Combined incubation of cadmium, docosahexaenoic and eicosapentaenoic acid results in increased uptake of cadmium and elevated docosapentaenoic acid content in hepatocytes in vitro.

BACKGROUND: Human hepatocellular cells Hep G2 were used to mimic and investigate the effect of the intake of cadmium (Cd(2+)) contaminated fish on cytotoxicity, fatty acid (FA) and phospholipid class composition. METHODS: Cells were incubated with a combination of Cd(2+) and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) fish specific FA. RESULTS: We measured a significant increased proportion of EPA and DHA in the treated cells compared to the control line confirming the uptake. While doses of 25 µM DHA showed to be toxic to the cells, repeated short term incubations (2 h) at lower doses resulted in an increased uptake of DHA. The resarzurin assay, evaluating cell viability, showed a significant decrease in cell viability between Cd(2+) incubation time and, further, the pre-incubation with BSA-FA complex resulted in significantly increased cell viability. On the metabolic level, increased concentrations of EPA and DHA resulted in an increased proportion of docosapentaenoic acid (DPA) which indicated an increased metabolism. Also 24-h combined incubations of 5 µM Cd(2+) and EPA and DHA showed a significant increase DPA in the total lipid fraction of the cells. In addition, incubation with 5 µM Cd(2+) for 24 h also decreased the total cardiolipin (CL) fraction from the identified phospholipids. CONCLUSIONS: We confirmed that the applied FA were taken up by the cells. A combination of EPA, DHA and Cd(2+) influenced lysosomal integrity, cell viability and lipid metabolism in the cells. The most important finding is that DHA and EPA reduced the detrimental effect of Cd(2+) on cell viability. The exact effects and kinetics behind our observations still need further evaluation.

Sustainable Delicacy: Variation in Quality and Sensory Aspects in Wild Boar (Sus scrofa) Meat and Comparison to Pork Meat-A Case Study.

The aim of this study was to evaluate quality and sensory variation in wild boar meat in comparison to pork. Meat quality in wild boar is expected to vary more compared to pork due to different feeding environment, age and gender. In order to be able to promote wild boar meat as a sustainable high-quality product, there is a need to evaluate the variation in meat quality attributes, including technological, compositional and sensory/texture aspects. We evaluated carcass characteristics, pH, colour, lipid profile and sensory aspects of wild boar meat of different age and gender and compared them with pork. Wild boars had lower carcass weight (p = <0.0001) and higher ultimate pH (p = 0.0063) compared to domestic pigs. Intramuscular fat content had a tendency to be higher in wild boar meat (p = 0.1010), as well as the proportion of nutritional valuable n-3 FA (p = 0.0029). The colour of pork was more pink (p = 0.0276) and pale (p = <0.0001) compared to meat from wild boar. Meat from wild boar gilts received the highest sensory scores. Based on these findings, we suggest that meat from younger animals could be sold in different cuts directly while meat from older animals might be more suitable for the production of sausage.

Oleaginous yeast Rhodotorula toruloides biomass effect on the metabolism of Arctic char (Salvelinus alpinus).

Sustainability issues arise when using fish oil and vegetable oils in fish feed production for aquaculture purposes. Microbial production of single cell oil is a potential alternative as a lipid ingredient in the production of fish feed. In this study, we replaced the vegetable oils with the oleaginous yeast R. toruloides biomass in the diet of Arctic char (S. alpinus) and investigated the effects on health and composition. Measurement of fish growth parameters showed a higher liver weight and hepatosomatic index in the experimental group of fish fed partly with yeast biomass compared to a control group fed a diet with vegetable oils. No significant differences in the lipid content of muscle and liver tissues were found. The fatty acid profiles in the muscle of both fish groups were similar while the experimental fish group had a higher amount of monounsaturated fatty acids in the liver. Histology of livers showed no significant difference in the number of lipid droplets. The size of hepatic lipid droplets seemed to be related to liver fat content. Quantification of metabolites in the liver revealed no differences between the fish groups while plasma metabolites involved in energy pathways such as alanine, 3-hydroxybutyrate, creatinine, serine, betaine, and choline were significantly higher in the experimental fish group.

