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Phospholipid Fatty Acid Profiles of Plasma and Erythrocyte Membranes in Dogs Fed with Commercial Granulated Food

Phospholipid Fatty Acid Profiles of Plasma and Erythrocyte Membranes in Dogs Fed with Commercial... Acta Veterinaria-Beograd 2023, 73 (1), 119-132 UDK: 636.7.085.3 DOI: 10.2478/acve-2023-0009 Research article PHOSPHOLIPID FATTY ACID PROFILES OF PLASMA AND ERYTHROCYTE MEMBRANES IN DOGS FED WITH COMMERCIAL GRANULATED FOOD # 1 #1 1 Tamara POPOVIĆ * , Jasmina DEBELJAK MARTAČIĆ , Biljana POKIMICA , 1 1 1 2 Branko RAVIĆ , Slavica RANKOVIĆ , Maria GLIBETIĆ , Predrag STEPANOVIĆ University of Belgrade, Institute for medical research, Laboratory for food and metabolism, Belgrade, Serbia; University of Belgrade, Faculty of Veterinary medicine, Associate Professor, Department of equine, small animal, poultry, and wild animal diseases, Belgrade, Serbia (Received 20 October 2022, Accepted 16 January 2023) Intake of long-chain n-3 polyunsaturated fatty acids (PUFA) benefi ts human and animal health. Our study aimed to analyze the long-chain n-3 PUFA content of two types of food and their effect on plasma and erythrocyte phospholipids of Belgian Shepherd dogs. A total of 10 dogs were fed commercial granulated food (Food 1), and another 10 were provided commercial Premium granulated food of high quality (Food 2). All the analyses were performed using gas-liquid chromatography. Our results showed that Food 1 contained more n-3 PUFA than Food 2, which was refl ected in higher n-3 PUFA in plasma and erythrocyte phospholipids. Because long- chain n-3 PUFA in phospholipids are precursors for antioxidative molecules, further studies should investigate the effects of the analyzed commercial granulated food rich in n-3 on oxidative stress parameters in dogs. Keywords: commercial foods, dogs, fatty acids profi les, n-3 fatty acids, phospholipids. INTRODUCTION In recent years, many pet owners have abandoned conventional, veterinary- recommended commercial diets in search of more “natural” and “homemade” choices [1]. Still, it is more common practice nowadays for owners to use commercialized labeled food for pets such as canines. Among other ingredients, polyunsaturated fatty acids (PUFA) are important for human and animal health. The conversion of short chain to long chain PUFA is rate-limiting and varies between species [2]. The balance of n-6/n-3 ratio in phospholipids, as well as the balance between reactive oxygen species and reactive nitric species on one side and antioxidative defense, on the other, is important for the normal physiological function of organisms. In dogs, fewer *Corresponding author: e-mail: poptam@gmail.com Authors have contributed equally to this work and, thus, share the fi rst authorship 119 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 infl ammatory mediators were produced when fed diets ratios (n-6/n-3) of 5:1 and 10:1 in comparison with being fed an n-6-rich diet with a fatty acid ratio of 100:1 [3]. Mammals are not able to synthesize fatty acids (FA) with double bonds at C-9, but are able to some extent to elongate and further saturate the aliphatic chain [4]. It is well known t hat diet could affect fatty acid profi les in plasma and erythrocyte membranes (EM) and that changing its composition causes changes in many parameters, which is confi rmed in animal and human studies [3]. Since the FA composition of the EM correlates with that of other cell membranes, the effect of dietary FA supplementation may be analyzed by studying EM. It is well known that plasma phospholipids profi les refl ect short-term changes in the diet while changes in erythrocyte membrane phospholipids refl ect long-term dietary habits (up to 3 months prior analysis). Comparing those two profi les could lead us to a conclusion about the diet habits of examined animals in general [5]. The FAs profi le of the serum phospholipids is related to the average dietary FAs intake during the last 3 to 6 weeks, while the composition of erythrocyte phospholipids depends on the dietary fat intake during the preceding months [5]. The FAs profi le in the tissues partly refl ects not only the dietary fat intake but also the effi ciency of FAs metabolism in the body [6]. The FAs profile of tissues and triglycerides (TG) is known to be influenced by many factors, including dietary intake, age, gender, and endogenous metabolism [7]. Considerable interest exists in the possible health benefits of increasing dietary intake of n-3 PUFA [8]. Three families of long-chain PUFAs with different biological roles exist, n-3, n-6 and n-9 and they are derived from the shortest non-synthesizable precursors: linoleic acid (LA) (18:2, n-6) and alpha-linolenic acid (ALA) (18:3, n-3). Humans can desaturate and elongate ALA, as a precursor of n-3 series, to eicosapentanoic acid (EPA) and docosahexanoic acid (DHA). This process is dependent on aging, presence and type of disease, inflammation processes, and other factors [4]. In rats, which are often used as animal models, the rate of conversion of ALA to DHA is high in the liver, although Δ5 and Δ6 desaturases are expressed in many other rodent tissues besides the liver [9]. There are few commercial pet dog foods with EPA and DHA concentrations adequate for the treatment of disease or some vulnerable conditions. Target ranges for EPA and DHA vary quite widely for different conditions but typically fall between 50 and 220 mg/kg body weight. Commercial diets with n-3 fatty acids typically provide less EPA and DHA than desirable and may be advertised as containing fl axseed or canola oil (rich in ALA) [10]. In fact, there are adverse effects associated with the use of n-3, and an increase in the concentration of EPA and DHA in commercial pet food (dogs) makes the topic important to revisit. Those effects include altered platelet function, gastrointestinal adverse effects, potential effects for nutrient excess, weight gain, altered immune function, and effects on glycemic control and insulin sensitivity. As far as dogs and specifi c abnormalities are concerned decreased epithelization of wounds after 5 120 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food days (n-6/n-3=0.3:1), increased plasma and urine thiobarbituric reactive substances (n-6/n-3=5.4:1), decreased plasma vitamin E (n-6/n-3=1.4:1), decreased skin and neutrophil leukotriene B4/increased leukotriene B5, lower delayed-type hypersensitivity response (n-6/n-3=1.4:1), decreased CD4+T lymphocyte count (n-6/n-3=1.4:1), decreased lymphocyte proliferation (EPA/DHA=0.8) [11]. Metabolically, however, fatty acid patterns of plasma phospholipid fractions again revealed a sparing effect of ALA on LA. It should be mentioned that a direct effect of ALA on improvements of skin and coat could not completely be ruled out in these studies while long-chain n-3 PUFAs from fi sh oil or other marine sources appear to be especially capable of modifying infl ammatory and immune responses [12]. The aim of the study was to analyze the fatty acid content of two different diets for dogs and to examine the effects of their daily consumption on the erythrocyte membrane and plasma phospholipid profi les. MATERIALS AND METHODS Animals This study which lasted for 12 weeks, was approved by the Ethics Commission of the Faculty of Veterinary Medicine are included dogs of the Belgian Shepherd (Malinoa) breed from two kennels (1 and 2). In kennels, we selected 10 dogs, (5 females and 5 males), age categories of 3 to 7 years, with body weight 30.2±2.2 kg. We measured weight gain monthly and it changed up to 1 kg/dog. Male dogs weight gain was slightly higher than females but without statistical difference. By the basic examination of the dogs in both kennels (blood pressure, pulse, temperature, breathing, skin condition, and skin cover), there was a constant absence of disease, otherwise, all dogs had neatly managed health cards. According to the constitution they fell into 3 categories, which means they have an ideal weight corresponding to this breed of dog. The dogs had their activities in the morning and in the evening for 60 minutes (walking, running), otherwise, these dogs are considered as working dogs. In kennel number 1, the dogs were fed commercial granular foods that normally satisfi es the standard nutritional needs of dogs (I). In kennel number 2, the dogs were fed Premium granulated food of high quality, this being the most sold dog food in Serbia (II). The amount of food (400 g/day/dog) was divided into two meals one in the morning and the other in the evening at the same time each day. Sample collection and analysis Granulated foods samples (I and II) (from four representative large markets) were analyzed. The primary sample was generated by mixing an equal portion of four samples taken from different markets. Five replicate samples of the composite sample were analyzed by standard laboratory methods to measure the concentration of 121 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 proteins, carbohydrates, lipids, minerals, and