Effects of aqueous extract of red propolis as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status

Effects of aqueous extract of red propolis as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effects of aqueous extract of red propolis as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status Patricia López Garcia, Lenita Stefani, Marcel Boiago, Denise Araujo, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1485683/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study aimed to determine the effects of aqueous extract of red propolis (RP) as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status. A total of 120 laying quails with 52 days of age were were randomly allotted to dietary treatments and six replicates of five birds each in completely randomized design. There were four dietary treatments as following: Control (TC), with enramycin (TE), 1g RP/kg of feed (RP1), 2g RP/kg of feed (RP2) in concetrated diet raised for 63 days. The results of productive performance such as percentage of egg production, feed intake per quail per day, average egg weight, egg mass, foods conversions showed no significant difference between treatments. The quails fed with red propolis in fresh eggs showed darker yolk (P < 0.05), with higher intensity of red and lower intensity of yellow; as less than 6.0 pH yolk; microbiological characteristics analyzed on the surface of eggs and feces of quails was lower and less than compared with other treatments. It is possible to conclude that red propolis showed promising results to be used as a feed additive for laying quails, since it maintained the productive performance of these animals and caused qualitative improvements in the physicochemical and microbiological characteristics of the eggs. Antimicrobial antioxidant laying quails lipid peroxidation performance red propolis Introduction Bee products appear as an alternative for antibiotics and promising food source in animal nutrition, due to their therapeutic substances (Carpes et al. 2007 ). Propolis is an apicultural product made with resins collected by Apis mellifera bees, from vegetative parts of plants or resinous exudates. In general, propolis is composed of 50% resin and balm, 30% wax, 10% essential and aromatic oils, 5% pollen and 5% various other substances (Burdock 1998 ; Fokt et al. 2010 ). Its color ranges from green, red to dark brown, and this variation is related to the geographical origin and the vegetation from which it is extracted (Salatino et al. 2011 ). The red propolis collected in the northeast region of Brazil has resinous exudates from the Dalbergia ecastophyllum plant, as a botanical source (Righi et al. 2011 ; Mendonça-Melo et al. 2017 ). It contains pterocarpans, isoflavonoids, chalcones and phenylpropanoids (Righi et al. 2011 ) as main constituents. It showed cytotoxic activity against various types of cancer cells, antibacterial, antifungal, anticariogenic, antioxidant, anti-inflammatory and antiproliferative activity (De Mendonça et al. 2015; Cavendish et al. 2015). In this context, this study aimed to evaluate whether the addition of aqueous extrat of red propolis to the diet of laying quails could improve productive performance, as well as physicochemical and microbiological quality of eggs. Raising quails has become a profitable activity due to its relevant characteristics, including their resistance to heat and diseases, low requirement for housing, and also due to the fact that quail eggs and meat are nutritious and tasty, presenting themselves as an additional meat protein option for humans (Umigi et al. 2012 ). Poultry farming has been using for many years some tools that have resulted in higher growth and higher yield, including the use of antibiotics as growth promoters (Pelicano et al. 2002 ). However, misuse of certain antimicrobials may promote the emergence of resistant bacteria (Garcia-Migura et al. 2014 ) and nowadays there is a search for more natural alternative additives, with similar animal performance (Zavarize et al. 2011 ). Materials And Methods The experiment was conducted in Japanese quails ( Coturnix japonica ) egg producing base at the Poultry Sector from UDESC (Chapecó, SC, Brazil), from 5 March to 9 May 2021. Animal welfare statement The animals used in this study and all experimental procedures were approved by the Committee of Ethics for Animal Use (CEUA) from Santa Catarina State University (Brazil) under protocol number 2307170920. All methods were in accordance with the approved guidelines and regulations. Experimental design and management The experiment was carried out at the Poultry Sector, the extraction of propolis and the analyzes were performed at the Laboratory of Molecular Biology, Immunology and Microbiology (LABMIM) both from the Animal Science Department of the Santa Catarina State University, in the city of Chapecó, Santa Catarina State, Brazil. One hundred twenty 52 day-old female Japanese quails were used as experimental animals and quails were fed a diet without addition of RP for 17 days to make the animals accustomed to the experimental diets, with water ad libitum . Then the quails were assigned to randomly distributed in four treatments, with six replications, with five quails per cage, totaling twenty-four experimental plots. The study lasted sixty three days, divided into three cycles of twenty one days each. The experimental quails were fed the basal ration was formulated according to the nutritional values and requirements established in the Brazilian Table of Poultry and Swine (Rostagno et al. 2017), with 2,750 Kcal of metabolizable energy per kg of ration and 19.9% of protein (Table 1). The treatments following: control treatment (CT): basal diet without antimicrobials and without red propolis; enramycin treatment (TE): 10 mg/kg of feed; red propolis (RP1): 1g of red propolis/kg of feed and red propolis (RP2): 2g of red propolis/kg of feed. The red propolis aqueous extract was mixed with the microingredients and later added to the vertical mixer. The lighting program used for the quails was 16 hours of light per day. Red propolis aqueous extract Red propolis was purchased from a commercial company located in the city of Canavieiras, Bahia State, Brazil. The extraction was performed according to the methodology adapted from Kubiliene et al. (2015), with the maceration of crude propolis with pistil and liquid nitrogen, inside a mortar until it becomes powder. In a beaker, 100 mL of distilled water, 10 grams of propolis and 20 grams of polyethylene glycol (PEG 400 PA) were added and the mixture was subjected to high pressure at 120°C for 5 minutes in an autoclave. After extraction, the extract was stored protected from light. The extract was homogenized and weighed for subsequent mixing with the feed. Productive P erformance parameters Daily quail mortality was observed and recorded in order to calculate animal viability, as well as feed intake per quail (g/quail.day). The number of eggs produced was recorded daily and the average production per quail was estimated, in addition to egg mass (g/quail.day). Feed conversion was evaluated by kg of feed consumed per kg of eggs produced and also per kg of dozens of eggs. Eggs were weighed in the last five days of each experimental period and the average egg weight was determined. P arameters in fresh eggs To assess egg quality, a sample consisting of two eggs from each experimental plot was collected and analyzed. Specific gravity was determined according to Barbosa et al. (2008). Texture analyzer equipment was used to measure egg shell strength and results were expressed as kgf. After breaking the eggs the following parameters were evaluated: height of the dense albumen measured with a tripod micrometer; Haugh unit (HU) according to Haugh (1937); yolk index, which was calculated by the ratio between the height and diameter of the yolk (mm), obtained with the micrometer and the caliper, respectively; yolk color was determined with the colorimetric fan (DSM) and with the colorimeter (Minolta CR-400), where the following universal colorimetric coordinates were observed: luminosity (L*), red intensity (a*) and yellow intensity (b*); the shell weight and percentages of shell, yolk and albumen; egg shell thickness was obtained with a caliper at three points and obtaining the arithmetic mean of the three measurements. The pH of the yolk and the albumen were measured with the digital phmeter (Testo 205). Parameters in stored eggs The eggs were weighed on a precision scale and stored under temperature and air humidity registered conditions. Twice per day, the temperature, maximum, minimum and the air humidity, maximum, minimum at the room storage were registered, during 21 days. After this period, the eggs were weighed where the weight loss after storage was calculated. The levels of lipid peroxidation in the yolk of these eggs were determined according to Giampietro et al. (2008), with the measurement of thiobarbituric acid reactive substances (TBARS), which are formed during the decomposition of lipid peroxides. Compound 1,1,3,3 tetramethoxypropane (TMP) was used as TBARS standard. The results were expressed in mg TMP/kg of yolk. Microbiological analysis Microbiological analysis were performed on the surface of eggs produced at the end of the second and third cycles. Each sample consisted of a group of five eggs per repetition. Egg surfaces were washed with peptone water according to the methodology adapted from Gentry & Quarles (1972). Quail feces samples, collected from four points of each experimental plot, were analyzed. Egg and feces samples were sown on Plate Count Agar (PCA) for total mesophilic aerobics and on Petrifilm® 6404 (3M) plates for total coliforms and Escherichia coli . Incubation was carried out at 37 ºC for 48 hours and the number of bacterial colonies present were expressed in colony-forming units per mL (CFU/mL). Statistical analysis Data were submitted to analysis of variance and means compared by Tukey test at 5% significance level (in cases of significant diferences). These statistical procedures were performed by the program Statistical Analysis System. Results And Discussion Productive performance Egg production, feed intake, average egg weight, egg mass, feed conversion (kg/kg and kg/dz) are shown in Table 2. No significant differences were observed in these variables compared to the control treatment (P>0.05). The results show that the red propolis aqueous extract did not influence the productive performance of the quails. Our findings are similar to those of Zeweil et al. (2016) while studying the effect of propolis extract (250 and 500 mg/kg) in the diet of Japanese quails, observed that the inclusion of propolis neither influence performance (body weight, laying rate, egg weight and egg mass) nor egg quality (egg weight, yolk and albumen percentage, albumen height, shell percentage and thickness, specific gravity and yolk color). Petrolli et al. (2014) using residue of green propolis extract, at the level of 1%, for broilers, also found no difference (P>0.05) for feed intake and feed conversion in the period from 1 to 21 days of age. Propolis can increase the beneficial microbiota and control pathogenic bacteria (Kacániová et al. 2012). The change in the dynamics of the intestinal microbiota is generally attributed to flavonoids, which have antibacterial activity and are present in the ethanol extract of propolis (Oldoni et al. 2011; Bueno-Silva et al. 2013; Frozza et al. 2013). Different results might be obtained between studies due to the variability and complexity of the composition of propolis, which changes according to the flora of each location, genetics of queen bees and even the time of year in which it is collection was done (Buriol et al. 2009). Another hypothesis that the addition of propolis did not show any difference in the analyzed variables may have been the fact that the breeding environment did not propose a sanitary challenge that would enable a more expressive response with the use of propolis. Since the birds did not have direct contact with the feces, as they were raised in cages with feces collecting trays, they were raised at an ideal density to develop their full potential (Albino & Barreto, 2003). Moreover, herbal medicines (propolis), probiotics, symbiotics and organic acids are classified as performance-enhancing additives (Brasil, 2004) and it is notorious how these additives cannot express positive results due to the lack of sanitary challenge in poultry studies, in different experimental variables (Silva et al. 2012; Bueno et al. 2012; Bastos-Leite et al. 2016). Because, good prophylactic conditions for raising animals and a minimum of stress (which is usually associated with nutritional, environmental or behavioral factors), do not present a sufficient increase in bacteria to cause an imbalance in intestinal health, compromising productive performance (Fukayama et al. 2005). It is known that red propolis aqueous extract has great potential to be used as a herbal additive in poultry feed, as red propolis has biologically active compounds (vestitol and neovestitol and biochanin A, liquiritigenin, formononetin and medicarpine) found only in this type of propolis. Trusheva et al. (2006) concluded the identification of new propolis constituents