Zootechnical additive based on resin acids and plant extracts for use in animal feed

Abstract

The present invention relates to a natural zootechnical additive for improving performance, composed of resin acids extracted from Pinus resin, with a minimum of 10% resin acids. This additive can be used in feed or water given to animals, or added to zootechnical facilities, with the aim of promoting improvements in zootechnical and economic performance by maintaining the integrity of the intestinal wall, improving intestinal eubiosis and, consequently, increasing the digestibility and absorption of nutrients in the diet, as well as improving the environment and animal welfare.

Description

"Animal feed additive based on residual acids and plant extracts for use in animal feed" Field of Invention

[0001] The present invention relates to a zootechnical performance-enhancing additive based on resin acids, which may or may not be produced in combination with eugenol (clove extract), gingerol (ginger extract), 1,8-cineole (rosemary extract) and carvacrol (oregano extract), together or separately, with application in the field of animal husbandry, aiming to improve the zootechnical performance of production animals and the health of companion animals, through administration via feed, or via water, or added to poultry litter, or by spraying in zootechnical facilities. Description of the State of the Art

[0002] Livestock feed additives that enhance performance focus on the intestinal health of animals, a topic widely discussed in studies and scientific articles. Intestinal health is defined as the absence or prevention of intestinal diseases, allowing the animal to perform its physiological functions and resist exogenous and endogenous stresses. Furthermore, it is directly related to the homeostasis of the gastrointestinal tract, including its structure, integrity, and overall functions. To achieve this balance, livestock feed additives are incorporated into the animals' feed or water, or into the livestock environment.

[0003] The intestinal ecosystem of animals, especially in production systems, is frequently challenged by pathogenic microorganisms, fungal toxins (mycotoxins), antinutritional factors, and other stressors. These challenges can trigger inflammatory responses and intestinal disorders, such as diarrhea, which affect nutrient digestibility and, consequently, the zootechnical and economic performance of the animals. The continuous presence of these stressors compromises the response. The immune system in animals suffers from a negative impact on their viability and productivity.

[0004] Currently, antimicrobials, mainly antibiotics, continue to be used as performance-enhancing additives in animal production in several countries. However, this practice has generated concerns among consumers, industry, and producers due to the associated risks, especially antimicrobial resistance, which represents a growing threat to both animals and humans. In light of this, companies, researchers, government agencies, and importing countries have been pressing for measures to ban the use of antimicrobials, oxides, and sulfates as performance-enhancing additives in animal production.

[0005] Studies show that once bacteria develop resistance to a particular antibiotic, they are likely to transfer this “resistance information” to a wide range of other bacteria (HARADA, K. et al. “Antimicrobial Susceptibility of Pathogenic Escherichia coli Isolated from Sick Cattle and Pigs in Japan”, Journal of Veterinary Medical Science, v.67 (10), p.999-1003, 2005).

[0006] Antimicrobial resistance occurs when pathogens evolve, becoming resistant to antibiotics, which renders these treatments ineffective. A UK government study published in 2016 projects that, without drastic changes in antibiotic use, deaths caused by antimicrobial resistance in humans could reach 10 million people per year by 2050 (Barbosa, FF; Bünsen, S.; “Pig production in times of antimicrobial restriction - a global view”, chapter 1, p.14-33, 2021, DOI 10.37885 / 210203382), as illustrated in Figure 1.

[0007] Since the early 1990s, an alternative has been used as a performance-enhancing additive in relation to antimicrobials for animals. The addition of zinc oxide in pharmacological dosages of 2500 to 4000 ppm (parts per million), as a performance-enhancing additive and for Controlling post-weaning diarrhea in piglets. Zinc oxide has an inverse relationship with E. coli in the gastrointestinal tract, effects on different immunological and metabolic processes, and direct effects on the gastrointestinal microbiota (Barbosa, FF; Bünsen, S.; “Pig production in times of antimicrobial restriction - a global view”, chapter 1, p.14-33, 2021, DOI 10.37885 / 210203382).

[0008] There is also evidence that it plays an important role in maintaining intestinal integrity at therapeutic dosages (LIU, P. et al. “Effect of dietary zinc oxide on morphological characteristics, mucin composition and gene expression in the colon of weaned piglets”, PLOS ONE, v.9(3), e91091 , 2014).

[0009] According to Barbosa & Bünsen (2021), pigs are entirely dependent on a continuous nutritional supply of zinc for various bodily functions, mainly enzymatic ones. Therefore, all pig feeds include zinc supplementation to meet nutritional requirements. The European Food Safety Authority (EFSA) currently suggests that a total level of 150 ppm of zinc in the feed is sufficient to meet the physiological needs of the animals.

[0010] The same agency warns that zinc concentrations above 150 ppm can cause significant environmental impacts and promote increased bacterial resistance. Being a heavy metal with low absorption, most of the zinc ingested by animals is excreted in their feces. This factor becomes even more critical given the pharmacological doses of 3000 to 4000 ppm currently used in piglet feed in Brazil. The progressive accumulation of this element in the soil and groundwater can result in adverse effects on plant crops, aquatic life, and drinking water quality.

[0011] The therapeutic use of zinc oxide (ZnO; 2500-4000 ppm) for piglets in the nursery phase has proven to be a highly effective strategy in controlling post-weaning diarrhea and, consequently, in its impact. Economic benefits generated on farms. On the other hand, the excessive use of this and other molecules, such as antimicrobials, exacerbates problems like bacterial resistance and environmental contamination, leading to their ban in some countries. In this scenario, the search for alternatives that can improve the intestinal health and performance of piglets in the post-weaning period has become increasingly relevant (López-Gálvez, G. et al., “Alternatives to antibiotics and trace elements (copper and zinc) to improve gut health and zootechnical parameters in piglets: A review”, Animal Feed Science and Technology, v.271, p.114727, 2021; Bonetti, A. et al., “Towards zero zinc oxide: feeding strategies to manage post-weaning diarrhea in piglets”, Animals, v.11, p.642, 2021; Canibe, N. et al., “A Review on preventive measures to reduce post-weaning diarrhea in piglets”, Animals, v.12, p.2585, 2022).

[0012] Among the alternatives, the use of phytogenic additives – compounds made up of bioactive substances derived from plants – can be used in the prevention and reduction of enteric challenges in the production of all animal species. The mode of action of these compounds is based on their role as a modulating agent, controlling the number and type of microorganisms present, and the integrity of the gastrointestinal tract of animals. According to a study by BURT, S. “Essential oils: their antibacterial properties and potential applications in foods - a review”, International Journal of Food Microbiology, v.94, p.223-253, 2004, the antimicrobial activity of phytogenics is not the result of a specific mode of action, but a combination on different cell targets. This includes membrane disruption by terpenes, terpenoids and phenolics, metal chelation by phenols and flavonoids, and protective effects against viral infections by certain alkaloids and coumarins (COWAN, MM).“Plant products as antimicrobial agents”, Clinical Microbiology Reviews, v.12, p.564-582, 1999). Furthermore, its support for the digestive process and its antioxidant and anti-inflammatory properties have been described in the literature.

[0013] In 2022, the European Union banned the use of zinc oxide in animal feed based on studies that concluded that the benefits in preventing diarrhea in post-weaning piglets did not outweigh the significant environmental risks associated with zinc pollution. Furthermore, the decision highlighted concerns that the use of this additive could contribute to the development of antimicrobial resistance. Other major pig producers, such as China and Vietnam, are also questioning its use. In China, since 2018, legislation has limited the maximum permitted concentration to 1800 ppm, only in the first two weeks post-weaning. Faced with this scenario, producers and the industry are seeking effective alternatives that guarantee the intestinal health of animals throughout the different phases of production, prioritizing natural and sustainable solutions that do not induce bacterial resistance, such as resin acids.

