1. Introduction
Shiitake mushrooms (Lentinula edodes) have remarkable health properties that have been used in Far Eastern medicine for centuries. In Asia, shiitake mushrooms have been cultivated for more than 1000 years and are now the third most cultivated mushroom variety in the world. Due to their health-promoting properties and easily digestible ingredients, they are referred to as the “elixir of life” in Asian countries [1,2]. These mushrooms, used as “functional foods”, have a long history in oriental folklore in the treatment of cancer, viral diseases, cardiovascular diseases, obesity, diabetes, respiratory diseases, liver disorders, fatigue and weakness [3]. Shiitake mushrooms are also called “medicinal mushrooms” because of their properties and their use as a dietary supplement to support patients during oncological therapy and radiotherapy [1,4,5].
Shiitake mushrooms are a source of two types of polysaccharide medicine: β-D-glucans (extracted from the mycelium) and fungal bodies [6]. Both components activate the immune system [7,8], have antioxidant properties [9] and significantly influence the regeneration of the body in studies based on humans and fish [10,11]. They also lower cholesterol levels, a process brought about by eritadenine, which can reduce low-density lipoprotein cholesterol by increasing the excretion of cholesterol through the faeces in humans [1]. Eritadenine is also a source of fatty acids with a high content of amino acids [12]. Shiitake mushrooms also contain significant amounts of vitamins, especially C, B1, B2 and B12 (niacin), which reduce stress responses and have calming effects [13]. They contain the highest amount of vitamin D among plant foods, with a presence of calcium [14,15].
In contrast to humans, the effects of shiitake mushrooms on animal health are scarce and mostly limited to laboratory animals such as rats [16] and mice [17] or poultry [18,19,20,21,22]. In rats, shiitake supplementation affected the growth of specific gut microbes, e.g., Clostridium and Bacteroides spp., compared to the control groups, and manipulation of the gut microbiota through the administration of L. edodes managed dyslipidemia [16]. Research on mice indicated significantly lowered serum total cholesterol, LDL cholesterol and triglycerides after mushroom supplementation, which could help in the regulation of lipid metabolism [17], and which was also suggested by the authors of a recent study of shiitake effects on dogs [23]. Several studies have confirmed the health-promoting effects of supplementation with shiitake mushroom on immune response modulation and stress reduction in poultry [18] and antibacterial, antiviral and antiseptic activity in chickens [19,20,21], as well as growth performance in broilers [22].
In our previous study on horses, supplementation with shiitake mushroom significantly influenced several hematology and blood biochemistry parameters and confirmed the beneficial effects of mushroom supplementation on horses [24]. However, there currently are no standard protocols to verify the efficacy of shiitake mushroom in equine nutritional supplementation. The lack of studies on the recommended daily dose of shiitake mushrooms in relation to blood parameters has limited research in this field. This requires more focused research on different daily doses and a standardized study group of horses to limit the physiological effects on blood parameters. The aim of the present study was therefore to investigate the effects of dietary supplementation with shiitake mushrooms (L. edodes; 30 g per day over a period of four months) on the hematological and biochemical blood parameters of young Thoroughbred racehorses in regular training.
2. Materials and Methods
Ethical approval was obtained from the Local Ethical Committee for Animal Experiments in Wroclaw, prior to data collection in June 2019 (resolution no. 036/2019/P1).
2.1. Animals
The study was conducted on a group of 20 three- to four-year-old Thoroughbred racehorses, with an average body weight of 454 ± 22 kg. The horses were clinically healthy and in good condition. They all had a similar fitness level and were trained daily for flat races at Partynice Racecourse (Poland). The daily training duration was the same for all horses. The animals were housed in individual boxes on straw, without access to pastures. All the boxes measured about 12.25 m2 and were cleaned six times a week. The animals were dewormed and vaccinated one month before the experiment according to standard veterinary practice. They were all fed a basal diet of oats and concentrates (muesli) (Oat Balancer Mix, Baileys Horse Feed, Braintree, Essex, UK—ingredient composition: micronized barley, soybean flakes, maize and soya, molasses, micronized wheat, distillers’ rains, soya oil, hulls of soybeans, cooked linseed, ScFOS (Digest Plus prebiotic) vitamins and minerals and roughage (meadow hay), in accordance with the requirements for racehorses, based on the Nutrient Requirements of Horses [25]. Water and mineral salt (Lisal, Klodawa, Poland) were provided as needed. Horses were fed three times a day, at 5.30 a.m., 12.00 p.m. and 5.00 p.m. The diet was followed for 6 months before the first samples were taken and continued throughout the study.
2.2. Experimental Procedure
The lyophilizate of shiitake mushrooms used in the study was produced on the Ecological Farm for the Cultivation of Chinese Mushrooms in Kalisz, Poland. The shiitake mushroom powder was analyzed and contained the following: dry weight—90.77%, ash—6.71 g/kg; protein—25.71 g/kg; fat—1.04 g/kg; fiber—9.05 g/kg; Mg—1.11 g/kg; Ca—0.28 g/kg; F—1.71 g/kg; N—19.95 g/kg; K—19.95 g/kg; Cu—6.98 mg/kg; Mn—17.41 mg/kg; Fe—82.75 mg/kg; Zn—66.64 mg/kg; and Se—871.85 ug/kg. The powder was granulated with additives so that 100 g of granulate contained 30 g of dried shiitake mushroom, 50 g of barley, 10 g of wheat bran, 4 g of corn, 4 g of lucerne and 2 g of molasses.
