1. Introduction
The performance of livestock is influenced by a variety of factors, including the type of production systems used, breed, age, sex, nutritional status, hormonal status, and environment [1]. The health and performance of animals are related to their nutritional status, which is reflected in metabolic products in the blood [2,3]. At the beginning of their lives, goat kids are monogastric animals or non-functional ruminants that become functional ruminants at around two months of age. Goat blood parameters can be influenced by the weaning processes and dynamics as well as the development of the liver, rumen, and immune system during the transition from pre-ruminant to ruminant [4]. It can also be a tool to check the quality of the feed, e.g., nutrient availability, mineral status, digestibility, and absorption by the animals [5]. Abdelsattar et al. [3] reported that the age of goats has a great influence on their blood profile, especially around the weaning. Thus, goat kids are very sensitive, especially when there are changes in diet, such as switching to a diet containing voluminous feeds and cereals with a gradual reduction in milk protein [6]. Goat kids are mostly known and valued for their meat, which is highly appreciated in Mediterranean Europe [7,8]. The meat of goat kids has a high nutritional value since it is rich in proteins and low in fat, with a significant proportion of beneficial fatty acids [9,10]. These attributes are what provide goat meat its potential for use in human diets and overall health. Furthermore, goat farming requires less input, due to natural adaptation to free-range farming, and yields lean red meat that is a healthier alternative [11]. These factors make goat kids’ meat a potentially sustainable supply of red meat.
Most of the livestock production systems worldwide are based on unsustainable feeding sources to maintain the need for proteins in animal nutrition, the most common of which is soybean meal [12]. Soybeans are an excellent source of protein and are often used in commercial feed rations to increase the crude protein content of the diet. As soybeans are in high demand, their increased cultivation and production as a crop are often associated with negative environmental impacts and increased use of natural resources, e.g., deforestation and soil depletion [13], a loss of biodiversity and natural habitats, long transport distances, high tillage, and fertilizer requirements [14]. Furthermore, using soybeans is costly in today’s global economy. Due to current feed imports, food production is no longer sustainable in many countries as the cost of food production rises [15]. In order to maintain a lower-cost protein supply, it is necessary to maintain the use of various alternative protein sources for animal nutrition. As reported by Klir et al. [16], soybeans, both extruded and as meal, can be completely replaced by pumpkin seed cake (PC, Cucurbita pepo L.) in the diet of lactating dairy goats due to the very high content of crude proteins and crude fats. Boldea et al. [17] included PC in the diet for lactating dairy goats and improved the fatty acids in their milk. Antunović et al. [18] found that PC is a high-quality feed that can partially replace soybean meal and is practicable in terms of good energy and protein balance in lamb’s serum, while lowering the serum NEFAs and BHB of lambs fed with 10% and 15% of PC in organic farming. In addition, Li et al. [19] completely replaced soybean meal with PC and dried distillers’ grains with a soluble mixture in dairy cows and concluded that this diet promoted antioxidant functions in dairy cows. Pumpkin is considered an agro-industrial by-product and sometimes industrial waste which has a very high potential as a nutraceutical [20] and has medicinal and pharmacological properties [21]. In addition, pumpkin is cultivated according to organic principles [22], so that it can be used as high-quality feed in organic animal farming. Moreover, PC, as feed for ruminants, is one of the local and low-cost feeds that meets the requirements of both farmers and customers [17].
On the other hand, extruded flaxseed (FS, Linum usitatissimum L.) has been used for over twenty years to enrich animal products with n-3 polyunsaturated fatty acids (PUFAs) in ruminant diets. Colonna et al. [23] reported that the saturated fatty acids in meat decreased, while monounsaturated fatty acids, PUFAs, and conjugated linoleic acid increased when goat kids were fed diets containing 3% flaxseed. Hao et al. [24] indicated that soybean meal can be partially replaced by flaxseed meal in the diets of fattening lambs in an optimal proportion of 12%, and increased the average daily weight gain. Ababakri et al. [25] observed increased serum cholesterol levels in ewes fed 10% FS in feed mixtures, while in the research by Nudda et al. [26], kidney and liver function parameters did not differ in the serum of dairy goats fed 180 g/day of FS in the diet. Alves Dutra et al. [27] reported that flaxseed added to the diet of Alpine goats affected the metabolic profile of the blood, with values still within the physiological interval, except for triglycerides. However, the available literature lacks information on the use of FS and PC in the diet of goat kids during their growth. As far as we know, there is no study on the influence of PC in the diet of goat kids on metabolic status. The clarification of these questions is of considerable scientific interest in the search for feed called nutraceuticals that can reduce the soybean content in the diet and at the same time improve the health status of growing goat kids on a small-scale goat farm.
The hypothesis here was that FS and PC added in the feed mixture as alternative protein sources have no adverse effects on the blood parameters and goat kids’ growth, which would be the novelty of the study. To test this hypothesis, the objective of this study was to evaluate two different feeding strategies for the inclusion of pumpkin seed cake and extruded flaxseed in the diet of Alpine goat kids and to research their effects on average daily weight gain, hematological parameters, and biochemical serum parameters (energy, protein and mineral status, and enzyme activities) during goat kids’ growth period.
2. Materials and Methods
This trial was carried out within the regulations of the Animal Protection Act of Croatia (NN 102/17, NN 32/19) and the Regulation on the Protection of Animals used for Scientific Purposes (NN 55/13, Declaration of Helsinki) and other relevant acts determining the welfare of farm animals as approved by the Bioethical Committee for the Animal Research of the Faculty of Agrobiotechnical Sciences Osijek (2158-94-02-24-18, 19 June 2024).
2.1. Animals and Management
At the small-scale goat farm in the Republic of Croatia, 31 French Alpine goat kids were used in the research. The experimental farm was located in the Slavonia part of Croatia (Marjančaci, Osijek-Baranja County, Croatia), characterized by a farm household system, using mainly family labor and using part of the products for family consumption and part for commercial uses (dairy and meat products). The goat kids were reared in a semi-intensive farming system, kept together with goats, until the age of two months, including a one-month weaning period. Each goat kid had given birth within seven days. The design of the experimental setup has been explained in Table 1. The experiment lasted for 55 days (from 32 to 87 days of kids’ age), and measurements were taken during growth: I—at the end of the suckling period (32 ± 3 days old); II—at the end of the weaning period (60 ± 3 days old); and III—in the post-weaning period (87 ± 3 days old). The following average body weights of goat kids were determined: 7.9 ± 1.5 kg, 12.8 ± 1.5 kg, and 16.2 ± 1.6 kg in the suckling, weaning, and post-weaning periods, respectively. The average daily weight gain (ADWG) of goat kids was determined as the difference between two consecutive weights from the suckling to the weaning period (32nd–60th day) and from the weaning to the post-weaning period (60th–87th day).