water. Fatty acid composition from the lipids was done by standard laboratory procedure, as described below. At the end of the study blood samples were taken from the (vena cephalica antebrachii), with the aid of EDTA vacuum blood collection tubes. Erythrocytes and plasma were separated and stored at a temperature of -80 C . After blood sampling by the routine method, analysis of erythrocytes and plasma was done by gas-liquid chromatography (GC). Nutritional analysis was carried out by an accredited chemical laboratory at the Institute of Public Health in Požarevac. Ash content was determined by the direct gravimetric method which includes ashing of the samples in an oven at 550 °C until a constant weight was attained. Moisture was determined gravimetrically [13]. Crude protein content was estimated based on the total nitrogen content of the sample determined by the Kjeldahl method (AOAC 955.04D) [13]. Crude fat content was determined gravimetrically (Soxhlet extraction, AOAC method [13]. Total carbohydrate content, crude “by difference”, was calculated by the following formula: total carbohydrate (%) = 100% - % (protein + ash + fat + moisture). The energy content of food was calculated based on determining content by the following formula: Energy value (estimated, kJ/100 g) = [4 x protein (%)] + [4 x carbohydrate (%)] + [9 x fat (%)]. Fatty acids extraction and analysis Isolation of lipids The method consists of homogenization of plasma with a 2:1 chloroform/methanol mixture. Washing of the mixture with a 5 times smaller volume of water or saline (0.9 g NaCl in 100 ml of water). The resulting mixture was separated into two phases. The lower phase was the total pure lipid extract. First the lipids present in the volume of 0.5 ml of erythrocytes were extracted with a mixture of chloroform isopropanol (7:11) following the prescribed procedure [14]. After that we isolated the phospholipids from the lipid subclasses by thin-layer chromatography on silica-gel plates using petroleum ether, diethyl ether and glacial acetic acid (87:12:1, by volume). In a modifi ed procedure by Christopherson and Glass [15], we made a direct transesterifi cation of fatty acids. Lipid extracts derived from hexane were evaporated under a nitrogen stream to full vapor and dry bottom of Eppendorf ’s tubes. After that, the residue was dissolved in 10µl hexane. We took 1 µl and injected it into a chromatograph. Methyl esters of fatty acids were analyzed by gas-liquid chromatography in a Shimadzu chromatograph GC 2014 (Kyoto, Japan) equipped with a fl ame ionization detector on an Rtx 2330 column (60mm x 0,25 mmID, fi lm thickness 0,2 µm, Restek, Bellefonte, PA, USA). Adequate separation was obtained over a 50 min period with an initial temperature of 140 C maintained for 5 0 0 min. The temperature was then increased to 220 C at a rate of 3 C/min and kept at the fi nal temperature for 20 min. The identifi cation of fatty acid methyl esters (FAME) 122 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food was made by comparing peak retention times with standard mixtures (PUFA-2 and/ or 37 FAMEs mix, Supelco, Bellefonte, PA, USA). Finally, the content of fatty acids, from 16:0 through 22:6n-3, was expressed as a percentage of the total content of identifi ed fatty acids. Statistical analysis Statistical analysis was done by one-way ANOVA test and Student t-test (signifi cance p<0.05) in SPSS. All results were expressed in percentages of a total of 100%. RESULTS Biochemical parameters Analyzed biochemical parameters such as triglycerides and plasma cholesterol did not show sta s cal signifi cance in the plasma of canines treated with two diff erent diets. Although there were some changes in LDL cholesterol, previously reported by Ravic et al [16]. Analyzed food There were differences in FA percentage between the examined foods. There was an equal percentage of palmitic (16:0) acid, but stearic acid was more abundant in Food 1 (p<0.01). Monosaturated FA, specially palmitoleic acid, was present at a higher percentage in Food 2 (p<0.01), while oleic acid (18:1 n-9) was detected almost in the same percentage. Linoleic acid (18:2 n-6) was also present in more than 10% in both Food 1 and Food 2. Long-chain PUFAs were present in a very low percentage with statistically signifi cant differences between Foods. ALA (18:3n3) was statistically signifi cant in a higher percentage in Food 1 (p<0.001), as well as EPA (p<0.001) and DHA (p<0.001) compared to Food 2 (Table 1). Table 1. Composition and FA patterns of the diets Fatty acids (%) Food 1 Food 2 16 :0 23.93± 0.15 23.13 ± 0.1 Palmitic acid 16 :1 2.47± 0.26 3.72 ± 0.07** Palmitoleic acid 18 :0 13.36± 0.26 9.26 ± 0.26** Stearic acid 18: 1,n 9 36.88± 0.55 40.61± 0.92 Oleic acid 18: 1,n 7 4.00± 0.21 3.46 ± 0.33 Vascenic acid 18: 2 15.13± 1.34 18.04 ± 0.26* Linoleic acid 123 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 cont. Table 1. 18: 3 n 3 2.99± 0.22 0.95 ± 0.02*** Linolenic acid 20: 3 0.07± 0.01 0.13± 0.02*** Eicosatrienoic acid 20: 4 0.45± 0.09 0.36± 0.12 Arachidonic acid 20: 5 0.22± 0.09 0.04 ± 0.02*** Eicosapentanoic acid 22 : 4 0.09± 0.01 0.13 ± 0.02** Docosatetranoic acid 22 : 5 0.07± 0.01 0.07 ± 0.01 Docosapentanoic acid 22: 6 0.31± 0.08 0.07 ± 0.01*** Docosahexanoic acid Proteins 27.29 (0.05) 25.51(0.04) Fats 16.44 (0.04) 15.22 (0.04) Carbohydrates 44.54 (0.22) 46.41 (0.24) Minerals 6.69 (0.01) 7.67 (0.01) Water 5.05 (0.02) 5.21 (0.03) Values (proteins, carbohydrates, fats, minerals and water) are expressed as % by 100g. Abbrrevations SD, Standard deviations. Fatty acids are expressed as % ± st. dev with signifi cance *p<0.05,**p<0.01,***p<0.001 Dog plasma and erythrocyte membrane phospholipid fatty acids It is well known that plasma phospholipids fatty acids profi les represent the short- term (up to several days) food intake, while erythrocytes present data composition of fatty acids for long term (last 3 months) food intake. In our study, we compare dogs’ FAs profi les in plasma and erythrocytes membrane phospholipids in two kennels (Table 2-4). In plasma phospholipids FA profi les in palmitic acid (16:0) were increased in Kennel 1(p<0.001) as well as stearic acid (18:0), results showed that dogs in Kennel 1 had a higher percentage of saturated fatty acids (SFA) in plasma phospholipids compared to Kennel 2. As far as monosaturated fatty acids, palmitoleic (16:1) was increased in Kennel 2 while oleic acid (18:1, n-9) was increased in Kennel 1 (p<0.01) compared to dogs fed Kennel 2. Linoleic acid (18:2) was increased (p<0.001) in dogs in Kennel 2 while polyunsaturated n-3 especially ALA (p<0.01) eicosapentaenoic (EPA) (p<0.001), docosapentaenoic (DPA) (p<0.001) and docosahexaenoic (DHA) (p<0.001) were increased in Kennel 1. In the erythrocyte membrane, phospholipids´ percentage of fatty acids has almost the same distribution and percentages as in plasma phospholipids between Kennels. Saturated fatty acids (PA and SA) (p<0.01) were in higher percentages in Kennel 1-fed dogs as well as monounsaturated FA (p<0.1). Percentage of EPA (p<0.001), DPA (p<0.01), and DHA (p<0.001) were increased in Kennel 1 in plasma phospholipids, 124 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food while ALA concentration did not signifi cantly change. These results go along with the plasma phospholipids FA distribution already mentioned. These results suggest that the differences are negligible between plasma and erythrocytes phospholipids fatty acids percentages. Ta ble 2. Plasma and erythrocyte phospholipids fatty acids profi les of dogs in Kennel 1 and 2. Fatty Acids (%) (plasma) Kennel 1 Kennel 2 16 :0 17.09 ± 1.29 14.55 ± 1.48 *** 16 :1 0.39 ± 0.12 0.68 ± 0.33 *** 18 :0 30.12 ± 1.50 29.81 ± 1.81 18: 1 ,n 9 5.76 ± 0.67 4.89 ± 0.45 ** 18: 1 ,n 7 1.96 ± 0.14 1.69 ± 0.23 * 18: 2 16.63 ± 2.12 24.52 ± 1.73 *** 18: 3 n 6 0.34 ± 0.06 0.40 ± 0.04 * 18:3 n 3 0.23 ± 0.01 0.15 ± 0.07 ** 20: 3 1.27 ± 0.11 1.86 ± 0.51 ** 20: 4 17.86 ± 2.27 17.71 ± 2.21 20: 5 2.26 ± 1.18 0.24 ± 0.03 *** 22 : 4 0.43 ± 0.01 0.82 ± 0.02 *** 22 : 5 2.38 ± 0.88 1.91 ± 0.51 *** 22: 6 3.00 ± 0.78 0.49 ± 0.02 *** Fatty Acids (%) (erythrocytes) Kennel 1 Kennel 2 16 :0 19.2 ± 0.69 17.15 ± 1.53 ** 16 :1 0.24 ± 0.038 0.22 ± 0.012 18 :0 30.20 ± 0.78 29.49 ± 1.68 18: 1 ,n 9 7.87 ± 0.24 7.09 ± 0.53 * 18: 1 ,n 7 2.03 ± 0.11 1.83 ± 0.12 * 18: 2 9.31 ± 1.04 13.82 ± 0.37 *** 18: 3 n 6 0.45 ± 0.06 0.42 ± 0.05 18:3 n 3 0.17 ± 0.04 0.20 ± 0.02 20: 3 2.51 ± 0.25 2.78 ± 0.35 20: 4 24.0 ± 0,96 23.20 ± 2.44 20: 5 1.48 ± 0.26 0.23 ± 0.04*** 22 : 4 0.96 ± 0.23 1.64 ± 0.22 *** 22 : 5 ± 0.22 0.73 ± 0.18 ** 22: 6 1.08 ± 0.21 0.23 ± 0.11 *** 125 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 Table 3. Estimated desaturase 9,6,5 activities in plasma and erythrocytes in dogs in Kennel 1 and 2. Estimated desaturase Kennel 1 Kennel 2 Kennel 1 Kennel 2 (D) activity plasma plasma erythrocytes erythrocytes D9 desaturase 0.19 ± 0.03 0.17 ± 0.02 0.25 ± 0.01*** 0.24 ± 0.02***# D6 desaturase 6.01 ± 0.20 5.04 ± 1.50 15.65 ± 5.42*** 13.43 ± 2.63 D5 desaturase 0.14 ± 0.07 0.01 ± 0.005### 0.06 ± 