in red Brazilian propolis, most of them having antibacterial, antimycotic and antiradical activities, is a further confirmation of the fact that propolis, independently of its plant source and chemical composition, present antimicrobial and antioxidant activity. Its result that propolis plays in the hive: it is the ‘chemical weapon’ of bees against pathogen microorganisms and the elements of weather. However, in different propolis types, different chemical constituents are responsible for the valuable activities (Bankova et al. 2005). Furthermore, as can be seen in this experiment, the inclusion of red propolis aqueous extract in the diet of the quails did not cause any negative effects such as mortality or on other studied variables and could be evaluated at different concentrations and with diets free of synthetic antimicrobials. Another study using very high dosage of red propolis in order to address its safety found that red propolis was unable to cause side effects (Reis et al. 2000). Berretta et al. (2017) also reported future perspectives for propolis in the health of the population and in the Brazilian and international market, especially because of its important biological activities and safety. Feeding the quails with 1g of red propolis aqueous extract showed the best feed conversion (kg of feed/kg of eggs), of 2.54 kg, although the difference was not statistically significant compared to the other treatments. Marieke et al. (2005) stated the improvement in feed, conversion rate could be due to the ability of propolis to improve nutrient digestibility and absorption as a result of sucrase, amylase and phosphatase activities. Quality of fresh eggs A significant difference (P<0.05) was observed in the results of the eggs analyzed in the following parameters: luminosity (L*), red intensity (a*), yellow intensity (b*) and yolk pH. In the other analyzed parameters: shell strength, specific gravity, Haugh unit, yolk index, colorimetric fan, yolk percentage, shell percentage, albumen percentage, shell thickness and albumen pH, no statistical difference were observed (Table 3). In summary, the yolk color of the eggs of quails fed with red propolis was darker, with greater intensity of red and lower intensity of yellow when compared to the control treatment, i.e., the eggs of quails fed diet without any additive had yolks of lighter color. Yolk color, as a sensory attribute, is considered an indicator of quality, and plays an important role in the acceptance of the egg by consumers, who associate the intense pigmentation of the yolk with higher nutritional value of the egg (Silva et al. 2000; Tocchini & Mercadante, 2001). Yolk color intensity is determined by the incorporation of xanthophylls (a group of carotenoid pigments) present in corn, particularly lutein and zeaxanthin, and depends on the levels of inclusion in the diet. However, other foods can change yolk color depending on their level of inclusion (Silva et al. 2000). Lee et al. (2001) stated that changes in yolk color can be observed when supplemental sources of carotenoids are added to the diet since carotenoid pigments are fat-soluble and therefore absorbed in the intestine along with the lipids. Reports also indicate that the inclusion of antioxidants in diets rich in unsaturated fatty acids, which are susceptible to oxidation, improves yolk pigmentation. In addition, Faitarone et al. (2016) stated that when dietary lipids produce peroxides, yolk pigmentation can be negatively affected due to the oxidation of carotenoids. The discoloration (oxidation) of carotene is induced by the oxidative degradation products of linoleic acid (Kumazawa et al. 2003). Berretta et al. (2017) stated that the antioxidant property of propolis is one of the most studied biological activities worldwide. During analyzes of Brazilian propolis, Wang et al. (2004) observed a strong inhibition of lipid peroxidation using rat liver homogenate at a concentration of 2 mg/mL, and this activity was related to the presence of flavonoids. However, it is known that, in addition to phenolic compounds, flavonoids are involved in the antioxidant activity of propolis. Thus, a number of phenolic compounds, including flavonoids, were evaluated against linoleic acid peroxidation in micellar solution. The results showed that polyphenols in general have greater activity than BHT (butylated hydroxytoluene), a well-known antioxidant (Bankosta et al. 2001). In a study using cell culture, artepillin C was proposed as a strong candidate responsible for the antilipoperoxidative activity of Brazilian propolis (Shimizu et al. 2004). The pH of the yolk of fresh eggs is usually around 6.0 and was significantly lower in the red propolis treatments compared to the control group. According to Sarcinelli et al. (2007) there is an increase in pH and connections between the molecules that make up the yolk membrane that surrounds the yolk loses selectivity and the water moves from the albumen to the yolk, increasing the size of the membrane that has already found fragile and thus, stretching it. Alkaline ions from the albumen can be exchanged with H+ ions present in the yolk with an increase in yolk pH. This pH variation could induce protein denaturation and increase yolk consistency (Shang et al. 2004). Egg quality after storage In the storage room the average temperatures registered were 22.1ºC with maximun 25.2ºC and minimun 18.7ºC and the average of the relative humidity of the air registered were 62.5% with maximun 71% and minimun 40%. Under these conditions, the lowest weight loss of eggs stored after 21 days was obtained in the group submitted to red propolis 1g and no significant diferences (P>0.05) were observed in these variables compared to the control and enramycin treatment (Table 4). The lowest levels of lipid peroxidation were in the egg yolks of quails subjected to treatment with enramycin and red propolis 1g, although it was not observed any significant difference (P>0.05) between birds fed with the control treatment and with the two levels of red propolis. Cabral et al. (2009) compared the antioxidant potential of different fractions obtained from a hydroethanolic extract of red propolis, through DPPH radical scavenging methods and oxidation inhibition of the β-carotene/linoleic acid system. The first method showed that the hexane fraction showed greater antioxidant activity (74.4%), while in the second method the chloroform fraction showed greater activity (64.8%). The study concluded that there is a greater correlation between the content of phenolic compounds and antioxidant activity by the oxidation of the β-carotene/linoleic acid system. According to Righi et al. (2011), active compounds such as isoflavones and chalcones have greater affinity for the organic phase. In a later study, the following compounds were isolated from the chloroform fraction: two isoflavonoids (vestitol and neovestitol) and a chalcone (isoliquiritigenin). Among these compounds, vestitol showed greater antioxidant activity compared to the others, through the β-carotene/linoleic acid model (Oldoni et al. 2011). Frozza et al. (2013) analyzed a hydroethanolic extract of red propolis and obtained strong enzymatic activity such as superoxide desmutase (SOD) and catalase (CAT), which are important in oxidative stress. Microbiological Analysis At the end of the third experimental cycle, a difference was observed in colony forming units (CFU) of total count of mesophilic aerobics on the surface of fresh and stored eggs compared to the second cycle (Table 5). Considering 100% presence of colonies in the second cycle, there was a representative decrease of colonies on the surface of the eggs of the group red propolis 1g at the end of the third cycle and in the stored eggs. A significant decrease was observed in the presence of CFU of mesophilic aerobics in the fecal samples of quails submitted to four different treatments throughout the experiment (Table 6). Considering 100% presence of colonies in the first cycle, there was a decrease in CFU in the analyzed feces. The count of total coliforms in the feces of quails submitted to different treatments at the end of the first cycle showed the presence of CFU (Table 7). The highest amount of CFU/mL was observed in the enramycin group and the lowest counts of CFU in the feces of the quails in the red propolis 1g group, i.e. 83.18% less when compared to the enramycin group. There were 70% fewer Escherichia coli colonies in the red propolis 1g group when compared to the enramycin group. Bueno-Silva et al. (2016) analyzed Brazilian red propolis, whose primary plant source was Dalbergia ecastophyllum . They studied the effect of the propolis collection period, its chemical composition and antibacterial activity. Seasonal variability was observed between the concentration of vestitol, neovestitol and isoliquiritigenin. The highest content of these ingredients and the antibacterial activity were recorded during the rainy season (period from January to May). In terms of antibacterial activity, the content of substances such as flavonoids and phenolic compounds is important (Inui et al. 2014; Górniak et al. 2019). However, depending on the solvent used, different biological activities are found (Przybyłek & Karpiski, 2019). Kubiliene et al. (2015) compared the composition and biological activities of propolis extracts prepared with an alternative non-alcoholic solvent mixture such as polyethylene glycol (PEG 400 PA), water and olive oil and concluded that propolis extraction with non-alcoholic solvents and the effects the high temperatures allow the most effective extraction of active compounds from propolis and that the non-ethanolic extracts of propolis have anti-radical and antimicrobial action. More studies should be developed by this research group in order to compare different extraction methods. Conclusion The red propolis aqueous extract supplementation as a feed additive in the diet of laying quails resulted in eggs with darker yolks, a desired characteristic for consumers, in addition to yolks with pH 6.0 which could impact shelf life. Moreover, red propolis 1g group showed antimicrobial action, caused lower weight loss in eggs stored for 21 days and lower lipid peroxidation in the yolk, indicating a potent antioxidant action, without affecting quail productive performance. In general, it is concluded that supplementing quails diet with red propolis 1g group can be an useful alternative to antibiotics in order to maintain the productive performance of quails, and improve physicochemical and microbiological quality of the eggs. Declarations Funding The authors thank the Brazilian National Council for Scientific and Technological Development - CNPq and the Scientific, Technological Research Support Foundation of Santa Catarina State – FAPESC and University of the State of Santa Catarina (UDESC), Chapecó, SC, Brazil. Competing Interests The authors have no relevant financial or non-financial interests to disclose. Conflict of interest The authors declare no competing interests. Author Contributions Patricia Rodrigues Antelo López Garcia conducted research and wrote the first draft of the manuscript. Authors: Patricia Rodrigues Antelo López Garcia, Marcel Manente Boiago, Denise Nunes Araujo, Lenita de Cássia Moura Stefani, contributed to the study conception and design. Material preparation, data collection and analysis were performed by Arieli Zibetti França, Bruno Milhoreto Sponchiado, Daniela Tomazi Nesi, Guilherme Luiz Deolindo, Maiara Rampazzo, Marcos José Migliorini, Paulo Vinicius de Oliveira Junior. All authors commented on previous versions of the manuscript, read and approved the final manuscript. Data availability The datasets generated and analysed during the current study are available from the corresponding author on reasonable request. Ethics approval At the State University of Santa Catarina, the ethics committee approved the project using animals in research, protocol number 2307170920. All names in the author list have been involved in various stages of experimentation or writing. References Albino, L.F.T., Barreto, S.L.T., 2003. Criação de codornas para produção de ovos e carnes. 1° ed. Viçosa, (Aprenda Fácil, Viçosa). Andreotti, R., Nicodemo, M.L.F., 2004. Uso de Antimicrobianos na produção de bovinos e desenvolvimento de resistência, Embrapa Gado de Corte, 144, 50. Bankosta, A.H., Tezuca, Y., Kadota, S., 2001. Recent progress in pharmacological research of propolis, Phytotherapy Research, 15, 561-571. Bankova, V., Christov, R., Popov, S., Marcucci, M.C., Tsvetkova, I., Kujumgiev, A., 1999. Antibacterial activity of essencial oils from Brazilian propolis, Fitoterapia, 70, 190-193. Bankova, V., 2005. Recent trends and important developments in propolis research, Evid Based Complementary Alternative Medicine, 2, 29-32. Barbosa, N.A.A., Sakomura, N.K., Menonça, M.O., Freitas, E.R., Fernandes, J.B.K., 2008. Qualidade de ovos comerciais provenientes de poedeiras comerciais armazenados sob diferentes tempos e condições de ambientes, Ars Veterinarian, 24, 127-133. Bastos-Leite, S.C., Alves, E.H.A., De Sousa, A.M., Goulart, C.D.E.C., Dos Santos, J.P.M., Silva, J.D.B., 2016. Ácidos orgânicos e óleos essenciais sobre o desempenho, biometria de órgãos digestivos e reprodutivos de frangas de reposição, Acta Veterinarian Brasilica, 10, 201-207. Berretta, A.A., 2017. Functional properties of Brazilian propolis: from chemical composition until the market. Superfood and Functional Food, An Overview of Their Processing and Utilization, Intech, chapter 4, 55-99. Brasil. Instrução Normativa nº 13, de 30 de novembro de 2004. Aprova o Regulamento Técnico sobre Aditivos para Produtos Destinados à Alimentação Animal. In: Brasília, DF, Presidência da República. 