Øystein Ahlstrøm

Intrapopulation Variability Shaping Isotope Discrimination and Turnover: Experimental Evidence in Arctic Foxes

Background Tissue-specific stable isotope signatures can provide insights into the trophic ecology of consumers and their roles in food webs. Two parameters are central for making valid inferences based on stable isotopes, isotopic discrimination (difference in isotopic ratio between consumer and its diet) and turnover time (renewal process of molecules in a given tissue usually measured when half of the tissue composition has changed). We investigated simultaneously the effects of age, sex, and diet types on the variation of discrimination and half-life in nitrogen and carbon stable isotopes (δ15N and δ13C, respectively) in five tissues (blood cells, plasma, muscle, liver, nail, and hair) of a top predator, the arctic fox Vulpes lagopus. Methodology/Principal Findings We fed 40 farmed foxes (equal numbers of adults and yearlings of both sexes) with diet capturing the range of resources used by their wild counterparts. We found that, for a single species, six tissues, and three diet types, the range of discrimination values can be almost as large as what is known at the scale of the whole mammalian or avian class. Discrimination varied depending on sex, age, tissue, and diet types, ranging from 0.3‰ to 5.3‰ (mean  = 2.6‰) for δ15N and from 0.2‰ to 2.9‰ (mean  = 0.9‰) for δ13C. We also found an impact of population structure on δ15N half-life in blood cells. Varying across individuals, δ15N half-life in plasma (6 to 10 days) was also shorter than for δ13C (14 to 22 days), though δ15N and δ13C half-lives are usually considered as equal. Conclusion/Significance Overall, our multi-factorial experiment revealed that at least six levels of isotopic variations could co-occur in the same population. Our experimental analysis provides a framework for quantifying multiple sources of variation in isotopic discrimination and half-life that needs to be taken into account when designing and analysing ecological field studies.

Chronic dietary exposure to environmental organochlorine contaminants induces thyroid gland lesions in Arctic foxes (Vulpes lagopus).

The impact of dietary organochlorine (OC) exposure on thyroid gland pathology was studied in farmed male Arctic foxes (Vulpes lagopus). The exposed group (n=16) was fed a diet based on wild minke whale (Balaenoptera acutorostrata) blubber as a main fat source in order to mimic the exposure to OC cocktails in the Artic environment. This resulted in an exposure of approximately 17 microg Sigma OC/kg day and a Sigma OC residue adipose tissue and liver concentration of 1700 and 4470 ng/gl.w., respectively, after 16 months of exposure. Control foxes (n=13) were fed a diet with pork (Sus scrofa) fat as a main fat source containing significantly lower OC concentrations. The food composition fed to the control and exposed group was standardized for nutrient contents. Four OC-related histopathological changes were found: (1) flat-epithelial-cell true thyroid cysts (TC) characterized by neutral content; (2) remnants of simple squamous epithelial-cell embryonic ducts containing neutral debris (EDN); (3) remnants of stratified squamous epithelial-cell embryonic ducts containing acid mucins often accompanied with debris of leukocyte inflammatory nature (EDM) and (4) disseminated thyroid C-cell hyperplasia (HPC). Of these, the prevalence of TC, EDN and HPC was significantly highest in the exposed group (chi(2) test: all p<0.04). The study shows that the OC mixture in minke whale blubber may cause development of thyroid gland cysts, C-cell hyperplasia and increase the prevalence of cystic remnants of embryonic ducts. The mechanism causing these effects could include endocrine disruption of the hypothalamus-pituitary-thyroid (HPT) axis, a disturbance of the calcium homeostasis/metabolism or energy metabolism or immune suppression. Because concentrations of OCs are higher in wild Arctic foxes, it is likely that these animals could suffer from similar OC-induced thyroid gland pathological and functional changes.