The horses participating in the study were randomly divided into two groups: the first, the supplemented group (G0) consisting of ten horses (five aged three years, five aged four years), and the second group was the control group (G1), which also included ten horses (five aged three years, five aged four years). The supplemented group received an addition of 100 g granulate containing 30 g of shiitake mushrooms once daily (during 12:00 feeding) throughout the entire study period (from March until June). The control group was given granules without the addition of L. edodes.
2.3. Blood Collection and Analysis
The protocol for blood sampling was the same as in our previous study [24]. Blood samples were collected four times, four weeks apart, starting on day 1 of treatment, followed by day 28, day 56 and day 84.
Blood was collected at rest, before morning feeding, by puncturing of the external jugular vein (vena jugularis externa) with a sterile BD Vacutainer® system including 20 G 1/2” needles. K2-EDTA tubes were used for hematological analysis, and plain tubes for serum biochemistry and fluoride tubes (sodium fluoride 15 mg/mL/EDTA 3.0 mg/mL) for determination of to assess lactic acid concentration (lactic acid, mmol/L) (BD Vacutainer Systems, Plymounth, UK). All blood samples were examined within a maximum period of 1.5 h after collection.
Blood hematological analyses were performed using a Sysmex XN-1000 hematology analyzer (Sysmex America, Inc., Lincolnshire, Illinois, IL, USA). The following parameters were assessed: white blood cell count (WBC, 109/L), absolute and differential distribution of neutrophils (NEU, 109/L, %), lymphocytes (LYM, 109/L, %), monocytes (MONO, 109/L, %), eosinophils (EOS, 109/L, %), and basophils (BASO, 109/L, %), red blood cell count (RBC, 1012/L), hemoglobin concentration (HGB, mmol/l), hematocrit (HCT, L/L,%), mean corpuscular volume (MCV, fL), mean corpuscular hemoglobin (MCH, fmol), mean corpuscular hemoglobin concentration, (MCHC, mmol/L) and platelet count (PLT, 1012/L).
The plain tubes were centrifuged (2800 rpm for 5 min) using a benchtop Rotanta 460 centrifuge (Andreas Hettich GmbH & Co. KG, Tuttlingen, Germany), with the serum aspirated. The levels of the following biochemical parameters were estimated: albumin (g/L), alkaline phosphatase (AP, U/L), aspartate aminotransferase (AST, U/L), total protein (g/L), total bilirubin (μmol/L), chlorides (mmol/L), cholesterol (mmol/L), creatine kinase (CK, U/L), phosphorus (P, mmol/L), glutamate dehydrogenase (GLDH, U/L), glucose (GLUC, mmol/L), gamma-glutamyl transferase (GGTP, U/L), creatinine (μmol/L), lactate dehydrogenase (LDH, U/L), magnesium (Mg, mmol/L), urea (mmol/L), potassium (K, mmol/L), sodium (Na, mmol/L), triglycerides (TGL, mmol/L), calcium (Ca, mmol/L), globulins (g/L) and albumin/globulin ratio (mmol/L). All the parameters were determined using a AU680 clinical chemistry analyzer (Backman Coulter Inc., Brea, CA, USA). Sysmex (Sysmex America, Inc., Lincolnshire, IL, USA) and Backman (Backman Coulter Inc., Brea, CA, USA) reagents, calibrators and standards were used for all the measurements. Only GLDH concentrations were determined using Randox diagnostic veterinary reagents (Randox, Crumlin, UK).
2.4. Data Analysis
All tests were performed with a significance level of α = 0.05. The Shapiro–Wilk and Kolmogorov–Smirnov tests of normality proved that the measurement did not originate from a Gaussian distribution, so non-parametric tests were used. The differences between the supplemented group (G0) and the control group (G1) were tested using the Mann–Whitney U significance test. In the following, each blood sampling was considered separately. Only the differences between the two groups were compared and the groups as such were considered independent. A Friedman ANOVA test was used to test the significance of differences in blood parameters measurements, between the G0 and G1 groups. For significant parameters, a post-hoc (Conover–Iman) test was used to test for the differences between individual days of sampling. Statistica software (v. 13.3, StatSoft Inc., Tulsa, OK, USA) was used for all calculations.
3. Results
3.1. Blood Hematological Analysis
The results of the comparisons of the hematological analysis between the supplemented and control groups are presented in Table 1. Statistically significant differences were found for MONO and MONO% at day 56. Considering the p-value of MONO% in this session, there was a significant difference between the supplemented group and the control group.
Friedman’s ANOVA was used to test the influence of shiitake mushroom supplementation throughout the test period. The same horses were compared across different days of blood sampling and, as such, the variables were considered as dependent. Significant differences were found in ten parameters between the days of blood sampling (Table 2). The largest difference was found for MCV (p = 0.000099).
Friedman’s ANOVA proved that parameters differed between sampling days but there was no indication between which days. To determine the differences between the two groups, the median parameters of the hematological blood analysis for the supplemented and control groups are shown in Figure 1. No clear patterns were observed for the blood parameters presented, nor were there any significant differences between the groups studied.
3.2. Blood Biochemical Analysis
An analysis of the differences in biochemical factors is shown in Table 3. Statistically significant differences in AP were found between the two groups, on all days of sampling. The largest differences were found on days 28 and 56. For glucose, there were statistical differences on days 28 and 56. Considering the p-value of glucose on day 1, it seems to be close to statistical validity. In addition, AST and lactic acid only differed significantly during individual days of blood sampling.