0.01*** 0.01 ± 0.003### Test signifi cance were used between groups, *statistical signifi cance between plasma and erythrocytes in both Kennels, # statistical signifi cance between plasma in both Kennels and erythrocytes in both Kennels, with *#p<005, **##p<0.01 and ***###p<0.001 Table 4. Percentage of overall n-3, n-6 and n-6/n-3 ratio in Kennel 1 and Kennel 2 in plasma and erythrocytes of examined dogs. Plasma K1 Plasma K2 Erythrocytes K1 Erythrocytes K2 N-3 7.18 ± 0.32 5.95 ±0.55 2.43±0.40 2.52 ± 0.33 N-6 36.50±1.33 33.23 ±0.11 35.48±1.42 34.52 ± 0.88 N6/N3 5.07±1.00 6.05 ±0.95 15.00 ± 3.64 16.7±0.43 K1: Kennel 1; K2: Kennel 2. DISCUSSION Generation of SFA, MUFA and PUFA fatty acids is connected with mediators such as prostaglandins, leukotrienes and others, which are able to infl uence metabolic changes in dogs in health and disease. The n-6/n-3 balance is important in everyday balanced diet and is recommended to be as low as it could be. Following up on intakes of linoleic acid, as n-6 family fatty acid, and its metabolic pathway to DGLA and arachidonic acid (AA) and cell membrane incorporation is of great importance as information of biochemical or clinical parameters for the evaluation of dog´s health [5]. Omega-6 linoleic and its transformation to ARA affect not only from the cell structural point but also affect the membrane response to stimuli, thus membrane fatty acid composition can be useful as information of pro and anti-infl ammatory predisposition of the canine organism [17]. By correlating plasma and erythrocyte phospholipids FA composition after the treatment, we investigated if FA from both plasma and erythrocytes could represent markers of dietary n-3 intake. Our discussion is more based on n-3 PUFA signifi cance and intake in canines. Certain results indicate a benefi cial effect of n-3 long-chain polyunsaturated fatty acids (LC PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on canine health. Several studies have examined the effects of fi sh oil, linseed oil, or combinations on plasma phospholipids fatty acids profi le in canine species [18], lymphocyte proliferation, or neutrophil composition in laboratory animals [19,20]. 126 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food The nutrient requirements of canine athletes who have a greater capacity for fat oxidation are unique concerning that canine metabolism is unique [10]. Relative to body mass size dogs metabolize free fatty acids at twice the rate observed in men. Consequently, the muscles are more adapted to use fat than human muscles [21]. Usual daily food requirements are water, energy, major nutrients, fi bers, minerals, and vitamins. High fat diet, fatty acids chain length and saturation affect a variety of issues from infl ammation to oxidative stress in animals [22,23]. However, there is very little information regarding optimal dietary fats intake for canine athletes [23]. Medium-chain triglycerides yield 8 carbon to 12 carbon-free fatty acids (coconut, palm oil) when digested and are directly transported through absorption into the blood, bound to albumin, to the liver via the portal circulation, leading to sparing of glycogen [24]. The fatty acid composition also infl uences detection in scent-trained dogs, hunting dogs, and service dogs. It is well known that polyunsaturated fatty acids may modestly improve performance in dogs that require activity as a part of their work [25]. The use of carbohydrates as a major dietary substrate is essential. Soluble fi bers may alter the large intestinal microfl ora which produces short-chain fatty acids. A small amount of soy fi ber and fructooligosaccharides are components of gastrointestinal veterinary diets. Defi ciencies in major minerals have been observed in dogs fed with non-traditional diets. It is recommended that bones should be ground into the diet to improve calcium and phosphorus balance. Vitamins (fat-soluble or water-soluble) are involved in cellular metabolism or coenzymes in the citric acid cycle. A study by Poel at al (2017) [26] was conduc ted to determine the stability of two preparations of GAA (granulated and crystallized) and CMH in a moist and a dry dog food formulation during production and storage. Most commercial dog foods contain vitamins above the minimum requirement. Diets high in PUFA (fi sh-based sled dog diets) should contain more vitamin E to prevent lipid peroxidation. In the study by Vessecchi et al., the study aimed to evaluate the macronutrient composition, fatty acids and amino acids profi le, and essential minerals contents of vegan pet foods available in the Brazilian market, and to assess their compliance with recommended allowances for dogs and cats [27]. Omega -3 fatty acids incorporat ed in cell membranes, especially EPA and DHA, decrease clinical signs of osteoarthritis in dogs. EPA serves as a substrate for the COX and LOX enzymes. Omega-6 FA are involved in physiological processes while infl ammation contributes to the formation of “proinfl ammatory” prostaglandins and leukotrienes. Omega-3 produces less infl ammatory prostanoids and 5-leukotrienes. Dogs have limited ability to convert ALA to DHA. That is the reason for diet EPA and DHA consumption directly. Diets that contain EPA and DHA have been recommended for degenerative joint diseases, aging in general, as anti-infl ammatory supplements, and in growing puppies [25]. Currently, omega-3 FAs are used in managing many diseases including neoplasia, dermatologic diseases, hyperlipidemia, cardiovascular, gastrointestinal, and orthopedic diseases [11]. There are few commercial pet foods with EPA and DHA adequate for the treatment of disease. Joint diets, renal diets 127 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 dermatologic conditions contain more omega-3 fatty acids than maintenance diets. Commercial diets with omega-3 provide less EPA and DHA than desirable and contain fl axseed instead. Because diets with ALA have different effects when compared with diets enriched in EPA and DHA composition information may very much contribute to results. Purushotman et al. study on beagles fed with fl axseed added to a basal diet (57% alfa-linolenic acid, ALA at the rate 100ml/kg for 3 weeks) showed that plasma ALA, EPA, and LA increased steadily and signifi cantly from 0-22 days while arachidonic acid showed no difference. Plasma DHA showed no signifi cant changes over time which agrees with previous studies in dogs [2]. Dunbar et al., concluded that hepatic conversion of DHA is slow in the canine and after its production from ALA it is likely to relocate to neurological tissue [29]. Extensive studies are required for clarifying this issue and to confi rm how different breeds metabolize PUFA. Waldron et al. confi rm earlier reports in their study on dogs fed a fi sh oil diet that ALA was converted to EPA and further elongated to DPA but was not converted to DHA in plasma phospholipids [18]. The benefi cial effect of n-3 FA due to their competition with n-6 for cellular membrane incorporation leads to the suggestion that n-6: n-3 ratio could be used as a dietary index to modulate cell composition and cellular function [30]. Still, the type and amount of n-3 PUFA, but not the n-6:n-3 ratio in diets, contribute to membrane n-3’s highly unsaturated fatty acid composition and further changes in cell function [18]. Stoeckel et al. (2011) concluded that in dogs an increase of dietary n-3 FA content leads to a rapid inclusion of n-3 into the erythrocyte membrane, regardless of whether the n-3 FA is offered as an enriched diet or as a normal diet supplemented with an n-3 FA additive [31]. The cardiovascular benefi ts of n-3 PUFA could originate from their ability to improve lipid metabolism and reduce the synthesis of proinfl ammatory eicosanoids derived from n-6 PUFA. At an adequate level of incorporation, EPA and DHA infl uence the membrane fl uidity as well as membrane protein-mediated reactions, generation of lipid-mediators, cell signaling, and gene expression in different cells [32]. Data on cats, together with the results of lipoprotein analysis, indicate possible disturbances in the hepatic transformation of LDL and VLDL, and a high risk of atherogenic events [33]. Dogs have the capacity to metabolize n-3 fatty acids and the effects of omega-3 fatty acids on the skin and coat, infl ammatory responses, and neurologic development in puppies are quite visible [34]. Concerning specifi c benefi cial effects that fi sh oil or EPA and DHA have on canines, it would be useful to add it to the examined food or as an everyday supplement to those dogs. Diagnostic tests performed in dogs provide valuable information [35] and our data can contribute to it. Our further examination and studies will address it, and we planned a study on the treatment of PUFA in canines (EPA +DHA). That study could be useful and give us a more complete conclusion about the signifi cance of EPA and DHA in canine feeding. 