Corrigendum: Oleaginous yeast Rhodotorula toruloides biomass effect on the metabolism of Arctic char (Salvelinus alpinus).

[This corrects the article DOI: 10.3389/fmolb.2022.931946.].

Microbial lipid production from crude glycerol and hemicellulosic hydrolysate with oleaginous yeasts.

BACKGROUND: Crude glycerol (CG) and hemicellulose hydrolysate (HH) are low-value side-products of biodiesel transesterification and pulp-and paper industry or lignocellulosic ethanol production, respectively, which can be converted to microbial lipids by oleaginous yeasts. This study aimed to test the ability of oleaginous yeasts to utilise CG and HH and mixtures of them as carbon source. RESULTS: Eleven out of 27 tested strains of oleaginous yeast species were able to grow in plate tests on CG as sole carbon source. Among them, only one ascomycetous strain, belonging to Lipomyces starkeyi, was identified, the other 10 strains were Rhodotorula spec. When yeasts were cultivated in mixed CG/ HH medium, we observed an activation of glycerol conversion in the Rhodotorula strains, but not in L. starkeyi. Two strains-Rhodotorula toruloides CBS 14 and Rhodotorula glutinis CBS 3044 were further tested in controlled fermentations in bioreactors in different mixtures of CG and HH. The highest measured average biomass and lipid concentration were achieved with R. toruloides in 10% HH medium mixed with 55 g/L CG-19.4 g/L and 10.6 g/L, respectively, with a lipid yield of 0.25 g lipids per consumed g of carbon source. Fatty acid composition was similar to other R. toruloides strains and comparable to that of vegetable oils. CONCLUSIONS: There were big strain differences in the ability to convert CG to lipids, as only few of the tested strains were able to grow. Lipid production rates and yields showed that mixing GC and HH have a stimulating effect on lipid accumulation in R. toruloides and R. glutinis resulting in shortened fermentation time to reach maximum lipid concentration, which provides a new perspective on converting these low-value compounds to microbial lipids.

Oleaginous yeasts respond differently to carbon sources present in lignocellulose hydrolysate.

BACKGROUND: Microbial oils, generated from lignocellulosic material, have great potential as renewable and sustainable alternatives to fossil-based fuels and chemicals. By unravelling the diversity of lipid accumulation physiology in different oleaginous yeasts grown on the various carbon sources present in lignocellulose hydrolysate (LH), new targets for optimisation of lipid accumulation can be identified. Monitoring lipid formation over time is essential for understanding lipid accumulation physiology. This study investigated lipid accumulation in a variety of oleaginous ascomycetous and basidiomycetous strains grown in glucose and xylose and followed lipid formation kinetics of selected strains in wheat straw hydrolysate (WSH). RESULTS: Twenty-nine oleaginous yeast strains were tested for their ability to utilise glucose and xylose, the main sugars present in WSH. Evaluation of sugar consumption and lipid accumulation revealed marked differences in xylose utilisation capacity between the yeast strains, even between those belonging to the same species. Five different promising strains, belonging to the species Lipomyces starkeyi, Rhodotorula glutinis, Rhodotorula babjevae and Rhodotorula toruloides, were grown on undiluted wheat straw hydrolysate and lipid accumulation was followed over time, using Fourier transform-infrared (FTIR) spectroscopy. All five strains were able to grow on undiluted WSH and to accumulate lipids, but to different extents and with different productivities. R. babjevae DVBPG 8058 was the best-performing strain, accumulating 64.8% of cell dry weight (CDW) as lipids. It reached a culture density of 28 g/L CDW in batch cultivation, resulting in a lipid content of 18.1 g/L and yield of 0.24 g lipids per g carbon source. This strain formed lipids from the major carbon sources in hydrolysate, glucose, acetate and xylose. R. glutinis CBS 2367 also consumed these carbon sources, but when assimilating xylose it consumed intracellular lipids simultaneously. Rhodotorula strains contained a higher proportion of polyunsaturated fatty acids than the two tested Lipomyces starkeyi strains. CONCLUSIONS: There is considerable metabolic diversity among oleaginous yeasts, even between closely related species and strains, especially when converting xylose to biomass and lipids. Monitoring the kinetics of lipid accumulation and identifying the molecular basis of this diversity are keys to selecting suitable strains for high lipid production from lignocellulose.