Differences were found between biochemical values on each day of blood sampling for most parameters (Table 4). The greatest difference was found for LDH, which increased significantly on day 56 and decreased on day 84 in both groups. The opposite trend was noted for albumin, which decreased on day 56 and increased on day 84 in the supplemented group. In addition, cholesterol levels decreased throughout the experiment, whereas GGTP increased. Furthermore, differences were found between the days of blood sampling in both the research and control groups.
Considering the median parameters of the blood biochemical analysis for all days of blood sampling (Figure 2), a significant variability can be observed. Significant variations in lactic acid, CK and AST and albumin values were observed in the control group, compared to the supplemented group. For the horses in the supplemented group, the median value of the lactic acid was at a stable low level compared to the control group, without a large jump on day 84. The median cholesterol in G0 decreased steadily and remained constant in G1. The levels of bilirubin, CK and AST were lower in G0 than in G1. A closer comparison of the p-values of these groups suggests that the control group differs more between days of blood collection than the supplemented group.
4. Discussion
The present study was conducted to determine the effects of L. edodes, added to commercial feed, on the hematological and biochemical blood parameters of young Thoroughbred racehorses in regular training. The results indicated single statistical differences among training days between the supplemented and control groups.
Differences in hematological parameters between the two groups were only found for MONO at day 56, where the values for MONO were within normal ranges for both the experimental and control groups [26]. Monocytes are a critical component of the innate immune system, capable of rapid transfer to sites of inflammation, phagocytosis, pathogens or allergens [27]. The results of previous studies conducted on different species of fish and broilers suggest that diets supplemented with L. edodes mushrooms enhance immune response [28,29]. Interestingly, in our previous study based on riding horses supplemented with shiitake mushrooms, they had also manifested elevated MONO values [24].
Concerning blood biochemical parameters, the supplemented groups of horses showed a statistically significant reduction in AP values on all days of sampling compared to the control group, whereas values for AP were within normal ranges for two groups [30]. The greatest differences were found on days 28 (value of median of 289 U/L) and 56 (value of median of 204 U/L). Furthermore, the overall decrease of AP from day 28 to day 84 was approximately 41%. The AP medians of the control group did not exhibit this tendency. Interestingly, we found very similar results in our previous study, where horses supplemented with shiitake mushrooms also had lower levels of AP [24]. AP enzyme production is routinely used as a marker of liver function. It was found that different types of infectious and inflammatory diseases caused an increase in AP, which is probably a secondary phenomenon in different types of infectious and inflammatory liver diseases and is related to increased hepatocellular metabolism, most likely caused by a general stress on the organism under different disease conditions [31,32].
The results of the present study showed significant differences in glucose between the two groups on the two days of blood sampling. Similar results were presented in our previous study [24], where horses supplemented with L. edodes had lower glucose concentrations. In particular, the biologically active polysaccharides (β-glucans) contained in the mushrooms can restore the pancreatic tissue function, causing better utilization of insulin, thus preventing high blood glucose levels [33]. The present study also indicated a significant decrease in AST in the supplemented group, but only once. AST is a liver enzyme that is released into the bloodstream when liver dysfunction occurs. A study with a group of mice diagnosed with hepatitis showed a significant improvement in liver histology, indicated by a decrease in AST serum levels, after the mice were fed mushroom extracts [34]. In the present study, a significant difference was also found between the two groups in lactic acid levels. Blood lactate is a metabolic end-product occurring during glycolysis of carbohydrates under anaerobic conditions, serving as a key energy source during intense exercise [35]. Lactic acid that accumulates in the blood during exercise lowers muscle pH, which can lead to acidosis [36].
Our study showed a tendency to lower cholesterol levels in the supplemented group, which is consistent with studies in school riding horses [24]. Previous studies have reported that eritadenine, a hypocholesterolemic factor isolated from shiitake mushrooms, suppresses the biosynthesis of cholesterol in the liver and lowers plasma cholesterol concentrations in rodents [37,38]. Another study based on rats indicated that administration of polysaccharides from shiitake mushrooms significantly reduced serum totalcholesterol level in rats [39], whereas a study based on healthy dogs given shiitake mushroom powder for four weeks indicated a decrease in total cholesterol levels [23].
Current studies indicate significantly lower levels of bilirubin in the supplemented group, as compared to the control group. Bilirubin has a tendency to evaluate during exercises due to intervascular hemolysis during exercises [40], or in response to increased erythrocyte fragility, functional disorders of bile duct [41] and damage to hepatocytes or functional disorders of the liver [42]. In our previous studies, we also observed bilirubin levels to be lower in the supplemented group with shitake mushrooms [24].
Studies in horses indicate an increase in albumin levels after acute exercise as well as during prolonged training schedule, most likely due to profuse sweating during exercise, causing a shift of fluid from the intravascular space to the interstitial and intracellular space. This shift between the fluid compartments of the body leads to a decrease in blood volume and hemoconcentration [43,44,45]. On the other hand, proteomic studies in horses suggest a decrease in albumin fragments after competition, returning to baseline levels after 48 h, which may also be related to exercise-induced damage to liver cell function and/or increased nutrient requirements during intense exercise [46]. There was a tendency for albumin to decrease in most of the supplemented groups, although a study using rainbow trout found no differences in albumin between the control and shiitake mushroom supplemented groups [47]. Similar results were found in a study on rats, where dietary supplementation with shiitake mushrooms had no effect on albumin levels [48]. In another study, the albumin concentration in the serum of rats was reduced by 10% after a shiitake mushroom diet [49].
