Original study Arch. Anim. Breed., 65, 135–144, 2022 Open Access https://doi.org/10.5194/aab-65-135-2022 © Author(s) 2022. This work is distributed under Archives Animal Breeding the Creative Commons Attribution 4.0 License. Long-term administration of a commercial supplement enriched with bioactive compounds does not affect feed intake, health status, and growth performances in beef cattle Marica Simoni1 , Arianna Goi2 , Erika Pellattiero2 , Alexandros Mavrommatis3 , Eleni Tsiplakou3 , Federico Righi1 , Massimo De Marchi2 , and Carmen L. Manuelian2 1 Department of Veterinary Science, University of Parma, Via del Taglio 10, 43126, Parma, Italy 2 Department of Agronomy, Food, Natural resources, Animals and Environment, University of Padova, Viale dell’Università 16, 35020, Legnaro, Italy 3 Laboratory of Nutritional Physiology and Feeding, Department of Animal Science, School of Animal Biosciences, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece Correspondence: Arianna Goi (arianna.goi@unipd.it) Received: 22 June 2021 – Revised: 14 February 2022 – Accepted: 7 March 2022 – Published: 1 April 2022 Abstract. Feed additives including natural bioactive compounds (BCs) in combination with vitamin E (VitE) and organic Se could mitigate animal stress associated with intensive livestock farming due to their anti- inflammatory and antioxidant properties. Yeast and yeast derivate are included in feed additives as probiotic products and digestion promoters. Scutellaria baicalensis is a source of bioactive compounds and has been tested in monogastrics, exhibiting many immunostimulating and hepato-protective activities. However, the liter- ature lacks information regarding S. baicalensis effects on beef cattle performance and health status. The aim of the present study was to evaluate the impact on beef cattle’s feed intake, health and oxidative status, and growth performances of the inclusion of a commercial supplement (CS) containing VitE, organic Se, yeast derivate, and S. baicalensis extract during the fattening and finishing period. A total of 143 Charolaise male cattle were allotted into 12 pens of 11–12 animals each and assigned to a control (463.9 ± 21.48 body weight – BW) or a treated (469.8 ± 17.91 BW) group. Each group included two replicates of three pens. The treated groups were supplemented with 20 g CS animal−1 d−1 . Feed intake was measured monthly on a pen base during two consec- utive days. Total mixed ration and fecal samples were collected at three time points (monthly, from November to February) and pooled by replicate for the analyses to monitor digestibility. Blood samples were individually collected at the beginning and at the end of the trial for oxidative status and metabolic profile determination. Final BW and carcass weight were individually recorded to calculate average daily gain, feed conversion ratio, and carcass yield. Similar feed digestibility between groups were observed during the whole experiment. Feed intake, growth performances, final body weight, average daily gain, feed conversion rate, oxidative status, and metabolic profile were not affected by the dietary inclusion of the tested CS indicating no detrimental effect of the treatment. Different doses of this product should be tested in the future in order to provide a more complete report on the product efficacy. Published by Copernicus Publications on behalf of the Research Institute for Farm Animal Biology (FBN). 136 M. Simoni et al.: Long-term administration of a commercial supplement 1 Introduction Króliczewska et al., 2004, 2008, 2017; Kwon et al., 2009). The dietary inclusion of a highly bioavailable form of S. Intensification of livestock farming can be associated with baicalensis extract obtained by enzymatic bio-conversion by several management, environmental, and nutritional stressors pure enzyme or whole cell culture showed comparable ef- that increase animals’ susceptibility to disease (Bhimte et fects with apramycin antibiotic on growth performance, nu- al., 2021; Endris et al., 2021; Fernandez-Novo et al., 2020). trient digestibility, fecal microbiota, fecal gas emission, and However, antibiotics use in livestock diets as growth promot- blood hematology and improved pigs’ average daily gain ers or to reduce the rumen dysfunctions is restricted, which (ADG) and final body weight (BW) (Dang et al., 2021). has led to the recruitment and study of new natural feed ad- Additionally, it has been demonstrated that the dietary sup- ditives to