Following kidding, the goat kids suckled colostrum; however, all goat kids were housed with goats until they were about 32 days old and suckled milk ad libitum. After 32 days of age, the goat kids were removed from their mothers before the evening milking and placed back in a pen with the goats after the morning milking. After the first sampling at 32 days of age, the goat kids were offered feed mixtures and a hay mixture of red clover and grass (Lolium multiflorium and Phleum pratense) ad libitum in an approximate ratio of 50:50 (feed mixture/hay). At the same time, suckling was restricted and only allowed throughout the day between morning and evening milking. The goat kids were completely weaned after 60 days and given feed mixtures and hay ad libitum. The average daily intake of feed mixture by goat kids was ~200 g/day/head during the post-weaning period.
The feed composition used in the study was consistent with that used by Klir et al. [16] and Klir Šalavardić et al. [28], since this trial is a part of wider research carried out with goats and their goat kids. The feed mixtures differed concerning the sources of protein and fat: the control feed mixture with soybean meal and extruded soybean; the feed mixture with 16% pumpkin seed cake (PC-16), which completely replaced soybean; and the feed mixture with 9% extruded flaxseeds (FS-9), which partially replaced soybean. In order to replace soybean as much as possible with PC and FS, without disturbing the metabolism of goat kids, firstly, a study of the chemical composition of PC and FS was carried out (Section 2.2). Then, PC and FS, together with other ingredients, were added in amounts that would balance the feed mixtures to obtain a norm for growing goat kids in terms of protein, fat, and energy content according to the National Research Council [29] (Table 2). It turned out that 16% PC and 9% FS were ideal for the feed mixtures, whose chemical composition was still within the prescribed norms. The chemical composition of the milk consumed by the kids was 3.43, 3.81, and 4.05% milk fat; 3.01, 2.98, and 3.22% protein; and 4.47, 4.40, and 4.29% lactose in goat’s milk in CON, PC-16, and FS-9, respectively [30].
2.2. Feed Analyses
Standard methods were used to determine the composition of feed [31]. Using the steam distillation unit for Kjeldahl nitrogen (Behr Labor-Technik GmbH, Düsseldorf, Germany) and the Kjeldahl method, crude protein concentrations were determined on the basis of the nitrogen content. The Universal Extractions System B-811 (Büchi, Flawil, Switzerland) was used to analyze crude fat concentrations. According to Menke et al. [32], the metabolic energy (ME, MJ/kg DM) of the feed samples was determined from the gas generation during a 24 h in vitro incubation period using the Hohenheim gas test. An inductively coupled plasma mass spectrometer (ICP-MS, Agilent 7500a, Agilent Technologies Inc., Santa Clara, CA, USA) was used to measure the concentrations of mineral elements (Ca, P, Mg, and Fe) in solutions containing digested plant materials. Gas chromatography was used to determine the fatty acid content of food according to the approach of the State Office for Agricultural Chemistry Baden-Württemberg (LaChemie P23-5-008, V. 01).
2.3. Blood Sampling and Analyses
Blood was collected from all animals by jugular venipuncture between 0700 and 0800 h into 10 mL vacuum tubes (Vacutube®, LT Burnik, Vodice, Slovenia) by the same trained professional at goat kids’ ages of 32, 60, and 87 days. The blood was sampled in the morning and completed in 1 min to avoid excessive stress. Within an hour of blood collection on each day, a tube from each animal was brought to the Central Agrobiotechnical Analytical Unit (Faculty of Agrobiotechnical Sciences, Osijek, Croatia) for hematology analyses and differential blood counts.
For hematology analysis, the blood of goat kids was sampled into the sterile vacuum tubes containing ethylenediaminetetraacetic acid (EDTA) as the anticoagulant. Before analysis, whole blood was mixed with the Coulter mixer (Coulter Electronics Ltd., Luton Bedfordshire, UK). In whole blood, within 2 h after sampling, the following hematological parameters were determined: the number of leukocytes, the number of erythrocytes, hemoglobin, and hematocrit (WBC, RBC, HGB, and HCT, respectively). The following RBC indices were examined: mean corpuscular volume, average hemoglobin content in erythrocytes, and mean hemoglobin concentration in erythrocytes (MCV, MCH, and MCHC, respectively). The hematological parameters were analyzed on an automatic three differential hematology analyzer (Sysmex PocH-100Iv, Sysmex Europe GmbH, Hamburg, Germany). Blood samples from EDTA tubes were collected to make blood smears on glass slides. The smears were stained according to the Pappenheim method. White blood cells, such as neutrophils (NEUTs) and lymphocytes (LYMs), were identified using a compound microscope (BX53, Olympus Corporation, Tokyo, Japan).
Serum was prepared from the remaining blood tubes using standard procedures. The serum for each animal was aliquoted into 2 mL vials within 2 h of blood collection, transported in dry ice, and stored at −80 °C until required. Blood serum biochemical parameters were determined, including concentrations of minerals (calcium, phosphorus-inorganic, magnesium, and iron) and concentrations of urea, glucose, total proteins, and albumin, as well as cholesterol, high density lipoprotein, low density lipoprotein, triglyceride, β-hydroxybutyrate, and non-esterified fatty acids (CHOL, HDL, LDL, TGCs, BHB, and NEFAs, respectively). In addition, the following enzymes activities were determined: alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl-transferase (ALT, AST, and GGT, respectively). Globulin was calculated as the difference between total protein and albumin. Analyses were obtained by Beckman Coulter analyzer (AU400, Brea, CA, USA). Superoxide dismutase activity was measured by the degree to which the xanthine oxidase and superoxide radicals inhibited the following reaction (Randox Laboratories, Crumlin, UK):
O2• + O2• + 2H SOD → O2 + H2O2
The glutathione peroxidase enzyme catalyzes the oxidation of glutathione by cumene hydroperoxide (Randox Laboratories, Crumlin, UK):
2GSH + ROOH GPx → ROH + GSSG + H2O
2.4. Statistical Analyses
Mean values for average daily weight gain and blood parameters were obtained by Proc MEANS for each parameter within each dietary treatment, during different growth periods. The statistical model of these analyses included the fixed effect of diet as a “between-subject factor” and goat kid’s growth effect as a “within-subject factor”. In the first step, a one-way Proc ANOVA was used to analyze the effect of dietary treatments for each growth period with the following model: Yij = μ + di + eij, where μ = overall mean, di = the fixed effect of diet (three treatments: i = CON, FS-9, and PC-16), and eij = residual error. In the second step, a repeated measures Proc ANOVA was used to analyze the effect of the growth period on blood parameters for each dietary treatment with the following model: Yij = μ + gi + eij, where μ = overall mean, gi = the fixed effect of the growth period (three periods: i = suckling, weaning, and post-weaning), and eij = residual error. Comparisons of mean values among dietary treatments and growth periods were performed using Tukey’s tests, while significant differences were declared at p < 0.05 and trends were declared at 0.05 < p ≤ 0.1. Standard errors of the mean were reported. Statistical analysis was performed using the statistical software SAS 9.4 [33].