[0014] In Brazil, the growth promoters authorized by MAPA (Ministry of Agriculture, Livestock and Supply) are mostly antibiotics for prophylactic use, but these are used in low doses (veterinary products). A minority of these are non-antibiotic products for zootechnical use in animal feed, as illustrated in Figure 3.

[0015] According to MAPA Normative Instruction 13 / 2004, "animal performance enhancers are chemically defined substances added to feed to improve productivity parameters." These parameters include feed intake, weight gain, and feed conversion, contributing to the health and well-being of animals. To be recognized as a performance enhancer, the product must undergo a series of efficacy, stability, and quality tests with regulatory bodies.

[0016] Faced with increasing restrictions on the use of zinc oxides and antimicrobial agents, the animal protein industry and producers are seeking effective, natural, and sustainable alternatives to ensure the intestinal health of animals.

[0017] In this context, the demand for natural and sustainably sourced additives has increased, since these compounds do not contaminate the meat or the environment. For the development of performance-enhancing additives, it is essential that nutritionists consider factors such as the reduction of undigested nutrients, the inhibition of the growth of pathogenic microorganisms, the modulation of beneficial microbiota, and the preservation of the integrity of the intestinal wall.

[0018] With the aim of solving these problems, the present invention was developed through the use of natural products such as oregano, ginger, rosemary, cloves, and resin acids extracted from pine. This approach provides a viable and promising alternative, ensuring the intestinal health of animals, preventing the development of bacterial resistance, and reducing the environmental impacts associated with the use of antimicrobial substances.

[0019] Based on Pinaceae extract, and possibly containing rosemary, and / or ginger, and / or clove, and / or oregano. The zootechnical additive of the invention is 100% natural and was developed to exert a modulating, antioxidant, and anti-inflammatory action in the gastrointestinal tract, promoting intestinal health, strengthening immunity, and improving the performance of production animals and the health of companion animals in different phases. This additive can be classified as a blend of resin acids extracted from plants, predominantly composed of silvic acid, and emerges as a natural, viable, and safe alternative to antimicrobials used as growth promoters, as well as to zinc oxide as performance enhancers.

[0020] Recent studies conducted in Europe have demonstrated the benefits of using resin acids in animal nutrition (HASAN, S. et al., 2019; UDDIN, M. et al., 2021; GUAN, X. et al., 2021), showing improvements in the intestinal microbiota, zootechnical performance, and immune response of animals. Based on these promising results, the present additive aims to... The objective is the partial and total replacement of zinc oxide and antibiotics as growth promoters.

[0021] The invention presents significant advantages compared to conventional livestock feed additives. Being a natural and sustainably extracted solution, it eliminates the need for antibiotics, chemical anticoccidial agents, as well as zinc and copper oxides and sulfates, thus reducing the risks related to antimicrobial resistance and environmental contamination. In addition to promoting the integrity of the intestinal epithelial barrier, it stimulates the growth of beneficial microbiota, improving digestion and nutrient absorption, which positively impacts livestock performance, resulting in better feed conversion, greater weight gain, and more efficient feed intake.

[0022] Unlike synthetic additives, the invention significantly reduces the excretion of polluting compounds into the environment, minimizing soil and water contamination. Its versatile application, both via feed and drinking water, as well as in livestock facilities, facilitates its incorporation into different animal production systems. In addition to the productive and environmental benefits, the invention meets the growing market demands for healthier and more sustainable solutions, aligning with the demands of consumers, industries, and regulatory bodies. Objectives of the Invention

[0023] The objective of the invention is to provide a natural and sustainable zootechnical performance-enhancing additive based on resin acids, with multiple applications in the intestinal tract of production and companion animals. This additive protects the intestinal epithelial barrier and promotes the growth of beneficial microbiota, and can completely replace antibiotics, chemical anticoccidial agents, copper and zinc oxides and sulfates. In this way, it contributes to reducing the development of resistant pathogenic microorganisms in both animals and humans, as well as decreasing the... excretion of these substances into the environment, mitigating environmental impacts related to their use. Brief Description of the Invention

[0024] The present invention relates to a zootechnical performance-enhancing additive based on resin acids, comprising a composition of resin acids extracted from Pinus resin, with a minimum of 10% and a maximum of 80% resin acids, which can be used via feed or water given to animals with the aim of improving the zootechnical and economic performance of animals, through the maintenance of the integrity of the intestinal wall, improvement of intestinal eubiosis and consequently improvement of the digestibility and absorption of nutrients from the diet. As the additive is added to the animal feed / food mixture, it can be an additive with a higher or lower concentration of resin acids, altering only the amount of additive to be added to the animal feed depending on the concentration of resin acids in the additive and animal species, which does not alter the efficiency or the final result.As well as its use in livestock facilities, reducing ammonia levels or improving the production environment, thus favoring animal performance.

[0025] The invention also includes the discovery of the use of resin acids as a performance enhancer and for improving the health of livestock. Brief Description of the Drawings

[0026] The present invention will be described in more detail below, with reference to the attached figures which, in a schematic and non-limiting way, represent examples of its embodiment. The drawings show: Figure 1 illustrates a graph of deaths attributable to different causes currently and due to antimicrobial resistance, projected for 2050; Figure 2 illustrates a graph of the inverse relationship between ZnO and E. coli in the animal gastrointestinal tract; Figure 3 illustrates the types of performance enhancers that exist; Figure 4 illustrates a graph of weight gain from 1 to 42 days of age of broiler chickens according to levels of inclusion of the zootechnical additive in the feed; Figure 5 illustrates a feed conversion graph for broiler chickens from 1 to 42 days of age according to the inclusion levels of the zootechnical additive in the rations; Figure 6 illustrates a graph of serum coloration according to treatments at 42 days of age in broiler chickens; Figure 7 illustrates a flowchart of the experiment conducted in the nursery phase of piglets. The process begins with the allocation of 240 piglets divided into three groups of 80 animals: (1) control group using zinc oxide (ZnO), (2) group treated with zootechnical additive (Pinaceae extract), and (3) group treated with a combination of the zootechnical additive and ZnO. Then, all groups undergo evaluation of zootechnical parameters according to a pre-established schedule, ending with the conclusion of the experiment. The arrows indicate the logical sequence of the steps; Figure 8 illustrates a graph of daily feed consumption by piglets in the first week of nursery according to the treatments studied; Figure 9 illustrates a graph showing the evolution of the number of piglets with diarrhea per day in the nursery according to the treatments studied; Figure 10 illustrates a graph of average fecal scores observed in the nursery phase of piglets according to the treatments studied; Figure 11 illustrates the fecal coloration of piglets observed in each of the treatments studied, where item A in Figure 11 represents: ZnO Control; item B in Figure 11 represents: zootechnical additive; item C in Figure 11 represents: zootechnical additive + ZnO; Figure 12 illustrates a graph showing the evolution of temperature in a daycare room throughout the experimental period. Detailed Description of the Invention

[0027] The present invention relates to the production of a zootechnical additive that improves the performance of all animals, the composition of which is based on resin acids extracted from pine resin. The resin is obtained through a resin tapping process from sustainably cultivated pine forests in various regions of the world where climatic conditions are favorable.

[0028] When added to animal feed or water, or to the animal production environment, the additive promotes improved zootechnical and economic performance of production and companion animals, as well as contributing to animal health and welfare.

[0029] At recommended doses, the additive can replace antibiotic growth promoters, as well as reduce the need for high concentrations of zinc and copper oxides in animal feed. Therefore, the invention aims at its application in the animal protein production sector, optimizing the zootechnical performance of animals.