Comparison of Quality Changes in Eurasian Perch (Perca fluviatilis L.) Fillets Originated from Two Different Rearing Systems during Frozen and Refrigerated Storage.

The current knowledge on how different Eurasian perch rearing systems impact the final fillet quality is scant. Therefore, two domestic storage conditions were investigated-10 months frozen (-20 °C) and 12 days refrigerated (+4 °C) storage conditions-in order to determine (i) how the choice of rearing system affects fillets quality during different processing conditions and (ii) if oxidative changes and other quality parameters were interactive. For the proposed idea, proteome analysis, oxidative changes, and some quality parameters were considered in this study. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) indicated a higher loss of protein in the frozen fillets from ponds (PF) than the fillets from recirculating aquaculture systems (RAS) (RF). Western blot showed a higher protein carbonyls level in RF compared to PF, which was confirmed by the total protein carbonyls during frozen storage. PF indicated less liquid loss, hardness, and oxidation progress than RF in both storage conditions. The biogenic amines index (BAI) in the fillets from either origin showed acceptable levels during storage at +4 °C. Furthermore, the n-3/n-6 ratio was similar for both fillets. The deterioration of fillets during frozen storage was mainly caused by formation of ice crystals followed by protein oxidation, while protein oxidation was the main concern during refrigerated storage confirmed by principal component analysis (PCA) analysis.

Fatty acid composition of black soldier fly larvae (Hermetia illucens) - Possibilities and limitations for modification through diet.

Black soldier fly larvae (Hermetia illucens; BSFL) can convert organic wastes into a nutrient-rich biomass suitable in animal feed, which could be a way to achieve more sustainable production of food. However, little is known about how the diet fed to BSFL affects their nutritional value, especially their fatty acid composition. In this study, BSFL were fed 11 diets based on four different organic waste sources (mussels, bread, fish and food waste). Fatty acid and proximate composition (dry matter, crude fat, crude protein and ash) were analysed in the diets, in two-week-old larvae and substrate residues. Larval weight, survival and feed conversion were also recorded. The diet was found to affect all parameters investigated. Irrespective of diet, the larval fat consisted mainly of lauric acid and other saturated fatty acids and these were found to be synthesised by the larvae. However, both the fatty acid composition of the substrate, and the larval weight were found to affect the fatty acid profile of the larvae. In general, larvae with a higher weight contained a higher percentage of saturated fatty acids and a lower percentage of unsaturated fatty acids, such as eicosapentaenoic (EPA) and docosahexaenoic acid (DHA). It was concluded that the possibilities to tailor the fatty acid composition of the BSFL through the diet are limited; thus, the BSFL fat may not be suitable to replace fish oil, but has potential of inclusion in other food, feed and fuel products.

Lipolysis and Oxidation in Ultra-High Temperature Milk Depend on Sampling Month, Storage Duration, and Temperature.

During storage, some factors (for example, storage duration and temperature) can affect milk stability and consumer acceptability. Thiobarbituric acid reactive substances (TBARSs), lipid classes, and fatty acid profiles in stored ultra-high temperature (UHT) milk were analyzed to assess the effects of storage time and temperature on lipid oxidation and lipolysis. With storage duration up to 12 months, the milk fat phase was separated and showed high levels of oxidation and lipolysis, manifested as increased levels of TBARS and free fatty acids. High oxidation levels decreased the percentage of unsaturated fatty acids (UFAs) in triacylglycerol and phospholipids. Higher storage temperatures (20, 30, and 37 °C) resulted in a higher degree of fat aggregation, oxidation, and lipolysis compared with refrigerated storage (4 °C). Additionally, sampling month of raw milk (May, July, and November) affected the lipid profiles of UHT milk during storage, with more UFA oxidized in July than in the other 2 months.

FT-NIR: a tool for rapid intracellular lipid quantification in oleaginous yeasts.