3. Results
3.1. Average Daily Weight Gain
Average daily weight gain is presented in Figure 1 from the suckling to weaning period (32nd–60th day) and from the weaning to post-weaning period (60th–87th day) of goat kids fed FS-9 and PC-16 compared to CON. It is evident that there were no differences observed in ADWG between different dietary treatments in both growth periods. However, the growth period affected ADWG in the PC-16 and CON groups (p < 0.05). In PC-16 and CON, ADWG decreased from the weaning to post-weaning period compared to the suckling to weaning growth period, while in FS-9, no differences were estimated.
3.2. Hematological Parameters
As presented in Table 3, no specific changes were observed in the majority of hematological parameters in goat kids’ blood as affected by dietary treatment. However, the WBC count was higher (p < 0.05) in goat kids fed FS-9 and PC-16 compared to CON diets during the weaning. The age of the goat kids affected some hematological parameters, like WBC, RBC, and HGB. The WBC count increased (p < 0.05) in all groups as kids were growing, specifically at post-weaning compared to suckling, while in CON, it increased compared to suckling and weaning periods. The RBC count increased only in the CON group post-weaning compared to the suckling period, while in PC-16 a tendency towards an increase was determined (p = 0.06). HGB increased (p < 0.05) in experimental groups during animal growth at the post-weaning period compared to suckling, while in the CON group a tendency towards an increase (p = 0.08) was observed.
In Figure 2, it is evident that diet influenced the LYM percentage between differently fed groups during the weaning period, when FS-9-fed kids had higher (p < 0.05) LYMs compared to PC-16, while the growth effect was noticeable in the PC-16 group by increasing LYMs post-weaning compared to the suckling period. The NEUT content was lower in FS-9 compared to PC-16 during weaning, while in PC-16, NEUTs decreased from the suckling to the post-weaning period.
3.3. Biochemical Parameters
In Table 4, biochemical parameters present mostly no significant changes as affected by FS-9 or PC-16 feeding. The concentration of BHB was the lowest (p < 0.05) in the serum of goat kids fed FS-9 compared to CON, while PC-16 feeding did not effect any changes in the post-weaning period. Growth affected a few parameters, like LDL being higher (p < 0.05) in serum post-weaning compared to the weaning period in the FS-9 group. The NEFA concentrations were lower (p < 0.05) at post-weaning and weaning compared to the suckling period in both experimental groups. Concentrations of BHB were higher (p < 0.05) in the suckling period compared to the weaning and post-weaning periods in both FS-9 and PC-16.
In all groups, the concentration of proteins increased (p < 0.05) during aging, specifically post-weaning, compared to the weaning or suckling periods. Similarity was determined in the concentration of globulin in experimental groups, where higher (p < 0.05) values were determined post-weaning compared to weaning or suckling, while in CON, globulin increased only at post-weaning compared to the weaning period (Table 5). Most minerals did not differ in goat kids’ serum as affected by different dietary treatments. Only Mg concentrations tended to increase (p = 0.05) with the inclusion of PC-16 in goat kids’ diets during the weaning period. It is evident that Mg concentration increased (p < 0.05) in the serum of goat kids fed PC-16 post-weaning compared to the weaning or suckling periods. Other minerals, like Ca, Fe, and P-inorganic, did not reveal any significant changes in the serum of goat kids during growth.
The activity of GGT was the lowest in the serum of goat kids fed PC-16 diets compared to CON, while FS-9 did not reveal any changes in the post-weaning period (Table 6). The AST activity tended to decrease (p = 0.07) with PC-16 in the post-weaning period, while the activity of GPx was the highest (p < 0.05) in the serum of FS-9 goat kids compared to PC-16 post-weaning. The GGT in the serum of CON goat kids was the highest (p < 0.05) post-weaning compared to the suckling period, and the GPx was higher in the serum of goat kids during weaning compared to the suckling or post-weaning periods, but only in the FS-9 group.
4. Discussion
4.1. Average Daily Weight Gain
Feeding goat kids FS-9 and PC-16 resulted in similar ADWG compared to CON from suckling to weaning and from the weaning to post-weaning period. This agrees with Mahouachi et al. [34], who concluded that extruded flaxseed (15%) can be used in lamb diets without adverse effects on average daily gains of lambs fed iso-caloric and iso-nitrogenous feed mixtures. Novoselec et al. [35] concluded that 7% pumpkin seed cake can be used as a protein source in lamb diets without affecting production characteristics such as the average daily gain of lambs. However, it can be seen from Figure 1 that ADWG decreased in PC-16 and CON goat kids after weaning, while there was no significant decrease in FS-9 goat kids. The results showed that the goat kids exhibited a very high growth intensity during the period from suckling to weaning. During this period, ADWG was achieved by both maternal milk consumption and supplemental feed [36,37], such as FS-9, PC-16, or CON feed mixtures.
According to a current study, 16% PC completely replaced the 15% extruded soybeans and 8.5% soybean meal in the feed mixtures for goat kids. This suggests that utilizing PC as an alternative protein source could reduce the high cost of soybeans in feed mixtures. Since PC is one of the affordable and locally available feeds in Europe, it may be financially advantageous for a small-scale goat farm to avoid the import of soybeans [17]. Since PC is primarily produced under organic conditions [22], it can also be used as a premium feed in organic animal farming, supporting sustainable production on small-scale goat farms. In addition, it is essential to maintain the quality of the product, the production output, and the overall health of the livestock [38]. However, 9% FS was combined with 15.7% soybean meal in feed mixtures for goat kids as an alternative source of fat and proteins. Although it is commonly recognized that FS is not very cost-effective for animal nutrition, it may help enhance the amount of functional compounds, including n-3 fatty acids, which adds additional value to animal products. Consequently, the increased cost of the produced functional foods is the only way to offset the cost of this feed mixture. Functional foods are more expensive than similar products that are not classified as “medical” healthy foods, according to Balogh et al. [39].
