Optimal bruk av planteproteinkonsentrater i ekstrudert fôr til kjøttetende fisk

The main objective of the research presented in this thesis was to evaluate multiple amino acidsupplemented plant protein concentrates as the main protein source in diets for carnivorous fish. This objective was approached experimentally by: 1) Determining the effects of using plant protein concentrates to provide 30 %, 50 %, and 95 % of total protein in extruded diets for rainbow trout and black sea bream on growth performance, feed utilization, and nutrient digestibility and retention; 2) Examining if combinations of different plant protein concentrates diminishes the negative effects of anti-nutritional factors associated with single plant protein sources when fed to carnivorous fish, thereby facilitating higher dietary plant protein inclusions; 3) Evaluating the effects of dietary inclusion of plant protein concentrates on fish physiology and health; 4) Assessing the efficiency of pre-treatment of plant protein concentrates with phytase to improve utilization of minerals and macronutrients in rainbow trout; 5) Quantifying the changes in loss of nutrients to the environment by replacing high-quality fish meal with untreated or dephytinized plant protein concentrates. The dietary plant protein concentrates used in this thesis were all supplemented with multiple limiting amino acids and taurine. Four experiments were conducted, and the results are reported in four papers. Paper I (Optimal inclusion of lupin and pea protein concentrates in extruded diets for rainbow trout (Oncorhynchus mykiss)) focuses on the response of rainbow trout to nine extruded diets. These diets included eight plant protein based diets formulated using four mixtures of lupin (LPC) and pea protein concentrates (PPC) (L/P ratio, 3:0, 2:1, 1:2 and 0:3) with two dietary inclusion levels (300 or 500 g plant protein kg -1 dietary protein) and one diet using LT-fish meal as the sole protein source (FM diet). The same nine diets, but with a lower level of oil coating, were also studied in black sea bream in Paper II (Mixtures of lupin and pea protein concentrates can efficiently replace high-quality fish meal in extruded diet for juvenile black sea bream (Acanthopagrus schlegeli)) In Paper III (Optimizing plant protein combinations in fish meal-free diets for rainbow trout (Oncorhynchus mykiss) by a mixture model) three plant protein premixtures (P-MIX, C-MIX, and S-MIX) were prepared by mixing four protein concentrates. These were pea protein concentrates (PPC), canola protein concentrate (CPC), potato protein concentrate, and soy protein concentrate (SPC). Seven plant protein based diets were formulated based on a mixture design using P-MIX, C-MIX, and S-MIX alone or in combinations to provide > 95% of the dietary protein. These diets were supplemented with 5% krill products as feeding attractant. One diet using LT-fish meal as the sole protein source (FM diet) was also produced. All diets were designed to contain equal amounts of digestible protein and digestible energy, and fed to rainbow trout. In Paper IV (Incubation of soy protein concentrate with phytase improves the nutritional value of a fish meal-free diet for rainbow trout (Oncorhynchus mykiss)), two of the diets from Paper III (the FM and SPC diets), and one additional experimental diet based on dephytinized SPC (DSPC diet) were fed to rainbow trout. In Papers I and II, the trout grew from 58 to 180 g during the 62 days of feeding, and the black sea bream grew from 13 to 46 g during 60 days. No significant differences in growth rate among dietary treatments were seen for either species. The only body composition parameter significantly affected by the diets in both experiments was ash, which could be ascribed to the presence of phytic acid in the plant protein concentrates. The highest inclusion of 500 g plant protein concentrates kg-1, and the diets with the highest L/P ratio resulted in higher feed intake (FI) in black sea bream. No significant effects of diet on feed intake were observed in rainbow trout. The average feed conversion ratio (FCR) was 0.72 g dry matter intake (g gain)-1 in rainbow trout, and 1.13 g g-1 in black sea bream. The diets with the highest L/P ratio resulted in higher FCR in both species, due to the higher content of non-starch polysaccharides in LPC than in PPC. In rainbow trout, the apparent digestibility of starch was reduced as a consequence of increasing dietary LPC at both inclusion levels, while the digestibility of lipid was increased with increasing dietary LPC only at high inclusion level. The diet with the highest inclusion of PPC resulted in reduced maltase activity in the intestines of both trout and sea bream. Trout fed the diet with the highest inclusion of PPC tended to have a slight decrease in mucosal fold height and a slight increase in fold fusion. The energy retentions did not