[0030] Livestock feed additives that enhance performance focus on the intestinal health of animals, a concept widely discussed in the scientific literature. Several authors, such as BRENES, A; ROURA, E. “Essential oils in poultry nutrition: Main effects and modes of action”, Animal Feed Science and Technology, v.158 (1-2), p.1-14, 2010; and CZECH, A; KOWALCZUK, E.; GRELA, E. “The effect of a herbal extract used in pig fattening on the animals' performance and blood components”, Annales UMCS, Zootechnica, v.27 (2), p.25-33, 2009, agree that intestinal health can be defined as the absence or prevention of intestinal diseases, allowing the animal to perform its physiological functions and resist exogenous and endogenous stress factors. Furthermore, gut health is described as a state of homeostasis in the gastrointestinal tract, ensuring the structural and functional integrity of this system.This is the main objective of adding zootechnical performance-enhancing additives to animal feed.

[0031] The invention establishes a minimum content of 10% resin acids in its composition. An example of a formulation suitable for administration via animal feed or drinking water includes: 30% Pinaceae extract (with approximately 80% resin acids), 10% oregano extract, 10% clove extract, 10% ginger extract, and 10% rosemary extract.

[0032] More specifically, the animal feed additive based on resin acids for use in animal feed, according to the present invention, comprises at least 10% by weight of Pinaceae extract, which constitutes a source of resin acids, but is not limited to this source of resin acids. Optionally, the additive may also comprise at least: 1% by weight of oregano extract (carvacrol), 1% by weight of clove extract (eugenol), 1% by weight of ginger extract (gingerol) and 1% by weight of rosemary extract (1,8-cineole).

[0033] The Pinaceae extract preferably contains at least 80% resin acids, which consist of a mixture of pimaric acid, dihydroabietic acid, isopimaric acid, palustric acid, neoabietic acid, and abietic acid.

[0034] Animal feed additives can be supplied in basic liquid, acidic liquid, or acidic solid form. In basic liquid form, they have a pH between 8 and 10; in acidic liquid form, they have a pH between 5 and 6.

[0035] The additive of the present invention can be added to water or incorporated into feed, being administered as part of animal feed in livestock facilities, in liquid or solid form. EXAMPLES

[0036] These examples serve as a proof of concept, highlighting the technical and functional advantages of the invention. However, it is important to note that these examples do not limit the scope of the present invention, allowing for various variations and applications without altering its main objective. Example 1: Evaluation of the chemical composition (organic molecules) of the starting material "Pinaceae Extract" powder.

[0037] Three samples of "Pinaceae Extract" powder, in solid form and pale yellow in color, were evaluated. To determine the composition, analyses were performed using Fourier Transform Infrared Spectroscopy (FTIR) and Hydrogen Nuclear Magnetic Resonance (HNMR). 1 H).

[0038] The Pinaceae Extract powder product has an average composition of 85.5% known resin acids and 14.5% other unidentified compounds (Table 1), but with indications that they may be acidic compounds derived from known resin acids. Regarding the known and identified resin acids, the samples are mainly composed of abietic acid, dehydroabietic acid, isopimaric acid, palustric acid, and pimaric acid. Table 1. Components of Pinaceae Extract. Percentage component in the sample* Abietic acid 27.3 Dehydroabietic acid 19.4 Isopimaric acid 10.4 Palustric acid 9.6 Pimaric acid 8.7 Levopimaric acid 4,8 Neoabietic Acid 3,7 Sandaracopimaric acid 1,6 Total resin acids identified 85.5 Another 14.5 *Average values ​​obtained from the analysis of 3 samples Example 2: Evaluation of the use of a zootechnical additive product as a performance enhancer for broiler chickens.

[0039] The experimental trial described below was conducted with the objectives of verifying the effects of using the dry extract of the phytogenic blend (zootechnical additive of the invention) on the performance, carcass characteristics and intestinal quality in broiler chickens raised in the conventional system.

[0040] To evaluate the effect of adding five levels of extract supplementation via feed, comparing it with an antibiotic growth promoter, with assessments of performance, carcass characteristics, intestinal morphology and quality, and cecal microbiome of poultry. 2.1. Materials and Methods

[0041] The experimental trial was conducted using one-day-old male chicks from a commercial hatchery, duly vaccinated against Marek's disease and Gumboro disease (vector-transmitted). Laboratory analyses were performed at the Animal Research Laboratories of the DZO, the Meat Technology Laboratory of the DCA, and specific analyses were sent to the Imunova Laboratory. 2.2. Birds and experimental procedures

[0042] A total of 960 one-day-old male broiler chicks were used, distributed into 48 experimental plots (2.0 x 1.5 m) containing 20 birds each, in a litter system (new wood shavings), with nipple drinkers and tubular feeders. The experimental design was completely randomized, with six treatments and eight replicates each.

[0043] The experimental diet was based on corn and soybean meal, formulated according to a four-ration feeding program distributed across the phases: starter (1-21 d), grower (22-35 d), and finisher (36-42 d). 2.3. Experimental treatments

[0044] The experimental treatments are presented in Table 2. For the positive control treatment, enramycin was used at 10 ppm up to 35 days of age and 6 ppm from 36 to 42 days of age in the birds.

[0045] Challenge proposed for treatments receiving the zootechnical additive with the blend of phytogenics (contains resin acids) - zootechnical additive: 5% nutritional reduction and 30 ppm of salinomycin as the active ingredient (half the recommendation). Table 2. Experimental treatments and their characteristics. T1 Positive control (PC): with enramycin (1-35d=10 ppm and 36-42d=6 ppm) T2 Negative control (NC): performance enhancer T3 Dose 1 - 50 mg / kg of the phytogenic blend T4 Dose 2 - 75 mg / kg of phytogenic blend T5 Dose 3 - 100 mg / kg of the phytogenic blend T6 Dose 3 - 150 mg / kg of the phytogenic blend Note: Positive and negative controls without challenge, and controls with the phytogenic blend with challenge. 2.4. Assessment measures

[0046] Performance at each stage and cumulative stages, as well as mortality, were evaluated. For intestinal health analysis, slides were prepared to assess villus height (V), crypt depth (C), and the V:C ratio in the middle portion of the jejunum and ileum at 42 days of age in the birds.

[0047] The integrity and absorptive capacity of the intestinal wall were observed by measuring goblet cells and by serum staining using a marker in the feed. The composition of the experimental baseline diets and nutritional levels are described in Table 3. Table 3. Composition of basal diets. . .. . zn / Initial Growth Final ng M ingredients (%) .. n . .. . .. . » ' (1-21 days) (22-35 days) (36-42 days) Corn, 7.8% 58,490 61,540 67,685 Soybean meal, 46% 36,000 33,000 27,000 Soybean oil 2,300 2,40 2,500 Limestone 0.800 0.800 0.650 Common salt 0.350 0.350 0.300 Sodium bicarbonate 0.150 0.150 0.200 Dicalcium phosphate 1.000 1.000 0.785 Choline chloride 0.030 0.030 0.030 DL-Methionine, 99% 0.350 0.300 0.280 L-Lysine HCI, 78% 0.200 0.160 0.260 L-Threonine, 98% 0.070 0.020 0.050 Vitamin premix 2 0.100 0.100 0.100 Mineral premix 1 0.100 0.100 0.100 Phytase, 10,000 FTU / g 0.010 0.010 0.010 Salinomycin, 12% 0.000 0.00 0.000 Enramycin, 8% 0.000 0.000 0.000 Techfeed 0.000 0.000 0.000 Inert 0.050 0.050 0.050 TOTAL 100,000 100,000 100,000 Nutritional levels Crude protein, % 21,500 19,000 18,000 Calcium, % 0.820 0.733 0.650 Available phosphorus, % 0.430 0.400 0.350 Sodium, % 0.180 0.200 0.195 Chlorine, % 0.210 0.180 0.170 Metabolizable energy, kcal / kg 2980 3030 3110 Digestible lysine, % 1.203 1.100 1.033 Digestible methionine, % 0.642 0.577 0.532 Digestible methionine + cystine, % 0.937 0.858 0.786 Digestible tryptophan, % 0.247 0.230 0.240 Digestible threonine, % 0.735 0.696 0.620 1 Supplementation per kilogram of feed: vitamin A, 12,000 IU; vitamin D3, 2,500 IU; vitamin E, 30 IU; vitamin B1, 2 mg; vitamin B6, 3 mg; calcium pantothenate, 10 mg; biotin, 0.07 mg; vitamin K3, 3 mg; folic acid, 1 mg; nicotinic acid, 35 mg; choline chloride, 100 mg; vitamin B12, 15 pg; selenium, 0.300 mg. 2 Supplementation per kg of feed: manganese, 80 mg; iron, 50 mg; zinc, 50 mg; copper, 10 mg; cobalt, 1 mg; iodine, 1 mg.