improve growth and feed efficiency (Balcells et plementation with S. baicalensis in piglets decreased diar- al., 2012; Dang et al., 2021). Hence, there is an interest in rhea frequency and increased the feed conversion ratio (C. testing natural bioactive compounds capable of expressing Huang et al., 2019). The inclusion of S. baicalensis extract anti-inflammatory and antioxidant activities to enhance live- or root also increased the final BW and feedstuff consump- stock health and performance as well as to improve prod- tion in broilers (Króliczewska et al., 2004, 2008; Kwon et al., uct quality (Righi et al., 2021, Tsiplakou et al., 2021, Pitino 2009) without affecting meat quality and chemical composi- et al., 2021, Manuelian et al., 2021). Bioactive compounds tion (Króliczewska et al., 2008). On the other hand, an ex- can be associated with minerals and vitamins that poten- cess of S. baicalensis root impaired the development of the tially enhance their action. Micronutrients such as Se, and immune organs in chickens (Króliczewska et al., 2017). especially its organic form, help to keep the correct func- In ruminants, some evidence indicates that S. baicalen- tionality of the immune system participating in the antiox- sis could interact with the rumen ecology, altering rumen idant defenses (Sgoifo Rossi et al., 2020) and are involved microbiome, i.e. increasing the prevalence of the phylum in the thyroid hormone metabolism affecting growth perfor- Firmicutes, Clostridia class, and Ruminococcaceae family mances and health status in a dose-dependent mode (Mehdi (Yausheva et al., 2019), which suggests a possible effect on and Dufrasne, 2016). Dietary vitamin E (VitE), a well-known diet digestibility. Recent literature reports that flavonoids, as antioxidant, modulates immune system function (Lewis et powerful antimicrobials, can increase ruminal volatile fatty al., 2019). Similar effects have been summarized for dry acids production, while reducing ammonia and methane pro- yeast, cell wall yeast, and other yeast derivatives (Broad- duction (Kalantar, 2018); however, in vitro results were not way et al., 2015; Shurson, 2018), which are added as sup- always consistent (Balcells et al., 2012). The dietary long- plements in animal feed due to their high nutritional value term supplementation (60 d) with S. baicalensis extract re- and their content in nutraceutical bioactive compounds (e.g. duced somatic cell counts and increased milk yield in early β-glucans, mannanoligosaccharides, nucleotides) that have lactating cows (Olagaray et al., 2019). However, few studies been demonstrated to improve animal health, immune status, have been conducted on ruminants (Olagaray et al., 2019). and growth performance (Shurson, 2018). Thus, S. baicalensis bioavailability and its effect on produc- Recently, Chinese medicinal herbs have attracted the at- tion, health, and metabolic profile need to be further explored tention of many researchers as possible sources of natural through in vivo trials, particularly in beef cattle, where, to the bioactive compounds in animals’ production. The roots of best of our knowledge, only one study tested S. baicalensis Scutellaria baicalensis have traditionally been used in Chi- (Song et al., 2014). This study demonstrated the positive ef- nese herbal medicine to treat liver and lung diseases, hy- fect of a mix of herbs including S. baicalensis on digestibility pertension, acute respiratory infections, acute gastroenteri- and growth performance in beef cattle when evaluated under tis, infantile diarrhea, and morning sickness (Zhao et al., heat-stress conditions. 2019; Nurul et al., 2011). The bioactive compounds isolated The aim of the present study was to evaluate the effect on from this plant are flavonoids, terpenoids volatile, oils, and beef cattle intake, growth performances, and metabolic pro- polysaccharides, which exhibit many activities and effects on file of the addition of a commercial supplement (CS) includ- the immune system and liver protection (Zhao et al., 2019). ing VitE, organic Se, and S. baicalensis during the fattening The four main flavonoids of S. baicalensis, namely baicalein, and finishing periods. baicalin, wogonoside, and wogonin, have demonstrated anti- tumor, antibacterial, antiviral, antioxidant, antihypertensive, 2 Materials and methods and hepato-protective effects when individually evaluated (T. Huang et al., 2019; Huynh et al., 2020; Olagaray et al., 2019; 2.1 Product tested Vergun et