Feeding level and diet quality influence trophic shift of c and n isotopes in nile tilapia (oreochromis niloticus (L.))

Many scientists use naturally occurring stable isotopes to reconstruct the diets of animals. However, isotopic ratios may be affected not only by the composition of the diet but also by the amount of food consumed. Thus, an experiment using tilapia (Oreochromis niloticus) was carried out to test the influence of feeding level on δ ¹³C and δ ¹⁵N of fish given a semi-synthetic wheat gluten/wheat starch based diet. In addition, the effect of diet quality was tested by comparing tilapia given this feed with tilapia fed a fish meal/wheat meal based diet. Forty-four tilapia were reared individually. After a prefeeding phase, fish were randomly assigned to five groups, four on the semi-synthetic diet at different feeding levels and one group on the fish meal/wheat meal based diet fed at the equivalent of the highest level of the semi-synthetic diet. The experiment lasted eight weeks. Proximate composition, gross energy content and δ ¹³C and δ ¹⁵N values were determined in feed and fish, for δ ¹³C separately in the lipids and the lipid-free matter. δ ¹³C in the lipids and the lipid-free matter and δ ¹⁵N of tilapia fed the semi-synthetic diet decreased significantly with increasing feeding rate. The absolute values of the trophic shift in fish fed the semi-synthetic wheat based diet were significantly higher than in fish fed the fish meal/wheat meal based diet. The different δ ¹³C and δ ¹⁵N values in tilapia fed the same diet at different feeding levels and the influence of feed quality on the trophic shift add to the uncertainty involved in the use of stable isotopes in ecological research.     

Dietary lipid content influences the activity of lipogenic enzymes in the liver and on whole body delta13C values of Nile tilapia, Oreochromis niloticus (L.)

The use of stable isotope techniques for the reconstruction of diets has increased over the last decade. However, isotopic ratios in an animal are not only affected by the composition of the feed, but also by the amount of feed consumed. An uncertainty of up to 1 per thousand for both delta13C and delta15N values has been observed when the feeding level is unknown. This may have substantial effects on the results of back-calculation. As the feeding level of animals is unknown in nature, an additional indicator for their nutritional status is needed. High feeding levels and a consequent surfeit of dietary energy lead to the synthesis of lipids. In order to test whether the level of lipogenesis could be used as an indicator, Nile tilapia (Oreochromis niloticus) were fed four isonitrogenous and isoenergetic wheat-based semi-synthetic diets with different lipid contents (2.0 %, 4.5 %, 9.5 % and 13.3 %) for eight weeks. Body composition, gross energy content and delta13C values in the lipids and the lipid-free material were determined in diets and fish bodies. The livers of three fish per feeding group were assayed for the activity of two lipogenic enzymes, ATP-citrate lyase and malic enzyme. There was a strong negative correlation between delta13C values in the lipids of the individual fish and the apparent lipid conversion. The activities of lipogenic enzymes decreased with rising lipid content in the diet. The delta13C values in the lipids decreased significantly with increasing specific activity for both enzymes. In this experiment where lipogenesis was influenced by the composition of the diet, it was possible to determine the exact value for the trophic shift in relation to the enzyme activities. Further experiments to investigate the use of enzyme activities in situations where the feeding level of an animal is unknown are recommended.     

Stable isotopes in animal ecology: the effect of ration size on the trophic shift of C and N isotopes between feed and carcass

The feeding ecology of living or extinct animal species is now frequently studied by analysis of stable isotope ratios in small quantities of carcass or remains such as bones or teeth. Although there are numerous papers on these applications in natural systems, the theoretical and experimental basis of this method is weak. In order to evaluate the effect of different feeding levels on the carbon and nitrogen trophic shift, an experiment was carried out in which fish (Nile tilapia, Oreochromis niloticus, initial weight 40.8 g) were fed for 4 weeks at three levels from slightly above maintenance to almost satiation. For each treatment, three fish were reared individually. The isotopic ratios of carbon and nitrogen in feeds and fish carcasses were determined and in the case of carbon this was done separately for lipids and lipid-free matter. The trophic shift was calculated at each feeding level from the delta13C and delta15N ratios of feed and fish. There was a significant trend towards higher values for the trophic shift at higher feeding rates in all fractions analysed. Although further research is required, it can be concluded that the effect of feeding level cannot be ignored when the diet of an animal has to be calculated from the ratios of isotopes in its body.     