[0048] Cecal material was collected from one bird per pen at 42 days of age for future analysis: one loop of cecum was placed in a freezer at -20°C to perform VFA analysis, and the other loop was placed at -80°C to perform microbiome analysis if it affects performance. These analyses were not included in the proposed budget.

[0049] At 42 days, one bird per batch was euthanized, and the thigh and drumstick were stored at -20°C for TBARS analysis (20 days of storage), and the breast was used for meat quality analysis (immediately). 2.5. Statistical analysis and presentation of results

[0050] ANOVA and Tukey's test were performed with a 5% significance level for parametric data, and Kruskal-Wallys' test for non-parametric data. The results are presented in a report for evaluation of the final results. 2.6. Results and Discussion 2.6.1. Performance

[0051] The performance results for broiler chickens from one to 21 days of age, according to the treatments studied, are presented in Table 4.

[0052] Feed intake was not influenced by the treatments (P>0.05), on the other hand, weight gain and feed conversion were similar for the positive control treatment and all others with the addition of the zootechnical additive (phytogenics) + challenge, and superior to the negative control without the addition of a performance enhancer (P<0.05). Table 4. Performance of broiler chickens according to experimental treatments (in ppm of phytogenics) for the phase from one to 21 days of age. Treatments, Consumption, Weight gain, Feed conversion, Kg / kg* 1.CP 1,186 0,956 a 1,240 a 2.CN 1,168 0,894 b 1,306 b 3.CN+50 ppm 1,184 0,943 a 1,256 a 4.CN+75 ppm 1,199 0,965 a 1,242 a 5.CN+100 ppm 1,180 0.947 a 1,245 a 6.CN+150 ppm 1,156 0,939 a 1,231 a P < 0.5190 0.0008 0.0048 CV, % 3.95 3.26 3 05 Error 0.0164 0.0108 Q 0135 *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05)

[0053] Considering the intermediate phase of 22 to 35 days of age of the birds (Table 5), no differences in consumption level were observed between treatments (P<0.05). For weight gain, the results were similar to the previous phase, with similar values ​​between the positive control treatments with the use of antibiotic performance enhancer and all others with the inclusion of the Zootechnical Additive + challenge, with the lowest value for the non-use of enhancer, with the feed without enhancer. In the case of feed conversion, the best values ​​(P<0.05) were observed for the positive control treatment and with the use of 100 and 150 ppm of phytogenics from the Zootechnical Additive. The lowest inclusion values ​​(50 and 75 ppm) resulted in feed conversion similar to the negative control (P>0.05), but also similar to the other treatments with a higher level of inclusion of the Zootechnical Additive with the phytogenic blend.These results indicate that with 100 ppm of these phytogenics, the result for feed conversion is similar to the use of the antibiotic growth promoter. Table 5. Performance of broiler chickens according to the treatments studied (in ppm of phytogenics) for the 22 to 35 day age phase of the birds. Treatments, Consumption, Weight gain, Feed conversion, Kg / kg* 1.CP 2.093 1,445 a 1,448 a 2.CN 2,026 1,315 b 1,543 b 3.CN+50 ppm 2.125 1.426 to 1.493 ab 4.CN+75 ppm 2.127 1.434 to 1.484 ab 5.CN+100 ppm 2,071 1,430 a 1,449 a 6.CN+150 ppm 2,075 1,448 a 1,434 a P < 0.2273 0.0030 0.0038 CV, % 4.29 4.87 3.78 Error 0.0316 0.0243 0.0197 *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05)

[0054] The results for the final rearing phase (36 to 42 days) are presented in Table 6 and reinforce the previous results regarding the level of feed consumption by the birds. For weight gain, the results were similar, where all treatments with performance enhancers, whether antibiotics or zootechnical additives, showed values ​​without statistical differences (P>0.05). On the other hand, regarding feed conversion, the best values ​​were observed for the positive control treatments and the use of the zootechnical additive at levels of 75 and 100 ppm of phytogenics. The highest level of inclusion of the zootechnical additive with the phytogenic blend resulted in a value similar to the negative control and the lowest level of supplementation of the zootechnical additive. Table 6. Performance of broiler chickens according to experimental treatments (in ppm of phytogenics) for the 36 to 42 day age phase of the birds. Treatments, Consumption, Weight gain, Feed conversion, Kg / kg* 1.CP 1,386 0,831 a 1,670 a 2.CN 1,384 0,741 b 1,871 b 3.CN+50 ppm 1.404 0.798 to 1.765 ab 4.CN+75 ppm 1,402 0,818 a 1,726 a 5.CN+100 ppm 1,427 0,875 a 1,635 a 6.CN+150 ppm 1.441 0.824 to 1.752 ab P < 0.1076 0.0008 0.0007 CV, % 3.29 6.66 5.580 Error 0.0163 0.0547 0.0356 *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05)

[0055] Evaluating the cumulative performance from one to 35 days (Table 7), the results indicate that the treatments studied do not influence (P>0.05) the level of feed consumption by the birds. For weight gain and feed conversion, the results indicate that the use of antibiotic performance enhancer or zootechnical additive results in better values, while the removal of the enhancer affected the performance of the birds with the worst indices (P<0.05). Table 7. Performance of broiler chickens according to experimental treatments (in ppm of phytogenics) for the period from one to 35 days of age of the birds. Treatments, Consumption, Weight gain, Feed conversion, Kg / kg of feed 1.CP 3.279 2.402 a 1 . 365 a 2.CN 3,193 2,208 b 1 ,446 b 3.CN+50 ppm 3,256 2,369 a 1,397 a 4.CN+75 ppm 3,326 2,399 a 1,386 a 5.CN+100 ppm 3,251 2,378 a 1,367 a 6.CN+150 ppm 3,230 2,387 a 1,353 a P < 0.2666 0.0001 0.0001 CV, % 3.73 3.41 2.66 Error 0.0430 0.0283 0.0130 *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05)

[0056] For the accumulated data for the entire experimental period (Table 8), it can be seen that feed consumption is not affected by the treatments studied (P>0.05), weight gain is similar with the use of the studied improvers, and feed conversion indicates that the use of a zootechnical additive with a blend of phytogenics at a dosage of 100 ppm of phytogenics can replace, with the same effect, the antibiotic used as a performance enhancer. The results were similar for the control treatments. positive and use of 100 and 150 ppm of phytogenics provided by the zootechnical additive. Table 8. Performance of broiler chickens according to experimental treatments (in ppm of phytogenics) for the period from one to 42 days of age of the birds. Treatments, Consumption, Weight gain, Feed conversion, Kg / kg of feed 1.CP 4,665 3,232 a 1,443 a 2.CN 4,576 2,949 b 1 ,552 c 3.CN+50 ppm 4.714 3.167 a 1.488 b 4.CN+75 ppm 4.728 3.212 to 1.470 ab 5.CN+100 ppm 4,678 3,252 a 1,438 a 6.CN+150 ppm 4,672 3,202 a 1,459 a P < 0.1882 0.0000 0.0000 CV, % 2.56 2.40 2.73 Error 0.0422 0.0269 0.0105 *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05)