significantly differ between the FM and plant protein diets in either species. Several of the plant protein-concentrate based diets resulted in similar or significantly higher nitrogen retentions in both species. The conclusion of the two papers was that any combination of essential amino acid (EAA)-fortified LPC and PPC can be efficiently used when total dietary plant protein inclusion is limited to 300 g kg-1. At higher inclusion, PPC seemed preferable. The reduced mineralization and tendencies of change in the intestinal physiology caused by the LPC and PPC require further attention. In Papers III and IV the trout grew from 61 to 214 g during 72 days of feeding, without significant differences in weight gain among dietary treatments. In both experiments, fish fed the plant protein based diets had significantly higher feed intake, FCR, and metabolic nitrogen (N) loss than those fed the FM diet. Digestibility of most nutrients other than mineral elements, and body composition did not significantly differ from the fish fed the FM diet. The digestibility of energy and retentions of both N and energy were significantly lower in trout fed the plant concentrate diets. Fish fed diets with P-MIX, containing protein from pea and potato, exhibited inflammatory changes of mild or moderate severity in the distal intestine. The mixture model predicted different optimal diet formulations based on different response criteria. A combination of P-MIX and C-MIX gave most efficient feed conversion. The digestibility of N and amino acids were maximized when S-MIX was used alone. The digestibility of lipid and energy were maximized by a combination of P-MIX and S-MIX. Retention of ingested N was most efficient when combining P-MIX and S-MIX, while the highest retention of digested N was obtained by a combination of P-MIX and C-MIX. Using CMIX alone supported the highest digestibility and retention of P, and whole-body concentrations of ash, P, Ca and Mg. The reason was that the CPC in C-MIX had been incubated with phytase by the producer. Dephytinization of the SPC also resulted in significant improvements in the utilization of dietary P, as well as Ca and Mg. In addition, dephytinization of the SPC resulted in lower FCR, and increased digestible energy concentration in the diet. Both metabolic losses of N and faecal loss of energy were higher for the plant protein diets than for the FM diet, while the faecal loss of P was higher for the FM diet. Dephytinization of SPC led to a reduction in faecal and metabolic loss of N and P, and faecal loss of energy. Considerable benefits both to secure the welfare of the fish and to minimize losses of P and N into water can thus be achieved by the use of dephytinized plant protein concentrates in fish feeds.Hovedmålet med forskningen presentert i denne avhandlingen var å undersøke anvendeligheten av ulike planteproteinkonsentrater anriket med flere begrensende aminosyrer, som hovedkilde i fôr til kjøttetende fisker. Følgende eksperimentelle tilnærminger ble benyttet for å nå dette målet: 1) Undersøke effekten på vekst, fôrutnyttelse, fordøyelighet og retensjon av næringsstoffer når planteproteinkonsentrater utgjør 30, 50 og 95 % av proteinet i ekstruderte fôr til både regnbueørret og black sea bream; 2) Finne ut hvorvidt kombinasjoner av ulike planteproteinkonsentrater kan redusere de negative effektene av antinæringsstoffer som forekommer i enkelte formidler fra planter, og derved gjøre det mulig å øke mengden planteprotein i fiskefôr; 3) Vurdere effekten av planteproteinkonsentrater på fiskens fysiologi og helse; 4) Måle effekten av å forbehandle planteproteinkonsentrater med fytase for å øke utnyttelsen av mineraler og hovednæringsstoffer hos regnbueørret; 5) Kvantifisere endringene i tap av næringsstoffer til miljøet som oppnås ved å bytte ut fiskemel av høy kvalitet som eneste kilde til protein i fôret med intakte eller fytasebehandlede planteproteinkonsentrater. Alle planteproteinkonsentratene som ble benyttet i dette arbeidet ble anriket med begrensende aminosyrer og taurin. Fire forsøk ble utført, og resultatene er rapportert i fire manuskript. Paper I (Optimal inclusion of lupin and pea protein concentrates in extruded diets for rainbow trout (Oncorhynchus mykiss)) fokuserer på responsen av ni ekstruderte for. Disse fôrene ble formulert ved å lage fire blandinger med lupin (LPC) og erteproteinkonsentrat (PPC) (L/P ratio, 3:0, 2:1, 1:2 og 0:3) og med to ulike nivåer av innblanding (300 eller 500 g planteprotein kg-1 fôrprotein), og et fôr med LT fiskemel som eneste proteinkilde (FM diet). I Paper II (Mixtures of lupin and pea protein concentrates can efficiently replace high-quality fish meal in extruded diet for juvenile black sea bream (Acanthopagrus schlegeli)), ble responser av de samme fôrene som ble benyttet i Paper I undersokt i black sea bream, men med lavere fettinnhold. I Paper III (Optimizing