Digestibility of diets with passion fruit by-product estimated through external and internal markers in dairy heifers

Mércia Regina Pereira de Figueiredo Eloisa de Oliveira Simões Saliba Geraldo Sérgio Carneiro Barbosa Juliana Cristina Nogueira Colodo Rebeca Agnes de Oliveira Almança Guilherme Rocha Moreira About the authors

ABSTRACT.

This study examined the influence of the external markers chromium oxide (CO), titanium dioxide (TD), isolated, purified and enriched lignin (LIPE®) and isolated, purified, enriched lignin in nanoparticles (NANOLIPE®) as well as the internal markers indigestible dry matter (iDM), indigestible neutral detergent fiber (iNDF) and indigestible acid detergent fiber (iADF) in diets with inclusion of passion fruit by-product for dairy heifers on the estimation of fecal output and nutrient digestibility. Sixteen Holstein × Zebu crossbred heifers at an average live weight of 363 ± 28 kg were randomly distributed in a completely randomized design where they received diets in which Tifton 85 (Cynodon sp.) hay was replaced with passion fruit by-product (0, 12, 24 and 36%, as-fed basis). The CO, LIPE®, NANOLIPE® and iNDF markers did not differ from the total collection method (p > 0.05) in the estimation of fecal output and nutrient digestibility. The TD and iDM markers overestimated, while iADF underestimated fecal output. Under the presented conditions, we recommend using the CO, LIPE®, NANOLIPE® and iNDF markers to estimate fecal output and nutrient digestibility in diets with inclusion of passion fruit by-product for dairy heifers.

Keywords:
fecal recovery; in situ; passion fruit by-product; total collection

Introduction

Intake and digestibility are variables of great importance in animal production systems, since they are directly related to nutrient uptake and, consequently, to whether the nutritional requirements of animals are being met. According to Mertens (1994Mertens, D. R. (1994). Regulation of forage intake. In. G. C. , Fahey Junir (Ed.), Forage quality, evaluation, and utilization (p. 450-493). Madison, WI: American Society of Agronomy.), animal performance is a direct reflection of digestible dry matter intake, with 60 to 90% of its variations resulting from changes in intake and 10 to 40% from changes in digestibility. These parameters can be estimated via total fecal collection and/or with the use of markers, which have exhibited promising results and an easy evaluation methodology. Markers are substances that are naturally present in or can be added to the animal diet, which characterizes them as internal or external, respectively. By using them, one can accurately estimate the nutrient digestibility and intake of the most diverse animal species and categories.

Although none of the substances used as markers fulfills all the ideal characteristics, many of them are sufficiently adequate to provide significant data (Berchielli, Garcia, & Oliveira, 2006Berchielli, T. T., Garcia, A. V., & Oliveira, S. G. (2006). Principais técnicas de avaliação aplicadas em estudo de nutrição. In T. T. Berchielli, A. V. Pires, & S. G. Oliveira (Orgs.), Nutrição de Ruminantes (p. 397-421). Jaboticabal, SP: Funep.). For this reason, the search for ideal markers is one of the subjects of great interest in the discovery of techniques that facilitate animal nutrition studies.

The seasonality of herbage production has been responsible, among other factors, for the reduced yields of herds. Coupled with the production cost invested in animal feed, this has aroused interest in the use of alternative feedstuffs. In this respect, by-products of fruits such as passion fruit have been constantly evaluated mainly in terms of their adequate level of inclusion in ruminant diets. In grass silage, these by-products have shown good results (Alves et al., 2015Alves, G. R., Fontes, C. A. A., Processi, E. F., Fernandes, A. M., Oliveira, T. S., & Glória, L. S. (2015). Performance and digestibility of steers fed by- product of fresh passion fruit or sorghum silage, with and without concentrate supplementation. Revista Brasileira de Zootecnia, 44(9), 314-320. doi: 10.1590/S1806-92902015000900002
https://doi.org/10.1590/S1806-9290201500...
; Bonfá et al., 2015Bonfá, C. S., Castro, G. H. F., Villela, S. D. J., Santos, R. A., Evangelista, A. R., Jayme, C. G., ... Barbosa, J. A. S. (2015). Silagem de capim- elefante adicionada de casca de maracujá. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 67(3), 801-808. doi: 10.1590/1678-4162-7982
https://doi.org/10.1590/1678-4162-7982...
; Lira Júnior et al., 2018Lira Júnior, W. B., Bezerra, S. B. L., Paula, T. A., Beelen, R. N., Amorim, P. L., & Beelen, P. M. G. (2018). Características de silagens de capim- elefante (Pennisetum purpureum) e casca de maracujá in natura. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 70(3), 905-912. doi: 10.1590/1678-4162-9410
https://doi.org/10.1590/1678-4162-9410...
).