4.2. Hematological Parameters
As shown in Table 3, no specific feeding-related changes were observed in most hematological parameters in the blood of goat kids, except for WBC count, which was higher in the blood of goat kids fed FS-9 and PC-16 during the weaning period. In the FS-9 group, this could be explained by the high level of n-3 PUFAs in the FS-9 group, which promoted lymphocyte responses. In the study by Abu El-Hamd et al. [40], the addition of flaxseed oil to the milk of Friesian calves (0.2 mL/kg live weight) during suckling increased the WBC count and improved the immune response of calves without adverse effects on hematological or biochemical parameters. According to Al-Zuhairy and Taher [41], feeding 5 or 10% flaxseed to chickens increased blood WBC counts at 40 days of age. These authors explained that the birds were in good condition and this was not so much a sign of disease but a sign possibly related to the immunomodulatory properties of the active components of flaxseed, like n-3 PUFAs. N-3 PUFAs have an impact on various immune mechanisms, such as lymphocyte proliferation, the production of cytokines by lymphocytes, and natural killer cell activity [42]. Gandra et al. [43] reported that phagocytosis-positive leukocytes were greater in prepartum and postpartum cows fed whole flaxseeds compared to a control group, concluding that n-3 PUFAs seem to have a greater effect on the activity of leukocytes when compared with n-6 PUFAs. These results are consistent with the results of the present research, since α-linolenic acid was the highest in the feed mixture of the FS-9 group (Table 1). Furthermore, membrane phospholipids act as substrates for the release of (non-esterified) PUFAs; these released PUFAs can function as transcription factor ligands, signaling molecules, or precursors for the biosynthesis of lipid mediators, which are involved in the regulation of numerous cell and tissue responses [44]. Lee and Kang [45] showed that n-3 PUFA treatment increased immune cell numbers such as WBCs and LYMs in 100-day-old male miniature pigs fed diets supplemented with n-3 PUFAs. Momeni-Pooya et al. [46] reported that supplementing the diet with n-3 PUFAs from flaxseed oil can be a strategy to improve the immune performance of calves fed barley-based starter diets. As reported by El-Saadany et al. [47], the addition of pumpkin seed oil to the poultry diet (0.5%) can enhance the physiological, antioxidative, and immunological status of birds, owing to its high polyphenol content. The slightly elevated WBC values observed in both experimental groups indicate that the immune system of the goat kids functioned well during the weaning. The values obtained in our study were within the laboratory reference values Jackson and Cockcroft [48] (4–13 × 109/L) obtained for goats. Consistent with the increase in WBCs in the FS-9 group of goat kids, LYM levels increased in the same group compared to PC-16, while PC-16 increased NEUTs compared to FS-9 during weaning. Lee et al. [49] observed a lower NEUT/LYM ratio in the blood of laying hens fed a basal diet containing 3.6% (w/w) FS product (17% α-linolenic acid), indicating its potential effect on alleviating inflammation and stress conditions in laying hens.
It is known that goat kids are very sensitive to changes in nutrition and rearing, especially at a young age. Therefore, the switch from milk to the feed mixtures’ proteins and forage in the diet needs to be gradual [6], and took 28 days in the current study. The WBCs increased in all groups from suckling until the post-weaning period. During the weaning, goat kids are usually under stress. It has been suggested that certain immune cell populations seek refuge in the bone marrow and adipose tissue during metabolic stress [50]. In the experimental groups of the present study, the effect of metabolic stress during weaning was not pronounced, which is also reflected in adequate ADWG. The variation in the total WBC changes from suckling to the post-weaning period reflects the adaptability process of the hematopoietic system to extrauterine life, which brings the WBC counts of young animals closer to those of adults [51] and the immune system becomes more functional [52]. It is evident that RBCs and HGB increased in all groups during the growth of goat kids, but the RBC count was only significant in CON, and a tendency towards an increase was observed in PC-16, while HGB increased in experimental groups during animal growth with a tendency towards an increase in CON. Souza et al. [52] found that lambs aged 30 to 60 days had higher levels of RBC (12.1–13.6 × 106 cells μL−1) and higher levels of HGB from 30 to 90 days (8.8–11.4 gdL−1) along with a decline at the age of 120 days. They explained that when the amount of solid foods consumed increased, so did the overall RBC count and HGB content. These findings are consistent with the results of the current study on goat kids’ growth.
4.3. Biochemical Parameters
When comparing the PC-16 or FS-9 group with the CON group, the concentrations of most blood serum parameters related to energy and protein status, as well as enzyme activities, remained unchanged. This indicates that PC-16 or FS-9 had no discernible negative effects on the health or nutritional status of the goat kids, making it a safe dietary ingredient. However, growth did affect some parameters in goat kids’ serum, e.g., serum LDL levels were increased after weaning compared to the weaning period but only in the FS-9 group. This implies that prolonged (>55 days) FS-9 feeding to goat kids may contribute to hyperlipidemic blood traits by increasing serum LDL, as reported in the goat study conducted by Alves Dutra et al. [27]. In this study, increasing flaxseed supplementation from 0 to 15% had a linear effect on LDL levels measured at both 40 and 60 days of the trial period. According to the authors, the effect of flaxseed on lipid concentrations in the plasma in this investigation can be explained by the fact that eating flaxseed increased the number of circulating lipoproteins. In our study, a trend of increasing CHOL concentration in the FS-9 group was reported from the suckling to the post-weaning period, which was slightly above the reference values (1–3 mmol/L) [48]. Most of the CHOL increase occurs in the LDL cholesterol fraction [53]. In a study with 30-month-old lambs fed 10% FS in concentrate mixture, Hossein Abadi et al. [54] discovered higher CHOL when compared to raw flaxseed and the control. According to Huerta et al. [55], some studies observed that supplementation with n-3 PUFAs could raise LDL concentration, while others have found no effect, concluding that the overall n-3 PUFAs’ effect on LDL remains unclear.
Regarding the biochemical parameters of energy status, the feeding effect was significant only in the post-weaning period, when feeding FS-9 resulted in decreased BHB compared to CON. Diet, stress, and age have all been shown to influence blood BHB and ketogenesis processes in goats [3]. Probably the lowest mobilization of body fat in the post-weaning period was in FS-9 goat kids, since the ADWG in FS-9 did not decrease as compared to CON or PC-16 from the weaning to the post-weaning period. It is very well known that increased circulating levels of NEFAs and BHB are a result of the increased mobilization of body fat reserves in response to a negative energy balance [56]. Thus, a decreased BHB and NEFAs in both experimental groups during the weaning and post-weaning period, compared to the suckling period, may alleviate the negative energy status of goat kids, during the weaning, more than commercial concentrate mixtures based on soybean. Mohapatra et al. [57] showed that the sudden separation of lambs from their mothers cause stress. However, in the present study, the transition from suckling to weaning took 28 days so that the goat kids had sufficient time to acclimatize to the feed mixture and cope with weaning stress at the age of two months. However, Khan et al. [58] observed that animals weaned gradually over 21 days had lower NEFA levels than animals weaned earlier, which is consistent with the present study. In addition, Qugley et al. [59] reported that NEFA concentrations in the plasma of calves decreased with increasing age (from 5 to 8 weeks). In cases where young animals have a negative energy balance, the BHB measurement in blood could indicate BHB produced in the liver [60]. In the present study, this probably occurred during the suckling period, when BHB levels were higher in all groups, although only significantly higher in the experimental groups than in the other periods. During the suckling period, goat kids are exclusively dependent on their mother’s milk and the fact that they were not suckling during the blood sampling probably led to stress, which could be reflected in a higher BHB value. In addition, there are no reference values for BHB in the serum of growing goat kids, which needs to be investigated further.
The total proteins in goat kids’ serum were lower during the suckling period compared to weaning and post-weaning, when goat kids consumed concentrate feed mixture rich in proteins. As reported by Yusuf et al. [61], an adequate supply of protein is a fundamental factor for the proper growth of goats. An adequate proportion of crude proteins and energy provided by the feed mixture (16.2% of crude protein and 13.2 MJ/kg of metabolizable energy) explains the higher serum total proteins in goat kids in the post-weaning period. Osman et al. [62] determined a similar concentration of total proteins in weaned goat kids, which was 61 ± 4.1 g/L. Lower values for total proteins and globulins at 30 days, according to Souza et al. [63], signify the beginning of the animal’s active production of immunoglobulins and the inference of the immunoglobulins’ passive degradation process as received via colostrum. After a few months, the increase in globulin concentration in serum mainly comes from the increase in gamma-globulin concentrations related to adaptation to environmental conditions, as at this age, animals already have a mature immune system, as determined in lambs by Santos et al. [64].