al., 2019), which suggests a potential beneficial ef- fect of the whole plant on the animal organism. The ingredients of the CS for young cattle tested in the In fact, the antioxidants and anti-inflammatory proper- present study (IAR4045 AXION BEEF SE; ITACA Europa ties of S. baicalensis have been tested in pigs (Dang et al., SA, Italy), as declared by the manufacturer, were calcium 2021), and in chickens (at 0.1 %, 0.5 %, and 1.5 % of the carbonate, yeast products, and S. baicalensis extract and in- diet) and broilers (at 0.1 %, 0.5 %, and 1.5 % of the diet; cluded 75 000 UI kg−1 of VitE, 20 mg kg−1 of hydroxylated Arch. Anim. Breed., 65, 135–144, 2022 https://doi.org/10.5194/aab-65-135-2022 M. Simoni et al.: Long-term administration of a commercial supplement 137 Table 1. Gross composition (mean) and detailed macromineral and the next 2 months and a finishing diet for the rest of the micromineral (mean ± standard deviation) characterization of the trial. The diets, whose compositions are reported in Table 2, tested commercial supplement. were formulated to meet the nutritional requirements sug- gested in the NRC (2016) for beef cattle. At the beginning Trait Content of the fattening period the pens were assigned to one of two Gross composition dietary groups: control (CON; age: 307.86 ± 22.87 d; BW: 465.15±21.42 kg) and treatment (CS; age: 304.86±22.89 d; DM∗ , % as fed 94.67 BW: 467.48 ± 19.56 kg), which received 20 g d−1 per animal Crude protein, % DM 2.77 of the CS product as a top dressing in the feedbunk accord- Ether extract, % DM 0.13 Crude fiber, % DM 1.27 ing to the manufacturer recommendations. Each thesis was Ash, % DM 86.75 replicated in two subgroups of three pens. Animals of both groups were slaughtered at the end of the trial on 6 and 13 Macromineral content g kg−1 March due to the schedule of the slaughterhouse. Ca 189.00 ± 3.00 Mg 4.55 ± 0.06 2.3 Sampling and measurements S 5.61 ± 0.05 P 0.69 ± 0.01 Feed intake was calculated on a pen basis once a month from Na 5.21 ± 0.02 November to February for two consecutive days by mea- K 2.82 ± 0.01 suring the difference between the amount of feed delivered Micromineral content mg kg−1 and residues. A sub-sample (1 kg) of the total mixed ration (TMR) offered to the animals was collected monthly from Zn 420 ± 10 November to February from each pen during the feed de- Cu 19.3 ± 0.6 livery and pooled by a subgroup of three pens (replicate) Fe 9420 ± 80 for their chemical analysis. Fecal samples were also col- Mn 194 ± 1 Se 17 ± 2 lected monthly from November to January from each pen Si 170 ± 10 and pooled by replicate for their posterior analysis. The fe- cal samples were collected from the rectum from at least five ∗ DM: dry matter. animals per pen to get a representative sample from each pen. Chemical composition of the TMR and fecal samples were determined as described in Simoni et al. (2021). Briefly, sam- analog of selenomethionine, and aromatic substances. The ples were dried at 55 ◦ C for 48 h (for TMR) and 72 h (for mineral content of the CS was determined through spec- feces) and then ground in a Cyclotec mill (Tecator, Hern- trophotometric analysis using an ICP-AES spectrophotome- don, VA, USA) to pass a 1 mm screen. The dry matter (DM) ter model ULTIMA 2 (HORIBA Jobin Yvon Inc., 3880 content was determined by drying the samples at 103 ◦ C Park Avenue, Edison, NJ, USA) after microwave miner- overnight, and the fiber fractions were analyzed and ex- alization performed through the Milestone system, model pressed as aNDFom (neutral detergent fiber assayed with a MLS-1200 MEGA (Gemini sustainable laboratory equip- heat-stable amylase and expressed exclusive of residual ash), ment, 7312 DG Apeldoorn, the Netherlands). Chemical com- ADFom (acid detergent fiber expressed exclusive of resid- position analysis of the CS was performed as described be- ual ash) and lignin(sa) (lignin determined by solubilization low in the “Sampling and measurements” section. Mineral of cellulose with sulfuric acid). All the fiber fractions were content and chemical composition are reported in Table 1. assayed in duplicate. The nitrogen (N) content was measured The manufacturer-recommended dose to be administered per by the combustion digestion of the samples at 900 ◦ C in ex- animal was 20 g d−1 . cess of oxygen by Dumatherm® (Gerhardt GmbH & Co, Königswinter, Germany). Ash content was determined by 2.2 Experimental design, animals, and housing ignition at 550 ◦ C. The starch content in the