Nitrogen and energy balance in growing mink (Mustela vison) fed different levels of bacterial protein meal produced with natural gas

Abstract The objective of this study was to estimate the effect of increasing the dietary content of bacterial protein meal (BPM) on energy and protein metabolism in growing mink kits. Sixteen male mink kits of the standard brown genotype were randomly fed one of four diets: A control (Diet I) based on high-quality fish meal, and three experimental diets in which 20% (Diet II), 40% (Diet III) and 60% (Diet IV) of the digested nitrogen (DN) was replaced with BPM. Nitrogen balance and respiration experiments (indirect calorimetry) were carried out when the animals were approximately 9.5, 14.5, 17.5, 23.5 and 28.5 weeks of age. The apparent digestibility of crude protein and energy decreased significantly with increasing dietary BPM. The retained nitrogen was 0.45, 0.54, 0.52 and 0.40 g/kg0.75 on Diets I, II, III and IV, respectively, the observed differences between diets being non-significant (p = 0.06). Heat production (HE) was between 645 and 665 kJ/kg0.75 on all diets (p = 0.78). Retained energy (RE) was approximately 150 – 160 kJ/kg0.75 on Diets I to III, whereas it was −11 kJ/kg0.75 on Diet IV, the differences being significant (p < 0.001). A lower feed intake and apparent digestibility of energy caused the negative RE on Diet IV. The amount of HE from oxidation of protein decreased from 32.7% on Diet I to 26.6% on Diet IV, and oxidation of fat increased from 53.8% on Diet I to 63.5% Diet IV. In conclusion, protein and energy metabolism remained unaffected when up to 40% of DN was derived from BPM.

Comparative nutrient digestibility of arctic foxes (Alopex lagopus) on Svalbard and farm-raised blue foxes (Alopex lagopus)

Arctic foxes from Svalbard (n=4) and farmed blue foxes (n=4) was used in a digestibility experiment with a high-carbohydrate feed to add more information to the nutritional physiology of the arctic fox, and to compare its digestive capacity with that of the farmed blue fox. The arctic fox has a diet containing mainly protein and fat from mammals and birds, while farmed blue foxes have been exposed to an omnivorous dietary regime for more than 80 generations. The experiment showed in general no difference in digestive capacity for protein and fat between the foxes (P>0.05), but for carbohydrates, including starch and glucose, the blue fox revealed higher digestibility values. The superior digestive capacity for carbohydrates in blue fox might be a result of a long-term selection of animals digesting dietary carbohydrates more efficiently, or that an early age exposition to dietary carbohydrates has given permanent improvement of the carbohydrate digestion in the gut.

Seasonal emaciation causes tissue redistribution and an increased potential for toxicity of lipophilic pollutants in farmed arctic fox (Vulpes lagopus)

Many Arctic animals carry high body burdens of organochlorine contaminants (OCs) as a result of long-range transport of persistent pollutants. It has been shown that seasonal mobilization of body fat in these species results in increased blood concentration of OCs. The authors investigated OC assimilation, tissue distribution, and biotransformation in farmed Arctic fox (Vulpes lagopus) continuously fed a diet containing contaminated minke whale blubber or lard (control) from 8 wk of age in August 2003, until sampling when they were at their fattest (in November 2004) and leanest (in June 2005). Markedly higher tissue (liver, adrenals, brain, and blood) OC levels were found in June than in November despite low exposure to OCs during emaciation, suggesting that OCs had been redistributed from adipose tissues to vital organs. There were no differences in the activities of hepatic biotransforming enzymes between exposed fat and control fat foxes, except for 16α-hydroxylation, which was higher in exposed fat foxes. In emaciated foxes, ethoxyresorufin activity was higher in exposed than in control foxes, indicating an enhanced potential for toxicity of OCs with emaciation. Lower activities of 6β- and 2β-hydroxylation were found in lean than in fat foxes, irrespective of OC treatment. The results show that emaciation increase the toxic potential of accumulated OCs and emphasize that body adiposity must be considered when time-trend analyses, risk assessments, and effect studies are designed. Environ Toxicol Chem 2013;32:1784-1792.