Summarizing the general results of the blood biochemical parameters, the supplemented group of horses was characterized by statistically significant but smaller differences in biochemical parameters throughout the entire study period.
It should be noted that there were some limitations to the present study. These include the short duration of the study, the small number of horses, the presence of different environmental variables and the lack of previous research on this topic. These are obstacles that should be considered in future research. However, the findings of the present study suggest that shiitake mushrooms have many health-promoting effects. Consequently, a larger study in horses with many more health-related investigations is warranted.
5. Conclusions
In conclusion, oral administration of L. edodes for four months at a daily dose of 30 g in young racehorses exercising regularly indicates an effect on their blood parameters in comparison to the untreated horses. The greatest effects of supplementation were found for monocytes and alkaline phosphatase. Further studies are needed to identify the causal effects of shiitake mushroom supplementation on various hematological parameters on larger groups of horses over a longer period of time.
Conceptualization, M.S.-D. and W.G.; methodology, M.S.-D. and P.Z.; validation, W.G. and M.K.; formal analysis, A.G. and W.G.; investigation, M.S.-D., W.G. and P.Z.; data curation, M.S.-D.; writing—original draft preparation, M.S.-D. and W.G.; writing—review and editing, N.Č.-K. and A.G.; visualization, W.G. and A.G.; project administration, M.K.; funding acquisition, M.S.-D. All authors have read and agreed to the published version of the manuscript.
The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Local Ethical Committee for Animal Experiments in Wroclaw, Poland (protocol code: 036/2019/P1, June 2019).
The data presented in this study are available on request from the corresponding author. The data are not publicly available for privacy reasons.
We would like to thank the Wroclawski Tor Wysigow Konnych Partynice, Wroclaw, Poland, for providing the horses.
Authors declare no conflict of interest.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Enlarge this image.Figure 1. Hematological blood analysis median values for the: (a) supplemented group (G0); (b) control groups (G1); day 1 (S1), day28 (S2), day 56 (S3), day 84 (S4).
Enlarge this image.Figure 2. The median values of biochemical blood analysis for: (a) supplemented group (G0); (b) control group (G1); day 1 (S1), day28 (S2), day 56 (S3), day 84 (S4).
p-values representing differences between the supplemented (G0) and control group (G1) in a blood hematological analysis.
| Parameter, Unit | Day 1 |
Day 28 |
Day 56 |
Day 84 |
|---|---|---|---|---|
| WBC, 109/L | 0.344705 | 0.969839 | 0.570606 | 0.623177 |
| NEU, 109/L | 0.495968 | 0.850107 | 0.909688 | 0.212295 |
| NEU% | 0.969850 | 0.969850 | 0.850107 | 0.140168 |
| LYM, 109/L | 0.570751 | 0.909722 | 0.384674 | 0.384674 |
| LYM% | 0.596425 | 0.596564 | 0.733634 | 0.241145 |
| MONO, 109/L | 0.255945 | 0.103590 | 0.037134 | 0.272496 |
| MONO% | 0.074563 | 0.103069 | 0.000986 | 0.211089 |
| EOS, 109/L | 0.539499 | 0.288100 | 0.969679 | 0.381614 |
| EOS% | 0.878922 | 0.136421 | 1.000000 | 0.320046 |
| BASO, 109/L | 0.907623 | 0.722023 | 1.000000 | 0.691548 |
| BASO% | 0.849094 | 0.969303 | 0.784078 | 0.412192 |
| RBC, 1012/L | 0.650025 | 0.622915 | 0.820397 | 0.705246 |
| HGB, 1012/L | 0.676782 | 0.676897 | 0.255945 | 0.495318 |
| HCT, L/L | 0.426660 | 1.000000 | 0.325570 | 0.405503 |
| MCV, fL | 0.427008 | 0.704503 | 0.677356 | 0.472510 |
| MCH, fmol | 1.000000 | 0.621995 | 0.704716 | 0.544441 |
| MCHC, mmol/L | 0.197243 | 0.849378 | 0.879559 | 0.969725 |
| PLT, 1012/L | 0.496130 | 0.733537 | 0.161184 | 0.544745 |
Results of Friedman’s ANOVA and post-hoc tests for the supplemented (G0) and control group (G1) in a blood hematological analysis.
| Parameter, Unit | p-Value of G0 | p-Value of G1 |
|---|---|---|
| BASO% | 0.01375 | 0.633762 |
| EOS, 109/L | 0.017324 | 0.098309 |
| EOS% | 0.00569 | 0.060406 |
| HTC, L/L | 0.019976 | 0.056037 |
| HGB, 1012/L | 0.03228 | 0.052612 |
| LYM, 109/L | 0.025522 | 0.032658 |
| MCHC, mmol/L | 0.007176 | 0.019034 |
| MCV, fL | 0.000099 | 0.000108 |
| PLT, 1012/L | 0.012166 | 0.948376 |
| RBC, 1012/L | 0.026264 | 0.024827 |
p-values representing differences between supplemented (G0) and control groups (G1) in a biochemical analysis.