TMR samples was determined by the polarimetric method. The undigested The study was carried out in a feedlot located in northern NDF (uNDF) was obtained by fermentation of 240 h in an in Italy (latitude 45.6705, longitude 12.4212). A total of 143 vitro batch system using rumen fluid collected at the slaugh- male Charolaise cattle imported from France were involved terhouse from five adult cows. The uNDF was used for the in the study and monitored throughout the fattening and fin- estimation of the in vivo apparent total-tract digestibility of ishing period for 159.7 d (SD, 3.5 d) from October 2019 to nutrients as described by Righi et al. (2017) and Simoni et al. the first half of March 2020. The animals were allotted to (2021). The mineral content of the TMR was determined by 12 pens, each one containing 11 or 12 animals, and were X-ray fluorescence according to Berzaghi et al. (2018). previously fed with an adaptation diet for the first month Individual beef BW was measured at the beginning and at (October 2019). Then, a fattening diet was administered for the end of the trial to calculate the average daily gain (ADG). https://doi.org/10.5194/aab-65-135-2022 Arch. Anim. Breed., 65, 135–144, 2022 138 M. Simoni et al.: Long-term administration of a commercial supplement Table 2. Ingredients (kg per animal as feed) and chemical compo- tory disease”, and “other health issues”. The specific antibi- sition (mean ± standard deviation) of fattening and finishing beef otic administered to each animal was also recorded (length cattle total mixed rations administered during the trial. of the treatment and drug used). Individual blood samples were simultaneously collected Trait Fattening diet Finishing diet during the official sanitary check at the beginning of Ingredient composition Kilogram per animal as feed the trial and during slaughter using 10 mL vacuum tubes containing lithium heparin (Terumo Venosafe 10 mL VF- Feedstuff1 1.4 1.4 109SHLTerumo Europe L.V., Leuven, Belgium). Plasma was Distiller 1.0 0.9 extracted from blood samples by centrifugation at 3000 g Corn gluten meal 0.5 0.4 for 15 min. The plasmatic α-tocopherol concentration was Alfalfa hay, 15 % of CP 0.3 0.3 Corn meal 3.0 determined by an iCheck fluorometer–spectrophotometer Cane molasses 0.7 0.8 (iCheck Vitamin E; BioAnalyt GmbH, Teltow, Germany). Wheat straw 0.4 0.4 The metabolic profile of plasma samples included bilirubin, Earlage2 1.8 1.8 aspartate aminotransferase (AST-GOT), gamma glutamyl- Dry beet pulp 0.6 0.5 transferase (GGT), creatine kinase, total protein, albumin, Corn silage3 7.0 6.8 urea, glucose, total cholesterol, triglycerides, and minerals Triticum silage4 1.0 1.0 (Ca, Mg, and P). Non-esterified fatty acids (NEFA) and β- hydroxybutyrate acid (BHBA) were also determined to mon- Chemical composition5 Mean ± standard deviation itor the energy balance of the animals. The total antioxidant Dry matter (DM), % of feed 59.99 ± 1.64 59.62 ± 1.41 capacity (FRAP: ferric reducing ability power; ABTS: 2,2’- CP, % DM 12.90 ± 0.39 12.93 ± 0.30 azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) and cata- aNDFom, % DM 30.38 ± 2.15 29.94 ± 1.00 lase (CAT) activity were determined spectrophotometrically ADFom, % DM 15.28 ± 1.31 13.01 ± 0.73 (GENESYS 180, Thermo Fisher Scientific, Massachusetts, Lignin (sa), % DM 2.75 ± 0.64 3.53 ± 1.21 USA) in plasma, aiming to assess the oxidative status of the Starch, % DM 32.96 ± 2.11 36.51 ± 2.30 animals (Tsiplakou et al., 2018). Ash, % DM 6.41 ± 0.72 6.34 ± 0.61 Ca, % DM 0.811 ± 0.07 0.943 ± 0.05 At the end of the trial, all cattle were transported to a com- P, % DM 0.289 ± 0.00 0.298 ± 0.01 mercial slaughterhouse. The BW at slaughter and the car- Mg, % DM 0.174 ± 0.00 0.174 ± 0.01 cass weight were recorded individually, and the proportion Cl, % DM 0.575 ± 0.10 0.427 ± 0.03 of carcass yield was calculated as the proportion of the car- K, % DM 0.836 ± 0.01 0.825 ± 0.02 cass weight on the BW at slaughter. Due to the Covid-19 Na, % DM 0.236 ± 0.04 0.218 ± 0.03 outbreak, it was not possible to perform measurements and S, % DM 0.218 ± 0.01 0.195 ± 0.02 sampling on organs and carcass except for the weight of the Fe, % DM 206.4 ± 35.21 232.2 ± 22.01 latter that was provided by the electronic scale of the slaugh- Cu, % DM 9.85 ± 0.17 11.3 ± 0.99 terhouse. Zn, % DM 78.23 ± 7.58 80.8 ± 6.10 Mn, % DM 40.85 ± 0.75 49.1 ± 4.23 Si, % DM 1.32 ± 0.06 1.45 ± 0.06 2.4 Statistical analyses DCAD6 , mEq kg−1 1.85 ± 0.72 6.43 ± 2.70 Sources of variation of performance traits were investigated 1 Feedstuff provided in a pelleted form and composed by rapeseed meal, soybean using the GLM procedure of SAS version 9.4 (SAS Inst. Inc., meal 46 % CP, sunflower meal, corn gluten meal, wheat middlings, wheat meal, calcium carbonate, sodium chloride, cane molasses, dry extract of Aspergillus oryzae, Cary, NC) according to the following linear model: Yucca schidigera, hydrolyzed lignocellulose, sodium bicarbonate, corn meal, magnesium oxide, yeasts. 