[0057] Considering the effects of increasing levels of the zootechnical additive and a regression analysis model, the results indicate that there is a maximum level of weight gain and a minimum level of feed conversion as a function of the levels of the new additive used. For weight gain, the maximum response level is equal to 92 ppm of the phytogenic blend of the zootechnical additive. For feed conversion, the inflection point occurred at 112 ppm. The average of the weight gain and feed conversion results indicates 102 ppm of the phytogenic blend of the zootechnical additive as the value that allows for the greatest weight gain and the best feed conversion in broiler chickens. 2.6.2. Casing characteristics

[0058] The results of the evaluation of carcass characteristics of male broiler chickens at 42 days of age are shown in Table 9. No different effects of the treatments on the carcass characteristics of the broiler chickens were observed (P>0.05). Table 9. Carcass and cut yields at 42 days of age according to treatments. Treatment Yield*, % RC RP RC+SC RD RA GA Liver 1 79.73 36.29 27.39 16.76 9.86 1.29 2.37 2 77.67 35.42 28.73 15.88 9.93 1.36 2.50 3 78.30 35.46 28.07 16.52 9.88 1.34 2.49 4 76.85 35.96 27.15 16.97 10.18 1.29 2.44 5 79.65 35.81 27.50 16.28 10.21 1.40 2.36 6 78.99 35.72 27.36 16.02 10.58 1.26 2.34 P< 0.4224 0.8472 0.2598 0.5799 0.2963 0.9566 0.9487 CV, % 4.08 4.06 5.20 8.30 6.91 25.45 17.10 *RC=carcass yield, RP=breast yield, RC+SC=thigh+drumstick yield, RD=back yield, RA=wing yield, GA=abdominal fat yield. 2.6.3. Carcass quality

[0059] Table 10 presents the pH, colorimetry, cooking loss (CL), and shear force (SF) data for the breasts of broiler chickens slaughtered at 42 days of age.

[0060] There was no effect of the treatments (P>0.05) on pH, luminosity, redness (a*) and yellowness (b*), cooking loss and shear force of broiler breast meat at 42 days of age (P>0.05). Table 10. Average values ​​of pH, luminosity (L), redness (a*), yellowness (b*) and drip loss (DLL, %) of broiler breast chickens slaughtered at 42 days of age according to the treatments studied (in ppm of phytogenics). pH L a* b* PPC FC Treatments (%) N 1.CP 5.90 66.74 8.26 13.16 16.97 14.68 2.CN 5.89 66.24 9.13 14.09 17.00 13.20 3.CN+50 ppm 5.93 66.66 8.46 13.80 16.00 12.90 4.CN+75 ppm 5.88 66.24 8.15 13.75 16.39 12.90 5.CN+100 ppm 5.90 64.31 8.64 13.30 18.13 14.68 6.CN+150 ppm 5.88 63.80 9.32 13.72 16.68 13.20 P < 0.9606 0.1757 0.3416 0.87994 0.0440 0.5338 CV, % 1 .72 4.74 14.31 12.12 10.59 19.28 2.6.4. Intestinal morphology

[0061] The intestinal morphology of broiler chickens at 42 days is shown in Tables 11 and 12, for villus height, crypt depth, and villus / crypt ratio of the jejunum and ileum, respectively. For the jejunum, no significant differences (P>0.05) were observed for villus height; however, the lowest crypt depth was observed for the positive control treatment with the use of enramycin as a growth promoter, and the highest depths were observed for the negative control treatments without additives and the use of 100 and 150 ppm of phytogenics from the zootechnical additive (P<0.05). Regarding the villus / crypt ratio, the lowest value was observed for the negative control treatment without additives (P<0.05), while the other treatments showed similar values ​​for this ratio (P>0.05). Table 11. Villus height, crypt depth, and villus / crypt ratio of the jejunum of broiler chickens at 42 days of age according to treatments (in ppm of phytogenics). Depth-to-Height Ratio Villous treatments, crypt PM, villus / crypt PM 1.CP 657.89 65.89 b 9.98 a 2.CN 619,77 74,21 a 8,36 b 3.CN+50 ppm 684.22 68.36 ab 10.06 a 4.CN+75 ppm 639.78 64.45 b 9.96 a 5.CN+100 ppm 678.31 71.33 a 9.56 a 6.CN+150 ppm 670.69 73.45 a 9.19 a P < 0.2217 0.0285 0.0003 CV, % 8.77 9.75 8.06

[0062] Regarding ileum measurements, significant differences (P<0.05) were observed only for villus height and villus / crypt ratio. Greater villus height was observed for the positive control treatment and the use of 150 ppm of the zootechnical additive phytogenics, while the lowest was for the negative control treatment (P<0.05). Evaluating the villus / crypt ratio, a lower value was observed for the negative control and higher values ​​for the treatments with enramycin and zootechnical additive (P<0.05). Table 12. Villus height, crypt depth, and villus / crypt ratio of the ileum of broiler chickens at 42 days of age according to treatments. Villus height, crypt depth, villus / crypt ratio 1.CP 815.38 to 80.81 10.09 to 2.CN 614.92 c 85.48 7.19 c 3.CN+50 ppm 725.62 ab 85.69 8.57 b 4.CN+75 ppm 697.61 b 79.99 8.94 ab 5.CN+100 ppm 709.79 b 74.13 9.66 a 6.CN+150 ppm 744.24 ab 82.16 9.08 a P < 0.0002 0.2111 0.000 CV, % 10.34 12.11 11.40 *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05) 2.6.5. Serum coloration

[0063] The results of serum staining at 42 days of age in the birds according to the treatments studied are presented in Table 13 and Figure 6. The serum staining results as an indicator of intestinal absorptive quality show significant differences in the effects of the experimental treatments (P<0.05). The poorest intestinal quality may be Observed for the negative control treatment, with greater physiological challenges (P<0.05). The use of enramycin at 10 ppm and the zootechnical additive at 100 ppm of phytogenics resulted in equal serum coloration indices (P>0.05). Figure 3 indicates an improvement in intestinal quality as increasing levels of the additive were added; conversely, with the highest level of use, this quality began to decrease, as can be observed. By deriving the equation, the maximum point can be observed as the use of the additive at 122.5 ppm. Performance results indicated an average for weight gain and feed conversion of 102 ppm. Thus, the optimal level of additive use could be 100 ppm of the blend of active ingredients, as indicated by the results obtained. Table 13. Serum color at 42 days of age according to treatments (in ppm of phytogenics). Treatments Serum coloration 1.CP 0.833 a 2.CN 0.516 c 3.CN+50 ppm 0.665 bc 4.CN+75 ppm 0.731 ab 5.CN+100 ppm 0.836 a 6.CN+150 ppm 0.753 ab P < 0.000 CV, 12.91% *Means followed by different letters in the column differ statistically by Tukey's test (P<0.05) 2.6.6. Conclusions and considerations

[0064] The animal feed additive with a blend of phytogenics does not alter feed intake by broiler chickens. In the initial phase, the inclusion of 50 ppm of the phytogenic blend (with resin acids) provides weight gain and feed conversion equivalent to the use of 10 ppm of enramycin as a performance enhancer. When considering the entire rearing phase, the inclusion of 100 ppm of the The blend of phytogenics results in zootechnical performance similar to that of enramycin. Furthermore, the use of the zootechnical additive does not influence carcass characteristics or breast meat quality, regardless of the inclusion level in the feed. However, its use improves intestinal morphology, showing results comparable to those obtained with enramycin. Intestinal quality is also improved with the inclusion of the zootechnical additive up to a level of 245 ppm of the phytogenic blend in the feed. Therefore, the zootechnical additive presents itself as an effective natural alternative for improving the performance of broiler chickens. Example 3: Evaluation of the use of a zootechnical additive product that improves performance with resin acids in nursery phase diets on animal performance in a high-performance pig production setting.

