Allometric scaling of fatty acyl chains in fowl liver,lung and kidney,but not in brain phospholipids

The phospholipid (PL) fatty acyl chain (FA) composition (mol%) was determined in the kidney, liver, lung and brain of 8 avian species ranging in body mass from 150 g (Japanese quail, Coturnix coturnix japonica) to 19 kg (turkey, Meleagris gallopavo). In all organs except the brain, docosahexaenoic acid (C22:6 n3, DHA) was found to show a negative allometric scaling (allometric exponent: B = ? 0.18; ? 0.20 and ? 0.24, for kidney, liver and lung, respectively). With minor inter-organ differences, smaller birds had more n3 FAs and longer FA chains in the renal, hepatic and pulmonary PLs. Comparing our results with literature data on avian skeletal muscle, liver mitochondria and kidney microsomes and divergent mammalian tissues, the present findings in the kidney, liver and lung PLs seem to be a part of a general relationship termed “membranes as metabolic pacemakers”. Marked negative allometric scaling was found furthermore for the tissue malondialdehyde concentrations in all organs except the brain (B = ? 0.17; ? 0.13 and ? 0.05, respectively). In the liver and kidney a strong correlation was found between the tissue MDA and DHA levels, expressing the role of DHA in shaping the allometric properties of membrane lipids.     

Sex, but not maternal protein or folic acid intake, determines the fatty acid composition of hepatic phospholipids, but not of triacylglycerol, in adult rats

The aim of the study was to investigate whether the protein and folic acid content of the maternal diet and the sex of the offspring alter the polyunsaturated fatty acid content of hepatic phospholipids and triacylglycerol (TAG). Pregnant rats were fed diets containing 18% or 9% protein with either 1 or 5mg/kg folic acid. Maternal diet did not alter hepatic lipid composition in the adult offspring. Data from each maternal dietary group were combined and reanalysed. The proportion of 18:0, 20:4n-6 and 22:6n-3 in liver phospholipids was higher in females than in males, while hepatic TAG composition did not differ between sexes. Delta5 Desaturase expression was higher in females than in males. Neither Delta5 nor Delta6 desaturase expression was related to polyunsaturated fatty acid concentrations. These results suggest that sex differences in liver phospholipid fatty acid composition may reflect primary differences in the specificity of phospholipid biosynthesis.     

Butyrophilin and xanthine oxidase occur in constant molar proportions in milk lipid globule membrane but vary in amount with breed and stage of lactation

Summary Butyrophilin and xanthine oxidase, major proteins of milk lipid globule membrane, both accounted for significantly higher percentages of total protein in membrane samples from Holstein than from Jersey animals. Both were high in membranes from animals in early lactation, both decreased in amount as lactation progressed to the midpoint, and then both rose in amount toward the end of lactation. In samples from both Holstein and Jersey animals, butyrophilin and xanthine oxidase were present in constant molar proportions of about 41. These proteins co-enriched together with low molecular weight GTP-binding proteins in a high salt and nonionic detergent insoluble fraction of milk lipid globule membrane. Butyrophilin and xanthine oxidase content of membranes was not related to milk lipid globule diameter, suggesting that these proteins alone may not be involved solely in anchoring the membrane to the lipid globule surface. However, the possibility that a complex composed in part of butyrophilin and xanthine oxidase serves an anchoring function remains a possibility.     

Determination of anandamide and other fatty acyl ethanolamides in human serum by electrospray tandem mass spectrometry

We developed a new selective liquid chromatography-electrospray ionization-tandem mass spectrometry method for the identification and quantification of anandamide (AEA), an endogenous cannabinoid receptor ligand, and other bioactive fatty acid ethanolamides (FAEs) in biological samples. Detection limit (0.025 pmol for AEA and 0.1 pmol for palmitoylethanolamide (PEA) and oleoylethanolamide (OEA)) and quantification limit (0.2 pmol for AEA and 0.4 pmol for OEA and PEA) were in the high fmol to low pmol range for all analytes. Linear correlations (r(2)=0.99) were observed in the calibration curves for standard AEA over the range of 0.025-25 pmol and for standard PEA and OEA over the range of 0.1-500 pmol. This method provides a time-saving and sensitive alternative to existing methods for the analysis of FAEs in biological samples.     