2 Earlage, 64 % DM, 58 % starch, 14 % NDF. yij kl = µ + feedingi + boxj (feeding)i + iBWk + εij kl , (1) 3 Corn silage, 34 % DM, 33 % starch, 44 % NDF. 4 Triticum silage, 30 % DM, 50 % NDF, 29 % NFC. where yij kl is the dependent variable; µ is the overall in- 5 Analyzed chemical composition – aNDFom: neutral detergent fiber assayed with a heat-stable amylase and expressed exclusive of residual ash; ADFom: acid detergent tercept of the model; feedingi is the fixed effect of the fiber expressed exclusive of residual ash; lignin(sa): lignin determined by ith CS supplementation (i = 1 and 2, where 1 means non- solubilization of cellulose with sulfuric acid; CP: crude protein. 6 DCAD: dietary cation–anion difference. supplemented and 2 means supplemented); boxj (feeding)i is the fixed effect of the j th box (j = 1 to 12) nested within the ith CS supplementation; iBWk is the fixed effect of the kth initial BW class (k = 1 to 3, where 1 = iBW ≤ 460 kg; The feed conversion ratio (FCR) was calculated at the be- 2 = 460 kg > iBW ≤ 475 kg; 3 = iBW > 475 kg); and εij kl ginning of the fattening period and at the end of the trial as a is the random residual ∼ N(0, σe2 ), where σe2 is the resid- ratio between the DM intake and the BW. The health status of ual variance. The feeding effect was tested using “box within the animals was assessed regularly by visual evaluation, and feeding” as the error term, whereas the other fixed effects pathologies detected were classified as “lameness”, “respira- were tested on the residual. Arch. Anim. Breed., 65, 135–144, 2022 https://doi.org/10.5194/aab-65-135-2022 M. Simoni et al.: Long-term administration of a commercial supplement 139 For the comparison of the slaughter weights, the same animals was acceptable and abnormal behavior was not ob- model was used but the initial body weight was introduced served, we decided to keep the treated animals within the as covariate in the model. study. The statistical analysis performed on the sum of health Sources of variation of the TMR and feces chemical com- issues did not show significant differences between groups. position as well as diet digestibility were investigated using A physiological increase in feed intake (on a DM ba- the GLM procedure of SAS according to the following linear sis) was observed moving from the fattening to the finish- model: ing period (P < 0.001). On average, daily overall intake was around 10 kg of DM per animal, moving from 9.88 ± 0.104 yij kl = µ + feedingi + boxj (feeding)i + Timek + εij kl , (2) to 10.45 ± 0.104 kg of DM per beef cattle in the fattening and finishing period, respectively (P < 0.001). No signifi- where yij kl is the dependent variable; µ is the overall in- cant differences were observed between feeding groups (Ta- tercept of the model; feedingi is the fixed effect of the ble 3). ith CS supplementation (i = 1 and 2, where 1 means non- A similar fecal composition was observed between the supplemented and 2 means supplemented); boxj (feeding)i is two dietary groups (P > 0.05). The average fecal DM was the fixed effect of the j th box (j = 1 to 4) nested within the 17.94 % (P = 0.451), while the average content of CP (P = ith CS supplementation; Timek is the fixed effect of the kth 0.928), aNDFom (P = 0.412), ADFom (P = 0.468), and time class (k = 1 to 4, where 1 = November, 2 = December; lignin(sa) (P = 0.143) was 17.2 %, 51.2 %, 25.7 %, and 3 = January, and 4 = February); and εij kl is the random 6.74 % of DM, respectively. Moreover, a similar apparent residual ∼ N(0, σe2 ), where σe2 is the residual variance. The total-tract digestibility of DM and NDF (% on DM) between feeding effect was tested using “box within feeding” as the groups was obtained (DM, 70.8 % and 73.59 % for CON error term, whereas the other fixed effects were tested on the and CS groups, respectively, P = 0.411; NDF: 50.42 % and residual. 