BACKGROUND: Lipid extraction for quantification of fat content in oleaginous yeasts often requires strong acids and harmful organic solvents; it is laborious and time-consuming. Therefore, in most cases just endpoint measurements of lipid accumulation are performed and kinetics of intracellular lipid accumulation is difficult to follow. To address this, we created a prediction model using Fourier-transform near-infrared (FT-NIR) spectroscopy. This method allows to measure lipid content in yeast. METHODS: The FT-NIR calibration sets were constructed from spectra of freeze-dried cells of the oleaginous yeasts Rhodotorula toruloides CBS 14, Lipomyces starkeyi CBS 1807 and Yarrowia lipolytica CBS 6114. The yeast cells were obtained from different cultivation conditions. Freeze-dried cell pellets were scanned using FT-NIR in the Multi Purpose Analyser (MPA) from Bruker. The obtained spectra were assigned corresponding to total fat content, obtained from lipid extraction using a modified Folch method. Quantification models using partial least squares (PLS) regression were built, and the calibration sets were validated on independently cultivated samples. The R. toruloides model was additionally tested on Rhodotorula babjevae DBVPG 8058 and Rhodotorula glutinis CBS 2387. RESULTS: The R 2 of the FT-NIR model for R. toruloides was 98%, and the root mean square error of cross-validation (RMSECV) was 1.53. The model was validated using a separate set of R. toruloides samples with a root mean square error of prediction (RMSEP) of 3.21. The R 2 of the Lipomyces model was 96%, with RMSECV 2.4 and RMSEP 3.8. The R 2 of the mixed model, including all tested yeast strains, was 90.5%, with RMSECV 2.76 and RMSEP 3.22, respectively. The models were verified by predicting the total fat content in newly cultivated and freeze-dried samples. Additionally, the kinetics of lipid accumulation of a culture were followed and compared with standard lipid extraction methods. CONCLUSIONS: Using FT-NIR spectroscopy, we have developed a faster, less laborious and non-destructive quantification of yeast intracellular lipid content compared to methods using lipid extraction.

Swimming at different temperatures: The lipid composition of sperm from three freshwater fish species determined by mass spectrometry and nuclear magnetic resonance spectroscopy.

The spawning behavior of different fish species is as diverse as their habitats. A lot of factors influence the (phospho)lipid composition of fish sperm, including the water temperature at which spawning takes place. Therefore, this study aimed on the elucidation of the phospholipid composition of sperm from three fish species from different orders (common carp - Cyprinus carpio, northern pike - Esox lucius and burbot - Lota lota) with different spawning temperatures by nuclear magnetic resonance spectroscopy (NMR), matrix-assisted laser desorption/ionization mass spectrometry and thin-layer chromatography (TLC) coupled to electrospray ionization mass spectrometry as well as gas chromatography. Next to the lipid composition that was different for carp, northern pike and burbot, regarding the moieties of the different (phospho)lipid classes (particularly sphingomyelin and acidic phospholipids) and the saturation degree of the fatty acyl residues, there were differences observed depending on the analytical method that was used. The results from TLC and NMR investigations differed regarding the amounts of the different phospholipids. Reasons for these discrepancies are discussed in detail.

The possible involvement of oxidative stress in the oocyte ageing process in goldfish Carassius auratus (Linnaeus, 1758).

Decreasing egg quality following oocyte ageing is a major restricting factor for the breeding programs. The mechanisms behind this process has not yet been clarified. To examine the possible involvement of oxidative stress in the oocyte ageing process, the relative mRNA abundance of specific transcripts were determined in oocytes collected from 6 females and incubated in vitro for 18 hours post stripping at 20 °C in goldfish Carassius auratus. During the 18 hour-post-stripping ageing of the oocytes, relative mRNA levels of candidate transcripts involved in oxidative injury, mitochondrial function and stress response, cell cycles, apoptosis, reproduction and germ line speciation and developmental competence were measured by real-time PCR. None of the relative mRNA abundance of the examined genes were significantly altered through oocyte ageing. In addition, the amount of thiobarbituric acid reactive substances (TBARS), an indicator of lipid peroxidation, did not change over time following stripping. The activity of the antioxidant enzymes also remained constant during oocyte ageing. The results of the current study indicated that oxidative stress unlikely plays a role as an initiator or promotor in the progress of oocyte ageing in goldfish.

Sperm motility and lipid composition in internally fertilizing ocellate river stingray Potamotrygon motoro.