plant protein combinations in fish meal-free diets for rainbow trout (Oncorhynchus mykiss) by a mixture model) ble tre blandinger av planteproteinkonsentrater (P-MIX, C-MIX og S-MIX) fremstilt ved å blande fire planteproteinkonsentrater. Disse var proteinkonsentrater fra erter (PPC), canola raps (CPC), potet og soyabønner (SPC). Sju fôr basert på planteproteiner ble formulert med utgangspunkt i et mixture design. P-MIX, C-MIX og S-MIX ble benyttet alene, eller i kombinasjon slik at de utgjorde mer enn 95 % av fôrets protein. Fôrene inneholdt også 5 % krillprodukter for å stimulere fôropptak. Et fôr som inneholdt LT fiskemel som eneste proteinkilde (FM diet) ble også produsert. Alle fôrene var formulert til å inneholde samme mengde fordøyelig protein og energi, og ble fôret til regnbueørret. I Paper IV (Incubation of soy protein concentrate with phytase improves the nutritional value of a fish meal-free diet for rainbow trout (Oncorhynchus mykiss)), ble to av fôrene fra Paper III (FM og SPC) og et forsøksfôr basert på SPC med redusert innhold av fytinsyre (DSPC diet) gitt til regnbueørret. I Papers I og II vokste ørreten fra 58 til 180 g i løpet av 62 fôringsdager, og black sea bream vokste fra 13 til 46 g i løpet av 60 dager. Ingen signifikante forskjeller i veksthastighet ble observert for noen av artene. Det eneste målet for kroppssammensetning som ble signifikant påvirket av fôrene, var askeinnholdet. Dette kan tilskrives innholdet av fytinsyre i plantekonsentratene. Det høyeste innblandingsnivaet av planteprotein i foret (500 g kg-1), og fôrene med høyest L/P ratio førte til økt fôropptak hos black sea bream. Ingen av fôrene gav signifikant effekt på fôropptak hos regnbueørret. Gjennomsnittet for fôrutnyttelse (FCR) hos regnbueørret var 0,72 g fôrtorrstoff (g tilvekst)-1, og 1.13 g g-1 hos black sea bream. Fôrene med høyest L/P ratio førte til høyere FCR hos begge arter, på grunn av at LPC inneholdt mer ufordøyelige polysakkarider enn de andre proteinkonsentratene. Fordøyelighet av stivelse hos regnbueørret ble redusert ved økende innblanding av LPC, mens fettfordøyeligheten økte med økende innhold av LPC ved høyeste innblandingsnivå av planteprotein i fôret. Fôret med mest PPC førte til redusert aktivitet av maltase i tarmen hos både ørret og bream. Ørret som fikk fôr med mest PPC tenderte til å ha noe lavere høyde på tarmtottene, og en svak økning i sammenslåtte tarmtotter. Retensjonen av energi var ikke signifikant forskjellig for FM og fôrene med planteprotein hos noen av artene. Flere av fôrene med planteproteinkonsentrat resulterte i tilsvarende eller høyere retensjon av nitrogen hos begge arter. Konklusjonen fra de to manuskriptene var at LPC og PPC, supplert med aminosyrer, utnyttes effektivt når total innblanding begrenses til 300 g (kg for)-1. Ved høyere innblanding virket det som PPC hadde fordeler. Den reduserte mineraliseringen og tendensene til endringer i tarmfysiologien forårsaket av LPC og PPC krever videre oppfølging. I Papers III og IV vokste ørreten fra 61 til 214 g i løpet av 72 fôringsdager. Det var ikke signifikante forskjeller mellom fôrene for veksthastighet. I begge forsøkene hadde fiskene som fikk fôr med planteproteiner signifikant høyere fôropptak, FCR, og metabolsk tap av nitrogen (N) enn de som fikk for med FM. Fordøyelighet av andre næringsstoffer enn mineraler, og kroppssammensetning var ikke signifikant forskjellig fra fisk som fikk fôr med FM. Fordøyelighet av energi, og retensjon av både N og energi var signifikant lavere hos ørret som fikk fôr med planteproteiner. Fisk som fikk fôr med P-MIX, som inneholdt protein fra erter og potet, hadde mild til moderat betennelse i baktarmen. Modellen for å analysere resultater fra mixture design gav ulike prediksjoner for optimal fôrformulering når ulike responskriteria ble benyttet. Kombinasjon av P-MIX og C-MIX førte til mest effektiv fôrutnyttelse. Fordøyelighet av N og aminosyrer var mest effektiv når S-MIX ble benyttet alene. Fordøyelighet av fett og energi ble maksimert ved en kombinasjon av P-MIX og S-MIX, mens høyest retensjon av fordøyd N ble oppnådd med å kombinere P-MIX og C-MIX. Bruk av C-MIX alene førte til høyest fordøyelighet og retensjon av P, og innhold av P, Ca, Mg og aske i fiskekroppen. Grunnen var at CPC i C-MIX hadde blitt innkubert med fytase av produsenten. Nedbrytning av fytinsyre i SPC førte også til signifikant forbedring i utnyttelsen av P, Ca og Mg. I tillegg førte hydrolyse av fytinsyre i SPC til lavere FCR, og økt konsentrasjon av fordøyelig energi i fôret. Innkubering av SPC med fytase førte til en klar reduksjon i tap av både P og N i feces og som metabolske tap, og reduserte også tap av energi i feces. Betydelige fordeler, både for å sikre fiskens velferd og redusere utslipp and P og N til vannet, kan oppnås ved å benytte planteproteinskonsentrater med redusert innhold av fytinsyre i fôr til fisk.Norges Forskningsrå