54.98 % for CON and CS groups, respectively, P = 0.498). Sources of variation of the blood parameters were inves- Performance traits retrieved at slaughter showed that di- tigated using the GLM procedure of SAS according to the etary supplementation with CS did not affect the BW at following linear model: slaughter (Table 3), which was on average 721 kg. The tested product did not affect carcass weight, carcass yield, ADG, yij kl = µ + feedingi + boxj (feeding)i + εij kl , (3) or FCR (Table 3), being on average 439.54 kg, 60.96 %, 1.59 kg d−1 , and 10.28 kg (kg BW)−1 , respectively. The di- where yij kl is the dependent variable; µ is the overall inter- etary supplementation with CS did not impair blood α- cept of the model; feedingi is the fixed effect of the ith CS tocopherol content, antioxidant status, CAT, or the metabolic supplementation (i = 1 and 2, where 1 = non supplemented; profile (Table 4). On average, blood α-tocopherol content, to- 2 = supplemented); boxj (feeding)i is the fixed effect of the tal antioxidant capacity (with FRAP and ABTS assays), and j th box (j = 1 to 4) nested within the ith CS supplementa- CAT activity were respectively 3.30 mg L−1 , 0.66 µM ascor- tion; and εij kl is the random residual ∼ N(0, σe2 ), where σe2 bic acid, 29.78 % inhibition, and 2.56 units mL−1 , whereas is the residual variance. The feeding effect was tested using NEFA, total cholesterol, and triglycerides content were 0.28, box within feeding as the error term, whereas the other fixed 2.65, and 0.17 mmol L−1 , respectively. Similar results be- effects were tested on the residual. tween groups were also observed for the liver parameters CK Sources of variation of FRAP, ABTS, and CAT were in- and BHBA, which average values where 388.38 IU L−1 and vestigated using the GLM procedure of SAS using the initial 0.26 mmol L−1 , respectively. value as covariate and the dietary treatment was the fixed ef- fect. For all the models, multiple comparison of least-square means was performed for the fixed effects using Bonferroni’s 4 Discussion test (P < 0.05). The frequency of health issues was compared between Despite the increasing interest of livestock producers and groups using the chi square test; the health issues were ex- feed additive suppliers in using natural feed additives to im- pressed as the sum of lameness, respiratory problems, and prove livestock health and performance as a consequence of other health problems recorded. Due to the low number of the ban in the European Union of growth promoters in 2006 animals that needed a treatment, these results are reported as (European Commission, 2003), in vivo trials information on total number of cases. their use is still lacking, in particular when referring to non- poultry species. Few studies have been conducted with S. 3 Results baicalensis, the majority having been on pigs (Dang et al., 2021; C. Huang et al., 2019) and chickens (Króliczewska et Few animals in both groups presented health issues during al., 2004, 2008, 2017; Kwon et al., 2009) and only one on the whole study (17 out of 143 beef cattle) that needed an- beef (Song et al., 2014). Therefore, the present study con- tibiotic treatment. Because the overall health status of these tributes to increasing the scientific knowledge on the inclu- https://doi.org/10.5194/aab-65-135-2022 Arch. Anim. Breed., 65, 135–144, 2022 140 M. Simoni et al.: Long-term administration of a commercial supplement Table 3. Average individual feed intake and growth performance traits (least-square mean and standard error of the mean – SEM) of control (CON) and commercial supplement (CS) groups. Trait CON CS SEM P value Intake, kg DM 10.22 10.14 0.131 0.658 Intake in the fattening period, kg DM 9.87 9.89 0.147 0.269 Intake in the finishing period, kg DM 10.58 10.39 0.147 0.269 Body weight at slaughter, kg 722.34 720.03 3.652 0.712 Carcass weight, kg 441.10 437.98 2.380 0.577 Carcass yield, %1 61.10 60.83 0.173 0.637 ADG2 1.60 1.58 0.019 0.709 FCR3 10.27 10.29 0.077 1.000 1 Calculated as percentage of live body weight at slaughter. 