Oral administration of uridylic acid increases plasma leptin, but suppresses glucose and non-esterified fatty acid concentrations in rats

Nucleic acids have been known to have biological effects on the digestive and immune systems, although less attention has been paid to the action on metabolism. In the present study, in order to investigate the effects of oral ingestion of uridylic acid (5'-uridine monophosphate, 5'-UMP) on hormonal and metabolic levels, we measured changes in the plasma concentrations of leptin, insulin, glucose, non-esterified fatty acids (NEFA), weights of the liver and abdominal fat and fat accumulation in the liver and M. gastrocnemius in male rats. Intragastric administration of 5'-UMP via a stomach tube at a dose of 44 mg/day for 7 days slightly (P=0.098) blunted the body weight gain without causing a significant change in food intake. The administration significantly reduced the plasma concentrations of glucose (P=0.004) and NEFA (P=0.004), whereas it significantly increased (P=0.03) plasma leptin concentration. The weights of perirenal (but not epididymal) fat (P=0.083) and the liver (P=0.061) were slightly increased. The triacylglyceride concentration in M. gastrocnemius was slightly increased (P=0.097), although the muscle weight was not significantly changed (P=0.197). In summary, acute oral administration of 5'-UMP was effective in the rat in reducing plasma concentrations of glucose and NEFA, an effect that was accompanied by an elevated plasma leptin concentration.     

Structural determinants for partitioning of lipids and proteins between coexisting fluid phases in giant plasma membrane vesicles

The structural basis for organizational heterogeneity of lipids and proteins underlies fundamental questions about the plasma membrane of eukaryotic cells. A current hypothesis is the participation of liquid ordered (Lo) membrane domains (lipid rafts) in dynamic compartmentalization of membrane function, but it has been difficult to demonstrate the existence of these domains in live cells. Recently, giant plasma membrane vesicles (GPMVs) obtained by chemically induced blebbing of cultured cells were found to phase separate into optically resolvable, coexisting fluid domains containing Lo-like and liquid disordered (Ld)-like phases as identified by fluorescent probes. In the present study, we used these GPMVs to investigate the structural bases for partitioning of selected lipids and proteins between coexisting Lo-like/Ld-like fluid phases in compositionally complex membranes. Our results with lipid probes show that the structure of the polar headgroups, in addition to acyl chain saturation, can significantly affect partitioning. We find that the membrane anchor of proteins and the aggregation state of proteins both significantly influence their distributions between coexisting fluid phases in these biological membranes. Our results demonstrate the value of GPMVs for characterizing the phase preference of proteins and lipid probes in the absence of detergents and other perturbations of membrane structure.     

Degradation of phospholipids and triacylglycerol,and accumulation of fatty acids in anoxic myocardial tissue,disrupted by freeze-thawing

Summary The degradation of lipids by endogenous hydrolytic activity has been studied in rat cardiac tissue deliberately damaged by freezing and thawing prior to storage under anoxic conditions.Aliquots of the freeze-thawed material were kept at 37°C under an atmosphere of N₂ up to 120 minutes. Triacylglycerol was hydrolyzed at a rate of 0.14 mol fatty acids per minute per gram dry weight of tissue. Hydrolysis of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was associated with proportional production of lyso PC and lyso PE, respectively. This finding indicates that the activity of lysophospholipase is negligible in autolyzing cardiac tissue. The rate of hydrolysis of PC and PE was found to be 0.10 and 0.06 mol per minute per gram dry weight of tissue. The observation that lyso PC and lyso PE mainly contained saturated and mono-unsaturated fatty acids indicates that phospholipase A₂ rather than A₁ is active in autolyzing cardiac tissue. The accumulation of fatty acids corresponded with the loss of triacylglycerol and phospholipids from the tissue during 120 minutes of autolysis.     

Changes in mammary uptake of free fatty acids,triglyceride, cholesterol and phospholipid in relation to milk synthesis during lactation in goats

Uptake of free fatty acids (FFA), triglycerides (TG), cholesterol (CHOL) and phospholipids (PL) was measured in both mammary glands of dairy goats during lactation. Arterial concentrations of TG, CHOL and PL as well as arterio-venous difference (AV) and extraction rate (E) for TG were highest in goats with the highest dietary feed intake. AV were linearly related to arterial concentrations for the four lipid classes, and arterial concentrations of CHOL were linearly related to output of lactose, protein and fat in milk. Arterial supply, and not mammary synthetic activity, is the main determinant of mammary FFA, TG and CHOL uptake.     