GGT activity decreased in the serum of goat kids fed a PC-16 diet compared to CON, while AST activity tended to decrease with PC-16, and ALT showed some numerical decrease in the same group during the post-weaning period. It is known that elevated levels of serum enzymes such as GGT, AST, and ALT are indications of liver injury. According to Makni et al. [65], pumpkin seeds are rich in dietary fiber, antioxidants, and unsaturated fatty acids, which are known to have hepatoprotective and anti-atherogenic properties. In an experiment with rats, Nkosi et al. [66,67] observed a hepatoprotective effect of pumpkin seed protein isolate in the diet by reducing the activity of AST and ALT in plasma. A similar decrease in AST activity in the plasma of dairy cows fed diets in which soybean meal was replaced by PC and dried distillers’ grains was found by Li et al. [19]. El-Saadany et al. [47] found that the addition of pumpkin seed oil to the poultry diet (0.5%) decreased AST and ALT activity in the plasma of laying hens, concluding that pumpkin seed oil can be added to laying hens diets to improve metabolism and liver functions. Additionally, PC is rich in polyphenol content [47], which may improve hepatic morphology and antioxidant capacity in the liver, as observed by He et al. [68] in the study with piglets fed dietary holly polyphenol extracts. The decrease in serum GGT and AST activity observed in the goat kids fed PC-16 in the present study suggests that replacing extruded soybeans and soybean meal with PC after feeding for 55 days may have a protective effect on liver cells in the post-weaning period.
Antioxidant enzymes like SOD, GPx, and catalase play a role in preserving an optimal level of antioxidants inside cells [69]. Superoxide dismutase, the first enzyme responding to the presence of oxygen radicals, was not affected by the diet. However, the response of GPx activity in serum was diet-dependent at the post-weaning period. Increased GPx activity in FS-9 compared to PC-16 may be due to the n-3 fatty acids in extruded flaxseed, as shown in Table 2, which indicates that the FS-9 diets are enriched in α-linolenic acid. Higher GPx activity in blood serum could also indicate that the body is ready to cope with reactive oxygen species [70], as determined in FS-9 goat kids during the post-weaning period in our study. Gangal [71] reviewed and explained that n-3 PUFAs regulate gene expressions and thereby influence signaling pathways by interacting with nuclear receptors, e.g., by binding to the peroxisomal proliferator-activated receptor gamma. As reported by Sembratowicz et al. [72], flaxseed oil has a beneficial effect by stimulating antioxidant defense mechanisms and reducing the severity of oxidative stress, which probably occurred in the FS-9 group of the present study compared to PC-16. Al-Maghadi et al. [73] suggested that flaxseed oil exerts an alleviating effect by increasing levels of antioxidant enzymes in the cells of the intestinal mucosa, resulting in improved defense against oxygen-free radicals. In addition, Śpitalniak-Bajerska et al. [70] found higher GPx activity in the serum of pre-weaning dairy calves fed milk replacer with ethyl esters of flaxseed oil (10 g/day) and lyophilized apples (25 g/day), which improved metabolic and oxidative functions. In our study, a positive effect of FS-9 diets was shown after 55 days of goat kid experimental feeding, which is worth further study. Barda et al. [74] conducted a study on rats with paw edema, who received topical treatment with flaxseed and pumpkin oils. The authors explained that the anti-inflammatory effect of the oils tested was found to be closely linked to their antioxidant properties and their bioactive compounds (PUFAs, vitamin E, and phytosterols). Indeed, the lipophilic aspect of these substances allowed them to diffuse into the cell membrane, which is rich in PUFAs and proteins [74]. In our study, the proportion of palmitic acid was higher in the PC-16 diet than in the FS-9 diet, as palmitic acid is predominant in pumpkin seed cake, together with oleic and linoleic acids. Palmitic acid promotes pro-inflammatory processes [75], while a higher inflammatory status was significantly correlated with lower levels of antioxidant enzymes [76] as observed in PC-16 compared to FS-9 serum in goat kids. Li et al. [77] demonstrated that treatment with palmitic acid promoted the formation of malondialdehyde and simultaneously reduced the activity of GPx determined in bovine endometrial cells. However, the GPx activity in the serum of PC-16 goat kids was similar to that of the CON group and was within normal physiological levels, as reported by Śpitalniak-Bajerska et al. [70].
Despite the promising outcomes regarding the use of alternative protein sources in the nutrition of growing goat kids, the complete replacement of soybean by PC-16 or the partial replacement by FS-9 in feed mixtures addresses some limiting factors in the present experimental setup. Lower sample size was probably a limitation that affected differences among treatments or growth periods, which were not significant but showed some tendencies. The main reason for the current sample size is that the experiment was done on the small-scale goat farm, which requires the production for family consumption and for commercial uses. This is why we tried to minimize the experiment’s risks on the farm, especially because goat kids were examined in a very sensitive period during growth. Another reason is that, since this trial is a part of wider research carried out with carefully selected lactating dairy goats, we used only their goat kids in the current experiment. The present study could give us some useful guidelines for further studies regarding PC-16 and FS-9 in diets for goat kids, which will comprise a larger sample size and a longer duration.
5. Conclusions
Regarding the blood parameters, which portray the health and nutritional status, PC-16 and FS-9, in isoproteic, isolipidic, and isoenergetic diets, can be used in goat kids’ nutrition as sources of proteins and fats on a small-scale goat farm without compromising goat kids’ growth. The current experimental setup demonstrated that pumpkin seed cake is a high-quality ruminant protein feed that contributes to the reduction of soybeans in diets. At the same time, PC-16 decreased GGT activity in serum, thus having a potential hepatoprotective role in the post-weaning period of goat kids. As compared to pumpkin seed cake diets, dietary FS-9 has a more beneficial impact on GPx activity, which is worth further study in which the overall antioxidant status of goat kids shall be studied. To justify these findings further, the meat quality, as well as profitability and environmental issues, like nitrogen excretions, should still be evaluated to study overall sustainability when using pumpkin seed cake and extruded flaxseed in goat kids’ diets on a small-scale goat farm.
Conceptualization, Ž.K.Š. and Z.A.; formal analysis, Ž.K.Š., J.N. and M.Đ.; investigation, Ž.K.Š., J.N. and M.Đ.; writing—original draft preparation, Ž.K.Š.; writing—review and editing, Ž.K.Š.; visualization, Ž.K.Š., J.N., M.Đ. and Z.A.; supervision, Z.A. All authors have read and agreed to the published version of the manuscript.
The animal study protocol was approved by the Institutional Review Board (or Ethics Committee) of the Faculty of Agrobiotechnical Sciences Osijek (protocol code 2158-94-02-24-18, 19 June 2024).