2 ADG: average daily gain. 3 FCR: feed conversion ratio. Table 4. Plasmatic oxidative status, and metabolic profile (least-square mean and standard error of the mean – SEM) of animals in the control (CON) and commercial supplement (CS) groups. Trait∗ CON CS SEM P value Oxidative status α-Tocopherol, mg L−1 3.11 3.54 0.101 0.202 FRAP, µM ascorbic acid 0.65 0.72 0.030 0.528 ABTS, inhibition % 29.7 29.5 0.229 0.768 CAT, units mL−1 2.76 2.55 0.099 0.303 Metabolic profile Total protein, g L−1 74.36 73.81 0.551 0.646 Albumin, g L−1 33.55 33.96 0.270 0.622 Globulin, g L−1 40.81 39.84 0.518 0.427 Albumin / globulin ratio 0.83 0.86 0.014 0.356 Urea, mmol L−1 2.90 2.81 0.087 0.543 NEFA, mmol L−1 0.31 0.23 0.013 0.158 Glucose, mmol L−1 6.48 6.41 0.162 0.635 Total cholesterol, mmol L−1 2.74 2.75 0.073 0.532 Triglycerides, mmol L−1 0.16 0.18 0.005 0.690 AST-GOT, IU L−1 78.86 80.64 1.158 0.938 GGT, IU L−1 24.17 25.15 0.571 0.395 Bilirubin, µmol L−1 1.18 0.78 0.064 0.223 Creatine kinase, IU L−1 365.43 439.73 27.270 0.307 Ca2+ , mmol L−1 2.57 2.62 0.027 0.196 P3+ , mmol L−1 2.65 2.63 0.035 0.730 Mg2+ , mmol L−1 1.02 1.01 0.019 0.607 BHBA, mmol L−1 0.27 0.27 0.007 0.861 ∗ FRAP: ferric reducing ability of plasma; ABTS: 2,2’-azino-di(3-ethylbenzthiazoline-6-sulfonic acid); CAT: catalase; NEFA: nonesterified fatty acids; AST-GOT: aspartate aminotransferase; GGT: gamma glutamyltransferase; BHBA: β -hydroxybutyric acid. sion of bioactive compounds from S. baicalensis during the quent legal restrictions to movement of people and to access fattening and finishing phases when rearing beef cattle. It to the slaughterhouse. should be mentioned that data collection on the organs and The chemical composition of the fattening and finishing carcass traits, including meat quality and composition, in or- diets in the present study were in line with those reported by der to completely evaluate the effects of the addition of CS other authors in Charolaise beef cattle (Cortese et al., 2019, was not possible due to the Covid-19 outbreak and the subse- Sgoifo Rossi et al., 2019). The NDF and CP levels were par- Arch. Anim. Breed., 65, 135–144, 2022 https://doi.org/10.5194/aab-65-135-2022 M. Simoni et al.: Long-term administration of a commercial supplement 141 ticularly similar to those reported by Cortese et al. (2019), logical status, blood biochemical parameters, nutrient appar- and starch level variation was comparable to those indicated ent digestibility, and growth performance under heat-stress by Sgoifo Rossi et al. (2019). The addition of the CS did not conditions (Song et al., 2014). In the present study, we did significantly modify the chemical composition of the diet. not find any of these effects when supplementing beef fat- This aspect is important to make comparable results from tening and finishing diets (Table 3) with CS, probably be- both groups, CON and CS. Moreover, feed intake observed in cause intake (Table 3), feed chemical composition, and feed the fattening and finishing phases is consistent with the phys- digestibility between groups were similar. No effect of the iological growth of the animals, and it is comparable to the organic Se was detected. This is in agreement with the liter- findings from both Cortese et al. (2019) and Sgoifo Rossi et ature, which explains the lack of visible effects of Se sup- al. (2019) on similar animals. Furthermore, the level of intake plementation in several studies on calves, cows, and bulls observed in our study, combined with the diet energy and (Mehdi and Dufrasne, 2016). protein amount, was the expected one to allow the relatively In our study, the metabolic profile results were within the high ADG observed. These results support that the addition normal range for beef animals (Alcantara et al., 2020) and of the product as a top dressing in the feedbunk did not im- similar between dietary groups. Also, plasmatic levels of pair the TMR palatability (Baumont, 1996). Our results are VitE, which were within the normal range for beef cattle in agreement with the ones reported by Swecker et al. (2008) (Hidiroglou et al., 1988), did not differ between groups even showing that parenteral administration of two doses of Se if the CS also included VitE. This agrees with Swecker et or/and VitE did not affect ADG in weaned beef calves. On al. (2008), who did not find VitE plasmatic differences ei- the other hand, Sgoifo Rossi et al. (2020) reported that the ther, despite administering the two doses of VitE or/and Se mineral supplementation with organic Se along with other to calves via parenteral, whereas O’Grady et al. (2001) found organic minerals (Zn, Cu, and Mn) improved BW and ADG an increase in plasma and in muscle α-tocopherol content in in Charolaise beef during the fattening period but achieved cattle fed 300 IU of α-tocopherol per kilogram of feed, with a similar FCR of those supplemented with minerals from an a decreased lipid and oxymyoglobin oxidation. The latter au- inorganic source. thor reported no differences in plasma Se content in cattle Although