Changes in plasma membrane lipids, aquaporins and proton pump of broccoli roots, as an adaptation mechanism to salinity

Salinity stress is known to modify the plasma membrane lipid and protein composition of plant cells. In this work, we determined the effects of salt stress on the lipid composition of broccoli root plasma membrane vesicles and investigated how these changes could affect water transport via aquaporins. Brassica oleracea L. var. Italica plants treated with different levels of NaCl (0, 40 or 80 mM) showed significant differences in sterol and fatty acid levels. Salinity increased linoleic (18:2) and linolenic (18:3) acids and stigmasterol, but decreased palmitoleic (16:1) and oleic (18:1) acids and sitosterol. Also, the unsaturation index increased with salinity. Salinity increased the expression of aquaporins of the PIP1 and PIP2 subfamilies and the activity of the plasma membrane H⁺-ATPase. However, there was no effect of NaCl on water permeability (P_f) values of root plasma membrane vesicles, as determined by stopped-flow light scattering. The counteracting changes in lipid composition and aquaporin expression observed in NaCl-treated plants could allow to maintain the membrane permeability to water and a higher H⁺-ATPase activity, thereby helping to reduce partially the Na⁺ concentration in the cytoplasm of the cell while maintaining water uptake via cell-to-cell pathways. We propose that the modification of lipid composition could affect membrane stability and the abundance or activity of plasma membrane proteins such as aquaporins or H⁺-ATPase. This would provide a mechanism for controlling water permeability and for acclimation to salinity stress.     

Human, but not rat, IRS1 targets to the plasma membrane in both human and rat adipocytes

Adipocytes are primary targets for insulin control of metabolism. The activated insulin receptor phosphorylates insulin receptor substrate-1 (IRS1), which acts as a docking protein for downstream signal mediators. In the absence of insulin stimulation, IRS1 in rat adipocytes is intracellular but in human adipocytes IRS1 is constitutively targeted to the plasma membrane. Stimulation of adipocytes with insulin increased the amount of IRS1 at the plasma membrane 2-fold in human adipocytes, but >10-fold in rat adipocytes, with the same final amount of IRS1 at the plasma membrane in cells from both species. Cross-transfection of rat adipocytes with human IRS1, or human adipocytes with rat IRS1, demonstrated that the species difference was due to the IRS1 protein and not the cellular milieus or posttranslational modifications. Chimeric IRS1, consisting of the conserved N-terminus of rat IRS1 with the variable C-terminal of human IRS1, did not target the plasma membrane, indicating that subtle sequence differences direct human IRS1 to the plasma membrane.     

Increased glucose oxidation in H-FABP null soleus muscle is associated with defective triacylglycerol accumulation and mobilization, but not with the defect of exogenous fatty acid oxidation

Heart-type fatty acid-binding protein (H-FABP) is a major fatty acid-binding factor in skeletal muscles. Genetic lack of H-FABP severely impairs the esterification and oxidation of exogenous fatty acids in soleus muscles isolated from chow-fed mice (CHOW-solei) and high fat diet-fed mice (HFD-solei), and prevents the HFD-induced accumulation of muscle triacylglycerols (TAGs). Here, we examined the impact of H-FABP deficiency on the relationship between fatty acid utilization and glucose oxidation. Glucose oxidation was measured in isolated soleus muscles in the presence or absence of 1 mM palmitate (simple protocol) or in the absence of fatty acid after preincubation with 1 mM palmitate (complex protocol). With the simple protocol, the mutation slightly reduced glucose oxidation in CHOW-muscles, but markedly increased it in HFD-muscles; unexpectedly, this pattern was not altered by the addition of palmitate, which reduced glucose oxidation in both CHOW- and HFD-solei irrespective of the mutation. In the complex protocol, the mutation first inhibited the synthesis and accumulation of TAGs and then their mobilization; with this protocol, the mutation increased glucose oxidation in both CHOW- and HFD-solei. We conclude: (i) H-FABP mediates a non-acute inhibition of muscle glucose oxidation by fatty acids, likely by enabling both the accumulation and mobilization of a critical mass of muscle TAGs; (ii) H-FABP does not mediate the acute inhibitory effect of extracellular fatty acids on muscle glucose oxidation; (iii) H-FABP affects muscle glucose oxidation in opposing ways, with inhibition prevailing at high muscle TAG contents.     