All extant groups of Elasmobranches have internal fertilization and the structure of the male reproductive organs is very specific: sperm passes from the internal organs via the cloaca, but the male copulating organ (clasper) is distant from the cloaca. This suggests that sperm can contact the surrounding medium before fertilization. Because of this involvement with the environment, external signaling in sperm motility activation could occur in these species even though their fertilization mode is internal. In this case, spermatozoa of Elasmobranches should hypothetically possess a specific structure and membrane lipid composition which supports physiological functions of the sperm associated with environmental tonicity changes occurring at fertilization. Additionally, sperm motility properties in these taxa are poorly understood. The current study examined sperm lipid composition and motility under different environmental conditions for the ocellate river stingray, Potamotrygon motoro, an endemic South America freshwater species. Sperm samples were collected from six mature males during the natural spawning period. Sperm motility was examined in seminal fluid and fresh water by light video microscopy. Helical flagellar motion was observed in seminal fluid and resulted in spermatozoon progression; however, when diluted in fresh water, spermatozoa were immotile and had compromised structure. Lipid class and fatty acid (FA) composition of spermatozoa was analyzed by thin layer and gas chromatography. Spermatozoa FAs consisted of 33 ±â€¯1% saturated FAs, 28 ±â€¯1% monounsaturated FAs (MUFAs), and 41 ±â€¯1% polyunsaturated FAs (PUFAs), and a high content of n-6 FAs (32 ±â€¯2%) was measured. These results allowed us to conclude that sperm transfer from P. motoro male into female should occur without coming into contact with the hypotonic environment so as to preserve potent motility. In addition, this unusual reproductive strategy is associated with specific spermatozoa structure and lipid composition. Low level of docosahexaenoic acid and relatively low PUFA/MUFA ratio probably account for the relatively low fluidity of freshwater stingray membrane and can be the main reason for its low tolerance to hypotonicity.

Alteration of mRNA abundance, oxidation products and antioxidant enzyme activities during oocyte ageing in common carp Cyprinus carpio.

Oocyte ageing is the most important factor affecting egg quality of several fish species after ovulation. Oxidative stress has been proposed as the initiator of the oocyte ageing process in other vertebrates. To identify the role of oxidative stress and apoptosis on the progress of oocyte ageing in the common carp Cyprinus carpio, changes in the relative mRNA abundance of selected transcripts were examined. The possible alteration in the oxidation status of the oocytes during ageing was also studied. In addition, the activity of antioxidant enzymes during oocyte ageing was evaluated. Oocytes from 6 females were incubated in vivo for 14 hours post-ovulation (HPO) and in vitro for 10 hours post-stripping (HPS) at 20°C before fertilization. Hatching rates were over 65% up to 4-6 HPO, finally dropping to 1.3% at 12-14 HPO.Hatching rates were over 65% up to 4-6 HPO, finally dropping to 1.3% at 12-14 HPO. Hatching rates were more than 70% for the eggs stored in vitro up to 6 HPS and then decreased to 21.3% at 10 HPS. The results demonstrated no significant changes in the relative mRNA levels of oxidative stress-related genes or genes involved in the cell cycle during the progress of oocyte ageing in common carp. Additionally, the amount of TBARS and carbonyls did not change as time elapsed following ovulation. The apoptosis-related genes however, were significantly altered following the prolonged time interval between ovulation and fertilization. The lack of response of both activities of antioxidant enzymes and oxidation products during oocyte ageing strengthens the conclusion that oxidative stress is unlikely to be a main factor determining the progress of oocyte ageing in common carp. However, an increase in the mRNA abundance of apoptosis-related genes demonstrates that apoptotic pathway might be involved in the progress of oocyte ageing.

Proteome analysis of xylose metabolism in Rhodotorula toruloides during lipid production.