The data presented in this study are available on request from the corresponding author.
The study was carried out within the research team Breeding-technological aspects of animal production (AnimTeh, No. 1126) at the Faculty of Agrobiotechnical Sciences Osijek. The authors wish to thank the farmers from the small-scale (family) goat farm “Ðurković”, who cared for animals.
The authors declare no conflicts of interest.
Footnotes
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![View Image - Figure 1. Average daily weight gain (ADWG) of growing goat kids fed diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11) from the suckling to the weaning period (32nd–60th day) and from the weaning to the post-weaning period (60th–87th day). c,d Means with different superscripts differ significantly at p [less than] 0.05 (growth period effect).](https://media.proquest.com/media/hms/THUM/2/Y24Ma?_a=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%3D%3D&_s=XwPGRebpYncXzgRMEaic5LxOvek%3D "View Image - Figure 1. Average daily weight gain (ADWG) of growing goat kids fed diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11) from the suckling to the weaning period (32nd–60th day) and from the weaning to the post-weaning period (60th–87th day). c,d Means with different superscripts differ significantly at p [less than] 0.05 (growth period effect).")
Enlarge this image.Figure 1. Average daily weight gain (ADWG) of growing goat kids fed diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11) from the suckling to the weaning period (32nd–60th day) and from the weaning to the post-weaning period (60th–87th day). c,d Means with different superscripts differ significantly at p [less than] 0.05 (growth period effect).
![View Image - Figure 2. Lymphocytes and neutrophils in growing goat kids fed diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11). a,b Means with different superscripts differ significantly at p [less than] 0.05 (diet effect). c,d Means with different superscripts differ significantly at p [less than] 0.05 (growth period effect).](https://media.proquest.com/media/hms/THUM/4/Y24Ma?_a=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%3D%3D&_s=R%2BsvV2i%2B43I%2B45bnHncrVh1H0Ic%3D "View Image - Figure 2. Lymphocytes and neutrophils in growing goat kids fed diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11). a,b Means with different superscripts differ significantly at p [less than] 0.05 (diet effect). c,d Means with different superscripts differ significantly at p [less than] 0.05 (growth period effect).")
Enlarge this image.Figure 2. Lymphocytes and neutrophils in growing goat kids fed diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11). a,b Means with different superscripts differ significantly at p [less than] 0.05 (diet effect). c,d Means with different superscripts differ significantly at p [less than] 0.05 (growth period effect).
Experimental design and composition of experimental diets offered to goat kids on a small-scale goat farm.
| Trait | Diets | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| Control | FS-9 | PC-16 | |||||||
| Goat kids (n) | 9 | 11 | 11 | ||||||
| Age of goat kids (days) | 32 | 60 | 87 | 32 | 60 | 87 | 32 | 60 | 87 |
| Growth period | Suckling | Weaning | Post-weaning | Suckling | Weaning | Post-weaning | Suckling | Weaning | Post-weaning |
| Production traits 1 | |||||||||
| Live body weight (kg) | 7.48 | 12.54 | 15.49 | 8.07 | 13.05 | 17.11 | 8.23 | 14.38 | 17.92 |
| ADWG, 32nd–87th day (g) | 145.64 | 164.21 | 163.77 | ||||||
| Feeding | |||||||||
| Suckling milk | Ad libitum | Restricted 2 | - | Ad libitum | Restricted 2 | - | Ad libitum | Restricted 2 | - |
| Feed mixture | - | 0% extruded flaxseed or pumpkin seed cake | - | 9% extruded flaxseed | - | 16% pumpkin seed cake | |||
| Ad libitum | ~200 g | Ad libitum | ~200 g | Ad libitum | ~200 g | ||||
| Hay | - | Ad libitum | - | Ad libitum | - | Ad libitum | |||
ADWG—average daily weight gain. 1 Klir Šalavardić et al. [
Dietary ingredients, chemical composition, and major fatty acid proportions of concentrate mixtures and hay used in the diets for goat kids.
| Ingredient, % | Concentrate Mixture | Hay | ||
|---|---|---|---|---|
| CON | FS-9 | PC-16 | ||
| Corn grain | 42.9 | 40.8 | 45.9 | |
| Barley grain | 8.0 | 8.0 | 9.0 | |
| Oat grain | 10.0 | 10.0 | 13.5 | |
| Wheat flour | 12.0 | 9.0 | 12.0 | |
| Extruded soybean | 15.0 | - | - | |
| Extruded linseed | - | 9.0 | - | |
| Pumpkin seed cake | - | - | 16.0 | |
| Alfalfa dehydrated | - | 4.0 | - | |
| Soybean meal (46% crude protein) | 8.5 | 15.7 | - | |
| Calcium carbonate | 1.6 | 1.5 | 1.6 | |
| Monocalcium phosphate | 0.5 | 0.5 | 0.5 | |
| Salt | 0.4 | 0.4 | 0.4 | |
| Pellet binder | 0.1 | 0.1 | 0.1 | |
| Mineral vitamin premix 1 | 1.0 | 1.0 | 1.0 | |
| Chemical composition, % | ||||
| DM (% fresh matter) | 87.6 | 87.4 | 87.3 | 92.5 |
| Crude protein | 16.2 | 16.2 | 16.3 | 11.0 |
| Crude fiber | 4.14 | 4.88 | 3.73 | 28.7 |
| Crude ash | 4.92 | 5.06 | 5.23 | 5.73 |
| Crude lipid | 5.64 | 5.83 | 5.63 | 1.33 |
| ME (MJ/kg DM) | 13.2 | 13.0 | 13.2 | 8.0 |
| Mineral composition (mg/kg DM) | ||||
| Ca | 8949 | 8108 | 8548 | 1572 |
| P | 7100 | 6147 | 6818 | 1410 |
| Mg | 2114 | 1944 | 2096 | 480 |
| Fe | 321 | 278 | 294 | 92.8 |
| Fatty acids (g/100 g FAME) | ||||
| C16:0 | 10.7 | 9.43 | 12.9 | 29.80 |
| C18:0 | 4.08 | 6.11 | 4.48 | 2.84 |
| C18:1 n-9 | 31.9 | 32.0 | 34.4 | 7.71 |
| C18:2 n-6 | 48.2 | 38.2 | 44.4 | 22.40 |
| C18:3 n-3 | 2.97 | 11.50 | 1.80 | 24.50 |
CON—control group; FS-9—feed mixture containing extruded flaxseed, PC-16—feed mixture containing pumpkin seed cake, DM—dry matter, ME—metabolizable energy, FAME—fatty acid methyl ester. 1 Mineral–vitamin premix: iron sulphate monohydrate 4000 mg, copper sulphate pentahydrate 800 mg, manganese oxide 3500 mg, zinc sulphate monohydrate 5000 mg, potassium iodide 80 mg, cobalt sulphate heptahydrate 20 mg, sodium selenite 15 mg, magnesium oxide 5000 mg, vitamin A 1,000,000 IU, vitamin D3 150,000 IU, α-tocopherol 1500 mg, vitamin K3 50 mg, vitamin B1 100 mg, vitamin B2 200 mg, vitamin B6 200 mg, vitamin B12 1 mg, niacin 1000 mg, Ca-pantothenate 500 mg, and choline chloride 10,000 mg.