128 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food Acknowledgments This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, Contract 451-03-9/2021-14/200015. Ethical approval The research related to the use of animals has been complied with all the relevant national regulations and institutional policies for the care and use of animals (Ministry of Agriculture and Forestry and Water Management - Veterinary Administration, number 323-07-00364/2017-05/6 date 13. 07. 2017., and approved by the Ethical comity of the Faculty of Veterinary Medicine, 17 June 2017 number 01-592 ). Authors’ contributions TP was involved in concept and design, drafting, and supervision. JDM, BP, and SR participated methodology and data collection. BR was involved in methodology, analysis, and interpretation of data. MG participated as project administration and funding acquisition. PS was involved in conceptualization funding acquisition. Declaration of confl icting interests The authors declare that they have no confl ict of interest with respect to the research, authorship, and/or publication of this article. Statement of Informed Consent The owner understood procedure and agrees that results related to investigation or treatment of their companion animals, could be published in Scientifi c Journal Acta Veterinaria-Beograd. REFERENCES 1. Schlesinger DP, Joffe DJ: Raw food diets in companion animals: a critical review, Can Vet J 2011, 52:50-54. 2. Purushotman D, Brown WY, Wu SB, Vanselow B: Evaluation of breed effects on n-3 PUFA metabolism with dietary fl axseed oil supplementation in dogs. Br J Nutr 2011, 106:139-141. 3. Avramovic N, Dragutinovic V, Krstic D, Colovic M, Trbovic A, De Luka S, Milovanovic I, Popovic T: The effects of omega 3 fatty acid supplementation on brain tissue oxidative status in aged Wistar rats. Hippokratia 2012, 16: 241-245. 4. Singer P, Jaeger, Voigt S, Thiel H: Defective desaturation and elongation of n-6 and n-3 fatty acids in hypertensive patients. Prostaglandins Leukot Med 1984, 15:159-165. 5. Popović T, Borozan S, Arsić A, Martačić JD, Vučić V, Trbović A, Mandić LJ, Glibetić M: Fish oil supplementation improved liver phospholipids fatty acid composition and parameters of oxidative stress in male Wistar rats. J. Anim Physiol Anim Nutr (Berlin) 2012, 96:1020-1029. 129 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 6. Ferrucci L, Cherubini A, Bandinelli S: Relationship of plasma polyunsaturated fatty acids to circulating inflammatory markers. J Clin Endocrinol Metab 2006, 91:439-446. 7. Oguzhan B, Sancho V, Acitores A, Villanueva-Peñacarrillo ML, Portois L, Chardigny JM, Sener A, Poel van der YA, Braun AU, Hendrix W, Bosch G: Stability of creatine monohydrate and guanidino acetic acid during manufacture (retorting and extrusion) and storage of dog foods. J Anim Physiol Anim Nutr 2019, 103:1242-1250. 8. Hagopian K, Weber KL, Hwee DT, Van Eenennaam AL, López-Lluch G, Villalba JM, Burón I, Navas P, German JB, Watkins SM, Chen Y, Wei A, McDonald RB, Ramsey JJ: Complex I-associated hydrogen peroxide production is decreased and electron transport chain enzyme activities are altered in n-3 enriched fat-1 mice. PLoS One 2010, 5:e12696. 9. Igarashi MF, Gao HW, Kim KMA, Bell JM, Rapoport SI: Dietary n-6 PUFA deprivation for 15 weeks reduces arachidonic acid concentrations while increasing n-3 PUFA concentrations in organs of post-weaning male rats. Biochim Biophys Acta 2009, 1791:132-139. 10. Hill R: The nutritional requirements of exercising dogs. J Nutr 1998, 128:2686-2690. 11. Lenox CE: Potential adverse effect of omega-3 fatty acids in dogs and cats. J Vet Intern Med 2013, 27:217-226. 12. Bauer JE: Responses of dogs to dietary omega-3 fatty acids. J Am Vet Med Assoc 2007, 231:1657-1661. 13. AOAC: Offi cial method of analysis: association of analytical chemists. 19th Edition, Washington DC; 2012, 121-130. 14. Rose HG, Oklander M: Improved procedure for the extraction of lipids from human erythrocytes. J Lipid Res 1965, 6:428-431. 15. Christopherson SW, Glass RLJ: Preparation of milk fat methyl esters by alcoholysis in an essentially nonalcoholic solution. Dairy Sci 1969, 52:1289-1290. 16. Ravić B, Debeljak-Martacić J, Pokimica B, Vidović N, Ranković S, Glibetić M, Stepanović P, Popović T: The effect of fi sh oil-based foods on lipid and oxidative status parameters in police dogs. Biomolecules 2022, 12(8), 1092. 17. Domingez T, Kizanpreat K, Burri L: Enhanced omega3 index after long-verus short chain omega 3 fatty acids supplementation in dogs. Vet Med Sci 2021, 7(2):370-377. 18. Waldron M, Hannah S, Bauer J: Plasma phospholipid fatty acid and ex vivo neutrophil responses are differencially altered in dogs fed fi sh and linseed-oil containing diets at the same n-6:n-3 fatty acid ratio. Lipids 2012, 47:425-434. 19. Varning K, Schmidt EB, Svaneborg N, Moller JM, Lervang HH, Grunnet N, Jersild C, Dyeberg J: The effect of n-3 fatty acid on neutrophil chemiluminescence. Scand J Clin Lab Invest 1995, 55:47-52. 20. Sperling RI: Effects of dietary fi sh oil on leukocyte leukotriene production and PAF generation and on neutrophil chemotaxis. World Rev Nutr Diet 1991, 66:391-400. 21. De Bruijne JJ, Altszuler N, Hampshire J, Visser TJ, Hackeng WH: Fat mobilization and plasma hormone levels in fasted dogs. Metabolism 1981, 30:190-194. 22. Emara E, El-Sayyad H, El-Ghaweet H: Bovine whey supplementation in a high-fat diet fed rats alleviated offspring’s cardiac injury. Maced Vet Rev 2022, 45:89-99. 23. Wakshlang J, Shmalberg J: Nutrition for working and service dogs. Vet Clin Small Anim 2014, 44:719-740. 24. Jeukendrup AE, Aldred S: Fat supplementation health and endurance performance. Nutrition 2004, 20:678-688. 130 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food 25. Angle TC, Wakshlag JJ, Gillette RS: The effects of exercise and diet on olfaction in detection dogs. J Nutr Sci 2014, 44:1-5. 26. Esther AHP, Denmark FO, Guido B, Wouter, HH, Antonious FB van der Poel: Retorting conditions affect palatability and physical characteristics of canned cat food. JNS 2017,6(e23):1-5. 27. Vessecchi R, Zafalon A, Risolia LW, Henrique T, Vendramini A, Rodrigues RBA, Pedrinelli V, Teixeira FA, Rentas MF, Perini MP, Alvarenga IC, Brunetto MA: Nutrional inadequacies in commercial vegan foods for dogs and cats. Plos ONE 2020, 15 (1):1-17. 28. Wandel S, Juni P, Tendal B: Effects of glucosamine, chondroitin or placebo in patients with osteoarthritis of the hip or knee: network meta-analysis. BMJ 2010, 341:4675. 29. Dunbar BL, Bigley KE, Bauer JE: Early and sustained enrichment of serum n-3 long chain polyunsaturated fatty acids in dogs fed a fl axseed supplemented diet. Lipids 2010, 45:1-10. 30. Harris, WS: The omega 6-omega 3 ratio and cardiovascular disease risk:uses and abuses. Curr Athero Rep 2006, 8:453-459. 31. Stoeckel K, Nielsen LH, Fuhrmann H, Bachmann L: Fatty acid patterns of dog erythrocyte membranes after feeding of a fi sh-oil based DHA-rich supplement with a base diet low in n-3 fatty acids versus a diet containing added n-3 fatty acids. Acta Vet Scand 2011, 24:53-57. 32. Cao J, Schwichtenberg KA, Hanson NQ, Tsai MY: Incorporation and clearance of omega-3 fatty acids in erythrocyte membranes and plasma phospholipids. Clin Chem 2006, 52 (12):2265-2272. 33. Chala I, Feshchenko DV, Dubava OA, Bakhur TI, Zghosinska OA, Rusak VS: Changes in the lipid profi le of neutered cats’ blood in cases of obesity and diabetes. Vet Arh 2021, 91 (6):635-645. 34. Calder PC, Aqoob P: Understanding omega-3 polyunsaturated fatty acids. Postgrad Med 2009, 121(6):148-157. 35. Gülersoy E, Ekici Y: Assessment of hematological and serum biochemistry parameters in dogs with acute diarrhea due to different etiologies. Maced Vet Rev 2022, 45(2):149-156. PROFILI FOSFOLIPIDA MASNIH KISELINA PLAZME I MEMBRANE ERITROCITA KOD PASA HRANJENIH KOMERCIJALNOM GRANULIRANOM HRANOM Tamara POPOVIĆ, Jasmina DEBELJAK MARTAČIĆ, Biljana POKIMICA, Branko RAVIĆ, Slavica RANKOVIĆ, Maria GLIBETIĆ, Predrag STEPANOVIĆ Unos dugolančanih n-3 polinezasićenih masnih kiselina (PUFA) koristi zdravlju ljudi i životinja. Naša studija je imala za cilj da analizira sadržaj dugolančanog n-3 PUFA u dve vrste hrane i njihov uticaj na fosfolipide plazme i eritrocita belgijskih ovčara. Ukupno 10 pasa je hranjeno komercijalnom granuliranom hranom (hrana 1), a još 10 je obezbeđeno komercijalnom premijum granuliranom hranom visokog kvaliteta (hrana 2). Sve analize su obavljene gasno-tečnom hromatografi jom. Naši rezultati su pokazali da hrana 1 sadrži više n-3 PUFA nego hrana 2, što se odrazilo na više n-3 PUFA u plazmi i eritrocitnim fosfolipidima pasa iz grupe 1. Pošto su dugolančani n-3 131 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 PUFA u fosfolipidima prekursori za antioksidativne molekule, dalje studije bi trebalo da istraže efekte analizirane komercijalne granularne hrane bogate n-3 na parametre oksidativnog stresa kod pasa. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Acta veterinaria de Gruyter

Phospholipid Fatty Acid Profiles of Plasma and Erythrocyte Membranes in Dogs Fed with Commercial Granulated Food

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de Gruyter