| Parameter, Unit | Day 1 |
Day 28 |
Day 56 |
Day 84 |
|---|---|---|---|---|
| Albumin, g/L | 0.677585 | 0.596702 | 0.520523 | 0.212295 |
| AP, U/L | 0.045155 | 0.014020 | 0.012612 | 0.037636 |
| AST, U/L | 0.241322 | 0.121225 | 0.017258 | 0.384674 |
| Total protein, g/L | 0.226477 | 0.820596 | 0.596702 | 0.405680 |
| Bilirubin, µmol/L | 0.212295 | 0.909722 | 1.000000 | 0.879829 |
| Chlorides, mmol/L | 0.733730 | 0.969850 | 0.241322 | 0.650148 |
| Cholesterol, mmol/L | 0.344705 | 0.241322 | 0.104111 | 0.104111 |
| CK, U/L | 0.384674 | 0.791337 | 0.850107 | 0.472676 |
| P, mmol/L | 0.762369 | 0.173618 | 0.909722 | 0.939743 |
| GLDH, U/L | 0.623177 | 0.850107 | 0.623177 | 0.762369 |
| Glucose, mmol/L | 0.096305 | 0.009109 | 0.025749 | 0.677585 |
| GGTP, U/L | 0.150928 | 0.762369 | 0.307490 | 0.241322 |
| Creatinine, µmol/L | 0.344705 | 0.307490 | 0.273037 | 0.650148 |
| LDH, U/L | 0.623177 | 0.075663 | 0.733730 | 0.449692 |
| Mg, mmol/L | 0.596702 | 0.879829 | 0.969850 | 0.075663 |
| Urea, mmol/L | 0.405680 | 0.969850 | 0.472676 | 0.449692 |
| K, mmol/L | 0.850107 | 0.596702 | 0.820596 | 0.791337 |
| Na, mmol/L | 0.969850 | 0.140466 | 0.449692 | 0.449692 |
| TGL, mmol/L | 0.705457 | 1.000000 | 0.069643 | 0.384674 |
| Ca, mmol/L | 0.570751 | 0.520523 | 0.405680 | 0.762369 |
| Globulins, g/L | 0.104111 | 0.545350 | 0.623177 | 0.520523 |
| Albumin/globulin ratio, mmol/L | 0.241322 | 0.405680 | 0.909722 | 1.000000 |
| Lactic acid, mmol/L | 0.256840 | 0.037636 | 0.879829 | 0.791337 |
Results of Friedman’s ANOVA and post-hoc test for the supplemented group (G0) and control group (G1) in a biochemical analysis.
| Parameter, Unit | p-Value |
p-Value |
|---|---|---|
| Albumin, g/L | 0.006038 | 0.041825 |
| AP, U/L | 0.001032 | 0.001032 |
| AST, U/L | 0.011694 | 0.008724 |
| Total protein, g/L | 0.00477 | 0.000655 |
| Bilirubin, µmol/L | 0.036403 | 0.228919 |
| Chlorides, mmol/L | 0.000515 | 0.008724 |
| Cholesterol, mmol/L | 0.004844 | 0.059153 |
| CK, U/L | 0.012858 | 0.53987 |
| P, mmol/L | 0.032312 | 0.322733 |
| GLDH, U/L | 0.022853 | 0.000921 |
| Glucose, mmol/L | 0.001032 | 0.130501 |
| GGTP, U/L | 0.01919 | 0.000059 |
| Creatinine, µmol/L | 0.008724 | 0.000522 |
| LDH, U/L | 0.001531 | 0.000822 |
| Mg, mmol/L | 0.0000009 | 0.046223 |
| Urea, mmol/L | 0.001156 | 0.017809 |
| K, mmol/L | 0.017626 | 0.012858 |
| Na, mmol/L | 0.003467 | 0.000293 |
| Ca, mmol/L | 0.003467 | 0.01848 |
| Globulins, g/L | 0.01919 | 0.016287 |
| Albumin/globulin ratio mmol/L | 0.017626 | 0.000293 |
| Lactic acid, mmol/L | 0.143604 | 0.000237 |
References
1. Bisen, P.S.; Baghel, R.K.; Sanodiya, B.S.; Thakur, G.S.; Prasad, G.B.K.S. Lentinus edodes: A macrofungus with pharmacological activities. Curr. Med. Chem.; 2010; 17, pp. 2419-2430. [DOI: https://dx.doi.org/10.2174/092986710791698495]
2. Chen, W.; Li, W.; Yang, Y.; Yu, H.; Zhou, S.; Feng, J.; Li, X.; Liu, Y. Analysis and evaluation of tasty components in the pileus and stipe of Lentinula edodes at different growth stages. J. Agric. Food Chem.; 2015; 63, pp. 795-801. [DOI: https://dx.doi.org/10.1021/jf505410a] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/25560993]
3. Elkhateeb, W.A.; Daba, G.M.; Thomas, P.W.; Wen, T.C. Medicinal mushrooms are a new source of natural therapeutic bioactive compounds. Egypt. Pharm. J.; 2019; 18, pp. 88-101.