Identification of prostamides,fatty acyl ethanolamines,and their biosynthetic precursors in rabbit cornea

Arachidonoyl ethanolamine (anandamide) and pros­taglandin ethanolamines (prostamides) are biologically active derivatives of arachidonic acid. Although available through different precursor phospholipids, there is considerable overlap between the biosynthetic pathways of arachidonic acid-derived eicosanoids and anandamide-derived prostamides. Prostamides exhibit physiological actions and are involved in ocular hypotension, smooth muscle contraction, and inflammatory pain. Although topical application of bimatoprost, a structural analog of prostaglandin F_2α ethanolamide (PGF_2α-EA), is currently a first-line treatment for ocular hypertension, the endogenous production of prostamides and their biochemical precursors in corneal tissue has not yet been reported. In this study, we report the presence of anandamide, palmitoyl-, stearoyl-, α-linolenoyl docosahexaenoyl-, linoleoyl-, and oleoyl-ethanolamines in rabbit cornea, and following treatment with anandamide, the formation of PGF_2α-EA, PGE₂-EA, PGD₂-EA by corneal extracts (all analyzed by LC/ESI-MS/MS). A number of N-acyl phosphatidylethanolamines, precursors of anandamide and other fatty acyl ethanolamines, were also identified in corneal lipid extracts using ESI-MS/MS. These findings suggest that the prostamide and fatty acid ethanolamine pathways are operational in the cornea and may provide valuable insight into corneal physiology and their potential influence on adjacent tissues and the aqueous humor.     

Proteolipid protein and DM-20 are synthesized by Schwann cells,present in myelin membrane,but they are not fatty acylated

Proteolipid protein (PLP) and DM-20 were intensely labeled after immunoprecipitation of total cellular proteins and myelin proteins labeled with [³⁵S]methionine in nerve slices. These results provided evidence that PLP and DM-20 are incorporated into the myelin membrane following their synthesis in Schwann cells. In contrast, PLP and DM-20 were not fatty acylated after incubation of the nerve slices with [³H]palmitic acid, however, PO glycoprotein and 24kDa protein were heavily fatty acylated. The lack of fatty acylation of PLP and DM-20 in the peripheral nervous system suggests that fatty acyltransferase responsible for their acylation is absent or non-functional in the peripheral nervous system.     

Stereoselective protein binding of tetrahydropalmatine enantiomers in human plasma,HSA, and AGP,but not in rat plasma

Tetrahydropalmatine (THP) is one of the active alkaloid ingredients of Rhizoma Corydalis. THP has a chiral center, and the stereoselective pharmacokinetics and tissue distribution have been reported. The aim of the present article is to study the stereoselective protein binding of THP using equilibrium dialysis followed by HPLC‐UV analysis. The results showed that THP stereoselectively binds to human serum albumin (HSA), α₁‐acid glycoprotein (AGP), and proteins in human plasma. The fraction binding of (+)‐THP was significantly higher than that of (?)‐THP, whereas such stereoselectivity was not found in rat plasma. The affinity of HSA and AGP to (+)‐THP, expressed as nK_A, were 9.0 × 10³ M^?1 and 2.34 × 10⁵ M^?1, respectively, which were notablely higher than to (?)‐THP, with the nK_A of 3.4 × 10³ M^?1 and 1.44 × 10⁵ M^?1, respectively. The binding site of HSA for (?)‐THP was Site I, whereas for (+)‐THP was both Site I and Site II. The F1/S variants of AGP were proved to be the key variants (?)‐ and (+)‐THP binding to both. Finally, the AGP binding drugs, such as mifepristone, were demonstrated to reduce the fraction binding of (?)‐ and (+)‐THP with pure AGP (1 mg/ml) but did not affect the fraction binding of both (?)‐ and (+)‐THP with proteins in human plasma. It can be concluded that protein binding of THP is species dependent and stereoselective, both HSA and AGP contribute to the stereoselective binding to THP enatiomers, and AGP binding drugs may not cause the drug–drug interaction on THP in healthy human plasma. Chirality, 2010. © 2009 Wiley‐Liss, Inc.     

Changes in the concentrations of glucose, non-esterified fatty acids, urea, insulin, cortisol and some mineral elements in the plasma of the primiparous sow before, during and after induced parturition.