Physical chromosome mapping of repetitive DNA sequences in Nile tilapia Oreochromis niloticus: evidences for a differential distribution of repetitive elements in the sex chromosomes

Repetitive DNAs have been extensively applied as physical chromosome markers on comparative studies, identification of chromosome rearrangements and sex chromosomes, chromosome evolution analysis, and applied genetics. Here we report the characterization of repetitive DNA sequences from the Nile tilapia (Oreochromis niloticus) genome by construction and screening of plasmid library enriched with repetitive DNAs, analysis of a BAC-based physical map, and hybridization to chromosomes. The physical mapping of BACs enriched with repetitive sequences and C(o)t-1 DNA (DNA enriched for highly and moderately repetitive DNA sequences) to chromosomes using FISH showed a predominant distribution of repetitive elements in the centromeric and telomeric regions and along the entire length of the largest chromosome pair (X and Y sex chromosomes) of the species. The distribution of repetitive DNAs differed significantly between the p arm of X and Y chromosomes. These findings suggest that repetitive DNAs have had an important role in the differentiation of sex chromosomes.     

Physicochemical properties of tilapia (Oreochromis niloticus) actomyosin subjected to high intensity ultrasound in low NaCl concentrations

Effects of high intensity ultrasound (HIU) on physicochemical properties of tilapia (Oreochromis niloticus) actomyosin in low NaCl concentrations were investigated. The protein content extracted in low NaCl concentrations (0.1–0.3 M NaCl) increased with increasing HIU intensity up to 20.62 W/cm² (p < 0.05). The effect of HIU on actomyosin extractability in high NaCl concentrations (0.6 and 1.2 M NaCl) was less obvious. Ca²⁺-ATPase activity and total sulfhydryl (SH) group content decreased in both 0.2 and 0.6 M NaCl. HIU showed more pronounced effect on oxidation of the SH groups in 0.6 M NaCl, while the reactive SH content at 0.2 M NaCl increased after a prolonged exposure to HIU, suggesting conformational changes induced by HIU. Surface hydrophobicity of actomyosin in 0.6 M NaCl increased with increasing ultrasonic intensity and exposure time to a higher degree than that in 0.2 M NaCl. A greater absolute value of the zeta potential of actomyosin subjected to HIU were also observed. The HIU treatments decreased the turbidity of actomyosin incubated at 40 and 60 °C. A drastic increase in the solubility of myosin heavy chain (MHC) and actin with 0.2 M NaCl were evident when HIU treatments were applied, but degradation of MHC occurred in both 0.2 and 0.6 M NaCl. Based on particle size and microstructure, actomyosin in 0.6 M NaCl underwent more disruption by HIU than that in 0.2 M NaCl. HIU induced protein unfolding and protein dissociation, enabling better extraction in a lower NaCl concentration.     

A dual 13C and 15N long-term labelling technique to investigate uptake and translocation of C and N in beech (Fagus sylvatica L.)

A continuous dual 13CO2 and 15NH4(15)NO3 labelling experimental set-up is presented that was used to investigate the C and N uptake and allocation within 3-year old beech (Fagus sylvatica L.) during one growing season. The C and N allocation pattern was determined after six, twelve and eighteen weeks of growth. The carbon uptake was distinctly different in the three phases examined: The first six weeks after budbreak were dedicated to leaf growth with a R/S (root to shoot) ratio of 0.14 for the new carbon. The second growth phase showed a balanced R/S ratio of C allocation and after week 13, the root compartment was the main carbon sink (R/S = 6.97). Nitrogen allocation was more basipetal as compared to carbon. In the second growth phase, R/S of Nnew was 5.57 but fell to 3.54 for the third growth phase probably due to formation of reserves in buds and stem.     