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© 2023 Tamara Popović et al., published by Sciendo
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1820-7448
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1820-7448
DOI
10.2478/acve-2023-0009
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Abstract

Acta Veterinaria-Beograd 2023, 73 (1), 119-132 UDK: 636.7.085.3 DOI: 10.2478/acve-2023-0009 Research article PHOSPHOLIPID FATTY ACID PROFILES OF PLASMA AND ERYTHROCYTE MEMBRANES IN DOGS FED WITH COMMERCIAL GRANULATED FOOD # 1 #1 1 Tamara POPOVIĆ * , Jasmina DEBELJAK MARTAČIĆ , Biljana POKIMICA , 1 1 1 2 Branko RAVIĆ , Slavica RANKOVIĆ , Maria GLIBETIĆ , Predrag STEPANOVIĆ University of Belgrade, Institute for medical research, Laboratory for food and metabolism, Belgrade, Serbia; University of Belgrade, Faculty of Veterinary medicine, Associate Professor, Department of equine, small animal, poultry, and wild animal diseases, Belgrade, Serbia (Received 20 October 2022, Accepted 16 January 2023) Intake of long-chain n-3 polyunsaturated fatty acids (PUFA) benefi ts human and animal health. Our study aimed to analyze the long-chain n-3 PUFA content of two types of food and their effect on plasma and erythrocyte phospholipids of Belgian Shepherd dogs. A total of 10 dogs were fed commercial granulated food (Food 1), and another 10 were provided commercial Premium granulated food of high quality (Food 2). All the analyses were performed using gas-liquid chromatography. Our results showed that Food 1 contained more n-3 PUFA than Food 2, which was refl ected in higher n-3 PUFA in plasma and erythrocyte phospholipids. Because long- chain n-3 PUFA in phospholipids are precursors for antioxidative molecules, further studies should investigate the effects of the analyzed commercial granulated food rich in n-3 on oxidative stress parameters in dogs. Keywords: commercial foods, dogs, fatty acids profi les, n-3 fatty acids, phospholipids. INTRODUCTION In recent years, many pet owners have abandoned conventional, veterinary- recommended commercial diets in search of more “natural” and “homemade” choices [1]. Still, it is more common practice nowadays for owners to use commercialized labeled food for pets such as canines. Among other ingredients, polyunsaturated fatty acids (PUFA) are important for human and animal health. The conversion of short chain to long chain PUFA is rate-limiting and varies between species [2]. The balance of n-6/n-3 ratio in phospholipids, as well as the balance between reactive oxygen species and reactive nitric species on one side and antioxidative defense, on the other, is important for the normal physiological function of organisms. In dogs, fewer *Corresponding author: e-mail: poptam@gmail.com Authors have contributed equally to this work and, thus, share the fi rst authorship 119 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 infl ammatory mediators were produced when fed diets ratios (n-6/n-3) of 5:1 and 10:1 in comparison with being fed an n-6-rich diet with a fatty acid ratio of 100:1 [3]. Mammals are not able to synthesize fatty acids (FA) with double bonds at C-9, but are able to some extent to elongate and further saturate the aliphatic chain [4]. It is well known t hat diet could affect fatty acid profi les in plasma and erythrocyte membranes (EM) and that changing its composition causes changes in many parameters, which is confi rmed in animal and human studies [3]. Since the FA composition of the EM correlates with that of other cell membranes, the effect of dietary FA supplementation may be analyzed by studying EM. It is well known that plasma phospholipids profi les refl ect short-term changes in the diet while changes in erythrocyte membrane phospholipids refl ect long-term dietary habits (up to 3 months prior analysis). Comparing those two profi les could lead us to a conclusion about the diet habits of examined animals in general [5]. The FAs profi le of the serum phospholipids is related to the average dietary FAs intake during the last 3 to 6 weeks, while the composition of erythrocyte phospholipids depends on the dietary fat intake during the preceding months [5]. The FAs profi le in the tissues partly refl ects not only the dietary fat intake but also the effi ciency of FAs metabolism in the body [6]. The FAs profile of tissues and triglycerides (TG) is known to be influenced by many factors, including dietary intake, age, gender, and endogenous metabolism [7]. Considerable interest exists in the possible health benefits of increasing dietary intake of n-3 PUFA [8]. Three families of long-chain PUFAs with different biological roles exist, n-3, n-6 and n-9 and they are derived from the shortest non-synthesizable precursors: linoleic acid (LA) (18:2, n-6) and alpha-linolenic acid (ALA) (18:3, n-3). Humans can desaturate and elongate ALA, as a precursor of n-3 series, to eicosapentanoic acid (EPA) and docosahexanoic acid (DHA). This process is dependent on aging, presence and type of disease, inflammation processes, and other factors [4]. In rats, which are often used as animal models, the rate of conversion of ALA to DHA is high in the liver, although Δ5 and Δ6 desaturases are expressed in many other rodent tissues besides the liver [9]. There are few commercial pet dog foods with EPA and DHA concentrations adequate for the treatment of disease or some vulnerable conditions. Target ranges for EPA and DHA vary quite widely for different conditions but typically fall between 50 and 220 mg/kg body weight. Commercial diets with n-3 fatty acids typically provide less EPA and DHA than desirable and may be advertised as containing fl axseed or canola oil (rich in ALA) [10]. In fact, there are adverse effects associated with the use of n-3, and an increase in the concentration of EPA and DHA in commercial pet food (dogs) makes the topic important to revisit. Those effects include altered platelet function, gastrointestinal adverse effects, potential effects for nutrient excess, weight gain, altered immune function, and effects on glycemic control and insulin sensitivity. As far as dogs and specifi c abnormalities are concerned decreased epithelization of wounds after 5 120 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food days (n-6/n-3=0.3:1), increased plasma and urine thiobarbituric reactive substances (n-6/n-3=5.4:1), decreased plasma vitamin E (n-6/n-3=1.4:1), decreased skin and neutrophil leukotriene B4/increased leukotriene B5, lower delayed-type hypersensitivity response (n-6/n-3=1.4:1), decreased CD4+T lymphocyte count (n-6/n-3=1.4:1), decreased lymphocyte proliferation (EPA/DHA=0.8) [11]. Metabolically, however, fatty acid patterns of plasma phospholipid fractions again revealed a sparing effect of ALA on LA. It should be mentioned that a direct effect of ALA on improvements of skin and coat could not completely be ruled out in these studies while long-chain n-3 PUFAs from fi sh oil or other marine sources appear to be especially capable of modifying infl ammatory and immune responses [12]. The aim of the study was to analyze the fatty acid content of two different diets for dogs and to examine the effects of their daily consumption on the erythrocyte membrane and plasma phospholipid profi les. MATERIALS AND METHODS Animals This study which lasted for 12 weeks, was approved by the Ethics Commission of the Faculty of Veterinary Medicine are included dogs of the Belgian Shepherd (Malinoa) breed from two kennels (1 and 2). In kennels, we selected 10 dogs, (5 females and 5 males), age categories of 3 to 7 years, with body weight 30.2±2.2 kg. We measured weight gain monthly and it changed up to 1 kg/dog. Male dogs weight gain was slightly higher than females but without statistical difference. By the basic examination of the dogs in both kennels (blood pressure, pulse, temperature, breathing, skin condition, and skin cover), there was a constant absence of disease, otherwise, all dogs had neatly managed health cards. According to the constitution they fell into 3 categories, which means they have an ideal weight corresponding to this breed of dog. The dogs had their activities in the morning and in the evening for 60 minutes (walking, running), otherwise, these dogs are considered as working dogs. In kennel number 1, the dogs were fed commercial granular foods that normally satisfi es the standard nutritional needs of dogs (I). In kennel number 2, the dogs were fed Premium granulated food of high quality, this being the most sold dog food in Serbia (II). The amount of food (400 g/day/dog) was divided into two meals one in the morning and the other in the evening at the same time each day. Sample collection and analysis Granulated foods samples (I and II) (from four representative large markets) were analyzed. The primary sample was generated by mixing an equal portion of four samples taken from different markets. Five replicate samples of the composite sample were analyzed by standard laboratory methods to measure the concentration of 121 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 proteins, carbohydrates, lipids, minerals, and