growth temperature from 15 °C to 5 °C. Around 4% (2944) of the genes were differentially expressed (DE) and only a few of the DE genes (533 upregulated; 206 downregulated) had significant matches to those in the SwissProt database. Most of the annotated DE genes were related to cell membrane composition (fatty acids, sterols, phosphatidylinositol), the membrane enzymes linked to cell energetics, and membrane structure (cytoskeletal proteins and enzymes). In RT2316-13, the synthesis of long-chain polyunsaturated fatty acids occurred through ω3- and ω6-pathways. Enzymes of the alternative pathways (Δ8-desaturase and Δ9-elongase) were also expressed. The upregulation of the genes coding for a Δ5-desaturase and a Δ5-elongase involved in the synthesis of EPA and DHA, explained the enrichment of total lipid with these two long-chain fatty acids at the low temperature. This molecular response has the potential to be used for producing microbial lipids with a fatty acids profile similar to that of fish oils.     

Behaviour and welfare of fattening lambs supplemented with varying sizes and types of straw

The aim was to evaluate the effect of size (long or chopped wheat straw), type (barley or wheat) and presence/absence (1 week) of straw supplementation on behaviour and welfare of fattening lambs. Sixteen lambs (65 days) were individually housed to be used in two simultaneous experiments (8 lambs each) for 5 weeks of fattening. After a habituation period, they were offered straw that differed either in size (Experiment 1) or type (Experiment 2), which was followed by a week without straw and then reallocation of straw. Maintenance behaviours, play and stereotypies were analysed using video recording. Physiological stress responses were measured using blood samples taken at the end of the week without straw and after the reallocation of the straw. A preference test was performed during the last week. Concentrate and roughage consumption, average daily gain and conversion index were estimated. Lambs provided with long straw spent more time eating forage, playing and performed fewer stereotypies (Experiment 1). Lambs given wheat straw increased their play behaviour compared with those given barley straw, without affecting stereotypies (Experiment 2). When the straw was removed, lambs performed more behaviours associated with stress; however, this was not reflected in blood stress indicators. Lambs clearly preferred long straw over chopped straw, but no clear preference was observed between wheat and barley straw. The amount of forage intake was consistent with the time spent eating forage, but the amount of concentrate intake did not vary with eating concentrate behaviour, since the amount of concentrate intake was higher when the straw was removed, especially in Experiment 2. Our results show that size more than type of straw supplement impact the behaviour of lambs. Long straw reduces the stereotypies and increases play. This may improve the welfare and performance of fattening lambs, adding ethical value to a highly appreciated product.