Ileal and total tract nutrient digestibility in blue foxes ( Alopex lagopus ) fed extruded diets containing different protein sources

This study compared apparent ileal and total tract digestibility of macronutrients and amino acids in blue foxes (Alopex lagopus) fed dry extruded diets. The control diet contained fish meal as the main protein source, while in the other diets 50% of the fish meal protein was replaced by not de-hulled solvent-extracted soybean meal, meat meal or bacterial protein meal. Ileal digestibility was obtained with animals modified with ileorectal anastomosis (IRA method). There were no significant differences in the average digestibility of CP, Lys, Trp, Val, Ala or Glu between IRA modified and intact animals. Average ileal digestibilities of His, Thr, Asp, Cys, Gly, Hyp, Pro, Ser, starch and total carbohydrates (CHO) were significantly lower compared with total tract values. Average ileal digestibility of Arg, Ile, Leu, Met, Phe, Tyr and crude fat was significantly higher than total tract digestibility. Digestibility measured in IRA modified animals showed less variability compared with values from intact animals. There were significant differences among diets in ileal as well as total tract digestibility of crude protein, most amino acids, starch and CHO, and in ileal digestibility of crude fat. Ileal crude protein digestibility of the diets ranged from 81.0 – 86.4% and total tract digestibility from 82.5 – 86.4%. Ileal amino acid digestibilities ranged from 70.1 (Asp) to 93.3% (Arg) and total tract values ranged from 72.8 (Cys) to 92.2% (Arg). Both ileal and total tract digestibilities of crude protein and most amino acids were significantly lower for diets containing meat meal or bacterial protein meal compared with the control fish meal-based diet and the diet with soybean meal. Although ileal digestibility may be more accurate than total tract digestibility in estimating the protein and amino acid supply in blue foxes, total tract digestibility may be acceptable because of numerically small differences between ileal and total tract digestibilities for protein and most amino acids. It was further concluded that ileal digestibilities of starch and CHO in the blue fox are lower than corresponding total tract digestibilities.

Mineral density and biomechanical properties of bone tissue from male Arctic foxes (Vulpes lagopus) exposed to organochlorine contaminants and emaciation

We investigated the impact from dietary OC (organochlorine) exposure and restricted feeding (emaciation) on bone mineral density (BMD; g hydroxy-apatite cm(-2)) in femoral, vertebrate, skull and baculum osteoid tissue from farmed Arctic blue foxes (Vulpes lagopus). For femur, also biomechanical properties during bending (displacement [mm], load [N], energy absorption [J] and stiffness [N/mm]) were measured. Sixteen foxes (EXP) were fed a wet food containing 7.7% OC-polluted minke whale (Balaenoptera acutorostrata) blubber in two periods of body fat deposition (Aug-Dec) and two periods of body fat mobilisation (Jan-July) in which the food contained less energy and only 2% blubber. SigmaOC food concentration in the food containing 7.7% whale blubber was 309 ng/g wet mass. This corresponded to a SigmaOC exposure of ca. 17 microg/kg body mass/d and a responding SigmaOC residue in subcutaneous adipose tissue of ca. 1700 ng/g live mass in the 8 EXP fat foxes euthanized after 16 months. A control group (CON) composed of 15 foxes were fed equal daily caloric amounts of clean pork (Sus scrofa) fat. After 16 months, 8 EXP and 7 CON foxes were euthanized (mean body mass=9.25 kg) while the remaining 8 EXP and 8 CON foxes were given restricted food rations for 6 months resulting in a body weight reduction (mean body mass=5.46 kg). The results showed that only BMD(skull) vs. BMD(vertebrae) were significantly correlated (R=0.68; p=0.03; n=10) probably due to a similar composition of trabecular and cortical osteoid tissue. No difference in any of the BMD measurements or femoral biomechanical properties was found between EXP and CON foxes although BMD baculum was 1.6-folds lower in the EXP group. However, lean summer foxes had significantly lower femoral biomechanical properties measured as displacement (mm), energy absorption (J) and time (s) biomechanical properties than fat winter foxes (all p<0.004). This indicates lower stiffness and softer bones from fasting which is in agreement with previous studies. Further, it should be kept in mind when studying bone tissues in Arctic mammals also in order to avoid confounding effects from body condition.