[0065] The objective of the evaluation was to develop concepts regarding the use and applicability of the technology in the nursery phase of high-performance commercial farms, as well as to identify the impacts of its inclusion in the diet of piglets throughout this entire stage. 3.1. Materials and Methods 3.1.1. Location and animals

[0066] The project was carried out at the Folhados Agricultural Farm, in the municipality of Patrocínio, Minas Gerais. The full-cycle farm currently has 2000 sows in its herd. For the experiment, 240 recently weaned piglets (DB90 females x Duroc males, DanBred Brasil) were used, which were housed in the nursery facilities, in collective cages with a capacity for 12 animals. The pens have partially slatted floors and a front thermal comfort zone, as well as a six-mouth feeder and a nipple drinker. The feed buckets containing the experimental diets were always kept in front of the corresponding study pen. 3.2. Experimental design

[0067] The pen formation and experimental design in the nursery followed a randomized block design, based on sex (mixed pens; 5 males and 5 females) and average weaning weight (pen for light animals: 5.616 ± 0.364; and pen for heavy animals: 7.082 ± 0.490). The animals were divided as follows: 10 animals per pen, comprising a total of 24 study pens. There were 8 pens for the ZnO Control group (n = 80), in which the animals received ZnO in their diet at the dosage commonly used by the farm; 8 pens for the zootechnical additive group (n = 80); and 8 pens for the zootechnical additive + ZnO group (n = 80); which included the use of the test product, without or with association with zinc oxide (half the dosage commonly used by the farm), respectively (Figure 3). For this study, two daycare rooms were needed, each consisting of 12 group cubicles (n = 120).The piglets in the experiment received all the relevant and routine zootechnical and sanitary management practices of the farm. 3.3. Nutrition in the nursery phase

[0068] The offering of the different experimental diets (treatments) began immediately after weaning and continued throughout the nursery period (45 days), associated with different rations provided during this phase (Pre-Starter; Starter I and II) (Table 14). The dates for feed changes were established based on the consumption of each pen (Pre-Starter: 4 kg; Starter I: 4 kg; and Starter II: until leaving the nursery). In the case of the ZnO Control group, the animals received ZnO at dosages of 3000 ppm (Pre-Starter), 2000 ppm (Starter I), and 1000 ppm (Starter II). In the case of the zootechnical additive + ZnO group, the animals received ZnO at dosages of 1500 ppm (Pre-Starter), 1000 ppm (Starter I), and 500 ppm (Starter II). In the case of the zootechnical additive and zootechnical additive + ZnO groups, the test product was used with the inclusion of 200 g / ton of feed, with a dose of 85 mg of resin acids / kg of feed. Table 14. Formulation for the different diets offered in the nursery phase. Ingredients (kg) Pre-Starter Starter I Starter II Corn grain (7.8% CP) 558,300 608,300 619,817 Soybean meal (45.5% CP) 200,000 250,000 300,000 Soybean oil 25,000 17,000 20,000 Limestone 37% - - 7,304 Dicalcium Phosphate 18.5% - - 12.879 Whey powder 110,600 27,600 Pre-Starter Concentrate 106,200 Initial Concentrate I - 97,100 Initial Concentrate II - 40,000 Table 15. Nutritional levels of the different diets offered in the nursery phase. Nutrients Pre-Starter Starter I Starter II Crude protein, % 18,815 19,339 19,507 Ether extract, % 5.225 4.895 5.435 Crude fiber, % 2.369 2.726 3.119 Mineral matter, % 4.810 4.789 Metabolizable energy, kcal - 3447.092 Lactose, % 8.111 2.028 Butyric acid 0.050 0.000 Citric acid 0.500 0.500 Calcium formate 0.300 0.300 Calcium, % 0.479 0.571 0.700 Total phosphorus, % 0.577 0.606 0.567 Sodium, % 0.360 0.300 0.225 Chlorine, % 0.681 0.529 0.316 Potassium, % 0.890 0.856 0.832 Total magnesium, % - - 0.157 Total sulfur, % - - 0.272 Available phosphorus, 0.458% Total copper, mg / kg 1 50,000 150,000 156,118 Total zinc, mg / kg 1 37,434 137,434 166,432 Total manganese, mg / kg 67.951 Total iron, mg / kg 310.423; total sludge, mg / kg 1.182 SID lysine, 1.295% SID methionine, 0.505% SID methionine + cystine, 0.785% SID tryptophan, 0.245% SID valine, 0.894% SID glutamine, 2.302% Lysine, 1.374% Methionine, 0.520% Methionine + Cystine, 0.834% Threonine, 0.913% Tryptophan, 0.261% Valina, 0.994% Glutamine, 3.384% Vitamin A, IU / kg 15,000,000 Vitamin D3, IU / kg 3900,000 Vitamin E, mg / kg 36,000 Vitamin B1, mg / kg 2,250 Vitamin B2, mg / kg 5,400 Vitamin B6, mg / kg 3,000 Vitamin B12, pg / kg 34,500 Total vitamin K3, mg / kg 3,900 Total biotin, pg / kg 292.932 Vitamin B5, mg / kg 13.502 Folic acid, mg / kg 2.399 3.4. Assessments in the nursery phase

[0069] The experiment was conducted during the nursery phase of piglets, with 240 animals equally distributed into three experimental groups, each containing 80 pigs. The first group, called the control group, received zinc oxide (ZnO) in the nursery diets, with progressively decreasing doses throughout the phases: 3000 ppm in the Pre-Starter phase, 2000 ppm in the Starter I phase, and 1000 ppm in the Starter II phase. The second group was treated with a zootechnical additive based on Pinaceae extract, included in the feed at a dosage of 200 g per ton of feed, corresponding to an intake of 85 mg of diterpenic acids per kilogram of feed. The third group received a combination of the additive based on Pinaceae extract with zinc oxide, the latter in reduced doses compared to the control group: 1500 ppm in the Pre-Initial phase, 1000 ppm in the Initial I phase, and 500 ppm in the Initial II phase.

[0070] The animals were weighed individually on days 0, 7, 14, 21, and 45 of housing (end of nursery). Feed consumption per pen was measured daily throughout the first week, and then on days 14, 21, and 45 of housing, based on feed intake and weighing of leftovers in the trough. In addition, daily feed intake (DFI), daily weight gain (DWG), and feed conversion ratio (FCR) were calculated for each evaluated period and total. At the end of the nursery, fecal samples were collected from all experimental groups (pool from each pen, divided into two collection cups, totaling 48 samples). The samples were collected directly from the rectal ampulla of the animals and sent refrigerated to the Laudo Laboratory (Uberlândia, MG). Daily, the maximum and minimum temperatures of the nursery rooms were recorded.Any occurrences of mortality and / or removal of animals from the experiment, as well as the observation of diarrhea, respiratory clinical signs, and drug treatments performed throughout the nursery period, were monitored and recorded on specific forms (Figure 7).

[0071] The diarrhea score and the prevalence of affected animals were measured daily and analyzed weekly using the average score and the total number of affected animals during the period. For this, the numerical fecal score was used, indicated by the numbers 0, 1, 2, and 3, which represent, respectively, normal, soft, pasty, and watery feces (MORES, N. et al., Risk factors in the maternity ward associated with diarrhea, mortality and low piglet performance. Concórdia: EMBRAPA - CNPSA, 1991. 4 p. (EMBRAPA - CNPSA. Technical Communication, 178).