2004. http://sistemasweb.agricultura.gov.br/sislegis/action/detalhaAto.do?method=visualizarAtoPortalMapa&chave=133040692. Accessed 21 Sept 2021 Bueno, R., Albuquerque, R., De Murarolli, V.D.A., Hernandez Aya, L.A., Raposo, R.D.A.S., Bordin, R.D.A., 2012. Efeito da influência de probiótico sobre a morfologia intestinal de codornas japonesas, Brazilian Journal Veterinarian Research Animal Science, 49, 111-115. Bueno-Silva, B., Alencar, S.M., Koo, H., Ikegaki, M., Silva, G.V.J., Napimoga, M.H., Rosalen, P.L., 2013. Anti-inflammatory and antimicrobial evaluation of neovestitol and vestitol isolated from Brazilian red propolis, Journal Agriculture Food Chemistry, 61, 4546-4550. Bueno-Silva, B., Marsola, A., Ikegaki, M., Alencar, S.M., Rosalen, P.L., 2016. The effect of seasons on Brazilian red propolis and its botanical source: chemical composition and antibacterial activity, Natural Products Research, 31, 1318-1324. Burdock, G.A., 1998. Review of the biological properties and toxicity of bee propolis (propolis), Food and Chemical Toxicology, 36, 347-363. Buriol, L., Finger, D., Schmid, E.M., Dos Santos, J.M.T., Da Rosa, M.R., Quináia, S.P., Torres, Y.R., 2009. Composição química e atividade biológica de extrato oleoso de própolis: uma alternativa ao extrato etanólicos, Química Nova, 32, 296-302. Cabral, I.S.R., Oldoni, T.L.C., Prado, A., Bezerra, R.M.N., De Alencar, S.M., Ikegaki, M., Rosalen, P.L., 2009. Composição fenólica, atividade antibacteriana e antioxidante da própolis vermelha brasileira, Química Nova, 32, 1523-1527. Carpes, S.T., Begnini, R., Alencar, S.M., Masson, M.L., 2007. Study of preparations of bee pollen extracts, antioxidant and antibacterial activity, Ciência e Agrotecnologia, 31, 1818-1825. Cavedish, R.L., 2015. Antinociceptive and anti-inflammatory effects of Brazilian red propolis extract and formononetin in rodents, Journal Ethnopharmacology, 173, 127-133. De Mendonça, I.C.G., 2015. Brazilian red propolis: phytochemical screening, antioxidant activity and effect against cancer cells, BMC Complementary Medicine and Therapies, 15, pp 357. Faitarone, A.B.G., Garcia, E.A., Roça, R.O., Andrade, E.M., Vercese, F., Pelícia, K., 2016. Yolk color and lipid oxidation of the eggs ofcommercial white layers fed diets supplemented with vegetable oils, Brazilian Journal of Poultry Science, 18, 9-16. Fokt, H., Pereira, A., Ferreira, A.M., Cunha, A., Aguiar, C., 2010. How do bees prevent hive infections? The antimicrobial properties of propolis, Current Research, Technology and Education. Topics in Applied Microbiology and Microbial Biotechnology, 1, 481–493. Frozza, C., 2013. Chemical characterization, antioxidant and cytotoxic activities of brazilian red propolis, Food and Chemical Toxicology, 52 , 137-142. Fukayama, E.H., Bertechini, A.G., Geraldo, A., Kato, R.K., Murgas, L.D.S., 2005. Extrato de oregano como aditivo em rações para frangos de corte, Revista Brasileira Zootecnia, 34, 2316-2326. Garcia-Migura, L., Hendriksen, R.S., Fraile, L., Aarestrup, F.M., 2014. Antimicrobial resistance of zoonotic and commensal bacteria in Europe: the missing link between consumption and resistance in veterinary medicine, Veterinary Microbiology, 170, 1-9. Gentry, R.F., Quarles, C.L., 1972. The measurement of bacterial contamination on egg shells, Poultry Science, 51, 930-933. Giampietro, A., Scatolini, A.M., Boiago, M.M., 2008. Estudo da metodologia de TBARS em ovos, Revista Avisite, 13, pp 18. Górniak, I., Bartoszewski, R., Króliczewski, J., 2019. Comprehensive review of antimicrobial activities of plant flavonoids, Phytochemistry Reviews, 18, 241–272. Inui, S., Hatano, A., Yoshino, M., Hosoya, T., Shimamura, Y., Masuda, S., Ahn, M.R., Tazawa, S., Araki, Y., Kumazawa, S., 2014. Identification of the phenolic compounds contributing to antibacterial activity in ethanol extracts of Brazilian red propolis, Natural Product Research, 28, 1293-1296. Kacániová, M., Rovná, K., Arpásová, H., Cubon, J., Hleba, L., Pochop, J., Kunová, S., Hascík, P., 2012. In vitro and in vivo antimicrobial activity of propolis on the microbiota from gastrointestinal tract of chickens, Journal of Environmental Science and Health, 47, 1665-1671. Kedzia, B., Holdrna-Kedzia, E., 2013. The antibiotic activity of native and European propolis. Postępy Fitoterapii, 2, 97-107. Kubiliene, L., Laugaliene, V., Pavilonis, A., Maruska, A., Majiene, D., Barcauskaite, K., Kunilus, R., Kasparaviciene, G., Savickas, A., 2015. Alternative preparation of propolis extracts: comparison of their composition and biological activities, BMC Complementary Alternative Medicine, 15, 1-7. Kumazawa, S., Yoneda, M., Shibata, I., Kanaeda, J., Hamasaka, T., Nakayama, T., 2003. Direct evidence for the plant origin of Brazilian propolis by the observation of honeybee behavior and phytochemical analysis, Chemical and Pharmaceutical Bulletin, 51, 740-742. Lee, B.D., Kim, D.J., Lee, S.J., 2001. Nutritive and economic values of high oil corn in layer diet, Poultry Science, 80, 1527-1534. Marieke, M., Blitterswijk, H., Leven, L., Kerkvliet, J., Waerd, J., 2005. Bee products (properties, processing and marketing), Nectar 42, 33-35. Mendonça-Melo, L., Mota. E., Lopez, B., Sawaya, A., Freitas, L., Jain, S., Batista, M., Araújo, E., 2017. Chemical and genetic similarity between Dalbergia ecastaphyllum and red propolis from the Northeastern Brazil, Journal Apiculture Research, 56, 1-8. Oldoni, T.L.C., Cabral, I.S.R., D’Arcea, M.A.B.R., Rosalen, P.L., Ikegaki, M., Nascimento, A.M., Alencar, S.M., 2011. Isolation and analysis of bioactive isoflavonoids and chalcone from a new type of Brazilian propolis, Separation and Purification Technology, 77, 208-213. Pelicano, E.R.L., Souza, P.A., Souza, H.B.A., 2002. Prebióticos e probióticos na nutrição de aves, Ciências Agrárias Saúde, 2(1), 59-64. Petrolli, T.G., Demeda, L., Zotti, C.A., Palhano, J., Simionatto, A.T., 2014. Utilização do resíduo do extrato de própolis verde como promotor de crescimento para frangos de corte, Enciclopédia Biosfera, 10, 1859-1868. Przybylek, I., Karpinski, T.M., 2019. Antibacterial properties of propolis, Molecules, 24, 2047. Reis, C.M., Carvalho, J.C.T., Caputo, L.R.G., 2000. Anti-inflammatory and antiulcer activity and subchronic toxicity of propolis ethanolic extract, Brazilian Journal of Pharmacognosy, 10, 43-49. Righi, A.A., Alves, T.R., Negri, G., Marques, L.M., Breyer, H., Salatino, A., 2011. Brazilian red propolis: unreported substances, antioxidant and antimicrobial activities, Journal Science Food Agriculture, 91, 2363-2370. Rostagno, H.S., 2017. Composição de alimentos e exigências nutricionais. Tabelas brasileiras para aves e suínos, 4ª ed., (Universidade Federal de Viçosa - MG), pp 488. Salatino, A., Fernandes-Silva, C.C., Righi, A.A., Salatino, M.L.F., 2011. Propolis research and the chemistry of plant products, Natural Product Research, 28, 925–936. Sarcinelli, M.F., Venturini, K.S., Silva, L.C., 2007. Características dos ovos. Universidade Federal do Espírito Santo, UFES, Boletim Técnico, PIE-UFES: 00707 http://www.agais.com/telomc/b00707_caracteristicas_ovos.pdf Accessed 02 Sept 2021. Shang, X.G., Wang, F.L., Li, D.F., Yin, J.D., Li, J.Y., 2004. Effects of dietary conjugated linoleic acid on the productivity of laying hens and egg quality during refrigerated storage, Poultry Science, 83(10), 1688-1695. Silva, J.D.T., Matos, A.D.A.S., Hada, F.H., Gravena, R.A., Marques, R.H., Moraes, V.M.B., 2012. Simbiótico e extratos naturais na dieta de codornas japonesas na fase de postura, Ciência Animal Brasilera, 13, 1-7. Silva, J.H.V., Albino, L.F.T., Godoi, M.J.S., 2000. Efeito do extrato de urucum na pigmentação da gema dos ovos, Revista Brasileira Zootecnia 29, 1435-1439. Shimizu, K., Ashida, H., Matsuura, Y., Kanazawa, K., 2004. Antioxidative bioavailability of artepillin C in Brazilian propolis, Archives of Biochemistry and Biophysics, 424, 181-188. Tocchini, L., Mercadante, A.Z., 2001. Extração e determinação, por CLAE, de bixina e norbixina em coloríficos, Ciência Tecnologia Alimentos, 21, 310-313. Trusheva, B., Popova, M., Bankova, V., Simova, S., Marcucci, M.C., Miorin, P.L., Pasin, F.R., Tsvetkova, I., 2006. Bioactive Constituents of Brazilian Red Propolis, Oxford University Press, 3, 249-254. Umigi, R.T., Barreto, S.L.T., Reis, R.S., Mesquita, R.M.F., Araújo, M.S., 2012. Níveis de treonina digestível para codorna japonesa na fase de produção, Arquivo Brasileiro Medicina Veternária Zootecnia, 64, 658-664. Wang, B.J., Lien, Y.H., Yu, Z.R. 2004. Supercritical fluid extractive fractionation - study of the antioxidant activities of propolis, Food Chemistry, 86, 237-243. Zavarize, K.C., Sartori, J.R.A., Pelícia, V.C.B., Pezzato, A.C.C., Araujo, P.C.D., Stradiotti, A.C.E., Madeira, L.A., 2011. Glutamina e nucleotídeos na dieta de frangos de corte criados no sistema alternativo, Archives Zootecnia, 60, 913-920. Zeweil, H.S., Zahran, S.M., Abd El-Rahman, M.H.A., Dosoky, W.M., Abu Hafsa, H., Moktar, A.A., 2016. Effect of using bee propolis as natural supplement on productive and physiological performance of japanese quail, Egypt Poultry Science Journal, 36, 161-175. Tables Table 1. Ingredient and nutrient composition of basal diet. Ingredients (%) Quantity Corn 58.7 Soybean meal 31.7 Soybean oil 0.18 Dicalcium phosphate 1.22 Calcium Carbonate 6.84 Sodium chloride (NaCl) 0.34 DL-Methionine (99%) 0.38 Lysine 0.24 Vitamin-mineral premix A 0.20 Nutrient composition (%) Quantity Crude Protein 19.9 Metabolizable energy (kcal/kg) 2.75 Calcium 3.00 Available Phosphorus 0.32 Digestible Lysine 1.11 Digestible Methionine 0.66 Methionine + digestible cysteine 0.98 A Product composition (kg): vitamin (vit.) A 5,000,000 IU; vit. D3 1,000,000 IU; vit. E 15,000 IU; vit. K3 1500 mg; vit. B1 1500 mg; vit. B2 3000 mg; vit. B6 2000 mg; vit. B12 7000 μg; folic acid 500 mg; nicotinic acid 15 g; pantothenic acid 7000 μg; choline 80 g; biotin 100 mg; Cu 10 g; Fe 50 g; I 1000 mg; Mn 80 g; Selenium 300 mg; Zn 70g. Table 2. Effects of red propolis feed supplementation on the productive performance of quails. Parameters CT TE RP1 RP2 P-Value Egg production 95.83 94.72 94.30 92.73 0.63 Feed consumption 27.29 26.54 26.50 26.09 0.32 Average egg weight 11.09 10.88 11.08 10.93 0.43 Eggs mass 10.65 10.34 10.47 10.16 0.45 Feed convertion (Kg/Kg) 2.56 2.58 2.54 2.58 0.78 Feed convertion (Kg/dz) 0.34 0.33 0.33 0.33 0.83 Viability 100 100 100 100 - CT: control treatment; TE: enramycin treatment; RP1: treatment with red propolis (1g/kg of feed); RP2: treatment with red propolis (2g/kg of feed). Feed conversion (kg/kg): kg of feed consumed per kg of produced egg; Feed conversion (kg/dz): kg of feed per dozen eggs produced. P-Value: P<0.05 indicating significance level by Tukey test at 95% confidence. Table 3. Effects of red propolis supplementation on quail egg quality. Parameter CT TE RP1 RP2 P-Value Eggshell strength 1,424.17 1,444.22 1,424.70 1,464.90 0.99 Specific gravity 1.064 1.067 1.067 1.067 0.64 Haugh unit 87.82 85.99 85.56 86.29 0.16 Yolk index 0.4708 0.4558 0.4775 0.4642 0.13 Colorimetric fan 5.75 5.96 5.92 5.69 0.86 Luminosity (L*) 65.629a 60.881b 59.428b 57.11b <0.0001 Intensity of red (a*) -9.96a -9.33ab -8.072b -7.92b 0.0095 Intensity of yellow (b*) 53.76a 49.07b 46.93b 46.43b <0.0001 Yolk % 31.49 31.86 32.12 32.63 0.69 Shell % 8.59 8.71 8.60 8.50 0.95 Albumen % 59.93 59.42 59.27 58.86 0.69 Shell thickness 0.17 0.18 0.18 0.19 0.10 Yolk pH 6.21a 6.05ab 5.88b 5.90b 0.0125 Albumen pH 8.58 8.59 8.59 8.60 0.90 CT: control treatment; TE: enramycin treatment; RP1: treatment with red propolis (1g/kg of feed); RP2: treatment with red propolis (2g/kg of feed). P-Value: P< 0.05 indicating significance level by Tukey test at 95% confidence. Means followed by different letters on the same line indicate a difference by Tukey's test at 5% significance (P<0.05). Table 4. Effects of red propolis supplementation for quails on egg weight and lipid peroxidation levels after 21 days of storage. Parameter CT TE RP1 RP2 P-Value AWL (g) 3.93ab 4.01ab 3.74b 4.14a 0.012 TBARS 0.32a 0.07b 0.12ab 0.29a 0.007 CT: control treatment; TE: enramycin treatment; RP1: treatment with red propolis (1g/kg of feed); RP2: treatment with red propolis (2g/kg of feed). AWL = average weight loss, P-Value: P<0.05 indicating significance level by Tukey test at 95% confidence. Means followed by different letters on the same line indicate a difference by Tukey's test at 5% significance (P<0.05). Table 5. Effects of red propolis on total count of mesophilic aerobics on the surface of quail eggs. Treatment Second cycle Third cycle Stored CFU/mL (%) CFU/mL (%) CFU/mL (%) TC 29 100 12 41.38 29 100.00 TE 12 100 10 83.33 4 33.33 RP1 42 100 4 9.50 4 9.52 RP2 3 100 2 66.66 8 266.67 TC: control treatment; TE: enramycin treatment; RP1: treatment with propolis (1g/kg of feed); RP2: treatment with propolis (2g/kg of feed). (%): percentage of colony presence considering 100% presence in the second cycle. CFU/mL: colony forming units per mL of sample. Table 6. Effects of red propolis on the total mesophilic aerobic counts in quail feces. Treatment First cycle Second cycle Third cycle CFU/mL (%) CFU/mL (%) CFU/mL (%) CT 136 100 2 1.47 9 6.61 TE 92 100 0 0 8 8.69 RP1 48 100 0 0 20 41.66 RP2 338 100 2 0.59 4 1.18 CT: control treatment; TE: enramycin treatment; RP1: treatment with propolis 1g/kg of feed; RP2: treatment with propolis 2g/kg of feed. (%): percentage of colony presence considering 100% presence in the first cycle. CFU/mL: colony forming units per mL of sample. Table 7. Counts of total and fecal coliforms in quail feces after the first cycle. Treatment TC (CFU/mL) EC (CFU/mL) CT 105 70 TE 327 110 RP1 55 33 RP2 196 102 CT: control treatment; TE: enramycin treatment; RP1: treatment with propolis 1g/kg of feed; RP2: treatment with propolis 2g/kg of feed. TC: total coliforms; EC: Escherichia coli ; CFU/mL: colony forming units per mL. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-1485683","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":106471487,"identity":"f5eaa193-a879-4b8a-9d8a-54a3aa114560","order_by":0,"name":"Patricia López Garcia","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7UlEQVRIiWNgGAWjYFACNgaGByCamfkgiObhI0pLAohmZ0s2AGlhI14LP4+aBJSPH5izH0t8kFBxR06+mYet8muOnQwbA/PDRzfwaLHsSTtskHDmmTFjM++x27LbkoEOYzM2zsGjxeBAeptEYtvhxGZmvrTbktuYgVp42KTxajn/vP0HUEt9GzOPWbHktnoitNxIO8YA1JLAA9TC+HHbYcJaLGc8S5YA+sVwBjNbsjTjtuM8bMwE/GLOn2b44UPFHXn5/sMHP/7cVm3Pz9788DFeh0GoA2CSmQdM4lGOoYXxBwHVo2AUjIJRMDIBAP5sRggXEyA1AAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-9899-3586","institution":"Universidade do Estado de Santa Catarina","correspondingAuthor":true,"prefix":"","firstName":"Patricia","middleName":"López","lastName":"Garcia","suffix":""},{"id":106471488,"identity":"100da9c3-71e7-4249-b216-2a21a4154421","order_by":1,"name":"Lenita Stefani","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Lenita","middleName":"","lastName":"Stefani","suffix":""},{"id":106471489,"identity":"2dcb43e2-650b-4702-831c-955ed49b0ee7","order_by":2,"name":"Marcel Boiago","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Marcel","middleName":"","lastName":"Boiago","suffix":""},{"id":106471490,"identity":"c244e773-c25c-4bf9-8d69-d00389cfad94","order_by":3,"name":"Denise Araujo","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Denise","middleName":"","lastName":"Araujo","suffix":""},{"id":106471491,"identity":"db12d745-5e22-4add-84e1-d3aac6674cbb","order_by":4,"name":"Arieli França","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Arieli","middleName":"","lastName":"França","suffix":""},{"id":106471492,"identity":"b2225b25-10f8-4fbc-9f09-9ab04806cb15","order_by":5,"name":"Bruno Sponchiado","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Bruno","middleName":"","lastName":"Sponchiado","suffix":""},{"id":106471493,"identity":"b1d30d40-342c-4385-a457-dd626e874084","order_by":6,"name":"Daniela Nesi","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Daniela","middleName":"","lastName":"Nesi","suffix":""},{"id":106471494,"identity":"0476be46-64d7-4540-85f8-f6b60b0a6bab","order_by":7,"name":"Guilherme Deolindo","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Guilherme","middleName":"","lastName":"Deolindo","suffix":""},{"id":106471495,"identity":"80623ea7-462c-4ec3-a783-0ea9fb494b52","order_by":8,"name":"Maiara Rampazzo","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Maiara","middleName":"","lastName":"Rampazzo","suffix":""},{"id":106471496,"identity":"4ca0f3db-e5d1-44dd-b6b3-fca996c54d88","order_by":9,"name":"Marcos Migliorini","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Marcos","middleName":"","lastName":"Migliorini","suffix":""},{"id":106471497,"identity":"cf95d74f-3c96-4190-ba01-1d416c79c5ce","order_by":10,"name":"Paulo de Oliveira Junior","email":"","orcid":"","institution":"University of Santa Catarina State: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Paulo","middleName":"de Oliveira","lastName":"Junior","suffix":""}],"badges":[],"createdAt":"2022-03-24 13:37:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1485683/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1485683/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":25889836,"identity":"b3babb32-7a28-467a-b854-42c30c3263ae","added_by":"auto","created_at":"2022-08-31 13:50:09","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":244002,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1485683/v1/238fcec8-3e5c-49ad-bc6b-3719bbe84d11.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eEffects of aqueous extract of red propolis as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBee products appear as an alternative for antibiotics and promising food source in animal nutrition, due to their therapeutic substances (Carpes et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Propolis is an apicultural product made with resins collected by \u003cem\u003eApis mellifera\u003c/em\u003e bees, from vegetative parts of plants or resinous exudates. In general, propolis is composed of 50% resin and balm, 30% wax, 10% essential and aromatic oils, 5% pollen and 5% various other substances (Burdock \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Fokt et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Its color ranges from green, red to dark brown, and this variation is related to the geographical origin and the vegetation from which it is extracted (Salatino et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe red propolis collected in the northeast region of Brazil has resinous exudates from the \u003cem\u003eDalbergia ecastophyllum\u003c/em\u003e plant, as a botanical source (Righi et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Mendon\u0026ccedil;a-Melo et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). It contains pterocarpans, isoflavonoids, chalcones and phenylpropanoids (Righi et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) as main constituents. It showed cytotoxic activity against various types of cancer cells, antibacterial, antifungal, anticariogenic, antioxidant, anti-inflammatory and antiproliferative activity (De Mendon\u0026ccedil;a et al. 2015; Cavendish et al. 2015). In this context, this study aimed to evaluate whether the addition of aqueous extrat of red propolis to the diet of laying quails could improve productive performance, as well as physicochemical and microbiological quality of eggs.\u003c/p\u003e \u003cp\u003eRaising quails has become a profitable activity due to its relevant characteristics, including their resistance to heat and diseases, low requirement for housing, and also due to the fact that quail eggs and meat are nutritious and tasty, presenting themselves as an additional meat protein option for humans (Umigi et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePoultry farming has been using for many years some tools that have resulted in higher growth and higher yield, including the use of antibiotics as growth promoters (Pelicano et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). However, misuse of certain antimicrobials may promote the emergence of resistant bacteria (Garcia-Migura et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) and nowadays there is a search for more natural alternative additives, with similar animal performance (Zavarize et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003eThe experiment was conducted in Japanese quails (\u003cem\u003eCoturnix japonica\u003c/em\u003e) egg producing base at the Poultry Sector from UDESC (Chapec\u0026oacute;, SC, Brazil), from 5 March to 9 May 2021.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAnimal welfare statement\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe animals used in this study and all experimental procedures were approved by the Committee of Ethics for Animal Use (CEUA) from Santa Catarina State University (Brazil) under protocol number 2307170920. All methods were in accordance with the approved guidelines and regulations.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;Experimental design and management\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe experiment was carried out at the Poultry Sector, the extraction of propolis and the analyzes were performed at the Laboratory of Molecular Biology, Immunology and Microbiology (LABMIM) both from the Animal Science Department of the Santa Catarina State University, in the city of Chapec\u0026oacute;, Santa Catarina State, Brazil.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eOne hundred twenty 52 day-old female Japanese\u003c/em\u003equails were used as experimental animals and quails were fed a diet without addition of RP for 17 days to make the animals accustomed to the experimental diets,\u0026nbsp;with water \u003cem\u003ead libitum\u003c/em\u003e. Then the quails were assigned to\u0026nbsp;randomly distributed in four treatments, with six replications, with five quails per cage, totaling twenty-four experimental plots. The study lasted sixty three days, divided into three cycles of twenty one days each. The experimental quails were fed the basal ration was formulated according to the nutritional values and requirements established in the Brazilian Table of Poultry and Swine (Rostagno et al. 2017), with 2,750 Kcal of metabolizable energy per kg of ration and 19.9% of protein (Table 1). The treatments following: control treatment (CT): basal diet without antimicrobials and without red propolis; enramycin treatment (TE): 10 mg/kg of feed; red propolis (RP1): 1g of red propolis/kg of feed and red propolis (RP2): 2g of red propolis/kg of feed. The red propolis aqueous extract was mixed with the microingredients and later added to the vertical mixer. The lighting program used for the quails was 16 hours of light per day.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eRed propolis aqueous extract\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eRed propolis was purchased from a commercial company located in the city of Canavieiras, Bahia State, Brazil. The extraction was performed according to the methodology adapted from Kubiliene et al. (2015), with the maceration of crude propolis with pistil and liquid nitrogen, inside a mortar until it becomes powder. In a beaker, 100 mL of distilled water, 10 grams of propolis and 20 grams of polyethylene glycol (PEG 400 PA) were added and the mixture was subjected to high pressure at 120\u0026deg;C for 5 minutes in an autoclave. After extraction, the extract was stored protected from light. The extract was homogenized and weighed for subsequent mixing with the feed.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eProductive P\u003c/em\u003e\u003cem\u003eerformance parameters\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eDaily quail mortality was observed and recorded in order to calculate animal viability, as well as feed intake per quail (g/quail.day). The number of eggs produced was recorded daily and the average production per quail was estimated, in addition to egg mass (g/quail.day). Feed conversion was evaluated by kg of feed consumed per kg of eggs produced and also per kg of dozens of eggs. Eggs were weighed in the last five days of each experimental period and the average egg weight was determined.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u003cem\u003earameters in fresh eggs\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eTo assess egg quality, a sample consisting of two eggs from each experimental plot was collected and analyzed. Specific gravity was determined according to Barbosa et al. (2008). Texture analyzer equipment was used to measure egg shell strength and results were expressed as kgf. After breaking the eggs the following parameters were evaluated: height of the dense albumen measured with a tripod micrometer; Haugh unit (HU) according to Haugh (1937); yolk index, which was calculated by the ratio between the height and diameter of the yolk (mm), obtained with the micrometer and the caliper, respectively; yolk color was determined with the colorimetric fan (DSM) and with the colorimeter (Minolta CR-400), where the following universal colorimetric coordinates were observed: luminosity (L*), red intensity (a*) and yellow intensity (b*); the shell weight and percentages of shell, yolk and albumen; egg shell thickness was obtained with a caliper at three points and obtaining the arithmetic mean of the three measurements. The pH of the yolk and the albumen were measured with the digital phmeter (Testo 205).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eParameters in stored eggs\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe eggs were weighed on a precision scale and stored under temperature and air humidity registered conditions. Twice per day, the temperature, maximum, minimum and the air humidity, maximum, minimum at the room storage were registered, during 21 days. After this period, the eggs were weighed where the weight loss after storage was calculated. The levels of lipid peroxidation in the yolk of these eggs were determined according to Giampietro et al. (2008), with the measurement of thiobarbituric acid reactive substances (TBARS), which are formed during the decomposition of lipid peroxides. Compound 1,1,3,3 tetramethoxypropane (TMP) was used as TBARS standard. The results were expressed in mg TMP/kg of yolk.