A Dual 13C and 15N Long Term Labelling Technique to Investigate Uptake and Translocation of C and N in Beech (Fagus sylvatica L.)

Abstract

A continuous dual ¹³CO₂ and ¹⁵NH₄ ¹⁵NO₃ labelling experimental set-up is presented that was used to investigate the C and N uptake and allocation within 3-year old beech (Fagus sylvatica L.) during one growing season. The C and N allocation pattern was determined after six, twelve and eighteen weeks of growth. The carbon uptake was distinctly different in the three phases examined: The first six weeks after budbreak were dedicated to leaf growth with a R/S (root to shoot) ratio of 0.14 for the new carbon. The second growth phase showed a balanced R/S ratio of C allocation and after week 13, the root compartment was the main carbon sink (R/S = 6.97).

Nitrogen allocation was more basipetal as compared to carbon. In the second growth phase, R/S of N_new was 5.57 but fell to 3.54 for the third growth phase probably due to formation of reserves in buds and stem.     

Effects of N-application on utilization of 15N and 13C and quality in two wheat cultivars

We studied N uptake and distribution in wheat, and the incorporation of nitrogen and carbon into gluten and non-gluten proteins using a double-labelling approach with ¹⁵N and ¹³C. Doses of N-fertilizer were split and applied at emergence, onset of stem elongation, and heading at rates of 280/140/140 mg N pot^?1, respectively simulating 90/45/45 kg N ha^?1. Five different combinations of N-fertilizations containing no or 10 % ¹⁵N were performed. The recovery of ¹⁵N added at the stages emergence, stem elongation or heading were 42, 60, and 64 %. Application of ¹⁵N at all three stages yielded in 51 % recovery. Remobilisation of straw N was greater for Golia. The ¹⁵N concentration in gluten proteins of Golia show higher values than Gönen. The ratio of ¹⁵N gluten/¹⁵N non-gluten proteins of Golia were higher, which implies a lower non-gluten protein activity during grain filling. The ¹³C concentration in gluten and non-gluten proteins did not differ between both cultivars.     

Effects of N-application on utilization of 15N and 13C and quality in two wheat cultivars

We studied N uptake and distribution in wheat, and the incorporation of nitrogen and carbon into gluten and non-gluten proteins using a double-labelling approach with 15N and 13C. Doses of N-fertilizer were split and applied at emergence, onset of stem elongation, and heading at rates of 280/140/140 mg N pot(-1), respectively simulating 90/45/45 kg N ha(-1). Five different combinations of N-fertilizations containing no or 10 % 15N were performed. The recovery of 15N added at the stages emergence, stem elongation or heading were 42, 60, and 64 %. Application of 15N at all three stages yielded in 51 % recovery. Remobilisation of straw N was greater for Golia. The 15N concentration in gluten proteins of Golia show higher values than Gonen. The ratio of 15N gluten/15N non-gluten proteins of Golia were higher, which implies a lower non-gluten protein activity during grain filling. The 13C concentration in gluten and non-gluten proteins did not differ between both cultivars.     

Stable isotopes of H,C and N in mice bone collagen as a reflection of isotopically controlled food and water intake*

²H/¹H ratios in animal biomass reflect isotopic input from food and water. A 10-week controlled laboratory study raised 48 mice divided in two generations (8 mothers Mus musculus and their offspring). The mice were divided into four groups based on the combination of ²H, ¹³C, ¹⁵N-enriched and non-enriched food and water. Glycine, the most common amino acid in bone collagen, carried the ²H, ¹³C, ¹⁵N-isotopic spike in food. ANOVA data analysis indicated that hydrogen in food accounted for ～81?% of the hydrogen isotope inventory in collagen whereas drinking water hydrogen contributed ～17?%. Air humidity contributed an unspecified amount. Additionally, we monitored ¹³C and ¹⁵N-enrichment in bone collagen and found strong linear correlations with the ²H-enrichment. The experiments with food and water indicate two biosynthetic pathways, namely (i) de novo creation of non-essential amino acids using hydrogen from water, and (ii) the integration of essential and non-essential amino acids from food. The lower rate of isotope uptake in mothers’ collagen relative to their offspring indicates incomplete bone collagen turnover after ten weeks. The variance of hydrogen stable isotope ratios within the same cohort may limit its usefulness as a single sample proxy for archaeological or palaeoenvironmental research.     