water. Fatty acid composition from the lipids was done by standard laboratory procedure, as described below. At the end of the study blood samples were taken from the (vena cephalica antebrachii), with the aid of EDTA vacuum blood collection tubes. Erythrocytes and plasma were separated and stored at a temperature of -80 C . After blood sampling by the routine method, analysis of erythrocytes and plasma was done by gas-liquid chromatography (GC). Nutritional analysis was carried out by an accredited chemical laboratory at the Institute of Public Health in Požarevac. Ash content was determined by the direct gravimetric method which includes ashing of the samples in an oven at 550 °C until a constant weight was attained. Moisture was determined gravimetrically [13]. Crude protein content was estimated based on the total nitrogen content of the sample determined by the Kjeldahl method (AOAC 955.04D) [13]. Crude fat content was determined gravimetrically (Soxhlet extraction, AOAC method [13]. Total carbohydrate content, crude “by difference”, was calculated by the following formula: total carbohydrate (%) = 100% - % (protein + ash + fat + moisture). The energy content of food was calculated based on determining content by the following formula: Energy value (estimated, kJ/100 g) = [4 x protein (%)] + [4 x carbohydrate (%)] + [9 x fat (%)]. Fatty acids extraction and analysis Isolation of lipids The method consists of homogenization of plasma with a 2:1 chloroform/methanol mixture. Washing of the mixture with a 5 times smaller volume of water or saline (0.9 g NaCl in 100 ml of water). The resulting mixture was separated into two phases. The lower phase was the total pure lipid extract. First the lipids present in the volume of 0.5 ml of erythrocytes were extracted with a mixture of chloroform isopropanol (7:11) following the prescribed procedure [14]. After that we isolated the phospholipids from the lipid subclasses by thin-layer chromatography on silica-gel plates using petroleum ether, diethyl ether and glacial acetic acid (87:12:1, by volume). In a modifi ed procedure by Christopherson and Glass [15], we made a direct transesterifi cation of fatty acids. Lipid extracts derived from hexane were evaporated under a nitrogen stream to full vapor and dry bottom of Eppendorf ’s tubes. After that, the residue was dissolved in 10µl hexane. We took 1 µl and injected it into a chromatograph. Methyl esters of fatty acids were analyzed by gas-liquid chromatography in a Shimadzu chromatograph GC 2014 (Kyoto, Japan) equipped with a fl ame ionization detector on an Rtx 2330 column (60mm x 0,25 mmID, fi lm thickness 0,2 µm, Restek, Bellefonte, PA, USA). Adequate separation was obtained over a 50 min period with an initial temperature of 140 C maintained for 5 0 0 min. The temperature was then increased to 220 C at a rate of 3 C/min and kept at the fi nal temperature for 20 min. The identifi cation of fatty acid methyl esters (FAME) 122 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food was made by comparing peak retention times with standard mixtures (PUFA-2 and/ or 37 FAMEs mix, Supelco, Bellefonte, PA, USA). Finally, the content of fatty acids, from 16:0 through 22:6n-3, was expressed as a percentage of the total content of identifi ed fatty acids. Statistical analysis Statistical analysis was done by one-way ANOVA test and Student t-test (signifi cance p<0.05) in SPSS. All results were expressed in percentages of a total of 100%. RESULTS Biochemical parameters Analyzed biochemical parameters such as triglycerides and plasma cholesterol did not show sta s cal signifi cance in the plasma of canines treated with two diff erent diets. Although there were some changes in LDL cholesterol, previously reported by Ravic et al [16]. Analyzed food There were differences in FA percentage between the examined foods. There was an equal percentage of palmitic (16:0) acid, but stearic acid was more abundant in Food 1 (p<0.01). Monosaturated FA, specially palmitoleic acid, was present at a higher percentage in Food 2 (p<0.01), while oleic acid (18:1 n-9) was detected almost in the same percentage. Linoleic acid (18:2 n-6) was also present in more than 10% in both Food 1 and Food 2. Long-chain PUFAs were present in a very low percentage with statistically signifi cant differences between Foods. ALA (18:3n3) was statistically signifi cant in a higher percentage in Food 1 (p<0.001), as well as EPA (p<0.001) and DHA (p<0.001) compared to Food 2 (Table 1). Table 1. Composition and FA patterns of the diets Fatty acids (%) Food 1 Food 2 16 :0 23.93± 0.15 23.13 ± 0.1 Palmitic acid 16 :1 2.47± 0.26 3.72 ± 0.07** Palmitoleic acid 18 :0 13.36± 0.26 9.26 ± 0.26** Stearic acid 18: 1,n 9 36.88± 0.55 40.61± 0.92 Oleic acid 18: 1,n 7 4.00± 0.21 3.46 ± 0.33 Vascenic acid 18: 2 15.13± 1.34 18.04 ± 0.26* Linoleic acid 123 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 cont. Table 1. 18: 3 n 3 2.99± 0.22 0.95 ± 0.02*** Linolenic acid 20: 3 0.07± 0.01 0.13± 0.02*** Eicosatrienoic acid 20: 4 0.45± 0.09 0.36± 0.12 Arachidonic acid 20: 5 0.22± 0.09 0.04 ± 0.02*** Eicosapentanoic acid 22 : 4 0.09± 0.01 0.13 ± 0.02** Docosatetranoic acid 22 : 5 0.07± 0.01 0.07 ± 0.01 Docosapentanoic acid 22: 6 0.31± 0.08 0.07 ± 0.01*** Docosahexanoic acid Proteins 27.29 (0.05) 25.51(0.04) Fats 16.44 (0.04) 15.22 (0.04) Carbohydrates 44.54 (0.22) 46.41 (0.24) Minerals 6.69 (0.01) 7.67 (0.01) Water 5.05 (0.02) 5.21 (0.03) Values (proteins, carbohydrates, fats, minerals and water) are expressed as % by 100g. Abbrrevations SD, Standard deviations. Fatty acids are expressed as % ± st. dev with signifi cance *p<0.05,**p<0.01,***p<0.001 Dog plasma and erythrocyte membrane phospholipid fatty acids It is well known that plasma phospholipids fatty acids profi les represent the short- term (up to several days) food intake, while erythrocytes present data composition of fatty acids for long term (last 3 months) food intake. In our study, we compare dogs’ FAs profi les in plasma and erythrocytes membrane phospholipids in two kennels (Table 2-4). In plasma phospholipids FA profi les in palmitic acid (16:0) were increased in Kennel 1(p<0.001) as well as stearic acid (18:0), results showed that dogs in Kennel 1 had a higher percentage of saturated fatty acids (SFA) in plasma phospholipids compared to Kennel 2. As far as monosaturated fatty acids, palmitoleic (16:1) was increased in Kennel 2 while oleic acid (18:1, n-9) was increased in Kennel 1 (p<0.01) compared to dogs fed Kennel 2. Linoleic acid (18:2) was increased (p<0.001) in dogs in Kennel 2 while polyunsaturated n-3 especially ALA (p<0.01) eicosapentaenoic (EPA) (p<0.001), docosapentaenoic (DPA) (p<0.001) and docosahexaenoic (DHA) (p<0.001) were increased in Kennel 1. In the erythrocyte membrane, phospholipids´ percentage of fatty acids has almost the same distribution and percentages as in plasma phospholipids between Kennels. Saturated fatty acids (PA and SA) (p<0.01) were in higher percentages in Kennel 1-fed dogs as well as monounsaturated FA (p<0.1). Percentage of EPA (p<0.001), DPA (p<0.01), and DHA (p<0.001) were increased in Kennel 1 in plasma phospholipids, 124 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food while ALA concentration did not signifi cantly change. These results go along with the plasma phospholipids FA distribution already mentioned. These results suggest that the differences are negligible between plasma and erythrocytes phospholipids fatty acids percentages. Ta ble 2. Plasma and erythrocyte phospholipids fatty acids profi les of dogs in Kennel 1 and 2. Fatty Acids (%) (plasma) Kennel 1 Kennel 2 16 :0 17.09 ± 1.29 14.55 ± 1.48 *** 16 :1 0.39 ± 0.12 0.68 ± 0.33 *** 18 :0 30.12 ± 1.50 29.81 ± 1.81 18: 1 ,n 9 5.76 ± 0.67 4.89 ± 0.45 ** 18: 1 ,n 7 1.96 ± 0.14 1.69 ± 0.23 * 18: 2 16.63 ± 2.12 24.52 ± 1.73 *** 18: 3 n 6 0.34 ± 0.06 0.40 ± 0.04 * 18:3 n 3 0.23 ± 0.01 0.15 ± 0.07 ** 20: 3 1.27 ± 0.11 1.86 ± 0.51 ** 20: 4 17.86 ± 2.27 17.71 ± 2.21 20: 5 2.26 ± 1.18 0.24 ± 0.03 *** 22 : 4 0.43 ± 0.01 0.82 ± 0.02 *** 22 : 5 2.38 ± 0.88 1.91 ± 0.51 *** 22: 6 3.00 ± 0.78 0.49 ± 0.02 *** Fatty Acids (%) (erythrocytes) Kennel 1 Kennel 2 16 :0 19.2 ± 0.69 17.15 ± 1.53 ** 16 :1 0.24 ± 0.038 0.22 ± 0.012 18 :0 30.20 ± 0.78 29.49 ± 1.68 18: 1 ,n 9 7.87 ± 0.24 7.09 ± 0.53 * 18: 1 ,n 7 2.03 ± 0.11 1.83 ± 0.12 * 18: 2 9.31 ± 1.04 13.82 ± 0.37 *** 18: 3 n 6 0.45 ± 0.06 0.42 ± 0.05 18:3 n 3 0.17 ± 0.04 0.20 ± 0.02 20: 3 2.51 ± 0.25 2.78 ± 0.35 20: 4 24.0 ± 0,96 23.20 ± 2.44 20: 5 1.48 ± 0.26 0.23 ± 0.04*** 22 : 4 0.96 ± 0.23 1.64 ± 0.22 *** 22 : 5 ± 0.22 0.73 ± 0.18 ** 22: 6 1.08 ± 0.21 0.23 ± 0.11 *** 125 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 Table 3. Estimated desaturase 9,6,5 activities in plasma and erythrocytes in dogs in Kennel 1 and 2. Estimated desaturase Kennel 1 Kennel 2 Kennel 1 Kennel 2 (D) activity plasma plasma erythrocytes erythrocytes D9 desaturase 0.19 ± 0.03 0.17 ± 0.02 0.25 ± 0.01*** 0.24 ± 0.02***# D6 desaturase 6.01 ± 0.20 5.04 ± 1.50 15.65 ± 5.42*** 13.43 ± 2.63 D5 desaturase 0.14 ± 0.07 0.01 ± 0.005### 0.06 ± 0.01*** 0.01 ± 0.003### Test signifi cance were used between groups, *statistical signifi cance between plasma and erythrocytes in both Kennels, # statistical signifi cance between plasma in both Kennels and erythrocytes in both Kennels, with *#p<005, **##p<0.01 and ***###p<0.001 Table 4. Percentage of overall n-3, n-6 and n-6/n-3 ratio in Kennel 1 and Kennel 2 in plasma and erythrocytes of examined dogs. Plasma K1 Plasma K2 Erythrocytes K1 Erythrocytes K2 N-3 7.18 ± 0.32 5.95 ±0.55 2.43±0.40 2.52 ± 0.33 N-6 36.50±1.33 33.23 ±0.11 35.48±1.42 34.52 ± 0.88 N6/N3 5.07±1.00 6.05 ±0.95 15.00 ± 3.64 16.7±0.43 K1: Kennel 1; K2: Kennel 2. DISCUSSION Generation of SFA, MUFA and PUFA fatty acids is connected with mediators such as prostaglandins, leukotrienes and others, which are able to infl uence metabolic changes in dogs in health and disease. The n-6/n-3 balance is important in everyday balanced diet and is recommended to be as low as it could be. Following up on intakes of linoleic acid, as n-6 family fatty acid, and its metabolic pathway to DGLA and arachidonic acid (AA) and cell membrane incorporation is of great importance as information of biochemical or clinical parameters for the evaluation of dog´s health [5]. Omega-6 linoleic and its transformation to ARA affect not only from the cell structural point but also affect the membrane response to stimuli, thus membrane fatty acid composition can be useful as information of pro and anti-infl ammatory predisposition of the canine organism [17]. By correlating plasma and erythrocyte phospholipids FA composition after the treatment, we investigated if FA from both plasma and erythrocytes could represent markers of dietary n-3 intake. Our discussion is more based on n-3 PUFA signifi cance and intake in canines. Certain results indicate a benefi cial effect of n-3 long-chain polyunsaturated fatty acids (LC PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on canine health. Several studies have examined the effects of fi sh oil, linseed oil, or combinations on plasma phospholipids fatty acids profi le in canine species [18], lymphocyte proliferation, or neutrophil composition in laboratory animals [19,20]. 126 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food The nutrient requirements of canine athletes who have a greater capacity for fat oxidation are unique concerning that canine metabolism is unique [10]. Relative to body mass size dogs metabolize free fatty acids at twice the rate observed in men. Consequently, the muscles are more adapted to use fat than human muscles [21]. Usual daily food requirements are water, energy, major nutrients, fi bers, minerals, and vitamins. High fat diet, fatty acids chain length and saturation affect a variety of issues from infl ammation to oxidative stress in animals [22,23]. However, there is very little information regarding optimal dietary fats intake for canine athletes [23]. Medium-chain triglycerides yield 8 carbon to 12 carbon-free fatty acids (coconut, palm oil) when digested and are directly transported through absorption into the blood, bound to albumin, to the liver via the portal circulation, leading to sparing of glycogen [24]. The fatty acid composition also infl uences detection in scent-trained dogs, hunting dogs, and service dogs. It is well known that polyunsaturated fatty acids may modestly improve performance in dogs that require activity as a part of their work [25]. The use of carbohydrates as a major dietary substrate is essential. Soluble fi bers may alter the large intestinal microfl ora which produces short-chain fatty acids. A small amount of soy fi ber and fructooligosaccharides are components of gastrointestinal veterinary diets. Defi ciencies in major minerals have been observed in dogs fed with non-traditional diets. It is recommended that bones should be ground into the diet to improve calcium and phosphorus balance. Vitamins (fat-soluble or water-soluble) are involved in cellular metabolism or coenzymes in the citric acid cycle. A study by Poel at al (2017) [26] was conduc ted to determine the stability of two preparations of GAA (granulated and crystallized) and CMH in a moist and a dry dog food formulation during production and storage. Most commercial dog foods contain vitamins above the minimum requirement. Diets high in PUFA (fi sh-based sled dog diets) should contain more vitamin E to prevent lipid peroxidation. In the study by Vessecchi et al., the study aimed to evaluate the macronutrient composition, fatty acids and amino acids profi le, and essential minerals contents of vegan pet foods available in the Brazilian market, and to assess their compliance with recommended allowances for dogs and cats [27]. Omega -3 fatty acids incorporat ed in cell membranes, especially EPA and DHA, decrease clinical signs of osteoarthritis in dogs. EPA serves as a substrate for the COX and LOX enzymes. Omega-6 FA are involved in physiological processes while infl ammation contributes to the formation of “proinfl ammatory” prostaglandins and leukotrienes. Omega-3 produces less infl ammatory prostanoids and 5-leukotrienes. Dogs have limited ability to convert ALA to DHA. That is the reason for diet EPA and DHA consumption directly. Diets that contain EPA and DHA have been recommended for degenerative joint diseases, aging in general, as anti-infl ammatory supplements, and in growing puppies [25]. Currently, omega-3 FAs are used in managing many diseases including neoplasia, dermatologic diseases, hyperlipidemia, cardiovascular, gastrointestinal, and orthopedic diseases [11]. There are few commercial pet foods with EPA and DHA adequate for the treatment of disease. Joint diets, renal diets 127 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 dermatologic conditions contain more omega-3 fatty acids than maintenance diets. Commercial diets with omega-3 provide less EPA and DHA than desirable and contain fl axseed instead. Because diets with ALA have different effects when compared with diets enriched in EPA and DHA composition information may very much contribute to results. Purushotman et al. study on beagles fed with fl axseed added to a basal diet (57% alfa-linolenic acid, ALA at the rate 100ml/kg for 3 weeks) showed that plasma ALA, EPA, and LA increased steadily and signifi cantly from 0-22 days while arachidonic acid showed no difference. Plasma DHA showed no signifi cant changes over time which agrees with previous studies in dogs [2]. Dunbar et al., concluded that hepatic conversion of DHA is slow in the canine and after its production from ALA it is likely to relocate to neurological tissue [29]. Extensive studies are required for clarifying this issue and to confi rm how different breeds metabolize PUFA. Waldron et al. confi rm earlier reports in their study on dogs fed a fi sh oil diet that ALA was converted to EPA and further elongated to DPA but was not converted to DHA in plasma phospholipids [18]. The benefi cial effect of n-3 FA due to their competition with n-6 for cellular membrane incorporation leads to the suggestion that n-6: n-3 ratio could be used as a dietary index to modulate cell composition and cellular function [30]. Still, the type and amount of n-3 PUFA, but not the n-6:n-3 ratio in diets, contribute to membrane n-3’s highly unsaturated fatty acid composition and further changes in cell function [18]. Stoeckel et al. (2011) concluded that in dogs an increase of dietary n-3 FA content leads to a rapid inclusion of n-3 into the erythrocyte membrane, regardless of whether the n-3 FA is offered as an enriched diet or as a normal diet supplemented with an n-3 FA additive [31]. The cardiovascular benefi ts of n-3 PUFA could originate from their ability to improve lipid metabolism and reduce the synthesis of proinfl ammatory eicosanoids derived from n-6 PUFA. At an adequate level of incorporation, EPA and DHA infl uence the membrane fl uidity as well as membrane protein-mediated reactions, generation of lipid-mediators, cell signaling, and gene expression in different cells [32]. Data on cats, together with the results of lipoprotein analysis, indicate possible disturbances in the hepatic transformation of LDL and VLDL, and a high risk of atherogenic events [33]. Dogs have the capacity to metabolize n-3 fatty acids and the effects of omega-3 fatty acids on the skin and coat, infl ammatory responses, and neurologic development in puppies are quite visible [34]. Concerning specifi c benefi cial effects that fi sh oil or EPA and DHA have on canines, it would be useful to add it to the examined food or as an everyday supplement to those dogs. Diagnostic tests performed in dogs provide valuable information [35] and our data can contribute to it. Our further examination and studies will address it, and we planned a study on the treatment of PUFA in canines (EPA +DHA). That study could be useful and give us a more complete conclusion about the signifi cance of EPA and DHA in canine feeding. 