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eMicrobiological analysis\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eMicrobiological analysis were performed on the surface of eggs produced at the end of the second and third cycles. Each sample consisted of a group of five eggs per repetition. Egg surfaces were washed with peptone water according to the methodology adapted from Gentry \u0026amp; Quarles (1972). Quail feces samples, collected from four points of each experimental plot, were analyzed. Egg and feces samples were sown on Plate Count Agar (PCA) for total mesophilic aerobics and on Petrifilm\u0026reg; 6404 (3M) plates for total coliforms and \u003cem\u003eEscherichia coli\u003c/em\u003e. Incubation was carried out at 37 \u0026ordm;C for 48 hours and the number of bacterial colonies present were expressed in colony-forming units per mL (CFU/mL).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eStatistical analysis\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eData were submitted to analysis of variance and means compared by Tukey test at 5% significance level (in cases of significant diferences). These statistical procedures were performed by the program Statistical Analysis System.\u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003cp\u003e\u003cem\u003eProductive performance\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eEgg production, feed intake, average egg weight, egg mass, feed conversion (kg/kg and kg/dz) are shown in Table 2. No significant differences were observed in these variables compared to the control treatment (P\u0026gt;0.05). The results show that the red propolis aqueous extract did not influence the productive performance of the quails. Our findings are similar to those of Zeweil et al. (2016) while studying the effect of propolis extract (250 and 500 mg/kg) in the diet of Japanese quails, observed that the inclusion of propolis neither influence performance (body weight, laying rate, egg weight and egg mass) nor egg quality (egg weight, yolk and albumen percentage, albumen height, shell percentage and thickness, specific gravity and yolk color). Petrolli et al. (2014) using residue of green propolis extract, at the level of 1%, for broilers, also found no difference (P\u0026gt;0.05) for feed intake and feed conversion in the period from 1 to 21 days of age.\u003c/p\u003e\n\u003cp\u003ePropolis can increase the beneficial microbiota and control pathogenic bacteria (Kac\u0026aacute;niov\u0026aacute; et al. 2012). The change in the dynamics of the intestinal microbiota is generally attributed to flavonoids, which have antibacterial activity and are present in the ethanol extract of propolis (Oldoni et al. 2011; Bueno-Silva et al. 2013; Frozza et al. 2013). Different results might be obtained between studies due to the variability and complexity of the composition of propolis, which changes according to the flora of each location, genetics of queen bees and even the time of year in which it is collection was done (Buriol et al. 2009). Another hypothesis that the addition of propolis did not show any difference in the analyzed variables may have been the fact that the breeding environment did not propose a sanitary challenge that would enable a more expressive response with the use of propolis. Since the birds did not have direct contact with the feces, as they were raised in cages with feces collecting trays, they were raised at an ideal density to develop their full potential (Albino \u0026amp; Barreto, 2003).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMoreover, herbal medicines (propolis), probiotics, symbiotics and organic acids are classified as performance-enhancing additives (Brasil, 2004) and it is notorious how these additives cannot express positive results due to the lack of sanitary challenge in poultry studies, in different experimental variables (Silva et al. 2012; Bueno et al. 2012; Bastos-Leite et al. 2016). Because, good prophylactic conditions for raising animals and a minimum of stress (which is usually associated with nutritional, environmental or behavioral factors), do not present a sufficient increase in bacteria to cause an imbalance in intestinal health, compromising productive performance (Fukayama et al. 2005).\u003c/p\u003e\n\u003cp\u003eIt is known that red propolis aqueous extract has great potential to be used as a herbal additive in poultry feed, as red propolis has biologically active compounds (vestitol and neovestitol and biochanin A, liquiritigenin, formononetin and medicarpine) found only in this type of propolis. Trusheva et al. (2006) concluded the identification of new propolis constituents in red Brazilian propolis, most of them having antibacterial, antimycotic and antiradical activities, is a further confirmation of the fact that propolis, independently of its plant source and chemical composition, present antimicrobial and antioxidant activity. Its result that propolis plays in the hive: it is the \u0026lsquo;chemical weapon\u0026rsquo; of bees against pathogen microorganisms and the elements of weather. However, in different propolis types, different chemical constituents are responsible for the valuable activities (Bankova et al. 2005). Furthermore, as can be seen in this experiment, the inclusion of red propolis aqueous extract in the diet of the quails did not cause any negative effects such as mortality or on other studied variables and could be evaluated at different concentrations and with diets free of synthetic antimicrobials. Another study using very high dosage of red propolis in order to address its safety found that red propolis was unable to cause side effects (Reis et al. 2000).\u0026nbsp;Berretta et al. (2017) also reported future perspectives for propolis in the health of the population and in the Brazilian and international market, especially because of its important biological activities and safety.\u003c/p\u003e\n\u003cp\u003eFeeding the quails with 1g of red propolis aqueous extract showed the best feed conversion (kg of feed/kg of eggs), of 2.54 kg, although the difference was not statistically significant compared to the other treatments. Marieke et al. (2005) stated the improvement in feed, conversion rate could be due to the ability of propolis to improve nutrient digestibility and absorption as a result of sucrase, amylase and phosphatase activities.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eQuality of fresh eggs\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eA significant difference (P\u0026lt;0.05) was observed in the results of the eggs analyzed in the following parameters: luminosity (L*), red intensity (a*), yellow intensity (b*) and yolk pH. In the other analyzed parameters: shell strength, specific gravity, Haugh unit, yolk index, colorimetric fan, yolk percentage, shell percentage, albumen percentage, shell thickness and albumen pH, no statistical difference were observed (Table 3).\u003c/p\u003e\n\u003cp\u003eIn summary, the yolk color of the eggs of quails fed with red propolis was darker, with greater intensity of red and lower intensity of yellow when compared to the control treatment, i.e., the eggs of quails fed diet without any additive had yolks of lighter color. Yolk color, as a sensory attribute, is considered an indicator of quality, and plays an important role in the acceptance of the egg by consumers, who associate the intense pigmentation of the yolk with higher nutritional value of the egg (Silva et al. 2000; Tocchini \u0026amp; Mercadante, 2001). Yolk color intensity is determined by the incorporation of xanthophylls (a group of carotenoid pigments) present in corn, particularly lutein and zeaxanthin, and depends on the levels of inclusion in the diet. However, other foods can change yolk color depending on their level of inclusion (Silva et al. 2000).\u003c/p\u003e\n\u003cp\u003eLee et al. (2001) stated that changes in yolk color can be observed when supplemental sources of carotenoids are added to the diet since carotenoid pigments are fat-soluble and therefore absorbed in the intestine along with the lipids. Reports also indicate that the inclusion of antioxidants in diets rich in unsaturated fatty acids, which are susceptible to oxidation, improves yolk pigmentation. In addition, Faitarone et al. (2016) stated that when dietary lipids produce peroxides, yolk pigmentation can be negatively affected due to the oxidation of carotenoids. The discoloration (oxidation) of carotene is induced by the oxidative degradation products of linoleic acid (Kumazawa et al. 2003).\u003c/p\u003e\n\u003cp\u003eBerretta et al. (2017) stated that the antioxidant property of propolis is one of the most studied biological activities worldwide. During analyzes of Brazilian propolis, Wang et al. (2004) observed a strong inhibition of lipid peroxidation using rat liver homogenate at a concentration of 2 mg/mL, and this activity was related to the presence of flavonoids. However, it is known that, in addition to phenolic compounds, flavonoids are involved in the antioxidant activity of propolis. Thus, a number of phenolic compounds, including flavonoids, were evaluated against linoleic acid peroxidation in micellar solution. The results showed that polyphenols in general have greater activity than BHT (butylated hydroxytoluene), a well-known antioxidant (Bankosta et al. 2001). In a study using cell culture, artepillin C was proposed as a strong candidate responsible for the antilipoperoxidative activity of Brazilian propolis (Shimizu et al. 2004).\u003c/p\u003e\n\u003cp\u003eThe pH of the yolk of fresh eggs is usually around 6.0 and was significantly lower in the red propolis treatments compared to the control group. According to Sarcinelli et al. (2007) there is an increase in pH and connections between the molecules that make up the yolk membrane that surrounds the yolk loses selectivity and the water moves from the albumen to the yolk, increasing the size of the membrane that has already found fragile and thus, stretching it. Alkaline ions from the albumen can be exchanged with H+ ions present in the yolk with an increase in yolk pH. This pH variation could induce protein denaturation and increase yolk consistency (Shang et al. 2004).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEgg quality after storage\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eIn the storage room the average temperatures registered were 22.1\u0026ordm;C with maximun 25.2\u0026ordm;C and minimun 18.7\u0026ordm;C and the average of the relative humidity of the air registered were 62.5% with maximun 71% and minimun 40%. Under these conditions, the lowest weight loss of eggs stored after 21 days was obtained in the group submitted to red propolis 1g and no significant diferences (P\u0026gt;0.05) were observed in these variables compared to the control and enramycin treatment (Table 4). The lowest levels of lipid peroxidation were in the egg yolks of quails subjected to treatment with enramycin and red propolis 1g, although it was not observed any significant difference (P\u0026gt;0.05) between birds fed with the control treatment and with the two levels of red propolis.\u003c/p\u003e\n\u003cp\u003eCabral et al. (2009) compared the antioxidant potential of different fractions obtained from a hydroethanolic extract of red propolis, through DPPH radical scavenging methods and oxidation inhibition of the \u0026beta;-carotene/linoleic acid system. The first method showed that the hexane fraction showed greater antioxidant activity (74.4%), while in the second method the chloroform fraction showed greater activity (64.8%). The study concluded that there is a greater correlation between the content of phenolic compounds and antioxidant activity by the oxidation of the \u0026beta;-carotene/linoleic acid system.\u003c/p\u003e\n\u003cp\u003eAccording to Righi et al. (2011), active compounds such as isoflavones and chalcones have greater affinity for the organic phase. In a later study, the following compounds were isolated from the chloroform fraction: two isoflavonoids (vestitol and neovestitol) and a chalcone (isoliquiritigenin). Among these compounds, vestitol showed greater antioxidant activity compared to the others, through the \u0026beta;-carotene/linoleic acid model (Oldoni et al. 2011). Frozza et al. (2013) analyzed a hydroethanolic extract of red propolis and obtained strong enzymatic activity such as superoxide desmutase (SOD) and catalase (CAT), which are important in oxidative stress.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eMicrobiological Analysis\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAt the end of the third experimental cycle, a difference was observed in colony forming units (CFU) of total count of mesophilic aerobics on the surface of fresh and stored eggs compared to the second cycle (Table 5). Considering 100% presence of colonies in the second cycle, there was a representative decrease of colonies on the surface of the eggs of the group red propolis 1g at the end of the third cycle and in the stored eggs.