The ErbB3 binding protein Ebp1 interacts with Sin3A to repress E2F1 and AR-mediated transcription

Ectopic expression of ebp1, a member of the PA2G4 family, inhibits the proliferation and induces the differentiation of human breast and prostate cancer cell lines. Ebp1 inhibits transcription of E2F1 and androgen receptor regulated genes such as prostate specific antigen (PSA) through its interactions with histone deacetylases (HDACs). To further understand Ebp1's interactions with other components of the transcriptional repression machinery, we examined the association of Ebp1 with the corepressor Sin3A. Ebp1 interacted with Sin3A both in vitro and in vivo as demonstrated by glutathione S-transferase (GST) pull-down and coimmunoprecipitation analysis. The C-terminal domain of Ebp1, responsible for its ability to repress transcription and arrest cell growth, was necessary and sufficient for binding Sin3A. The C-terminal domain of Sin3A, containing the paired amphipathic domain 4 and the HDAC interacting domain, bound Ebp1. Recombinant Sin3A bound Ebp1 directly, but recombinant HDAC2 failed to bind Ebp1. Chromatin immunoprecipitation (ChIP) and DNA affinity precipitation analysis demonstrated that Ebp1 and Sin3A associate at the PSA and E2F1 promoters. Functionally, Sin3A enhanced the ability of Ebp1 to repress transcription of androgen receptor (AR) and E2F1 regulated genes. These results demonstrate that Ebp1 participates in transcriptional regulation via its interaction with the Sin3–HDAC