The concentrations of selected metabolites, minerals and hormones relative to parturition were studied in 12 primiparous sows. Blood sampling was performed on day -5, 0 and +5 relative to the farrowing day. On the day of parturition (d 0), samples were taken every hour from 07.00 to 24.00 hours. All sows had an indwelling catheter in the jugular vein, and the evolution of glucose, insulin, urea, non-esterified fatty acids (UEFA), calcium (Ca), phosphorus (P), magnesium (Mg) and cortisol were studied. The concentrations of NEFA, cortisol and P were significantly higher at d 0 than at d -5 or d +5, whereas the Mg level was lower. During the expulsion of foetuses, NEFA and cortisol levels increased (+18 and +30%, respectively), and they decreased immediately after the birth of the last piglet, to reach the initial levels observed before farrowing (around 700 microEq.L-1 and 110 ng.mL-1, respectively). Glucose and insulin levels remained unchanged during the expulsion of the piglets (105 ng.dL-1 and 5 microIU.mL-1, respectively), but they both increased immediately after the birth of the last animal. During the expulsion of the foetuses, the Ca concentration remained unchanged (93 mg.L-1), whereas the P level increased (+9%) and the Mg concentration decreased (-7.4%). These data suggested that parturition induces large variations in the concentrations of plasma metabolites that may affect its normal process.     

Changes in fatty acid composition of lipids from birds, rodents, and preschool children exposed to lead

Chronic treatment with inorganic lead (Pb) has been shown to increase the proportion of arachidonic acid (ArA), as well as the arachidonate/linoleate (ArA/LA) ratio, in the fatty acids of lipids from a variety of avian tissues. Changes in two fatty acid-mediated phenomena, peroxidation of membrane lipids and synthesis of eicosanoid cytokines, are associated with this enhanced ArA content. The authors are not aware of any reports in the literature in which these effects of Pb have been described for any animals other than birds. In the current study, the authors investigated the effect of Pb on lipid metabolism in three species: avian, rodent, and human. The group of children identified as suffering environmental Pb exposure were from a Pb-surveillance program and had blood Pb concentrations (PbB) averaging 23 μg/dL. Turkey poults fed 100 ppm dietary Pb as Pb acetate-trihydrate for 19 d had a PbB of 46 μg/dL. Gastric intubation of rats with 80 mg Pb/kg/d for 10 d resulted in a PbB of 74 μg/dL. We analyzed fatty acid composition of whole blood from children, poults, and virgin rats. Low-dose (nongrowth inhibitory) Pb exposure resulted in significantly increased ArA concentration and ArA/LA ratio in blood from all species. Also analyzed were plasma and liver of poults, virgin rats, and pregnant rats and their fetuses. In plasma and liver from Pb-treated poults and virgin rats, ArA and the ArA/LA ratio were again enhanced. Pb intoxication also affected ω3 composition, increasing the concentrations of all long-chain ω3 fatty acids of fetuses from Pb-treated pregnant dams. The authors propose that altered fatty acid metabolism may be responsible for some indications of Pb poisoning. Possible consequences mediated through lipid peroxidation and production of ArA-derivative eicosanoids are considered.     

Simultaneous quantification of alprazolam, 4- and alpha-hydroxyalprazolam in plasma samples using liquid chromatography mass spectrometry

A sensitive and specific method was developed for quantification of alprazolam and its two metabolites 4-hydroxyalprazolam and alpha-hydroxyalprazolam in plasma. The work up procedure was solid phase extraction. Liquid chromatography-mass spectrometry (LC-MS) was used for separation, detection and quantification of the analytes. The limit of quantitation (LOQ) was 0.05 ng/mL for alprazolam and the two metabolites. The extraction recovery was more than 82% for alprazolam and its metabolites. The within- and between-assay coefficients of variation were in the range of 1.9-17.9%. The method was used for determination of the pharmacokinetics parameters of alprazolam and its two metabolites in healthy Caucasian subjects who ingested 1mg of alprazolam.     

Combined effects of oleic,linoleic and linolenic acids on lactation performance and the milk fatty acid profile in lactating dairy cows