\u003c/p\u003e\n\u003cp\u003eA significant decrease was observed in the presence of CFU of mesophilic aerobics in the fecal samples of quails submitted to four different treatments throughout the experiment (Table 6). Considering 100% presence of colonies in the first cycle, there was a decrease in CFU in the analyzed feces. The count of total coliforms in the feces of quails submitted to different treatments at the end of the first cycle showed the presence of CFU (Table 7). The highest amount of CFU/mL was observed in the enramycin group and the lowest counts of CFU in the feces of the quails in the red propolis 1g group, i.e. 83.18% less when compared to the enramycin group. There were 70% fewer \u003cem\u003eEscherichia coli\u003c/em\u003e colonies in the red propolis 1g group when compared to the enramycin group.\u003c/p\u003e\n\u003cp\u003eBueno-Silva et al. (2016) analyzed Brazilian red propolis, whose primary plant source was \u003cem\u003eDalbergia ecastophyllum\u003c/em\u003e. They studied the effect of the propolis collection period, its chemical composition and antibacterial activity. Seasonal variability was observed between the concentration of vestitol, neovestitol and isoliquiritigenin. The highest content of these ingredients and the antibacterial activity were recorded during the rainy season (period from January to May).\u003c/p\u003e\n\u003cp\u003eIn terms of antibacterial activity, the content of substances such as flavonoids and phenolic compounds is important (Inui et al. 2014; G\u0026oacute;rniak et al. 2019). However, depending on the solvent used, different biological activities are found (Przybyłek \u0026amp; Karpiski, 2019). Kubiliene et al. (2015) compared the composition and biological activities of propolis extracts prepared with an alternative non-alcoholic solvent mixture such as polyethylene glycol (PEG 400 PA), water and olive oil and concluded that propolis extraction with non-alcoholic solvents and the effects the high temperatures allow the most effective extraction of active compounds from propolis and that the non-ethanolic extracts of propolis have anti-radical and antimicrobial action. More studies should be developed by this research group in order to compare different extraction methods.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe red propolis aqueous extract supplementation as a feed additive in the diet of laying quails resulted in eggs with darker yolks, a desired characteristic for consumers, in addition to yolks with pH 6.0 which could impact shelf life. Moreover, red propolis 1g group showed antimicrobial action, caused lower weight loss in eggs stored for 21 days and lower lipid peroxidation in the yolk, indicating a potent antioxidant action, without affecting quail productive performance. In general, it is concluded that supplementing quails diet with red propolis 1g group can be an useful alternative to antibiotics in order to maintain the productive performance of quails, and improve physicochemical and microbiological quality of the eggs.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cem\u003eFunding\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe authors thank the Brazilian National Council for Scientific and Technological Development - CNPq and the Scientific, Technological Research Support Foundation of Santa Catarina State \u0026ndash; FAPESC and University of the State of Santa Catarina (UDESC), Chapec\u0026oacute;, SC, Brazil.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eCompeting Interests\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eConflict of interest\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAuthor Contributions\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003ePatricia Rodrigues Antelo L\u0026oacute;pez Garcia conducted research and wrote the first draft of the manuscript. Authors: Patricia Rodrigues Antelo L\u0026oacute;pez Garcia, Marcel Manente Boiago, Denise Nunes Araujo, Lenita de C\u0026aacute;ssia Moura Stefani, contributed to the study conception and design. Material preparation, data collection and analysis were performed by Arieli Zibetti Fran\u0026ccedil;a, Bruno Milhoreto Sponchiado, Daniela Tomazi Nesi, Guilherme Luiz Deolindo, Maiara Rampazzo, Marcos Jos\u0026eacute; Migliorini, Paulo Vinicius de Oliveira Junior. All authors commented on previous versions of the manuscript, read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eData availability\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and analysed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEthics approval\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAt the State University of Santa Catarina, the ethics committee approved the project using animals in research, protocol number 2307170920. All names in the author list have been involved in various stages of experimentation or writing.\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003eAlbino, L.F.T., Barreto, S.L.T., 2003. Cria\u0026ccedil;\u0026atilde;o de codornas para produ\u0026ccedil;\u0026atilde;o de ovos e carnes. 1\u0026deg; ed. Vi\u0026ccedil;osa, (Aprenda F\u0026aacute;cil, Vi\u0026ccedil;osa).\u003c/p\u003e\n\u003cp\u003eAndreotti, R., Nicodemo, M.L.F., 2004. Uso de Antimicrobianos na produ\u0026ccedil;\u0026atilde;o de bovinos e desenvolvimento de resist\u0026ecirc;ncia, Embrapa Gado de Corte, 144, 50.\u003c/p\u003e\n\u003cp\u003eBankosta, A.H., Tezuca, Y., Kadota, S., 2001. Recent progress in pharmacological research of propolis, Phytotherapy Research, 15, 561-571.\u003c/p\u003e\n\u003cp\u003eBankova, V., Christov, R., Popov, S., Marcucci, M.C., Tsvetkova, I., Kujumgiev, A., 1999. Antibacterial activity of essencial oils from Brazilian propolis, Fitoterapia, 70, 190-193.\u003c/p\u003e\n\u003cp\u003eBankova, V., 2005. Recent trends and important developments in propolis research, Evid Based Complementary Alternative Medicine, 2, 29-32.\u003c/p\u003e\n\u003cp\u003eBarbosa, N.A.A., Sakomura, N.K., Menon\u0026ccedil;a, M.O., Freitas, E.R., Fernandes, J.B.K., 2008. Qualidade de ovos comerciais provenientes de poedeiras comerciais armazenados sob diferentes tempos e condi\u0026ccedil;\u0026otilde;es de ambientes, Ars Veterinarian, 24, 127-133.\u003c/p\u003e\n\u003cp\u003eBastos-Leite, S.C., Alves, E.H.A., De Sousa, A.M., Goulart, C.D.E.C., Dos Santos, J.P.M., Silva, J.D.B., 2016. \u0026Aacute;cidos org\u0026acirc;nicos e \u0026oacute;leos essenciais sobre o desempenho, biometria de \u0026oacute;rg\u0026atilde;os digestivos e reprodutivos de frangas de reposi\u0026ccedil;\u0026atilde;o, Acta Veterinarian Brasilica, 10, 201-207.\u003c/p\u003e\n\u003cp\u003eBerretta, A.A., 2017. Functional properties of Brazilian propolis: from chemical composition until the market. Superfood and Functional Food, An Overview of Their Processing and Utilization, Intech, chapter 4, 55-99.\u003c/p\u003e\n\u003cp\u003eBrasil. Instru\u0026ccedil;\u0026atilde;o Normativa n\u0026ordm; 13, de 30 de novembro de 2004. Aprova o Regulamento T\u0026eacute;cnico sobre Aditivos para Produtos Destinados \u0026agrave; Alimenta\u0026ccedil;\u0026atilde;o Animal. In: Bras\u0026iacute;lia, DF, Presid\u0026ecirc;ncia da Rep\u0026uacute;blica. 2004. http://sistemasweb.agricultura.gov.br/sislegis/action/detalhaAto.do?method=visualizarAtoPortalMapa\u0026amp;chave=133040692. Accessed 21 Sept 2021\u003c/p\u003e\n\u003cp\u003eBueno, R., Albuquerque, R., De Murarolli, V.D.A., Hernandez Aya, L.A., Raposo, R.D.A.S., Bordin, R.D.A., 2012. Efeito da influ\u0026ecirc;ncia de probi\u0026oacute;tico sobre a morfologia intestinal de codornas japonesas, Brazilian Journal Veterinarian Research Animal Science, 49, 111-115.\u003c/p\u003e\n\u003cp\u003eBueno-Silva, B., Alencar, S.M., Koo, H., Ikegaki, M., Silva, G.V.J., Napimoga, M.H., Rosalen, P.L., 2013. Anti-inflammatory and antimicrobial evaluation of neovestitol and vestitol isolated from Brazilian red propolis, Journal Agriculture Food Chemistry, 61, 4546-4550.\u003c/p\u003e\n\u003cp\u003eBueno-Silva, B., Marsola, A., Ikegaki, M., Alencar, S.M., Rosalen, P.L., 2016. The effect of seasons on Brazilian red propolis and its botanical source: chemical composition and antibacterial activity, Natural Products Research, 31, 1318-1324.\u003c/p\u003e\n\u003cp\u003eBurdock, G.A., 1998. Review of the biological properties and toxicity of bee propolis (propolis), Food and Chemical Toxicology, 36, 347-363.\u003c/p\u003e\n\u003cp\u003eBuriol, L., Finger, D., Schmid, E.M., Dos Santos, J.M.T., Da Rosa, M.R., Quin\u0026aacute;ia, S.P., Torres, Y.R., 2009. Composi\u0026ccedil;\u0026atilde;o qu\u0026iacute;mica e atividade biol\u0026oacute;gica de extrato oleoso de pr\u0026oacute;polis: uma alternativa ao extrato etan\u0026oacute;licos, Qu\u0026iacute;mica Nova, 32, 296-302.\u003c/p\u003e\n\u003cp\u003eCabral, I.S.R., Oldoni, T.L.C., Prado, A., Bezerra, R.M.N., De Alencar, S.M., Ikegaki, M., Rosalen, P.L., 2009. Composi\u0026ccedil;\u0026atilde;o fen\u0026oacute;lica, atividade antibacteriana e antioxidante da pr\u0026oacute;polis vermelha brasileira, Qu\u0026iacute;mica Nova, 32, 1523-1527.\u003c/p\u003e\n\u003cp\u003eCarpes, S.T., Begnini, R., Alencar, S.M., Masson, M.L., 2007. Study of preparations of bee pollen extracts, antioxidant and antibacterial activity, Ci\u0026ecirc;ncia e Agrotecnologia, 31, 1818-1825.\u003c/p\u003e\n\u003cp\u003eCavedish, R.L., 2015. Antinociceptive and anti-inflammatory effects of Brazilian red propolis extract and formononetin in rodents, Journal Ethnopharmacology, 173, 127-133.\u003c/p\u003e\n\u003cp\u003eDe Mendon\u0026ccedil;a, I.C.G., 2015. Brazilian red propolis: phytochemical screening, antioxidant activity and effect against cancer cells, BMC Complementary Medicine and Therapies, 15, pp 357.\u003c/p\u003e\n\u003cp\u003eFaitarone, A.B.G., Garcia, E.A., Ro\u0026ccedil;a, R.O., Andrade, E.M., Vercese, F., Pel\u0026iacute;cia, K., 2016. Yolk color and lipid oxidation of the eggs ofcommercial white layers fed diets supplemented with vegetable oils, Brazilian Journal of Poultry Science, 18, 9-16.\u003c/p\u003e\n\u003cp\u003eFokt, H., Pereira, A., Ferreira, A.M., Cunha, A., Aguiar, C., 2010. How do bees prevent hive infections? The antimicrobial properties of propolis, Current Research, Technology and Education. Topics in Applied Microbiology and Microbial Biotechnology, 1, 481\u0026ndash;493.\u003c/p\u003e\n\u003cp\u003eFrozza, C., 2013. Chemical characterization, antioxidant and cytotoxic activities of brazilian red propolis, Food and Chemical Toxicology, 52\u003cstrong\u003e,\u003c/strong\u003e 137-142.\u003c/p\u003e\n\u003cp\u003eFukayama, E.H., Bertechini, A.G., Geraldo, A., Kato, R.K., Murgas, L.D.S., 2005. Extrato de oregano como aditivo em ra\u0026ccedil;\u0026otilde;es para frangos de corte, Revista Brasileira Zootecnia, 34, 2316-2326.\u003c/p\u003e\n\u003cp\u003eGarcia-Migura, L., Hendriksen, R.S., Fraile, L., Aarestrup, F.M., 2014. Antimicrobial resistance of zoonotic and commensal bacteria in Europe: the missing link between consumption and resistance in veterinary medicine, Veterinary Microbiology, 170, 1-9.\u003c/p\u003e\n\u003cp\u003eGentry, R.F., Quarles, C.L., 1972. The measurement of bacterial contamination on egg shells, Poultry Science, 51, 930-933.\u003c/p\u003e\n\u003cp\u003eGiampietro, A., Scatolini, A.M., Boiago, M.M., 2008. Estudo da metodologia de TBARS em ovos, Revista Avisite, 13, pp 18.\u003c/p\u003e\n\u003cp\u003eG\u0026oacute;rniak, I., Bartoszewski, R., Kr\u0026oacute;liczewski, J., 2019. Comprehensive review of antimicrobial activities of plant flavonoids, Phytochemistry Reviews, 18, 241\u0026ndash;272.\u003c/p\u003e\n\u003cp\u003eInui, S., Hatano, A., Yoshino, M., Hosoya, T., Shimamura, Y., Masuda, S., Ahn, M.R., Tazawa, S., Araki, Y., Kumazawa, S., 2014. Identification of the phenolic compounds contributing to antibacterial activity in ethanol extracts of Brazilian red propolis, Natural Product Research, 28, 1293-1296.\u003c/p\u003e\n\u003cp\u003eKac\u0026aacute;niov\u0026aacute;, M., Rovn\u0026aacute;, K., Arp\u0026aacute;sov\u0026aacute;, H., Cubon, J., Hleba, L., Pochop, J., Kunov\u0026aacute;, S., Hasc\u0026iacute;k, P., 2012. In vitro and in vivo antimicrobial activity of propolis on the microbiota from gastrointestinal tract of chickens, Journal of Environmental Science and Health, 47, 1665-1671.\u003c/p\u003e\n\u003cp\u003eKedzia, B., Holdrna-Kedzia, E., 2013. The antibiotic activity of native and European propolis. Postępy Fitoterapii, 2, 97-107.\u003c/p\u003e\n\u003cp\u003eKubiliene, L., Laugaliene, V., Pavilonis, A., Maruska, A., Majiene, D., Barcauskaite, K., Kunilus, R., Kasparaviciene, G., Savickas, A., 2015. Alternative preparation of propolis extracts: comparison of their composition and biological activities, BMC Complementary Alternative Medicine, 15, 1-7.\u003c/p\u003e\n\u003cp\u003eKumazawa, S., Yoneda, M., Shibata, I., Kanaeda, J., Hamasaka, T., Nakayama, T., 2003. Direct evidence for the plant origin of Brazilian propolis by the observation of honeybee behavior and phytochemical analysis, Chemical and Pharmaceutical Bulletin, 51, 740-742.\u003c/p\u003e\n\u003cp\u003eLee, B.D., Kim, D.J., Lee, S.J., 2001. Nutritive and economic values of high oil corn in layer diet, Poultry Science, 80, 1527-1534.\u003c/p\u003e\n\u003cp\u003eMarieke, M., Blitterswijk, H., Leven, L., Kerkvliet, J., Waerd, J., 2005. Bee products (properties, processing and marketing), Nectar 42, 33-35.