Investigation of level energies and B(E2) values for rotation-aligned bands in Hg isotopes

High spin states in ^{191, 192, 193, 195, 197, 199}Hg were investigated by observing γ-rays and conversion electrons in the compound reactions ${}^{\mathrm{192,\; 194,\; 198}}\text{Pt(α, x}\text{n}\text{)}\text{and}{}^{192}\text{Pt}\text{(}{}^3\text{He}\text{, 4}\text{n}\text{)}$. In ¹⁹⁷Hg the decoupled band built on the $\text{13}\text{2}{}^{\mathrm{+}}$ state and the semi-decoupled negative-parity band are observed up to $\text{I}{}^{\mathrm{\pi }}\text{=}\text{41}\text{2}{}^{\mathrm{+}}\text{and}\text{33}\text{2}{}^{\mathrm{?}}$, respectively. A careful investigation of ¹⁹⁹Hg revealed no new high spin states above the previously known levels with $\text{I}{}^{\mathrm{\pi }}\text{=}\text{25}\text{2}{}^{\mathrm{+}}\text{and}\text{31}\text{2}$^?. Half-lives were determined for the 10⁺, 7^?, 8^? and 16^? states in ¹⁹²Hg, the if$\text{33}\text{2}{}^{\mathrm{+}}$$\text{states in}{}^{\mathrm{191,193}}\text{Hg and the}$ (frc states in ^{191, 193, 195, 197}Hg. The systematics of the level energies and B(E2) values for the positive-parity ground and $\text{13}\text{2}{}^{\mathrm{+}}$ bands and the negative-parity semi-decoupled bands in ^190–200Hg is discussed.     

Investigation on the influence of ultrasonic pretreatment on color,quality and antioxidant attributes of microwave dried Inula viscosa (L.)

Impact of various ultrasound pretreatment and microwave drying parameters on the qualitative and antioxidant characteristics of Inula viscosa (L.) was investigated in this study. The leaves of Inula viscosa (L.) were sonicated for 10, 20, and 30 min in an ultrasonic bath (37 kHz, 150 Watts). Microwave drying was done at three distinct times (1, 3, and 5 min) and with three different microwave power levels (100, 180, and 300 Watts). Microwave dried samples were tested for color characteristics (L*, a*, b*), chlorophyll, carotenoid, total phenol, and antioxidant content. All dried samples were prepared by infusing them in hot water as tea, and the sensorial properties of teas including odor, color, aroma, and overall acceptability were evaluated by panelists. For 10, 20, and 30 min of ultrasound pretreatment, the L* values of leaves varied from 37.70 to 49.76, 34.97 to 46.25, and 27.88 to 43.34, respectively. The total carotenoid concentration ranged from 0.12 to 0.32 mg/g DW, while the total chlorophyll content was from 0.44 to 0.94 mg/g DW. The antioxidant activity of Inula viscosa (L.) leaves that were dried at 300 Watts for 5 min did not change significantly as a result of ultrasound pretreatment. There was a significant positive correlation between aroma and TPC, as well as between color and overall acceptability. The darkest-colored teas were deemed preferable by the panelists.     

Study of plastic deformation and proton irradiation influence on phase transitions in C18N9T stainless steel

Transmission and post-nuclear-reaction emission Mössbauer spectroscopy was used for investigation of radiation effects and plastic deformation influence on C18N9T steel. The rolled foil was annealed in 10^?6 Torr vacuum at 870 K during 2 hours and was irradiated by 30 MeV protons at current density of 0.8μ A up to doses of 5×10¹⁷ cm^?2 on the isochronous cyclotron at the liquid nitrogen cooling of the target. The Mössbauer spectra obtained before and after the sample irradiation in transmission geometry are identical. The emission spectrum is analogous to the spectrum of the plastically deformed sample. It is concluded that during irradiation in the steel the processes take place which are similar to those at plastic deformation and connected with redistribution of the alloy component atoms.     