4. Li, X.; Zhang, H.; Xu, H. Analysis of chemical components of shiitake polysaccharides and its anti-fatigue effect under vibration. Int. J. Biol. Macromol.; 2009; 45, pp. 377-380. [DOI: https://dx.doi.org/10.1016/j.ijbiomac.2009.07.005]
5. Gargano, M.L.; Griensven, L.J.L.D.; Isikhuemhen, O.S.; Lindequist, U.; Venturella, G.; Wasser, S.P.; Zervakis, G.I. Medicinal mushrooms: Valuable biological resources of high exploitation potential. Plant Biosyst.; 2017; 151, pp. 548-565. [DOI: https://dx.doi.org/10.1080/11263504.2017.1301590]
6. Chibata, I.; Okumura, K.; Takeyama, S.; Kotera, K. Lentinacin: A new hypocholesterolemic substance in Lentinus edodes. Experientia; 1969; 25, pp. 1237-1238. [DOI: https://dx.doi.org/10.1007/BF01897467] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/5391883]
7. Carbonero, E.R.; Ruthes, A.C.; Freitas, C.S.; Utrilla, P.; Galvez, J.; Silva, E.V.D.; Sassaki, G.L.; Gorin, P.A.J.G. Chemical and biological properties of a highly branched β-glucan from edible mushroom Pleurotus sajor-caju. Carbohydr. Polym.; 2012; 90, pp. 814-819. [DOI: https://dx.doi.org/10.1016/j.carbpol.2012.06.005] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/22840006]
8. Liu, Y.; Zhang, J.; Tang, Q.; Yang, Y.; Guo, Q.; Wang, Q.; Wu, D.; Cui, S.W. Physicochemical characterization of a high molecular weight bioactive β-D-glucan from the fruiting bodies of Ganoderma lucidum. Carbohydr. Polym.; 2014; 101, pp. 968-974. [DOI: https://dx.doi.org/10.1016/j.carbpol.2013.10.024]
9. Choi, Y.; Lee, S.M.; Chun, J.; Lee, H.B.; Lee, J. Influence of heat treatment on the antioxidant activities and polyphenolic compounds of shiitake (Lentinus edodes) mushroom. Food Chem.; 2006; 99, pp. 381-387. [DOI: https://dx.doi.org/10.1016/j.foodchem.2005.08.004]
10. Li, W.; Wang, J.; Hu, H.; Li, Q.; Liu, Y.; Wang, K. Functional polysaccharide Lentinan suppresses human breast cancer growth via inducing autophagy and caspase-7-mediated apoptosis. J. Funct. Foods; 2018; 45, pp. 75-85. [DOI: https://dx.doi.org/10.1016/j.jff.2018.03.024]
11. Ren, G.; Xu, L.; Lu, T.; Zhang, Y.; Wang, Y.; Yin, J. Protective effects of lentinan on lipopolysaccharide induced inflammatory response in intestine of juvenile taimen (Hucho taimen, Pallas). Int. J. Biol. Macromol.; 2019; 121, pp. 317-325. [DOI: https://dx.doi.org/10.1016/j.ijbiomac.2018.09.121] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/30248420]
12. Longvah, T.; Deosthale, Y.G. Compositional and nutritional studies on edible wild mushroom from northeast India. Food Chem.; 1998; 63, pp. 331-334. [DOI: https://dx.doi.org/10.1016/S0308-8146(98)00026-0]
13. Mattila, P.; Suonpää, K.; Piironen, V. Functional properties of edible mushrooms. Nutrition; 2000; 16, pp. 694-696. [DOI: https://dx.doi.org/10.1016/S0899-9007(00)00341-5]
14. Keegan, R.J.; Lu, Z.; Bogusz, J.M.; Williams, J.E.; Holick, M.F. Photobiology of vitamin D in mushrooms and its bioavailability in humans. Dermatoendocrinology; 2013; 5, pp. 165-176. [DOI: https://dx.doi.org/10.4161/derm.23321]
15. Li, S.; Wang, A.; Liu, L.; Tian, G.; Wei, S.; Xu, F. Evaluation of nutritional values of shiitake mushroom (Lentinus edodes) stipes. J. Food Meas. Charact.; 2018; 12, pp. 2012-2019. [DOI: https://dx.doi.org/10.1007/s11694-018-9816-2]
16. Anwar, H.; Suchodolski, J.S.; Ullah, M.I.; Hussain, G.; Shabbir, M.Z.; Mustafa, I.; Sohail, M.U. Shiitake culinary-medicinal mushroom, Lentinus edodes (agaricomycetes), supplementation alters gut microbiome and corrects dyslipidemia in rats. Int. J. Med. Mushrooms; 2019; 21, pp. 79-88. [DOI: https://dx.doi.org/10.1615/IntJMedMushrooms.2018029348] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/30806258]
17. Yang, H.; Hwang, I.; Kim, S.; Hong, E.J.; Jeung, E.B. Lentinus edodes promotes fat removal in hypercholesterolemic mice. Exp. Ther. Med.; 2013; 6, pp. 1409-1413. [DOI: https://dx.doi.org/10.3892/etm.2013.1333] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/24255670]
18. Lee, S.H.; Lillehoj, H.S.; Jang, S.I.; Kim, D.K.; Ionescu, C.; Bravo, D. Effect of dietary curcuma, capsicum, and lentinus, on enhancing local immunity against Eimeria acervulina infection. J. Poult. Sci.; 2010; 47, pp. 89-95. [DOI: https://dx.doi.org/10.2141/jpsa.009025]
19. Pang, F.H.; Xie, M.Q.; Ling, H.H. The investigation of immunodulator tested for the results on the control of a coccidial infection. Chin. J. Vet. Parasitol.; 2000; 8, pp. 1-3.
20. Yu, J.G.; Zhu, L.Y. The use of Astragalus polysaccharide against infectious bursa disease in chickens. J. Trad. Chin. Vet. Med.; 2000; 6, pp. 3-4.