\u003c/p\u003e\n\u003cp\u003eMendon\u0026ccedil;a-Melo, L., Mota. E., Lopez, B., Sawaya, A., Freitas, L., Jain, S., Batista, M., Ara\u0026uacute;jo, E., 2017. Chemical and genetic similarity between \u003cem\u003eDalbergia ecastaphyllum\u003c/em\u003e and red propolis from the Northeastern Brazil, Journal Apiculture Research, 56, 1-8.\u003c/p\u003e\n\u003cp\u003eOldoni, T.L.C., Cabral, I.S.R., D\u0026rsquo;Arcea, M.A.B.R., Rosalen, P.L., Ikegaki, M., Nascimento, A.M., Alencar, S.M., 2011. Isolation and analysis of bioactive isoflavonoids and chalcone from a new type of Brazilian propolis, Separation and Purification Technology, 77, 208-213.\u003c/p\u003e\n\u003cp\u003ePelicano, E.R.L., Souza, P.A., Souza, H.B.A., 2002. Prebi\u0026oacute;ticos e probi\u0026oacute;ticos na nutri\u0026ccedil;\u0026atilde;o de aves, Ci\u0026ecirc;ncias Agr\u0026aacute;rias Sa\u0026uacute;de, 2(1), 59-64.\u003c/p\u003e\n\u003cp\u003ePetrolli, T.G., Demeda, L., Zotti, C.A., Palhano, J., Simionatto, A.T., 2014. Utiliza\u0026ccedil;\u0026atilde;o do res\u0026iacute;duo do extrato de pr\u0026oacute;polis verde como promotor de crescimento para frangos de corte, Enciclop\u0026eacute;dia Biosfera, 10, 1859-1868.\u003c/p\u003e\n\u003cp\u003ePrzybylek, I., Karpinski, T.M., 2019. Antibacterial properties of propolis, Molecules, 24, 2047.\u003c/p\u003e\n\u003cp\u003eReis, C.M., Carvalho, J.C.T., Caputo, L.R.G., 2000. Anti-inflammatory and antiulcer activity and subchronic toxicity of propolis ethanolic extract, Brazilian Journal of Pharmacognosy, 10, 43-49.\u003c/p\u003e\n\u003cp\u003eRighi, A.A., Alves, T.R., Negri, G., Marques, L.M., Breyer, H., Salatino, A., 2011. Brazilian red propolis: unreported substances, antioxidant and antimicrobial activities, Journal Science Food Agriculture, 91, 2363-2370.\u003c/p\u003e\n\u003cp\u003eRostagno, H.S., 2017. Composi\u0026ccedil;\u0026atilde;o de alimentos e exig\u0026ecirc;ncias nutricionais. Tabelas brasileiras para aves e su\u0026iacute;nos, 4\u0026ordf; ed., (Universidade Federal de Vi\u0026ccedil;osa - MG), pp 488.\u003c/p\u003e\n\u003cp\u003eSalatino, A., Fernandes-Silva, C.C., Righi, A.A., Salatino, M.L.F., 2011. Propolis research and the chemistry of plant products, Natural Product Research, 28, 925\u0026ndash;936.\u003c/p\u003e\n\u003cp\u003eSarcinelli, M.F., Venturini, K.S., Silva, L.C., 2007. Caracter\u0026iacute;sticas dos ovos. Universidade Federal do Esp\u0026iacute;rito Santo, UFES, Boletim T\u0026eacute;cnico, PIE-UFES: 00707 http://www.agais.com/telomc/b00707_caracteristicas_ovos.pdf Accessed 02 Sept 2021.\u003c/p\u003e\n\u003cp\u003eShang, X.G., Wang, F.L., Li, D.F., Yin, J.D., Li, J.Y., 2004. Effects of dietary conjugated linoleic acid on the productivity of laying hens and egg quality during refrigerated storage, Poultry Science, 83(10), 1688-1695.\u003c/p\u003e\n\u003cp\u003eSilva, J.D.T., Matos, A.D.A.S., Hada, F.H., Gravena, R.A., Marques, R.H., Moraes, V.M.B., 2012. Simbi\u0026oacute;tico e extratos naturais na dieta de codornas japonesas na fase de postura, Ci\u0026ecirc;ncia Animal Brasilera, 13, 1-7.\u003c/p\u003e\n\u003cp\u003eSilva, J.H.V., Albino, L.F.T., Godoi, M.J.S., 2000. Efeito do extrato de urucum na pigmenta\u0026ccedil;\u0026atilde;o da gema dos ovos, Revista Brasileira Zootecnia 29, 1435-1439.\u003c/p\u003e\n\u003cp\u003eShimizu, K., Ashida, H., Matsuura, Y., Kanazawa, K., 2004. Antioxidative bioavailability of artepillin C in Brazilian propolis, Archives of Biochemistry and Biophysics, 424, 181-188.\u003c/p\u003e\n\u003cp\u003eTocchini, L., Mercadante, A.Z., 2001. Extra\u0026ccedil;ão e determina\u0026ccedil;ão, por CLAE, de bixina e norbixina em coloríficos, Ciência Tecnologia Alimentos, 21, 310-313.\u003c/p\u003e\n\u003cp\u003eTrusheva, B., Popova, M., Bankova, V., Simova, S., Marcucci, M.C., Miorin, P.L., Pasin, F.R., Tsvetkova, I., 2006. Bioactive Constituents of Brazilian Red Propolis, Oxford University Press, 3, 249-254.\u003c/p\u003e\n\u003cp\u003eUmigi, R.T., Barreto, S.L.T., Reis, R.S., Mesquita, R.M.F., Ara\u0026uacute;jo, M.S., 2012. N\u0026iacute;veis de treonina digest\u0026iacute;vel para codorna japonesa na fase de produ\u0026ccedil;\u0026atilde;o, Arquivo Brasileiro Medicina Vetern\u0026aacute;ria Zootecnia, 64, 658-664.\u003c/p\u003e\n\u003cp\u003eWang, B.J., Lien, Y.H., Yu, Z.R. 2004. Supercritical fluid extractive fractionation - study of the antioxidant activities of propolis, Food Chemistry, 86, 237-243.\u003c/p\u003e\n\u003cp\u003eZavarize, K.C., Sartori, J.R.A., Pel\u0026iacute;cia, V.C.B., Pezzato, A.C.C., Araujo, P.C.D., Stradiotti, A.C.E., Madeira, L.A., 2011. Glutamina e nucleot\u0026iacute;deos na dieta de frangos de corte criados no sistema alternativo, Archives Zootecnia, 60, 913-920.\u003c/p\u003e\n\u003cp\u003eZeweil, H.S., Zahran, S.M., Abd El-Rahman, M.H.A., Dosoky, W.M., Abu Hafsa, H., Moktar, A.A., 2016. Effect of using bee propolis as natural supplement on productive and physiological performance of japanese quail, Egypt Poultry Science Journal, 36, 161-175.\u003c/p\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 1. Ingredient and nutrient composition of basal diet.\u003c/p\u003e\n\u003ctable width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eIngredients (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003eQuantity\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eCorn\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e58.7\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eSoybean meal\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e31.7\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eSoybean oil\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.18\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eDicalcium phosphate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e1.22\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eCalcium Carbonate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e6.84\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eSodium chloride (NaCl)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.34\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eDL-Methionine (99%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.38\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eLysine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.24\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eVitamin-mineral premix \u003csup\u003eA\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.20\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eNutrient composition (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003eQuantity\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eCrude Protein\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e19.9\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eMetabolizable energy (kcal/kg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e2.75\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eCalcium\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e3.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eAvailable Phosphorus\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.32\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eDigestible Lysine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e1.11\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eDigestible Methionine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.66\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66%\"\u003e\n\u003cp\u003eMethionine + digestible cysteine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"33%\"\u003e\n\u003cp\u003e0.98\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003eA\u003c/sup\u003e Product composition (kg): vitamin (vit.) A 5,000,000 IU; vit. D3 1,000,000 IU; vit. E 15,000 IU; vit. K3 1500 mg; vit. B1 1500 mg; vit. B2 3000 mg; vit. B6 2000 mg; vit. B12 7000 \u0026mu;g; folic acid 500 mg; nicotinic acid 15 g; pantothenic acid 7000 \u0026mu;g; choline 80 g; biotin 100 mg; Cu 10 g; Fe 50 g; I 1000 mg; Mn 80 g; Selenium 300 mg; Zn 70g.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 2. Effects of red propolis feed supplementation on the productive performance of quails.\u003c/p\u003e\n\u003ctable width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003eParameters\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003eTE\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003eRP1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003eRP2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eP-Value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003eEgg production\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e95.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e94.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e94.30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e92.73\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.63\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003eFeed consumption\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e27.29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e26.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e26.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e26.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.32\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003eAverage egg weight\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e11.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e10.88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e11.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e10.93\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.43\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003eEggs mass\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e10.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e10.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e10.47\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e10.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.45\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003eFeed convertion (Kg/Kg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e2.56\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e2.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e2.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e2.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.78\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003eFeed convertion (Kg/dz)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e0.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e0.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e0.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e0.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.83\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"43%\"\u003e\n\u003cp\u003e\u0026nbsp;Viability\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"10%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eCT: control treatment; TE: enramycin treatment; RP1: treatment with red propolis (1g/kg of feed); RP2: treatment with red propolis (2g/kg of feed). Feed conversion (kg/kg): kg of feed consumed per kg of produced egg; Feed conversion (kg/dz): kg of feed per dozen eggs produced. P-Value: P\u0026lt;0.05 indicating significance level by Tukey test at 95% confidence.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 3. Effects of red propolis supplementation on quail egg quality.\u003c/p\u003e\n\u003ctable width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eParameter\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eTE\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eRP1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eRP2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eP-Value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003e\u003cem\u003eEggshell strength\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1,424.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1,444.22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1,424.70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1,464.90\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eSpecific gravity\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1.064\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1.067\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1.067\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e1.067\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.64\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eHaugh unit\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e87.82\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e85.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e85.56\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e86.29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.16\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eYolk index\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.4708\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.4558\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.4775\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.4642\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eColorimetric fan\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e5.75\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e5.