Hematological parameters in the whole blood of goat kids fed with diets containing soybean (CON, n = 9), extruded flaxseed (FS-9, n = 11), and pumpkin seed cake (PC-16, n = 11).
| Parameters | Growth Period | Diet | SEM | p Value 1 | ||
|---|---|---|---|---|---|---|
| CON | FS-9 | PC-16 | ||||
| WBCs (×109 L) | Suckling | 6.44 d | 8.49 d | 8.76 d | 0.530 | 0.185 |
| Weaning | 6.61 b,d | 9.84 a,c,d | 9.54 a,c,d | 0.455 | 0.005 | |
| Post-weaning | 10.76 c | 12.09 c | 12.58 c | 0.563 | 0.468 | |
| p value 2 | 0.011 | 0.003 | 0.027 | |||
| RBCs (×1012 L) | Suckling | 10.18 d | 11.35 | 11.85 | 0.503 | 0.442 |
| Weaning | 11.63 c,d | 12.00 | 13.03 | 0.505 | 0.556 | |
| Post-weaning | 13.54 c | 13.43 | 14.53 | 0.393 | 0.481 | |
| p value 2 | 0.017 | 0.232 | 0.064 | |||
| HGB (g/L) | Suckling | 85.86 | 83.20 d | 86.38 d | 3.568 | 0.927 |
| Weaning | 85.14 | 90.91 c,d | 96.38 c,d | 3.115 | 0.410 | |
| Post-weaning | 103.0 | 103.9 c | 110.3 c | 2.715 | 0.538 | |
| p value 2 | 0.076 | 0.038 | 0.013 | |||
| HCT (L/L) | Suckling | 0.485 | 0.540 | 0.408 | 0.032 | 0.231 |
| Weaning | 0.558 | 0.560 | 0.501 | 0.043 | 0.824 | |
| Post-weaning | 0.467 | 0.599 | 0.417 | 0.043 | 0.186 | |
| p value 2 | 0.720 | 0.851 | 0.323 | |||
| MCV (fL) | Suckling | 49.90 | 52.39 | 38.01 | 4.829 | 0.444 |
| Weaning | 56.77 | 60.33 | 40.74 | 5.422 | 0.302 | |
| Post-weaning | 48.20 | 61.55 | 36.31 | 5.200 | 0.117 | |
| p value 2 | 0.804 | 0.730 | 0.919 | |||
| MCH (pg) | Suckling | 8.02 | 7.38 | 7.33 | 0.178 | 0.050 |
| Weaning | 7.41 | 7.73 | 7.45 | 0.129 | 0.545 | |
| Post-weaning | 7.63 | 7.83 | 7.60 | 0.109 | 0.638 | |
| p value 2 | 0.111 | 0.203 | 0.562 | |||
| MCHC (g/L) | Suckling | 201.6 | 174.6 | 216.3 | 13.652 | 0.439 |
| Weaning | 159.9 | 165.1 | 214.8 | 14.990 | 0.290 | |
| Post-weaning | 194.0 | 158.5 | 241.9 | 15.001 | 0.079 | |
| p value 2 | 0.570 | 0.897 | 0.609 | |||
CON—control group; FS-9—feed mixture containing extruded flaxseed, PC-16—feed mixture containing pumpkin seed cake; RBCs—erythrocytes, WBCs—leukocytes, HGB—hemoglobin, HCT—hematocrit, MCH—average hemoglobin content in erythrocytes, MCV—mean corpuscular volume, MCHC—mean hemoglobin concentration in erythrocytes. 1 Diet effect. 2 Growth period effect. a,b Row means with different superscripts differ significantly at p < 0.05 (diet effect). c,d Column means with different superscripts differ significantly at p < 0.05 (growth period effect).
Biochemical parameters of energy status in the serum of goat kids fed with diets containing soybean (CON, n = 9), extruded flaxseed (n = 11), and pumpkin seed cake (n = 11).
| Parameters, mmol/L | Growth Period | Diet | SEM | p Value 1 | ||
|---|---|---|---|---|---|---|
| CON | FS-9 | PC-16 | ||||
| Glucose | Suckling | 3.22 | 3.34 | 4.17 | 0.213 | 0.163 |
| Weaning | 3.80 | 3.94 | 4.29 | 0.195 | 0.621 | |
| Post-weaning | 3.94 | 3.94 | 4.03 | 0.108 | 0.926 | |
| p value 2 | 0.314 | 0.316 | 0.749 | |||
| Cholesterol | Suckling | 2.54 | 2.60 | 2.41 | 0.159 | 0.896 |
| Weaning | 2.27 | 2.21 | 2.40 | 0.130 | 0.830 | |
| Post-weaning | 2.66 | 3.05 | 2.86 | 0.191 | 0.740 | |
| p value 2 | 0.689 | 0.084 | 0.474 | |||
| TGCs | Suckling | 0.423 | 0.422 | 0.339 | 0.029 | 0.430 |
| Weaning | 0.291 | 0.323 | 0.384 | 0.033 | 0.569 | |
| Post-weaning | 0.397 | 0.395 | 0.454 | 0.037 | 0.780 | |
| p value 2 | 0.282 | 0.387 | 0.455 | |||
| HDL | Suckling | 1.49 | 1.58 | 1.36 | 0.066 | 0.384 |
| Weaning | 1.32 | 1.30 | 1.38 | 0.061 | 0.856 | |
| Post-weaning | 1.48 | 1.54 | 1.51 | 0.066 | 0.944 | |
| p value 2 | 0.633 | 0.088 | 0.654 | |||
| LDL | Suckling | 0.856 | 0.823 c,d | 0.900 | 0.094 | 0.949 |
| Weaning | 0.813 | 0.765 d | 0.846 | 0.071 | 0.893 | |
| Post-weaning | 0.998 | 1.33 c | 1.15 | 0.124 | 0.583 | |
| p value 2 | 0.788 | 0.035 | 0.433 | |||
| NEFAs | Suckling | 1.37 | 1.50 c | 1.69 c | 0.188 | 0.819 |
| Weaning | 0.590 | 0.242 d | 0.185 d | 0.077 | 0.111 | |
| Post-weaning | 0.443 | 0.324 d | 0.113 d | 0.103 | 0.485 | |
| p value 2 | 0.104 | <0.001 | <0.001 | |||
| BHB | Suckling | 0.463 | 0.396 c | 0.498 c | 0.033 | 0.436 |
| Weaning | 0.283 | 0.191 d | 0.210 d | 0.029 | 0.451 | |
| Post-weaning | 0.373 a | 0.229 b,d | 0.268 a,b,d | 0.023 | 0.036 | |
| p value 2 | 0.197 | <0.001 | 0.004 | |||
CON—control group; FS-9—feed mixture containing extruded flaxseed, PC-16—feed mixture containing pumpkin seed cake; HDL—high-density lipoprotein, LDL—low-density lipoprotein, TGCs—triglycerides, NEFAs—non-esterified fatty acids, BHB—β-hydroxybutyrate, SEM—standard error of mean. 1 Diet effect. 2 Growth period effect. a,b Row means with different superscripts differ significantly at p < 0.05 (diet effect). c,d Column means with different superscripts differ significantly at p < 0.05 (growth period effect).