[0072] Respiratory clinical signs (cough / sneeze) were counted daily and analyzed by the total number of affected animals and the occurrence of signs. The methodology was adapted from SONCINI, RA; MADUREIRA JUNIOR, SE. Sanitary monitoring. In: SOBESTIANSKY, J.; WENTZ, I.; SILVEIRA, PRS; SESTI, AC (Ed.) Intensive Swine Production: production, management and herd health. Brasília: EMBRAPA, 1998: 1) Enter the pen and move the animals for one minute; 2) Wait one minute; 3) Count coughs and sneezes simultaneously and identify the affected animals; 4) Move the animals again; 5) Make another count; 6) Move the animals a third time; 7) Make a third count. The animal is considered affected if it presents respiratory clinical signs in at least one of the counts. 3.5. Data Collection and Record Keeping

[0073] Data collection took place daily, using paper forms for recording information. These forms were later updated in Excel files. 3.6. Statistical Methods

[0074] The data were subjected to analysis of variance (ANOVA) and the means were compared using Tukey's test, through the Minitab 19 statistical program. Differences between means were considered statistically significant when the P-value < 0.05 (5%) and as tending to be significant when the P-value < 0.10 (10%). 3.7. Results

[0075] In the first week of nursery, a higher feed consumption was observed in the animals of the zootechnical additive and zootechnical additive + ZnO groups (P < 0.05) (Table 16 and Figure 8). This result suggests a greater attractiveness of the diet when containing the test product and a lower inclusion of ZnO. Table 16. Daily feed consumption in the first week of nursery according to the treatments studied. Treatment p Additive Control Additive v â| O Zootechnical ZnO + ZnO r (n = 8) (n = 8) (n = 8) Consumption 0.00 0.00 , „ , x _ , 0.113 average / piglet, kg b 0.132 a 0.130 a 4 3 SEM, standard error of the mean; ZnO, zinc oxide. a b Different lowercase letters on the same line indicate a significant difference at the 5% level according to Tukey's test.

[0076] No treatment effects were observed for the other performance variables evaluated throughout the study (P > 0.05) (Table 17), which demonstrates the possibility of partially or totally replacing ZnO with the zootechnical additive without any loss of productivity. Table 17. Performance of piglets in the nursery according to the treatments studied. Treatment C _ont .ro .le Z ,nO _ A . d ..it..ivo zoot .é,cn .ico Zootechnical additive P + Z _nO EPM va .lor (n = 8 ) (n = 8) (n = 8) Prohibited Piglet weight, kg 6.206 6.203 6.187 0.153 0.701 Week 1 Piglets, n 10,000 10,000 10,000 0,000 Piglet weight, kg 6.705 6.702 6.747 0.148 0.892 CDR, kg 0.097 0.113 0.112 0.004 0.135 GPD. kg 0.071 0.071 0.080 0.007 0.794 AC 1 .537 2.280 2.760 0.523 0.639 Week 2 Piglets, n 9,875 9,750 9,750 0,085 0,821 Piglet weight, kg 8.752 8.656 8.737 0.188 0.850 CDR . kg 0 335 0.332 0.341 0.006 0.796 GPD, kg 0.292 0.279 0.284 0.007 0.648 CA 1 .157 1 .195 1 .204 0.019 0.588 Entry - 14 days CDR, kg 0.216 0.223 0.226 0.004 0.542 GPD, kg 0.182 0.175 0.182 0.005 0.833 CA 1 .210 1 .280 1 .253 0.025 0.564 Week 3 Piglets, n 9,750 9,750 9,750 0,090 1,000 Piglet weight, kg 11.483 11.256 11.365 0.191 0.575 CDR. kg 0.517 0.504 0.515 0.007 0 682 GPD, kg 0.390 0.371 0.375 0.007 0.513 CA 1 .336 1 .359 1 .374 0.021 0.675 Entry - 21 days CDR, kg 0.316 0.317 0.323 0.005 0.803 GPD, kg 0.251 0.241 0.247 0.004 0.584 AC 1 .261 1 .316 1 .312 0.013 0.153 Exit Piglets, n 9,625 9,750 9,625 0,115 0,883 Piglet weight, kg 26.420 26.846 27.196 0.468 0.632 CDR . kg 0.931 0.971 0.949 0.025 0.814 GPD, kg 0.622 0.650 0.660 0.013 0.383 AC 1 .515 1 .490 1 .440 0.038 0.764 Total CDR, kg 0.644 0.666 0.657 0.015 0.838 GPD, kg 0.449 0.459 0.467 0.008 0.619 CA 1 .446 1 .448 1 .407 0.029 0.849 CA, feed conversion ratio; CDR, daily feed intake; SEM, standard error of the mean; GPD, daily weight gain; ZnO, zinc oxide.

[0077] Considering the different weight categories into which the animals were divided for housing in the nursery phase (light piglet pens and heavy piglet pens) as a study factor, a higher CDR was observed in the light piglets in the zootechnical additive + ZnO group, when compared to the light animals in the ZnO Control group (P < 0.05) in the first week of housing (Table 18). This result reinforces the greater attractiveness of the diet when containing the test product and a lower inclusion of ZnO, as already mentioned, especially for weaned animals with lower weight. Although an interaction effect was also observed for the GPD variable in the first week (P < 0.05), the statistical methods used in this study were not able to indicate differences between the means. However, It can be observed that light animals in the zootechnical additive + ZnO group showed the greatest numerical gain during the period, suggesting better performance and utilization of the diet by these animals (Table 18). Table 18. Performance of piglets in the first week of nursery according to the factors studied. Treatment P value P pp e s =no A _J Additive - . , „ Additive i l n n i i r c i i the a i l Cont. ro e. ZnO zooté ,cnico zootécnico . M.é.d ..ia EPM PP*T (n = 4) + ZnO ( " = 4) (n = 4) Prohibited Lightweights 5.465 5.467 5.464 5.466 b 0.002 Piglet weight, kg - 0.000 0.743 Heavy vehicles 6,946 6,939 6,910 6,932 a 0.018 Week 1 Light 10,000 10,000 10,000 10,000 0,000 Heavy 10,000 10,000 10,000 10,000 0,000 Lightweight 5,850 B 6,061B 6,226 B 6,046 0,072 Piglet weight, kg - 0.000 0.017 Heavy 7,560 A 7,342 A 7,268 A 7,390 0,067 Light 0.082 B 0.115 AB 0.126 A 0.107 0.007 CDR, kg . 0.993 0.010 Heavyweights 0.112 AB 0.112 AB 0.098 AB 0.107 0.005 Light 0.055 A 0.085 A 0.109 A 0.083 0.010 GPD, kg - 0.162 0.020 Heavyweights 0.088 A 0.057 A 0.051 A 0.065 0.009 Light 1,743 1,404 1,221 1,456 0,146 CA . 0.177 0.410 Heavy vehicles 1,332 3,160 4,300 2,930 1,010 CA, feed conversion ratio; CDR, daily feed intake; SEM, standard error of the mean; GPD, daily weight gain; P, weight; P*T, interaction between weight and treatment factors; ZnO, zinc oxide. AB Different capital letters in the same main line indicate a significant difference at the 5% level by Tukey's test.