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e5.92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e5.69\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.86\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eLuminosity (L*)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e65.629a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e60.881b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e59.428b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e57.11b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eIntensity of red (a*)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e-9.96a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e-9.33ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e-8.072b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e-7.92b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.0095\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eIntensity of yellow (b*)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e53.76a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e49.07b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e46.93b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e46.43b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eYolk %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e31.49\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e31.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e32.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e32.63\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.69\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eShell %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.71\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.95\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eAlbumen %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e59.93\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e59.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e59.27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e58.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.69\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eShell thickness\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.18\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.18\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eYolk pH\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e6.21a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e6.05ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e5.88b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e5.90b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.0125\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"32%\"\u003e\n\u003cp\u003eAlbumen pH\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e8.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.90\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eCT: control treatment; TE: enramycin treatment; RP1: treatment with red propolis (1g/kg of feed); RP2: treatment with red propolis (2g/kg of feed). P-Value: P\u0026lt; 0.05 indicating significance level by Tukey test at 95% confidence. Means followed by different letters on the same line indicate a difference by Tukey's test at 5% significance (P\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 4. Effects of red propolis supplementation for quails on egg weight and lipid peroxidation levels after 21 days of storage.\u003c/p\u003e\n\u003ctable width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eParameter\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eTE\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eRP1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003eRP2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eP-Value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eAWL (g)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e3.93ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e4.01ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e3.74b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e4.14a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e0.012\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eTBARS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e0.32a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e0.07b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e0.12ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e0.29a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e0.007\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eCT: control treatment; TE: enramycin treatment; RP1: treatment with red propolis (1g/kg of feed); RP2: treatment with red propolis (2g/kg of feed). AWL = average weight loss, P-Value: P\u0026lt;0.05 indicating significance level by Tukey test at 95% confidence. Means followed by different letters on the same line indicate a difference by Tukey's test at 5% significance (P\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 5. Effects of red propolis on total count of mesophilic aerobics on the surface of quail eggs.\u003c/p\u003e\n\u003ctable width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eTreatment\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"24%\"\u003e\n\u003cp\u003eSecond cycle\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"25%\"\u003e\n\u003cp\u003eThird cycle\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"27%\"\u003e\n\u003cp\u003eStored\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"15%\"\u003e\n\u003cp\u003eCFU/mL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003eCFU/mL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"11%\"\u003e\n\u003cp\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003eCFU/mL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eTC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"15%\"\u003e\n\u003cp\u003e29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"11%\"\u003e\n\u003cp\u003e41.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e100.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eTE\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"15%\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"11%\"\u003e\n\u003cp\u003e83.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e33.33\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eRP1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"15%\"\u003e\n\u003cp\u003e42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"11%\"\u003e\n\u003cp\u003e9.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e9.52\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"21%\"\u003e\n\u003cp\u003eRP2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"15%\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"11%\"\u003e\n\u003cp\u003e66.66\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"14%\"\u003e\n\u003cp\u003e8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"13%\"\u003e\n\u003cp\u003e266.67\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eTC: control treatment; TE: enramycin treatment; RP1: treatment with propolis (1g/kg of feed); RP2: treatment with propolis (2g/kg of feed). (%): percentage of colony presence considering 100% presence in the second cycle. CFU/mL: colony forming units per mL of sample.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 6. Effects of red propolis on the total mesophilic aerobic counts in quail feces.\u003c/p\u003e\n\u003ctable\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"106\"\u003e\n\u003cp\u003eTreatment\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"158\"\u003e\n\u003cp\u003eFirst cycle\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"140\"\u003e\n\u003cp\u003eSecond cycle\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"162\"\u003e\n\u003cp\u003eThird cycle\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"106\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eCFU/mL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"63\"\u003e\n\u003cp\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003eCFU/mL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"56\"\u003e\n\u003cp\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eCFU/mL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"67\"\u003e\n\u003cp\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"106\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e136\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"63\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"56\"\u003e\n\u003cp\u003e1.47\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"67\"\u003e\n\u003cp\u003e6.61\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"106\"\u003e\n\u003cp\u003eTE\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"63\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"56\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"67\"\u003e\n\u003cp\u003e8.69\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"106\"\u003e\n\u003cp\u003eRP1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"63\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"56\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"67\"\u003e\n\u003cp\u003e41.66\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"106\"\u003e\n\u003cp\u003eRP2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e338\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"63\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"56\"\u003e\n\u003cp\u003e0.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"67\"\u003e\n\u003cp\u003e1.18\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eCT: control treatment; TE: enramycin treatment; RP1: treatment with propolis 1g/kg of feed; RP2: treatment with propolis 2g/kg of feed. (%): percentage of colony presence considering 100% presence in the first cycle. CFU/mL: colony forming units per mL of sample.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 7. Counts of total and fecal coliforms in quail feces after the first cycle.\u003c/p\u003e\n\u003ctable width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"37%\"\u003e\n\u003cp\u003eTreatment\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003eTC (CFU/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003eEC (CFU/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"37%\"\u003e\n\u003cp\u003eCT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e105\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"37%\"\u003e\n\u003cp\u003eTE\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e327\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e110\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"37%\"\u003e\n\u003cp\u003eRP1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e55\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e33\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"37%\"\u003e\n\u003cp\u003eRP2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e196\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"31%\"\u003e\n\u003cp\u003e102\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eCT: control treatment; TE: enramycin treatment; RP1: treatment with propolis 1g/kg of feed; RP2: treatment with propolis 2g/kg of feed. TC: total coliforms; EC: \u003cem\u003eEscherichia coli\u003c/em\u003e; CFU/mL: colony forming units per mL.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Antimicrobial, antioxidant, laying quails, lipid peroxidation, performance, red propolis","lastPublishedDoi":"10.21203/rs.3.rs-1485683/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1485683/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study aimed to determine the effects of aqueous extract of red propolis (RP) as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status. A total of 120 laying quails with 52 days of age were were randomly allotted to dietary treatments and six replicates of five birds each in completely randomized design. There were four dietary treatments as following: Control (TC), with enramycin (TE), 1g RP/kg of feed (RP1), 2g RP/kg of feed (RP2) in concetrated diet raised for 63 days. The results of productive performance such as percentage of egg production, feed intake per quail per day, average egg weight, egg mass, foods conversions showed no significant difference between treatments. The quails fed with red propolis in fresh eggs showed darker yolk (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), with higher intensity of red and lower intensity of yellow; as less than 6.0 pH yolk; microbiological characteristics analyzed on the surface of eggs and feces of quails was lower and less than compared with other treatments. It is possible to conclude that red propolis showed promising results to be used as a feed additive for laying quails, since it maintained the productive performance of these animals and caused qualitative improvements in the physicochemical and microbiological characteristics of the eggs.\u003c/p\u003e","manuscriptTitle":"Effects of aqueous extract of red propolis as an alternative to synthetic antimicrobial for quails enhanced egg quality and maintained health status","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-05-18 19:19:02","doi":"10.21203/rs.3.rs-1485683/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5d8de79b-9d1e-45b5-84ed-7e121d401b0b","owner":[],"postedDate":"May 18th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-08-31T13:50:05+00:00","versionOfRecord":[],"versionCreatedAt":"2022-05-18 19:19:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1485683","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1485683","identity":"rs-1485683","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}