128 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food Acknowledgments This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, Contract 451-03-9/2021-14/200015. Ethical approval The research related to the use of animals has been complied with all the relevant national regulations and institutional policies for the care and use of animals (Ministry of Agriculture and Forestry and Water Management - Veterinary Administration, number 323-07-00364/2017-05/6 date 13. 07. 2017., and approved by the Ethical comity of the Faculty of Veterinary Medicine, 17 June 2017 number 01-592 ). Authors’ contributions TP was involved in concept and design, drafting, and supervision. JDM, BP, and SR participated methodology and data collection. BR was involved in methodology, analysis, and interpretation of data. MG participated as project administration and funding acquisition. PS was involved in conceptualization funding acquisition. Declaration of confl icting interests The authors declare that they have no confl ict of interest with respect to the research, authorship, and/or publication of this article. Statement of Informed Consent The owner understood procedure and agrees that results related to investigation or treatment of their companion animals, could be published in Scientifi c Journal Acta Veterinaria-Beograd. REFERENCES 1. Schlesinger DP, Joffe DJ: Raw food diets in companion animals: a critical review, Can Vet J 2011, 52:50-54. 2. Purushotman D, Brown WY, Wu SB, Vanselow B: Evaluation of breed effects on n-3 PUFA metabolism with dietary fl axseed oil supplementation in dogs. Br J Nutr 2011, 106:139-141. 3. Avramovic N, Dragutinovic V, Krstic D, Colovic M, Trbovic A, De Luka S, Milovanovic I, Popovic T: The effects of omega 3 fatty acid supplementation on brain tissue oxidative status in aged Wistar rats. Hippokratia 2012, 16: 241-245. 4. Singer P, Jaeger, Voigt S, Thiel H: Defective desaturation and elongation of n-6 and n-3 fatty acids in hypertensive patients. Prostaglandins Leukot Med 1984, 15:159-165. 5. Popović T, Borozan S, Arsić A, Martačić JD, Vučić V, Trbović A, Mandić LJ, Glibetić M: Fish oil supplementation improved liver phospholipids fatty acid composition and parameters of oxidative stress in male Wistar rats. J. Anim Physiol Anim Nutr (Berlin) 2012, 96:1020-1029. 129 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 6. Ferrucci L, Cherubini A, Bandinelli S: Relationship of plasma polyunsaturated fatty acids to circulating inflammatory markers. J Clin Endocrinol Metab 2006, 91:439-446. 7. Oguzhan B, Sancho V, Acitores A, Villanueva-Peñacarrillo ML, Portois L, Chardigny JM, Sener A, Poel van der YA, Braun AU, Hendrix W, Bosch G: Stability of creatine monohydrate and guanidino acetic acid during manufacture (retorting and extrusion) and storage of dog foods. J Anim Physiol Anim Nutr 2019, 103:1242-1250. 8. Hagopian K, Weber KL, Hwee DT, Van Eenennaam AL, López-Lluch G, Villalba JM, Burón I, Navas P, German JB, Watkins SM, Chen Y, Wei A, McDonald RB, Ramsey JJ: Complex I-associated hydrogen peroxide production is decreased and electron transport chain enzyme activities are altered in n-3 enriched fat-1 mice. PLoS One 2010, 5:e12696. 9. Igarashi MF, Gao HW, Kim KMA, Bell JM, Rapoport SI: Dietary n-6 PUFA deprivation for 15 weeks reduces arachidonic acid concentrations while increasing n-3 PUFA concentrations in organs of post-weaning male rats. Biochim Biophys Acta 2009, 1791:132-139. 10. Hill R: The nutritional requirements of exercising dogs. J Nutr 1998, 128:2686-2690. 11. Lenox CE: Potential adverse effect of omega-3 fatty acids in dogs and cats. J Vet Intern Med 2013, 27:217-226. 12. Bauer JE: Responses of dogs to dietary omega-3 fatty acids. J Am Vet Med Assoc 2007, 231:1657-1661. 13. AOAC: Offi cial method of analysis: association of analytical chemists. 19th Edition, Washington DC; 2012, 121-130. 14. Rose HG, Oklander M: Improved procedure for the extraction of lipids from human erythrocytes. J Lipid Res 1965, 6:428-431. 15. Christopherson SW, Glass RLJ: Preparation of milk fat methyl esters by alcoholysis in an essentially nonalcoholic solution. Dairy Sci 1969, 52:1289-1290. 16. Ravić B, Debeljak-Martacić J, Pokimica B, Vidović N, Ranković S, Glibetić M, Stepanović P, Popović T: The effect of fi sh oil-based foods on lipid and oxidative status parameters in police dogs. Biomolecules 2022, 12(8), 1092. 17. Domingez T, Kizanpreat K, Burri L: Enhanced omega3 index after long-verus short chain omega 3 fatty acids supplementation in dogs. Vet Med Sci 2021, 7(2):370-377. 18. Waldron M, Hannah S, Bauer J: Plasma phospholipid fatty acid and ex vivo neutrophil responses are differencially altered in dogs fed fi sh and linseed-oil containing diets at the same n-6:n-3 fatty acid ratio. Lipids 2012, 47:425-434. 19. Varning K, Schmidt EB, Svaneborg N, Moller JM, Lervang HH, Grunnet N, Jersild C, Dyeberg J: The effect of n-3 fatty acid on neutrophil chemiluminescence. Scand J Clin Lab Invest 1995, 55:47-52. 20. Sperling RI: Effects of dietary fi sh oil on leukocyte leukotriene production and PAF generation and on neutrophil chemotaxis. World Rev Nutr Diet 1991, 66:391-400. 21. De Bruijne JJ, Altszuler N, Hampshire J, Visser TJ, Hackeng WH: Fat mobilization and plasma hormone levels in fasted dogs. Metabolism 1981, 30:190-194. 22. Emara E, El-Sayyad H, El-Ghaweet H: Bovine whey supplementation in a high-fat diet fed rats alleviated offspring’s cardiac injury. Maced Vet Rev 2022, 45:89-99. 23. Wakshlang J, Shmalberg J: Nutrition for working and service dogs. Vet Clin Small Anim 2014, 44:719-740. 24. Jeukendrup AE, Aldred S: Fat supplementation health and endurance performance. Nutrition 2004, 20:678-688. 130 Popović et al.: Phospholipid fatty acid profi les of plasma and erythrocyte membranes in dogs fed with commercial granulated food 25. Angle TC, Wakshlag JJ, Gillette RS: The effects of exercise and diet on olfaction in detection dogs. J Nutr Sci 2014, 44:1-5. 26. Esther AHP, Denmark FO, Guido B, Wouter, HH, Antonious FB van der Poel: Retorting conditions affect palatability and physical characteristics of canned cat food. JNS 2017,6(e23):1-5. 27. Vessecchi R, Zafalon A, Risolia LW, Henrique T, Vendramini A, Rodrigues RBA, Pedrinelli V, Teixeira FA, Rentas MF, Perini MP, Alvarenga IC, Brunetto MA: Nutrional inadequacies in commercial vegan foods for dogs and cats. Plos ONE 2020, 15 (1):1-17. 28. Wandel S, Juni P, Tendal B: Effects of glucosamine, chondroitin or placebo in patients with osteoarthritis of the hip or knee: network meta-analysis. BMJ 2010, 341:4675. 29. Dunbar BL, Bigley KE, Bauer JE: Early and sustained enrichment of serum n-3 long chain polyunsaturated fatty acids in dogs fed a fl axseed supplemented diet. Lipids 2010, 45:1-10. 30. Harris, WS: The omega 6-omega 3 ratio and cardiovascular disease risk:uses and abuses. Curr Athero Rep 2006, 8:453-459. 31. Stoeckel K, Nielsen LH, Fuhrmann H, Bachmann L: Fatty acid patterns of dog erythrocyte membranes after feeding of a fi sh-oil based DHA-rich supplement with a base diet low in n-3 fatty acids versus a diet containing added n-3 fatty acids. Acta Vet Scand 2011, 24:53-57. 32. Cao J, Schwichtenberg KA, Hanson NQ, Tsai MY: Incorporation and clearance of omega-3 fatty acids in erythrocyte membranes and plasma phospholipids. Clin Chem 2006, 52 (12):2265-2272. 33. Chala I, Feshchenko DV, Dubava OA, Bakhur TI, Zghosinska OA, Rusak VS: Changes in the lipid profi le of neutered cats’ blood in cases of obesity and diabetes. Vet Arh 2021, 91 (6):635-645. 34. Calder PC, Aqoob P: Understanding omega-3 polyunsaturated fatty acids. Postgrad Med 2009, 121(6):148-157. 35. Gülersoy E, Ekici Y: Assessment of hematological and serum biochemistry parameters in dogs with acute diarrhea due to different etiologies. Maced Vet Rev 2022, 45(2):149-156. PROFILI FOSFOLIPIDA MASNIH KISELINA PLAZME I MEMBRANE ERITROCITA KOD PASA HRANJENIH KOMERCIJALNOM GRANULIRANOM HRANOM Tamara POPOVIĆ, Jasmina DEBELJAK MARTAČIĆ, Biljana POKIMICA, Branko RAVIĆ, Slavica RANKOVIĆ, Maria GLIBETIĆ, Predrag STEPANOVIĆ Unos dugolančanih n-3 polinezasićenih masnih kiselina (PUFA) koristi zdravlju ljudi i životinja. Naša studija je imala za cilj da analizira sadržaj dugolančanog n-3 PUFA u dve vrste hrane i njihov uticaj na fosfolipide plazme i eritrocita belgijskih ovčara. Ukupno 10 pasa je hranjeno komercijalnom granuliranom hranom (hrana 1), a još 10 je obezbeđeno komercijalnom premijum granuliranom hranom visokog kvaliteta (hrana 2). Sve analize su obavljene gasno-tečnom hromatografi jom. Naši rezultati su pokazali da hrana 1 sadrži više n-3 PUFA nego hrana 2, što se odrazilo na više n-3 PUFA u plazmi i eritrocitnim fosfolipidima pasa iz grupe 1. Pošto su dugolančani n-3 131 Acta Veterinaria-Beograd 2023, 73 (1), 119-132 PUFA u fosfolipidima prekursori za antioksidativne molekule, dalje studije bi trebalo da istraže efekte analizirane komercijalne granularne hrane bogate n-3 na parametre oksidativnog stresa kod pasa.

Journal

Acta veterinariade Gruyter

Published: Mar 1, 2023

Keywords: commercial foods; dogs; fatty acids profiles; n-3 fatty acids; phospholipids

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