S. baicalensis has been suggested to modify ru- fed 0.3 mg kg−1 of organic Se and also no differences in the men microbiome increasing Firmicutes phylum, Clostridia susceptibility to lipid and oxymyoglobin oxidation in pres- class, and the Ruminococcaceae family in an in vivo exper- ence or absence of VitE. The amount of Se supplemented iment (Yausheva et al., 2019), in our study we did not see in the present study was probably too low to see an effect; an effect on diet digestibility which might be related to the moreover, since blood Se is bound to globulins (α- and β-), low doses of bioactive compounds, minerals, and vitamins in albumin, and low- and very low-density lipoproteins (Mehdi the tested product. In fact, there is an absence of differences and Dufrasne, 2016) that were not affected by CS supple- in the estimated total tract apparent digestibility of nutrients mentation, the Se transport itself would not be enhanced of the animals. Moreover, the absence of differences in di- in this case. The β-glucans in the yeast cell wall may im- gestibility is in contrast with conclusions of Shurson (2018) prove immune competence in young animals; they can ad- in his review on yeast and yeast derivatives’ effects on ru- sorb or bind toxins, viruses, and several pathogenic bacte- minant digestion. In fact, the latter author reported that in ria, while other components of the yeast cell wall (e.g. man- many studies performed on ruminants the supplementation nanoligosaccharides) serve as prebiotics and have antioxi- with yeast or yeast products improved fiber intake and diges- dant and anti-mutagenic effects (Shurson, 2018). Similarly, tion, enhancing cellulolytic and fibrolytic growth. Se yeast supplements enhanced antioxidant status, immune The supplementation with yeast and their derivatives responses, and anti-radicalic activity (Mehdi and Dufrasne, demonstrated to improve growth performances in several 2016; Sgoifo Rossi et al., 2017), while VitE as an exoge- studies conducted on ruminants (Broadway et al., 2015; nous antioxidant improves cellular membrane stability. Sev- Shurson, 2018). On the other hand, pigs supplemented with eral flavonoids have been demonstrated to have the capacity S. baicalensis extract improved ADG, final BW (Dang et al., to trap free radicals, including reactive nitrogen species and 2021), and FCR (C. Huang et al., 2019) and reduced diar- reactive oxygen species (ROS) as well as chelating metals, rhea frequency (C. Huang et al., 2019). In broilers, greater whereas phenolic acids act as antioxidants mainly by scav- final BW has been reported. However, this improvement enging free radicals (Manuelian et al., 2021). Furthermore, can be related to the higher intake observed in the studies baicalin included in S. baicalensis at high doses induced a (Króliczewska et al., 2004, 2008; Kwon et al., 2009). It is reduction in ROS production associated with increased intra- worth mentioning that an excess of S. baicalensis root has cellular concentrations of catalase in bovine mammary ep- been reported to reduce the development of the immune or- ithelial cells (Perruchot et al., 2019). Despite the promising gans in chickens (Króliczewska et al., 2017). The only study effects of this bioactive compounds, in the present study there conducted in beef revealed the positive effect of a mix of were no differences between the two groups, neither in the herbs including S. baicalensis on digestibility and growth total antioxidant capacity nor in catalase activity of blood performances in beef cattle’s body temperature, endocrino- plasma. https://doi.org/10.5194/aab-65-135-2022 Arch. Anim. Breed., 65, 135–144, 2022 142 M. Simoni et al.: Long-term administration of a commercial supplement 5 Conclusions Review statement. This paper was edited by Manfred Mielenz and reviewed by two anonymous referees. The results of the present study revealed that the supplemen- tation with a CS that includes VitE, organic Se, yeast deriva- tive, and S. baicalensis added as a top dressing during the References fattening and finishing period did not impair beef cattle feed Alcantara, U. A. A., Rebouças, R. A., Assis, R. N. de, Souza, L. M. intake and digestibility. Moreover, supplementation did not de, Oliveira Filho, E. F. de, Mendonça, C. L. de, Soares, P. C., have a detrimental or beneficial impact on growth perfor- and Afonso, J. A. B.: Metabolic profile of cattle receiving “Max mances. 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