BACKGROUND: Rhodotorula toruloides is a promising platform organism for production of lipids from lignocellulosic substrates. Little is known about the metabolic aspects of lipid production from the lignocellolosic sugar xylose by oleaginous yeasts in general and R. toruloides in particular. This study presents the first proteome analysis of the metabolism of R. toruloides during conversion of xylose to lipids. RESULTS: Rhodotorula toruloides cultivated on either glucose or xylose was subjected to comparative analysis of its growth dynamics, lipid composition, fatty acid profiles and proteome. The maximum growth and sugar uptake rate of glucose-grown R. toruloides cells were almost twice that of xylose-grown cells. Cultivation on xylose medium resulted in a lower final biomass yield although final cellular lipid content was similar between glucose- and xylose-grown cells. Analysis of lipid classes revealed the presence of monoacylglycerol in the early exponential growth phase as well as a high proportion of free fatty acids. Carbon source-specific changes in lipid profiles were only observed at early exponential growth phase, where C18 fatty acids were more saturated in xylose-grown cells. Proteins involved in sugar transport, initial steps of xylose assimilation and NADPH regeneration were among the proteins whose levels increased the most in xylose-grown cells across all time points. The levels of enzymes involved in the mevalonate pathway, phospholipid biosynthesis and amino acids biosynthesis differed in response to carbon source. In addition, xylose-grown cells contained higher levels of enzymes involved in peroxisomal beta-oxidation and oxidative stress response compared to cells cultivated on glucose. CONCLUSIONS: The results obtained in the present study suggest that sugar import is the limiting step during xylose conversion by R. toruloides into lipids. NADPH appeared to be regenerated primarily through pentose phosphate pathway although it may also involve malic enzyme as well as alcohol and aldehyde dehydrogenases. Increases in enzyme levels of both fatty acid biosynthesis and beta-oxidation in xylose-grown cells was predicted to result in a futile cycle. The results presented here are valuable for the development of lipid production processes employing R. toruloides on xylose-containing substrates.

Nutritional value of several commercially important river fish species from the Czech Republic.

Proximate and fatty acid (FA) composition of seven freshwater fish species from the Czech Republic were examined. Moreover, the index of atherogenicity (IA) and the index of thrombogenicity (IT) were calculated from the obtained data. These two indices along with the total content of the essential n-3 FAs, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) as well as the ratio of n-6/n3 FAs, provide good indicators for the nutritional value of the fish. The species had been selected owing to the limited amount of information about their nutritional composition available. Furthermore, they are not typically subject to aquaculture, being almost exclusively obtained by angling. The protein content was relatively stable in all species (17.1 ± 1.55 to 19.2 ± 2.20 g/100 g). The content of carbohydrates ranged from 0.02 ± 0.1 to 0.99 ± 0.0 g/100 g and ash from 1.08 ± 0.20 to 2.54 ± 1.57 g/100 g. As expected, a high variability was observed in the fat content (0.74 ± 0.04 to 4.04 ± 0.81 g/100 g) and the FA composition, as well as the contents of EPA and DHA. IA and IT were close to the values stated for the Eskimo diet, indicating a high nutritional value with a positive effect for human health.

Spermatozoa quality and sperm lipid composition in intensively cultured and wild burbot (Lota lota).

The aim of this study was to compare the spermatozoa quality parameters in spermatozoa of RAS (Recirculating Aquaculture System; RAS group) cultured (commercial pellets) and natural condition cultured (WILD group) burbot Lota lota (live prey, Pseudorasbora parva). Seven of nine fish of the RAS group produced sperm, with sperm from only four of the fish having a motility of >5%. Sperm were collected from all nine fish of the WILD group, and sperm of six of the fish from the WILD group had motility of about 100% and three had sperm with 50% to 60% motility. Spermatozoa from the RAS group had a delay in activation compared to the WILD group. Fish from the RAS group also had a lesser volume of sperm (1.8 ± 1.2 mL) collected compared to the WILD group (3.6 ± 1.2 mL). Compared to the RAS group, sperm of the WILD group had a greater proportion of saturated fatty acids (SFA), as well as the phospholipid, phosphatidylethanolamine. The findings indicate that fish grown in natural conditions may be more suitable as broodstock. Ongoing research to develop methods of enhancing reproductive performance of burbot broodstock cultured in RAS is needed to investigate whether the quality of sperm can be improved by adjusting environmental conditions, diet, or combination of these factors.

Oleaginous yeast as a component in fish feed.