Vibrational Excitation of N2(B,C) and N+2(B) States in N2 and N2 – H2 Flowing Microwave Discharges

The ro‐vibrational spectra of N₂ microwave discharges have been analysed by emission spectroscopy. It is deduced the rotational and vibrational temperatures of N₂ states. The characteristic of vibrational temperature Θ₁ of the N₂ (X, v) ground state has been specifically determined.It has been found that the N₂ (C, B, v') and N⁺₂ (B, v') radiative states are directly excited by electron collisions on the N₂ (X,v) ground state at a N₂ gas pressure of 0.1 Torr (discharge tube of 5 mm I.D, microwave power 100 Watt) with a Θ₁ value near 10⁴ K. At higher gas pressure up to 5 Torr, the N₂ (C, v') states remain alone to be mainly excited by electron collisions on N₂ (X, v). It is considered the excitations of the N₂ (B, v') states by collisions of electrons and N₂ (X,v > 4) vibrational molecules on the N₂ (A) metastable states.With x < 9% H₂ into N₂, it is observed an increase of N₂, 2^nd pos intensity, resulting of an increase of high energy electrons. Inversely, the N₂, 1^st pos intensity decreased, partly following the decrease of low energy electrons (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

The Influence of Feeding Frequency on Nitrogen Turnover and Whole Body Protein Synthesis in Adult Rats (Estimation using [14C]- and [15N]- Labelled Leucine)

Abstract

Male Wistar rats (17 wks. old, body weight ～400 g), fitted with an intra gastric cannula and with a catheter in the vena jugularis were divided into 3 groups and given a marginal ration of the feeding solution Nutrison Standard (1g protein and 350 kJ ME per day). Group 1 had ad lib. access to the drinking bottle, the groups 2 and 3 were pair fed by gastric infusion, splitted up into 2 greater meals for group 2 respectively into 6 smaller meals for group 3. After adaptation all animals get an i.p. injection of doubly labelled tracer solution (200μl) containing 2.5mg L-[¹⁵N]leucine (72 atom% ¹⁵N) combined with either [1-¹⁴C]- or [U-¹⁴C] leucine (37 kBq).

The course of ¹⁴CO₂ expiration was estimated by breath test over 4h in intervals of 15 min and the course of urinary ¹⁵N excretion over 24h in intervals of 45 resp. 90 min. An infusion of saline (0.9% 5ml/h) into the vena jugularis was used to provoke sustained urine production of the animals during the experiment.

From the parameters of the excretion curves of breath ¹⁴CO₂ resp. urine ¹⁵N (cumulative end value) and from the N balance the portions of leucine-C and leucine-N used for protein synthesis, transamination decarboxylation and total oxidation as well as the kinetic parameters for whole body protein metabolism were computed.

The following conclusions were drawn:

6 x feeding regime produces a small but measurable amino acid economy effect in comparison to 2 x feeding regime.

Protein gain for 2 x feeding group was significant smaller than for 6 x feeding group, though protein synthesis rate was higher, but was overcompensated by a greater increase of protein breakdown rate for the 2 x feeding group. Energy storage in form of fat and glycogen built from decarboxylation was unaffected by feeding frequency. The amount of leucine oxidized for heat production was 4% higher for the 6 x feeding group. Transamination rate for leucine was estimated to 8–15%. Absolute values for protein flux, protein synthesis and protein breakdown may be overestimated or underestimated because the metabolism of [¹⁵N] leucine does not exactly agree with that of total N; but the proportions of them and therefore also the conclusions will be true. Better results for absolute values will be obtained using a mixture of ¹⁵N labelled AA, ¹⁵N labelled protein or hydrolysate of ¹⁵N labelled protein (yeast) as the tracer source.     