In addition to contributing to reducing environmental contamination caused by improper disposal, the use of industrial by-products in animal feed serves as an alternative source of nutrients for the period of food scarcity, constituting an important fiber source. The use of a non-forage fiber source such as passion fruit by-product in ruminant diets contributes to the supply of nutrients and may lower the diet cost, representing a viable option in regions where it is available. Studies have focused on the evaluation of markers in different diets because the varied results obtained may be related to the animal diet.

Therefore, this study proposes to examine the effectiveness of the external markers chromium oxide (CO), titanium dioxide (TD), LIPE® and NANOLIPE® and the internal markers indigestible dry matter (iDM), indigestible neutral detergent fiber (iNDF) and indigestible acid detergent fiber (iADF) in the estimation of fecal output and nutrient digestibility in Tifton 85 grass hay-based diets with increasing inclusion levels of passion fruit by-product for dairy heifers.

Material and methods

This study was carried out in accordance with the Ethical Principles of Animal Experimentation adopted by the Ethics Committee on Animal Experimentation (CETEA/UFMG) (approval no. 225/2015).

The experiment was carried out in the facilities of the Center for Agrarian Education and Development of Florestal at the Federal University of Viçosa, located in Florestal - MG, Brazil, from May to August 2012. Sixteen Holstein × Zebu crossbred heifers at an average weight of 363 ± 28 kg were identified, weighed, dewormed and housed individually in cement-floored stalls.

The diets were based on Tifton 85 grass hay, corn meal and passion fruit by-product. The chemical composition of the feedstuffs and the average nutrient levels determined in the diets are shown in Tables 1 and 2.

Table 1
Chemical composition (%) and gross energy levels (kcal g-1) of the ingredients of the diets supplied in the experiment (DM basis).

The experimental period was 17 days, which consisted of 12 days of adaptation to diets, management and facilities plus five days of experimental collections. Feed was supplied daily, in two meals: 50% at 07h00 and 50% at 16h00. Orts were collected daily in the morning and weighed and the amount of feed provided was adjusted to allow 10% orts.

The diets and orts were dried for 72 h at 55ºC in an air oven and crushed to 1 mm through a knife mill. Chemical analytical methods (Association Official Analytical Chemists International [AOAC], 2012Association Official Analytical Chemists International [AOAC]. (2012). Official Methods of Analysis (19th ed.). Arlington, VA: AOAC International.) were used to determine the dry matter (DM; method 934.01), mineral matter (MM; Method 942.05), crude protein (CP; method 984.13) and ether extract (EE; method 920.39) contents. For neutral detergent fiber (NDF; method INCT-CA F-001/1), acid detergent fiber (ADF; method INCT-CA F-003/1), total and non-fibrous carbohydrates and lignin (method INCT-CA F-007/1), the analysis procedures proposed by Detmann et al. (2012Detmann, E., Souza, M. A., Valadares Filho, S. C., Queiroz, A. C., Berchielli, T. T., Saliba, E. O. S., ... Azevedo, J. A. G. (2012). Métodos para análises de alimentos: INCT - Ciência Animal. Visconde do Rio Branco, MG: Suprema.) were adopted. Gross energy was determined by combustion in an adiabatic calorimeter (PARR 6300). In vitro dry matter digestibility (IVDMD) was determined by the method proposed by Tilley and Terry (1963Tilley, J. M. A., & Terry, R. A. (1963). A two-stage technique for the in vitro digestion of forage crops. Journal of the British Grassland Society, 18(2), 104-111. doi: 10.1111/j.1365-2494.1963.tb00335.x
https://doi.org/10.1111/j.1365-2494.1963...
).