This study investigates the replacement of vegetable oil (VO) in aquaculture feed for Arctic char (Salvelinus alpinus) with oil produced by the oleaginous yeast Lipomyces starkeyi grown in lignocellulose (wheat straw) hydrolysate. VO is extensively used to partially replace fish oil in aquaculture feed, which can be seen as non-sustainable. VO itself is becoming a limited resource. Plant oils are used in many different applications, including food, feed and biodiesel. Its replacement in non-food applications is desirable. For this purpose, yeast cells containing 43% lipids per g dry weight were mechanically disrupted and incorporated into the fish feed. There were no significant differences in this pilot study, regarding weight and length gain, feed conversion ratio, specific growth rate, condition factor and hepatosomatic index between the control and the yeast oil fed group. Fatty and amino acid composition of diet from both groups was comparable. Our results in fish demonstrate that it is possible to replace VO by yeast oil produced from lignocellulose, which may broaden the range of raw materials for food production and add value to residual products of agriculture and forestry.

Biomarker response, health indicators, and intestinal microbiome composition in wild brown trout (Salmo trutta m. fario L.) exposed to a sewage treatment plant effluent-dominated stream.

Concerns about the effect of sewage treatment plant (STP) effluent on the health of freshwater ecosystems have increased. In this study, a unique approach was designed to show the effect of an STP effluent-dominated stream on native wild brown trout (Salmo trutta L.) exposed under fully natural conditions. Zivny stream is located in South Bohemia, Czech Republic. The downstream site of Zivny stream is an STP-affected site, which receives 25% of its water from Prachatice STP effluent. Upstream, however, is a minimally polluted water site and it is considered to be the control site. Native fish were collected from the upstream site, tagged, and distributed to both upstream and downstream sites. After 30, 90, and 180days, fish were recaptured from both sites to determine whether the downstream site of the Zivny stream is associated with the effects of environmental pollution. Several biomarkers indicating the oxidative stress and antioxidant enzyme activities, cytochrome P450 activity, xenoestrogenic effects, bacterial composition, and lipid composition were investigated. Additionally, polar chemical contaminants (pharmaceuticals and personal care products (PPCPs)) were quantified using polar organic chemical integrative samplers (POCIS). Fifty-three PPCPs were detected in the downstream site; 36 of those were constantly present during the 180-day investigation period. Elevated hepatic 7-benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylase (BFCOD) (after 90days) and blood plasma vitellogenin concentrations in males were detected in fish downstream of the STP effluent during all sampling events. An increase in the fishes' total fat content was also observed, but with low levels of ω-3 fatty acid in muscle tissue. Two bacterial taxa related to activated sludge were found in the intestines of fish from downstream. Our results show that Prachatice STP is a major source of PPCPs in the Zivny stream, which has biological consequences on fish physiology.

The effects of sewage treatment plant effluents on hepatic and intestinal biomarkers in common carp (Cyprinus carpio).

Sewage treatment plants (STPs) are one of the major source of pharmaceuticals and personal care products in the aquatic environment. Generally, the effects of individual chemicals on fish are studied under laboratory conditions, which leads to results that are potentially not realistic regarding the effects of these chemicals under environmental conditions. Therefore, in this study, common carps were held in exposed pond that receive water from STP effluents for 360 days under natural conditions. Elimination of xenobiotics starts in the fish intestine, in which the microbial community strongly influences its function. Moreover, the fish intestine functions as crucial organ for absorbing lipids and fatty acids (FA), with consequent transport to the liver where their metabolism occurs. The liver is the primary organ performing xenobiotic metabolism in fish, and therefore, the presence of pollutants may interact with the metabolism of FA. The catalytic activity of CYP1A and CYP3A-like enzymes, their gene expression, FA composition and intestinal microbiome consortia were measured. The catalytic activity of enzymes and their gene and protein expression, were induced in hepatic and intestinal tissues of fish from the exposed pond. Also, fish from the exposed pond had different compositions of FA than those from the control pond: concentration of 18:1 n-9 and 18:2 n-6 were significantly elevated and the longer chain n-3 FA 20:5 n-3, 22:5 n-3 and 22:6 n-3 were significantly lowered. There were clear differences among microbiome consortia in fish intestines across control and exposed groups. Microbiome taxa measured in exposed fish were also associated with those found in STP activated sludge. This study reveals that treated STP water, which is assumed to be clean, affected measured biomarkers in common carp.