The potential combined effects of oleic, linoleic and linolenic acids supplementation on lactation performance and the milk fatty acid (FA) profile in dairy cows have not been well investigated. Our objective was to examine the effects of supplementation with a combination of these FA as well as the effects of removing each from the combination on lactation performance and the milk FA profile in dairy cows. Eight Holstein cows (101±11 days in milk) received four intravenously infused treatments in a 4×4 Latin square design, and each period lasted for 12 days which consisted of 5 days of infusion and 7 days of recovery. The control treatment (CTL) contained 58.30, 58.17 and 39.96 g/day of C18: 1 cis-9; C18: 2 cis-9, cis-12; and C18: 3 cis-9, cis-12, cis-15, respectively. The other three treatments were designated −−C18: 1 (20.68, 61.17 and 41.72 g/day of C18: 1 cis-9; C18: 2 cis-9, cis-12; and C18: 3 cis-9, cis-12, cis-15, respectively), −C18: 2 (61.49, 19.55 and 42.13 g/day of C18: 1 cis-9; C18: 2 cis-9, cis-12; and C18: 3 cis-9, cis-12, cis-15, respectively) and −C18: 3 (60.89, 60.16 and 1.53 g/day of C18: 1 cis-9; C18: 2 cis-9, cis-12; and C18: 3 cis-9, cis-12, cis-15, respectively). Dry matter intake and lactose content were not affected by the treatments, but the milk protein content was lower in cows treated with −C18: 2 than that in CTL-treated cows. Milk yield as well as milk fat, protein and lactose yields were higher in cows treated with −C18: 3 than the yields in CTL-treated cows, and these yields increased linearly as the unsaturation degree of the supplemental FA decreased. Compared with the CTL treatment, the −C18: 2 treatment decreased milk C18: 2 cis-9 content (by 2.80%) and yield (by 22.12 g/day), and the −C18: 3 treatment decreased milk C18: 3 cis-9, cis-12, cis-15 content (by 2.72%) and yield (by 22.33 g/day). In contrast, removing C18: 1 cis-9 did not affect the milk content or yield of C18: 1 cis-9. The −C18: 2-treated cows had a higher C18: 1 cis-9 content and tended to have a higher C18: 1 cis-9 yield than CTL-treated cows. The yields of C8: 0, C14: 0 and C16: 0 as well as <C16: 0 tended to increase linearly as the unsaturation degree of the supplemental FA decreased (P=0.06, 0.07, 0.07 and 0.09, respectively). These results indicated that supplementation with C18 unsaturated FA might not independently affect the lactation performance and the milk FA profile of dairy cows.     

In thrombin stimulated human platelets Citalopram, Promethazine, Risperidone, and Ziprasidone, but not Diazepam, may exert their pharmacological effects also through intercalation in membrane phospholipids in a receptor-independent manner

Intercalation of drugs in the platelet membrane affects phospholipid-requiring enzymatic processes according to the drugs’ intercalation capability. We investigated effects of Promethazine, Citalopram, Ziprasidone, Risperidone, and Diazepam on phospholipase A₂ (PLA₂) and polyphosphoinositide (PPI) metabolism in thrombin-stimulated human platelets. We also examined effects of the drugs on monolayers of glycerophospholipids using the Langmuir technique. Diazepam did not influence PLA ₂ activity, had no effects on PPI cycle, and caused no change in mean molecular area of phospholipid monolayers. The remaining psychotropic drugs affected these parameters in different ways and levels of potency suggesting that they act by being intercalated between the molecules of adjacent membrane phospholipids, thus causing changes in substrate availability for phospholipid-hydrolyzing enzymes (PLA₂ and Phospholipase C). We show that several psychotropic drugs can also have other cellular effects than receptor antagonism. These effects may be implicated in the psychotropic effects of the drugs and/or their side effects.     

Changes in the Montagu’s Harrier Circus pygargus diet in Eastern Poland across decades promote insects and reptilians,but not birds and rodents

We investigated temporal changes in diet composition of the Montagu''s Harrier Circus pygargus breeding in natural habitat (calcareous peat bog) in SE Poland. We characterized diet composition in a three‐year period (2007–2009), based on pellet analyses. We investigated whether diet composition was affected by years or stage of breeding. We compared diet of the studied population between 2000s and 1990s and with other populations. We found that the food of the studied population was dominated by insects and mammals (by number) and mammals and birds (by biomass). Biomass and abundance of main prey items differed between studied years because of different air temperatures. We found some interannual differences in contribution of some prey items including higher number of thermophilic prey (insects and amphibians) in warmer years. Comparison of pellet composition in the 1990s and 2000s revealed significant increase in the abundance of thermophilic prey (insects and reptiles) and decrease of mammals including Microtus voles and birds. Those changes may be linked to habitat changes in areas neighboring peat bogs and climate change‐induced changes in prey communities. The studied population was able to respond to changes in foraging habitats and prey composition by opportunistic foraging on easily available prey. The diet of the studied population is the most similar to the geographically closest populations foraging in similar habitats and characterized by high contribution of insects.