[0078] For the other phases evaluated in the nursery, only isolated effects of the animals' initial weight were observed, naturally expected due to the blocking performed at the beginning of the study: heavier animals presented greater live weight, CDR, GPD and CA (P < 0.05) (Table 19). Table 19. Piglet performance during the nursery phase (week 2 to week 3) according to the factors studied. Treatment P value Weight A Additive Initial Control ZnO IVO animal husbandry Média EPM (n _ = 4 xx) zootechnician + Z _nO PP*T < = 4 > („ = 4) Week 2 Lightweight 9,750 9,750 9,750 9,750 0,131 Piglets, n. 0.660 0.821 Heavy vehicles 10,000 9,750 9,750 9,833 0,112 Lightweight 7,728 7,869 8,045 7,881 b 0.096 Piglet weight, kg - 0.000 0.197 Heavy vehicles 9,776 9,442 9,430 9,549 a 0.112 Light 0.316 0.314 0.321 0.317 b 0.007 CDR, kg . 0.002 0.989 Heavy 0.354 0.351 0.362 0.356 a 0.007 Light 0.268 0.258 0.260 0.262 b 0.006 GPD, kg - 0.001 0.961 Heavy 0.317 0.300 0.309 0.308 a 0.009 Light 1,181 1,220 1,234 1,211 0,024 CA . 0.198 0.988 Heavy 1,133 1,171 1,174 1,159 0,027 Entry - 14 days Light 0.199 0.214 0.223 0.212 b 0.006 Heavy 0.233 0.232 0.230 0.231 a 0.005 Light 0.162 0.172 0.184 0.172 0.007 GPD, kg . 0.190 0.228 Heavy 0.202 0.179 0.180 0.187 0.008 Light 1,246 1,259 1,216 1,240 0,033 CA . 0.787 0.510 Heavy 1,174 1,300 1,290 1,255 0,039 Week 3 Lightweight 9,500 9,750 9,750 9,667 0,142 Piglets, n 0.398 0.487 Heavy vehicles 10,000 9,750 9,750 9,833 0,112 Lightweights 10.544 10.375 10.716 10.545 b 0.110 Piglet weight, kg. 0.000 0.371 Heavy vehicles 12,421 12,137 12,014 12,191 a 0.130 Light 0.491 0.481 0.507 0.493 b 0.008 CDR, kg . 0.008 0.474 Heavy 0.543 0.528 0.523 0.531 a 0.010 Light 0.402 0.358 0.382 0.381 0.011 GPD, kg . 0.810 0.294 Heavy 0.378 0.385 0.369 0.377 0.008 Light 1,231 1,346 1,330 1,302 b 0.026 CA - 0.006 0.110 Heavy 1,442 1,372 1,418 1,411 a 0.024 Entry - 21 days Light 0.296 0.303 0.318 0.306 b 0.005 CDR. kg . 0.008 0.351 Heavy 0.337 0.330 0.327 0.331 a 0.006 Light 0.242 0.234 0.250 0.242 0.005 GPD, kg . 0.317 0.418 Heavy 0.261 0.247 0.243 0.250 0.006 Light 1,229 1,297 1,274 1,267 b 0.019 CA . 0.025 0.811 Heavy 1,294 1,335 1,349 1,326 a 0.015 Exit Lightweight 9,250 9,750 9,750 9,583 0,149 Piglets, n - 0.489 0.225 Heavy 10,000 9,750 9,500 9,750 0.179 p eso Light 24,390 25,292 25,600 25,094 b 0.473 , ... , . 0,000 0,805 piglet, kg Weighed 28,450 28,399 28,792 28,548 a 0.388 Light 0.867 0.955 0.908 0.910 0.034 CDR, kg - 0.132 0.755 Heavy 0.995 0.988 0.989 0.990 0.033 Light 0.577 0.622 0.620 0.606 b 0.017 GPD, kg . 0.003 0.806 Heavy 0.668 0.678 0.699 0.682 a 0.013 Light 1,541 1,530 1,464 1,511 0,070 CA . 0.493 0.986 Heavy 1,490 1,451 1,416 1,452 0,034 Total Light 0.601 0.651 0.633 0.628 0.020 CDR, kg . 0.081 0.731 Heavy 0.688 0.681 0.680 0.683 0.019 Light 0.421 0.441 0.447 0.436 b 0.010 GPD, kg - 0.007 0.812 Heavy 0.478 0.477 0.486 0.480 a 0.009 Light 1,453 1,473 1,414 1,447 0,054 CA . 0.697 0.965 Heavy 1,440 1,422 1,400 1,421 0,026 CA, feed conversion ratio; CDR, daily feed intake; SEM, standard error of the mean; GPD, daily weight gain; P, weight; P*T, interaction between weight and treatment factors; ZnO, zinc oxide

[0079] Assessed daily using fecal scoring, animals presenting with diarrhea and the average fecal score obtained for each experimental group throughout the study are illustrated in Figures 9 and 10.

[0080] Furthermore, a difference in fecal color was observed between the feces of the different experimental groups. As illustrated in Figure 11, it can be seen that the feces produced by the animals that received the zootechnical additive diet (item B of Figure 11) and the zootechnical additive + ZnO diet (item C of Figure 11) were darker when compared to the ZnO Control group (item A of Figure 11). 11). Here, given the known effect of ZnO as a bleaching agent for the fecal bolus produced, it is easy to understand the increased pigmentation of feces when diets are used in which the micromineral is no longer present, or is present in reduced doses.

[0081] Furthermore, it is not possible to rule out some effect of the tested product on the coloration of the feces, which, however, should not be confused with the presence of blood in the material. Unlike what has been observed for diets containing blood meal or even hemoglobin in their composition, which result in pigmentation of the animals, pens, and feeders due to darkened feces, similar situations were not reported for the present study. For verification purposes, tests attesting to the absence of occult blood in the feces of animals fed the diet containing the zootechnical additive were performed with samples from three pens in the study, as reproduced in Table 20. Table 20. Occult blood test in stool.

[0082] Regarding the need for medication during the nursery phase, only two animals required medication during this period. The causes were dermatitis (Product group; 1 animal) and arthritis (Positive Control group; 1 animal). Table 21 below shows the records for mortality during the nursery phase, as well as any animals removed from the study. Table 21. Animals that died or were removed from the study. Treatment Output of the Additive Zootechnical Additive ZnO Control Causes Zootechnical Experiment + ZnO (n = 80) (n = 80) (n = 80) Death, No. 121 Unknown Removal, No. 202 Waste Total, n (%) 3 (3.75) 2 (2.5) 3 (3.75) ZnO, zinc oxide.

[0083] Finally, Figure 12 illustrates the daily evolution of maximum and minimum temperatures in the nursery room. The extremes recorded were 11 and 32°C, both from the 38th day of placement. In this final nursery period, the largest temperature ranges of the study (17°C) were also observed, recorded on days 38 and 44. 3.8. Final Considerations

[0084] This study concludes that the zootechnical additive with resin acids is capable of acting as a performance enhancer, partially or totally replacing the therapeutic use of ZnO for piglets in the nursery phase, with equal efficiency and without any detriment to the health and performance of the animals. Furthermore, the product shows excellent acceptance by newly weaned piglets, positively impacting feed intake in the first week of the nursery phase.

[0085] It should be noted that, although the present invention has been described with respect to the accompanying drawings, it may undergo modifications and adaptations by those skilled in the art, depending on the specific situation, but provided that they remain within the inventive scope defined herein.

Claims

1. CLAIMS 1. ANIMAL FEED ADDITIVE BASED ON RESINIC ACIDS, characterized by comprising at least 10% by weight of Pinaceae extract.

2. ADDITIVE, according to claim 1, characterized in that it further comprises at least: - 1% by weight of oregano extract (carvacrol); - 1% by weight of clove extract (eugenol); - 1% by weight of ginger extract (gingerol); and - 1% by weight of rosemary extract (1,8-cineole).

3. ADDITIVE, according to claims 1 and 2, characterized in that the Pinaceae extract is one of the sources of resin acids and contains at least 80% resin acids.

4. ADDITIVE, according to claims 1 to 3, characterized in that the resin acids are a blend of pimaric acid, dihydroabietic acid, isopimaric acid, palustric acid, neoabietic acid and pimaric acid.

5. ADDITIVE, according to claims 1 to 4, characterized by being in basic liquid, acidic liquid or acidic solid form.

6. ADDITIVE, according to claim 5, characterized in that the basic liquid form has a pH between 8 and 10.

7. ADDITIVE, according to claim 5, characterized in that the acidic liquid form has a pH between 5 and 6.

8. USE OF THE ZOOTECHNICAL ADDITIVE IN ANIMAL FEED, as defined in any one of claims 1 to 7, characterized by the fact that it is added to water or feed and used as part of animal feed.

9. USE, according to claim 8, characterized by being added to livestock facilities in liquid or solid form.

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