21. Hearst, R.; Nelson, D.; McCollum, G.; Millar, B.C.; Maeda, Y.; Goldsmith, C.E.; Rooney, P.J.; Loughrey, A.; Rao, J.R.; Moore, J.E. An examination of antibacterial and antifungal properties of constituents of Shiitake (Lentinula edodes) and Oyster (Pleurotus ostreatus) mushrooms. Complement. Ther. Clin. Pract.; 2009; 15, pp. 5-7. [DOI: https://dx.doi.org/10.1016/j.ctcp.2008.10.002]
22. Guo, F.C.; Williams, B.A.; Kwakkel, R.P.; Li, H.S.; Li, X.P.; Luo, J.Y. Effects of mushroom and herb polysaccharides, as alternatives for an antibiotic, on the cecal microbial ecosystem in broiler chickens. Poult. Sci.; 2004; 83, pp. 175-182. [DOI: https://dx.doi.org/10.1093/ps/83.2.175] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/14979567]
23. Kusaba, A.; Okada, Y.; Ueno, H.; Yamamoto, I.; Mori, Y.; Tanaka, N.; Arai, T.; Kawasumi, K. Effects of supplementation with Shiitake powder, Lentinula edodes, on anti-oxidative activities and energy/lipid metabolism in healthy dogs. Res. Sq.; 2022; [DOI: https://dx.doi.org/10.21203/rs.3.rs-1199968/v1]
24. Soroko, M.; Górniak, W.; Zielińska, P.; Górniak, A.; Śniegucka, K.; Nawrot, K.; Korczyński, M. Effect of Lentinula edodes on morphological and biochemical blood parameters of horses. Animals; 2022; 12, 1106. [DOI: https://dx.doi.org/10.3390/ani12091106]
25. National Research Council (NRC). Nutrient Requirements of Horses; 6th ed. National Academy Press: Washington, DC, USA, 2007.
26. Satué, K.; Hernández, A.; Muńoz, A. Physiological factors in the interpretation of equine hematological profile. Hematology-Science and Practice; Laurie, C. IntechOpen: London, UK, 2012; pp. 573-596.
27. Guilliams, M.; Mildner, A.; Yona, S. Developmental and functional heterogeneity of monocytes. Immunity; 2018; 49, pp. 595-613. [DOI: https://dx.doi.org/10.1016/j.immuni.2018.10.005] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/30332628]
28. Willis, W.L.; Wall, D.C.; Isikhuemhen, O.S.; Ibrahim, S. Effect of different mushrooms fed to eimeria-challenged broilers on rearing performance. Int. J. Poult. Sci.; 2012; 11, pp. 433-437. [DOI: https://dx.doi.org/10.3923/ijps.2012.433.437]
29. Baba, E.; Uluköy, G.; Öntaş, C. Effects of feed supplemented with Lentinula edodes mushroom extract on the immune response of rainbow trout, Oncorhynchus mykiss, and disease resistance against Lactococcus garvieae. Aquaculture; 2015; 448, pp. 476-482. [DOI: https://dx.doi.org/10.1016/j.aquaculture.2015.04.031]
30. Munoz, A.; Riber, C.; Trigo, P.; Castejon, F. Age and gender related variations in hematology, clinical biochemistry, and hormones in Spanish fillies and colts. Res. Vet. Sci.; 2012; 93, pp. 943-949. [DOI: https://dx.doi.org/10.1016/j.rvsc.2011.11.009]
31. Thorén-Tolling, K. Characterization of Equine Alkaline Phosphate Isoenzymes by Chemical Inhibition and Agaros Gel Electrophoresis. J. Vet. Med.; 1988; 35, pp. 1-12. [DOI: https://dx.doi.org/10.1111/j.1439-0442.1988.tb00001.x]
32. Trumble, T.N.; Brown, M.P.; Merritt, K.A.; Billinghurst, R.C. Joint dependent concentrations of bone alkaline phosphatase in serum and synovial fluids of horses with osteochondral injury: An analytical and clinical validation. Osteoarthr. Cartil.; 2008; 16, pp. 779-786. [DOI: https://dx.doi.org/10.1016/j.joca.2007.11.008]
33. Xiao, C.; Wu, Q.P.; Tan, J.B.; Cai, W.; Yang, X.B.; Zhang, J.M. Inhibitory effects on alpha-glucosidase and hypoglycemic effects of the crude polysaccharides isolated from 11 edible fungi. J. Med. Plants Res.; 2011; 5, pp. 6963-6967.