Determination of the isotopic shift of the first resonance line in Cs(I) spectrum for the isotopes 131Cs, 132Cs,and 136Cs

The isotope shifts of the resonance line $\text{λ = 8521}\text{A}\text{?}$ of ¹³¹Cs, ¹³²Cs, and ¹³⁶Cs with respect to ¹³³Cs were determined by a new scanning technique to be $\text{v}\text{?}\text{(}{}^{133}\text{Cs}\text{)-}\text{v}\text{?}\text{(}{}^{131}\text{Cs}\text{) = +1.70(40)}\text{mK}$, $\text{v}\text{?}\text{(}{}^{133}\text{Cs}\text{)-}\text{v}\text{?}\text{(}{}^{132}\text{Cs}\text{) = ?0.95(80)}\text{mK}$, $\text{v}\text{?}\text{(}{}^{136}\text{Cs}\text{)-}\text{v}\text{?}\text{(}{}^{133}\text{Cs}\text{) = ?4.0(3.0)}\text{mK}$. In accordance with results from other Cs isotopes and neighbouring elements they show the change 〈δr²〉 in the second radial moment of the nuclear charge distribution to be a factor of 5 to 10 smaller than expected from the model of uniform charge distribution. Integral isotope shifts over the isotopic chains of Xe, Cs, and Ba were analyzed in terms of collective models. Assuming an isotopic shift discrepancy of 0.5 for the nuclear volume shift it was possible to derive deformation parameters $\text{〈β}{}^2\text{〉}{}^{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ in quantitative agreement with those from B(E2) measurements.     

Comparative study of compound components in N(N, α) and C(O, α) reactions

Experimental spectra of the reaction ¹⁴N(¹⁴N, α) were measured at 111 MeV. Continuum spectra extending to high excitation energy of ²⁴Mg do not show any structure which is completely at variance with the reaction ¹²C(¹⁶O, α). A compound evaporation calculation reproduces the smooth components of the spectra of both reactions. The same theoretical prediction, however, seems to contradict the conjecture that the compound process yields the broad peaks in the ¹²C(¹⁶O, α) reaction.     

Direct mass measurements of neutron-rich ~(86)Kr projectile fragments and the persistence of neutron magic number N=32 in Sc isotopes

In this paper, we present direct mass measurements of neutron-rich86 Kr projectile fragments conducted at the HIRFL-CSR facility in Lanzhou by employing the Isochronous Mass Spectrometry(IMS) method. The new mass excesses of52-54 Sc nuclides are determined to be-40492(82),-38928(114),-34654(540) ke V, which show a significant increase of binding energy compared to the reported ones in the Atomic Mass Evaluation 2012(AME12).In particular,53 Sc and54Sc are more bound by 0.8 Me V and 1.0 Me V, respectively. The behavior of the two neutron separation energy with neutron numbers indicates a strong sub-shell closure at neutron number N =32 in Sc isotopes.     

Effect of different ratios of wheat to corn flour in the diet on the development and isotopic composition (delta13C, delta15N) of the red flour beetle Tribolium castaneum

The back-calculation of the diet is a common application of stable isotopes in animal ecology. The method is based on a predictable relation between the isotopic signature of the diet and the animal's tissues. Frequently, the assumption of a constant difference in isotopic signatures (trophic shift) is made. Carbon isotopic ratios of C(3) and C(4) plants differ by approximately 10 per thousand, making wheat (C(3)-plant) and corn (C(4)-plant) ideal materials for isotopic studies in nutritional ecology and especially for testing the back-calculation method. In this experiment, red flour beetles, Tribolium castaneum, were reared on wheat flour, corn flour and three different mixtures thereof, either in pure flour or with the addition of yeast inoculum or yeast grains. Development of T. castaneum on these experimental diets was monitored, and isotopic signatures of carbon and nitrogen in emerging adults were analysed. The values of trophic shift of C and N isotopes for wheat and corn flour were different, and the values for the mixtures did not correspond to those expected from a linear mixing model. The latter can be taken as an indication that the tiny larvae of T. castaneum may be capable of differentiating between particles of wheat and corn flour, making this animal model unsuitable for testing the back-calculation method.