Fecal output was estimated using the external markers chromium oxide (CO), titanium dioxide (TD), isolated, purified and enriched lignin (LIPE®) and isolated, purified and enriched lignin in nanoparticles (NANOLIPE®) and the internal markers indigestible dry matter (iDM), indigestible neutral detergent fiber (NDF) and indigestible acid detergent fiber (iADF). The CO and TD markers were mixed with the diet at the dose of 10 g animal-1 day-1, for an adaptation period of seven days followed by five days of feces collection. The LIPE® and NANOLIPE® markers were administered orally, in the form of capsules, at the dose of 500 mg animal-1, for an adaptation period of two days (Lima, Graça, Borges, Saliba, & Simão, 2008Lima, J. B. P., Graça, D. S., Borges, A. L. C. C., Saliba, E. O. S., & Simão, S. M. B. (2008). Uso do óxido crômico e do LIPE® na estimativa do consumo de matéria seca por bezerros de corte. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 60(5), 1205-1212. doi: 10.1590/S0102-09352008000500023
https://doi.org/10.1590/S0102-0935200800...
) followed by five days of total fecal collection for LIPE® and one day of adaptation followed by one day of total fecal collection for NANOLIPE® (Figueiredo et al., 2019Figueiredo, M. R. P., Saliba, E. O. S., Barbosa, G. S. S. C., Silva, F. A., Nunes, A. N., Silva, C. R. M., & Moreira, G. R. (2019). Use of indigestible markers to estimate the apparent dry matter digestibility of diets containing a cocoa by-product. Semina: Ciências Agrárias, 40(6), 2771-2782. doi: 10.5433/1679-0359.2019v40n6p2771
https://doi.org/10.5433/1679-0359.2019v4...
).

Table 2
Percentage (%) and chemical compositions and mean gross energy (kcal g-1) of diets containing increasing levels of passion fruit by-product (DM basis).

The internal markers iDM, iNDF and iADF were obtained from samples of feed, orts and feces, which were packed in non-woven fabric bags (“TNT”; 100 g cm-2) measuring 4 × 5 cm and incubated in the rumen of two adult cattle for 264h, as suggested by Casali et al. (2008Casali, A. O., Detmann, E., Valadares Filho, S. C., Pereira, J. C., Henriques, L. T., Freitas, S. G., & Paulino, M. F. (2008). Influência do tempo de incubação e do tamanho de partículas sobre os teores de compostos indigestíveis em alimentos e fezes bovinas obtidos por procedimentos in situ. Revista Brasileira de Zootecnia, 37(2), 335-342. doi: 10.1590/S1516-35982008000200021
https://doi.org/10.1590/S1516-3598200800...
), to obtain the indigestible fractions in situ.

After 264 h of in situ incubation, the bags were removed from the rumen and washed in running water, dried in a forced-air oven and weighed and the residue was used to determine iDM. The iNDF and iADF markers were determined after washing the bags with neutral and acid detergent solutions, respectively (in a non-sequential manner), oven-drying and weighing, and the residue was used to determine the indigestible fractions.