Comparative apparent total tract digestibility of major nutrients and amino acids in dogs ( Canis familiaris ), blue foxes ( Alopex lagopus ) and mink ( Mustela vison )

Major nutrient and amino acid digestibilities were evaluated in experiments with dogs ( Canis familiaris ), blue foxes ( Alopex lagopus ) and mink ( Mustela vison ) using seven extruded diets containing different protein sources. Four dogs ( German shepherd ), four blue foxes and four mink were offered each of the experimental diets to determine apparent total tract digestibility. Average digestibility of crude protein (CP) in blue foxes and mink was similar, and lower than the corresponding value in dogs (0·834 and 0·831 v. 0·864) ( P P P P P P P > 0·05) for several non-essential amino acids. Average digestibility of crude fat was high and similar among the species studied: 0·968, 0·967 and 0·959 in dogs, blue foxes and mink, respectively. Average digestibility of starch ranged from 0·971 in mink to 0·998 in dogs; significantly higher in dogs than in blue foxes and mink, and significantly higher in blue foxes than in mink ( P

Dietary Contaminant Exposure Affects Plasma Testosterone, but not Thyroid Hormones, Vitamin A, and Vitamin E, in Male Juvenile Arctic Foxes (Vulpes lagopus)

Levels of persistent organic pollutants (POP), such as polychlorinated biphenyls (PCB), are high in many Arctic top predators, including the Arctic fox (Vulpes lagopus). The aim of this study was to examine possible endocrine-disruptive effects of dietary POP exposure in male juvenile Arctic foxes in a controlled exposure experiment. The study was conducted using domesticated farmed blue foxes (Vulpes lagopus) as a model species. Two groups of newly weaned male foxes received a diet supplemented with either minke whale (Baleneoptera acutorostrata) blubber that was naturally contaminated with POP (exposed group, n = 5 or 21), or pork (Sus scrofa) fat (control group, n = 5 or 21). When the foxes were 6 mo old and had received the 2 diets for approximately 4 mo (147 d), effects of the dietary exposure to POP on plasma concentrations of testosterone (T), thyroid hormones (TH), thyroid-stimulating hormone (TSH), retinol (vitamin A), and tocopherol (viramin E) were examined. At sampling, the total body concentrations of 104 PCB congeners were 0.1 ± 0.03 μg/g lipid weight (l.w.; n = 5 [mean ± standard deviation]) and 1.5 ± 0.17 μg/g l.w. (n = 5) in the control and exposed groups, respectively. Plasma testosterone concentrations in the exposed male foxes were significantly lower than in the control males, being approximately 25% of that in the exposed foxes. There were no between-treatment differences for TH, TSH, retinol, or tocopherol. The results suggest that the high POP levels experienced by costal populations of Arctic foxes, such as in Svalbard and Iceland, may result in delayed masculine maturation during adolescence. Sex hormone disruption during puberty may thus have lifetime consequences on all aspects of reproductive function in adult male foxes.

Effects of lactic acid fermentation and gamma irradiation of barley on antinutrient contents and nutrient digestibility in mink (Mustela vison) with and without dietary enzyme supplement.

Abstract The experiment was conducted to study the effects of fermentation of barley, using two different strains of lactic acid bacteria, a Lactobacillus plantarum/pentosus strain isolated from spontaneously fermented rye sourdough (AD2) and a starch-degrading Lactobacillus plantarum (AM4), on contents of mixed-linked (1 → 3) (1 → 4)-β-glucans, α-amylase inhibitor activity, inositol phosphates, and apparent digestibility of macronutrients in mink. Effects of fermentation were compared with effects of gamma irradiation (γ-irradiation: 60Co γ-rays at 25 kGy). The diets were fed to mink with and without a supplementary enzyme preparation. Both lactic acid fermentation and γ-irradiation followed by soaking and incubation, reduced concentrations of soluble β-glucans, phytate and α-amylase inhibitor activity. Dietary enzyme supplementation increased significantly digestibility of crude protein, fat, starch and crude carbohydrate (CHO). Fermentation of the barley increased digestibility of starch and CHO. Fermentation with lactic acid bacteria AD2 resulted in higher starch and CHO digestibility than strain AM4, and had greater effect than γ-irradiation, soaking and incubation. The highest digestibility of starch and CHO was obtained after AD2 fermentation followed by enzyme supplementation. It is concluded that both lactic acid fermentation of barley and enzyme supplementation have positive nutritional implications in the mink by limiting the effects of antinutrients and improving digestibility and energy utilization.