Biochemical parameters of protein and mineral status in the serum of goat kids fed with diets containing soybean (CON, n = 9), extruded flaxseed (n = 11), and pumpkin seed cake (n = 11).
| Parameters (g/L) | Growth Period | Diet | SEM | p Value 1 | ||
|---|---|---|---|---|---|---|
| CON | FS-9 | PC-16 | ||||
| Urea (mmol/L) | Suckling | 4.15 | 4.90 | 4.06 | 0.236 | 0.254 |
| Weaning | 3.96 | 3.86 | 3.74 | 0.261 | 0.953 | |
| Post-weaning | 4.72 | 4.57 | 3.77 | 0.297 | 0.413 | |
| p value 2 | 0.614 | 0.271 | 0.793 | |||
| Proteins | Suckling | 53.21 d | 53.48 d | 57.11 d | 0.855 | 0.129 |
| Weaning | 53.53 d | 54.45 d | 53.64 d | 0.831 | 0.883 | |
| Post-weaning | 61.48 c | 64.42 c | 64.31 c | 0.786 | 0.303 | |
| p value 2 | 0.010 | <0.001 | <0.001 | |||
| Albumin | Suckling | 28.84 | 30.81 | 29.23 | 0.472 | 0.178 |
| Weaning | 29.07 | 28.75 | 29.09 | 0.378 | 0.766 | |
| Post-weaning | 30.45 | 30.44 | 31.00 | 0.362 | 0.787 | |
| p value 2 | 0.305 | 0.081 | 0.151 | |||
| Globulin | Suckling | 24.37 c,d | 22.67 d | 27.88 d | 0.940 | 0.057 |
| Weaning | 24.07 d | 25.70 d | 24.55 d | 0.679 | 0.605 | |
| Post-weaning | 31.03 c | 33.98 c | 33.31 c | 0.712 | 0.266 | |
| p value 2 | 0.032 | <0.001 | <0.001 | |||
| Ca (mmol/L) | Suckling | 2.36 | 2.45 | 2.54 | 0.029 | 0.053 |
| Weaning | 2.41 | 2.45 | 2.38 | 0.020 | 0.352 | |
| Post-weaning | 2.48 | 2.41 | 2.25 | 0.083 | 0.589 | |
| p value 2 | 0.191 | 0.680 | 0.415 | |||
| Mg (mmol/L) | Suckling | 0.931 | 0.967 | 0.880 d | 0.018 | 0.112 |
| Weaning | 0.869 | 0.945 | 0.960 d | 0.015 | 0.050 | |
| Post-weaning | 0.959 | 0.990 | 1.06 c | 0.020 | 0.144 | |
| p value 2 | 0.094 | 0.552 | <0.001 | |||
| Fe (μmol/L) | Suckling | 28.64 | 26.32 | 27.09 | 2.863 | 0.952 |
| Weaning | 30.67 | 26.68 | 33.58 | 2.567 | 0.333 | |
| Post-weaning | 25.48 | 23.26 | 22.69 | 1.259 | 0.710 | |
| p value 2 | 0.554 | 0.740 | 0.123 | |||
| P-inorganic (mmol/L) | Suckling | 3.06 | 3.15 | 3.10 | 0.078 | 0.906 |
| Weaning | 3.12 | 3.08 | 3.22 | 0.064 | 0.669 | |
| Post-weaning | 3.49 | 2.96 | 3.17 | 0.087 | 0.057 | |
| p value 2 | 0.187 | 0.491 | 0.809 | |||
CON—control group; FS-9—feed mixture containing extruded flaxseed, PC-16—feed mixture containing pumpkin seed cake; SEM—standard error of mean. 1 Diet effect. 2 Growth period effect. c,d Column means with different superscripts differ significantly at p < 0.05 (growth period effect).
Activities of serum enzymes of goat kids fed with diets containing soybean (CON, n = 9), extruded flaxseed (n = 11), and pumpkin seed cake (n = 11).
| Enzyme (U/L) | Growth Period | Diet | SEM | p Value 1 | ||
|---|---|---|---|---|---|---|
| CON | FS-9 | PC-16 | ||||
| Aspartate aminotransferase (AST) | Suckling | 71.17 | 71.38 | 68.73 | 2.525 | 0.905 |
| Weaning | 97.15 | 80.72 | 74.57 | 5.984 | 0.377 | |
| Post-weaning | 101.06 | 82.81 | 77.19 | 3.974 | 0.072 | |
| p value 2 | 0.223 | 0.383 | 0.369 | |||
| Alanine aminotransferase (ALT) | Suckling | 11.76 | 18.78 | 20.45 | 2.885 | 0.498 |
| Weaning | 16.27 | 17.73 | 14.07 | 2.514 | 0.841 | |
| Post-weaning | 25.38 | 20.54 | 18.14 | 3.610 | 0.771 | |
| p value 2 | 0.251 | 0.927 | 0.694 | |||
| γ-glutamyl transferase (GGT) | Suckling | 34.23 d | 37.52 | 36.20 | 1.823 | 0.791 |
| Weaning | 43.97 c,d | 43.63 | 41.56 | 2.079 | 0.894 | |
| Post-weaning | 48.40 a,c | 39.52 a,b | 38.65 b | 1.606 | 0.040 | |
| p value 2 | 0.009 | 0.400 | 0.507 | |||
| Superoxide dismutase | Suckling | 0.340 | 0.402 | 0.464 | 0.066 | 0.803 |
| Weaning | 0.383 | 0.501 | 0.609 | 0.071 | 0.524 | |
| Post-weaning | 0.769 | 0.617 | 0.716 | 0.089 | 0.789 | |
| p value 2 | 0.209 | 0.323 | 0.610 | |||
| Glutathione peroxidase (GPx) | Suckling | 960.1 | 1061.4 d | 914.7 | 62.739 | 0.610 |
| Weaning | 1011.1 | 1104.2 c | 893.6 | 63.785 | 0.396 | |
| Post-weaning | 700.2 a,b | 809.7 a,d | 600.8 b | 35.716 | 0.039 | |
| p value 2 | 0.154 | 0.008 | 0.119 | |||
CON—control group; FS-9—feed mixture containing extruded flaxseed, PC-16—feed mixture containing pumpkin seed cake; SEM—standard error of mean. 1 Diet effect. 2 Growth period effect. a,b Row means with different superscripts differ significantly at p < 0.05 (diet effect). c,d Column means with different superscripts differ significantly at p < 0.05 (growth period effect).
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