34. Drori, A.; Shabat, Y.; Ya’acov, A.B.; Danay, O.; Levanon, D.; Zolotarov, L.; Ilan, Y. Extracts from Lentinula edodes (Shiitake) edible mushrooms enriched with vitamin D exert an anti-inflammatory hepatoprotective effect. J. Med. Food; 2016; 19, pp. 383-389. [DOI: https://dx.doi.org/10.1089/jmf.2015.0111] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/27027234]
35. Coco, M.; Buscemi, A.; Guerrera, C.S.; Di Corrado, D.; Cavallari, P.; Zappalà, A.; Di Nuovo, S.; Parenti, R.; Maci, T.; Razza, G. Effects of a bout of intense exercise on some executive functions. Int. J. Environ. Res. Public Health; 2020; 17, 898. [DOI: https://dx.doi.org/10.3390/ijerph17030898] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/32024008]
36. Westerblad, H.; Bruton, J.D.; Katz, A. Skeletal muscle: Energy metabolism, fiber types, fatigue and adaptability. Exp. Cell Res.; 2010; 316, pp. 3093-3099. [DOI: https://dx.doi.org/10.1016/j.yexcr.2010.05.019]
37. Shimada, Y.; Yamakawa, A.; Morita, T.; Sugiyama, K. Effects of dietary Eritadenine on the liver microsomal ⊿6-desaturase activity and its mRNA in rats. Biosci. Biotechnol. Biochem.; 2003; 67, pp. 1258-1266. [DOI: https://dx.doi.org/10.1271/bbb.67.1258]
38. Fukada, S.; Setoue, M.; Morita, T.; Sugiyama, K. Dietary Eritadenine suppresses guanidinoacetic acid-induced hyperhomocysteinemia in rats. J. Nutr.; 2006; 136, pp. 2797-2802. [DOI: https://dx.doi.org/10.1093/jn/136.11.2797]
39. Xu, C.; Yan, Z.H.; Hong, Z.J.; Jing, G. The pharmacological effect of polysaccharides from Lentinus edodes on the oxidative status and expression of VCAM-1mRNA of thoracic aorta endothelial cell in high-fat-diet rats. Carbohydr. Polym.; 2008; 74, pp. 445-450. [DOI: https://dx.doi.org/10.1016/j.carbpol.2008.03.018]
40. Rose, R.J.; Hodgson, D.R.; Sampson, D.; Chan, W. Changes in plasma biochemistry in horses competing in a 160 km endurance ride. Aust. Vet. J.; 1983; 60, pp. 101-105. [DOI: https://dx.doi.org/10.1111/j.1751-0813.1983.tb05905.x]
41. Wagner, M.; Halilbasic, E.; Marschall, H.U.; Zollner, G.; Fickert, P.; Langner, C.; Zatloukal, K.; Denk, H.; Trauner, M. CAR and PXR agonists stimulate hepatic bile acid and bilirubin detoxification and elimination pathways in mice. Hepatology; 2005; 42, pp. 420-430. [DOI: https://dx.doi.org/10.1002/hep.20784]
42. Delvescovo, B.; Tomlinson, J.; DeNotta, S.; Hodge, E.; Bookbinder, L.; Mohammed, H.O.; Divers, T.J. Bile acids, direct bilirubin and gamma-glutamyltransferase as prognostic indicators for horses with liver disease in the Eastern United States: 82 cases (1997–2019). J. Equine Vet. Sci.; 2021; 105, 103729. [DOI: https://dx.doi.org/10.1016/j.jevs.2021.103729]
43. Fazio, F.; Assenza, A.; Tosto, F.; Casella, S.; Piccione, G.; Caola, G. Training and haematochemical profile in Thoroughbreds and Standardbreds: A longitudinal study. Livest. Sci.; 2011; 141, pp. 221-226. [DOI: https://dx.doi.org/10.1016/j.livsci.2011.06.005]
44. Piccione, G.; Arfuso, F.; Marafioti, S.; Giannetto, C.; Giudice, E.; Fazio, F. Different training schedules influence serum electrophoretic protein profile in the athletic horse. J. Equine Vet. Sci.; 2015; 35, pp. 856-859. [DOI: https://dx.doi.org/10.1016/j.jevs.2015.08.008]
45. Assunçăo, P.; Barbosa, T.; Yonezawa, L.; Barbosa, L.; Watanabe, M.; Kohayagawa, A.; Schmidt, E. Acute-phase protein profile in horses subjected to different exercise protocols. Can. J. Vet. Res.; 2019; 83, pp. 272-278.
46. Scoppetta, F.; Tartaglia, M.; Renzone, G.; Avellini, L.; Gaiti, A.; Scaloni, A.; Chiaradia, E. Plasma protein changes in horse after prolonged physical exercise: A proteomic study. J. Proteom.; 2012; 75, pp. 4494-4504. [DOI: https://dx.doi.org/10.1016/j.jprot.2012.04.014]
47. Baba, E.; Uluköy, G. Effects of Dietary Supplemented Shiitake Mushroom Extract on Growth, Non-specific Immune Parameters and in-vitro Resistance against Aeromonas hydrophila in Rainbow Trout (Oncorhynchus mykiss). J. Limnol. Fish. Res.; 2022; 8, pp. 28-36. [DOI: https://dx.doi.org/10.17216/limnofish.898170]
48. Yoon, K.N.; Alamm, N.; Lee, J.S.; Cho, H.J.; Kim, H.Y.; Shim, M.J.; Lee, M.W.; Lee, T.S. Antihyperlipidemic effect of dietary Lentinus edodes on plasma, feces and hepatic tissues in hypercholesterolemic rats. Mycobiology; 2011; 39, pp. 96-102. [DOI: https://dx.doi.org/10.4489/MYCO.2011.39.2.096] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/22783084]
49. Yu, S.; Wu, X.; Ferguson, M.; Simmen, R.; Cleves, M.; Simmen, F.; Fang, N. Diets containing shiitake mushroom reduce serum lipids and serum lipophilic antioxidant capacity in rats. J. Nutr.; 2016; 146, pp. 2491-2496. [DOI: https://dx.doi.org/10.3945/jn.116.239806] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/27798348]
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Details
; Górniak, Wanda 2
; Zielińska, Paulina 3
; Górniak, Aleksander 2
; Čebulj-Kadunc, Nina 4
; Korczyński, Mariusz 5
1 Institute of Animal Breeding, Wroclaw University of Environmental and Life Sciences, Chelmonskiego 38C, 51-630 Wroclaw, Poland
2 Department of Automotive Engineering, Mechanical Faculty, Wroclaw University of Science and Technology, Na Grobli 13, 50-421 Wroclaw, Poland
3 Department of Surgery, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, Plac Grunwaldzki 51, 50-366 Wroclaw, Poland
4 Institute of Preclinical Sciences, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, 1000 Ljubljana, Slovenia
5 Department of Animal Nutrition and Feed Management, Wroclaw University of Environmental and Life Sciences, Chelmonskiego 38C, 51-630 Wroclaw, Poland