Evaluation of conventional or hydrolyzed stickwater from food-grade skipjack tuna by-product in diet for hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

acceptedVersio

Impact of morphology on polaron delocalization in a semicrystalline conjugated polymer

We investigate the delocalization of holes in the semicrystalline conjugated polymer poly(2,5-bis(3-alkylthiophene-2-yl)thieno[3,2-b]thiophene) (PBTTT) by directly measuring the hyperfine coupling between photogenerated polarons and bound nuclear spins using electron nuclear double resonance spectroscopy. An extrapolation of the corresponding oligomer spectra reveals that charges tend to delocalize over 4.0–4.8 nm with delocalization strongly dependent on molecular order and crystallinity of the PBTTT polymer thin films. Density functional theory calculations of hyperfine couplings confirm that long-range corrected functionals appropriately describe the change in coupling strength with increasing oligomer size and agree well with the experimentally measured polymer limit. Our discussion presents general guidelines illustrating the various pitfalls and opportunities when deducing polaron localization lengths from hyperfine coupling spectra of conjugated polymers

Alterations in cell growth and signaling in ErbB3 binding protein-1 (Ebp1) deficient mice

<p>Abstract</p> <p>Background</p> <p>The ErbB3 binding protein-1 (Ebp1) belongs to a family of DNA/RNA binding proteins implicated in cell growth, apoptosis and differentiation. However, the physiological role of Ebp1 in the whole organism is not known. Therefore, we generated <it>Ebp1</it>-deficient mice carrying a gene trap insertion in intron 2 of the <it>Ebp1 (pa2g4) </it>gene.</p> <p>Results</p> <p>Ebp1^-/-mice were on average 30% smaller than wild type and heterozygous sex matched littermates. Growth retardation was apparent from Day 10 until Day 30. IGF-1 production and IGBP-3 and 4 protein levels were reduced in both embryo fibroblasts and adult knock-out mice. The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells. Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway. In addition, the expression or activation of proliferation related genes such as AKT and the androgen receptor, previously demonstrated to be affected by Ebp1 expression <it>in vitro</it>, was altered in adult tissues.</p> <p>Conclusion</p> <p>These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific. The Ebp1^-/-mouse line represents a new <it>in vivo </it>model to investigate Ebp1 function in the whole organism.</p

The ratio between digestible protein and digestible energy affects accumulation and depuration of geosmin and 2-methylisoborneol (2-MIB) in Japanese seabass (Lateolabrax japonicus) raised in a recirculated aquaculture system

acceptedVersio

Hydrolysates of whole forage-fish and Pacific krill are useful to reduce fish meal in practical diets for largemouth bass (Micropterus salmoides), and dietary fish hydrolysate suppresses expressions of intestinal oligopeptide transporter and taurine transporter genes

publishedVersio

Lithosphere thinning beneath west North China Craton: Evidence from geochemical and Sr-Nd-Hf isotope compositions of Jining basalts

This study shows lithosphere evolution history in the west North China Craton (NCC) from the early Cretaceous to Quaternary by studying the major element, trace element and Sr-Nd-Hf isotope compositions in Jining basalts of 119.6-108.6. Ma, 23.5-21.9. Ma and 1.3-0.11. Ma.The early Cretaceous basalts (119.6-108.6Ma) display enriched characteristics with high contents of incompatible elements, high 87Sr/86Sri, low εNd(t) and low εHf(t). These basalts resulted from partial melting of ancient metasomatized lithospheric mantle, and we consider the 119.6-108.6Ma magmatism as indicating lithosphere thinning in the west NCC. Although the Pacific slab seen seismically in the mantle transition zone beneath eastern China is no older than 60Ma, there exists convincing evidence for the presence of the Paleo-Pacific slab in the transition-zone in the Mesozoic. Thus we propose that the water released from the transition-zone slab hydrated the overlying lithosphere and further converted the base of the lithosphere into asthenosphere. This is the most likely mechanism responsible for the lithosphere thinning in the west NCC and the petrogenesis of the Jining 119.6-108.6Ma basalts.The Jining 23.5-21.9Ma basalts also have high contents of incompatible elements, but they display high εNd(t), high εHf(t) and variably low 87Sr/86Sri. We propose that these Miocene basalts were derived from the asthenosphere with contributions from ancient metasomatized lithospheric mantle during melt ascent. The Jining Quaternary basalts (1.3-0.11Ma) represent the melt of upwelling asthenosphere with low 87Sr/86Sri, high εNd(t) and high εHf(t). Upwelling and decompression melting of the eastward flowing asthenosphere from beneath western plateaus to beneath eastern hilly plains in the Cenozoic is the most plausible mechanism for the petrogenesis of Jining Cenozoic basalts (both of 23.5-21.9Ma and 1.3-0.11Ma), but the Jining 1.3-0.11Ma basalts must have been produced beneath even thinner lithosphere.Taken together geophysical studies and our petrological and geochemical studies of all these three episodes of the Jining basalts, we propose that the lithosphere in the west NCC has been thinning since the early Cretaceous and the thinning continues to the present

New insights into β-glucan-enhanced immunity in largemouth bass Micropterus salmoides by transcriptome and intestinal microbial composition

β-glucan is widely used in aquaculture due to its immunostimulatory effects, but the specific effect and potential regulatory mechanism on largemouth bass (Micropterus salmoides) are still unclear. Here, we evaluated the effects of β-glucan on growth, resistance to Aeromonas schubertii, intestinal health, and transcriptome of largemouth bass to reveal the potential regulators, metabolic pathways, and altered differential microbiota. Four experimental diets were designed with β-glucan supplementation levels of 0 (control), 100 (LA-100), 200 (MA-200), and 300 (HA-300) mg kg-1, and each diet was fed to largemouth bass (79.30 ± 0.50 g) in triplicate for 70 days, followed by a 3-day challenge experiment. Results showed that different β-glucan supplementations had no significant effects on growth performance and whole-body composition. Fish fed a diet with 300 mg kg-1 β-glucan significantly increased the activity of lysozyme than those fed diets with 0 and 100 mg kg-1 β-glucan. In addition, the survival rate of largemouth bass in β-glucan supplementation groups was significantly higher than the control group at 12- and 24-h challenge by Aeromonas schubertii. Transcriptome analysis showed that a total of 1,245 genes were differentially expressed [|log2(fold change)| ≥1, q-value ≤0.05], including 109 immune-related differentially expressed genes (DEGs). Further analysis revealed that significantly upregulated and downregulated DEGs associated with immunity were mapped into 12 and 24 pathways, respectively. Results of intestinal microflora indicated that fish fed a diet with 300 mg kg-1 β-glucan had higher bacterial richness and diversity as evaluated by Sobs, Chao, Ace, and Simpson indices, but no significant differences were found in the comparison groups. Furthermore, 300 mg kg-1 β-glucan significantly increased the relative abundance of Mycoplasma and decreased Proteobacteria (mainly Escherichia-Shigella and Escherichia coli) and Bacillus anthracis in largemouth bass intestinal microflora. The findings of this study provided new insights that will be valuable in future studies to elucidate the mechanism of immunity enhancement by β-glucan