Fecal output (FO) as estimated by the markers was determined by the following formula:

FO= Marker intake (g) Marker content in feces (%) (DM 105 º C )

The LIPE® and NANOLIPE® markers were analyzed by Near Infrared Spectroscopy using a FTIV - 800 (Varian) instrument, following Saliba et al. (2015Saliba, E. O. S., Faria, E. P., Rodriguez, N. M., Moreira, G. R., Sampaio, I. B. M., Saliba, J. S., & Borges, A. L. C. C. (2015). Use of infrared spectroscopy to estimate fecal output with marker Lipe. International Journal of Food Science, Nutrition and Dietetics, 4(001), 1-10. doi: 10.19070/2326-3350-SI04001
https://doi.org/10.19070/2326-3350-SI040...
) and Moss et al. (2017Moss, P. C. B., Rezende, A. S. C., Saliba, E. O., Lana, Â. M. Q., Moura, R. S., Cassou, F., & Alves, G. E. S. (2017). Validação do Nanolipe ® como método para determinar a digestibilidade aparente dos nutrientes pelos equinos. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 69(3), 687-694. doi: 10.1590/1678-4162-8270
https://doi.org/10.1590/1678-4162-8270...
), respectively. Chromium oxide was determined as suggested by Detmann et al. (2012Detmann, E., Souza, M. A., Valadares Filho, S. C., Queiroz, A. C., Berchielli, T. T., Saliba, E. O. S., ... Azevedo, J. A. G. (2012). Métodos para análises de alimentos: INCT - Ciência Animal. Visconde do Rio Branco, MG: Suprema.) (INCT method - CA M-006/1). The TD content was determined according to Myers, Ludden, Nayigihugu, and Hess (2004Myers, W. D., Ludden, P. A., Nayigihugu, V., & Hess, B. W. (2004). Technical note: a procedure for the preparation and quantitative analysis of samples for titanium dioxide. Journal of Animal Science, 82(1), 179-183. doi:10.2527/2004.821179x
https://doi.org/0.2527/2004.821179x...
).

The apparent digestibility coefficients of nutrients as obtained by the external and internal marker methods were calculated as proposed by Silva and Leão (1979Silva, J. F. C., & Leão, M. I. (1979). Fundamentos de nutrição de ruminantes. Piracicaba, SP: Livroceres.):

Digestibility (%)= 100 - 100 x % of marker in feed % of marker in feces

The equation proposed by Lanzetta et al. (2009Lanzetta, V. A. S., Rezende, A. S. C., Saliba, E. O. S., Lana, Â. M. Q., Rodriguez, N. M., & Moss, P. C. B. (2009). Validação do Lipe® como método para determinar a digestibilidade dos nutrientes em eqüinos. Revista Brasileira de Zootecnia, 38(1), 69-74. doi: 10.1590/S1516-35982009000100009
https://doi.org/10.1590/S1516-3598200900...
) was applied to calculate the fecal recovery rate of each marker:

Fecal recovery rate = Fecal output by marker Fecal output by total fecal collection x 100

The experiment was laid out in a completely randomized split-plot design where the plots were the treatments (diets) and the markers represented the subplots. Data analysis was performed using SISVAR (Statistical Analysis and Experimental Design Program) software (Ferreira, 2011Ferreira, D. F. (2011). Sisvar: a computer statistical analysis system. Ciência e Agrotecnologia, 35(6), 1039-1042. doi: 10.1590/S1413-70542011000600001
https://doi.org/10.1590/S1413-7054201100...
). Fecal output as well as the apparent digestibility and fecal recovery coefficients were subjected to analysis of variance and means were compared by the SNK test (p < 0.05).

Results and discussion

The external markers CO, LIPE®, NANOLIPE® and the internal marker iNDF did not differ from the total collection method (p > 0.05) in the estimation of fecal output. Divergent results were obtained for the other evaluated markers. The external marker TD and the internal marker iDM overestimated fecal output by 27.12 and 34.51%, respectively, whereas the internal marker iADF underestimated it by almost 41% (Table 3). The fecal recovery rates of the iNDF and CO markers were 0.54% and 5.75% higher than observed. With LIPE® and NANOLIPE®, however, fecal recovery rate was 7.4% and 6% lower, respectively.

Table 3
Mean fecal output values (kg DM day-1), standard error of the mean (SEM) and fecal recovery rate (FRR) as obtained by total fecal collection and estimated using external and internal markers.

The results obtained with the external markers CO, LIPE® and NANOLIPE® and the internal marker iNDF for the digestibility of DM, CP and NDF did not differ (p > 0.05) from the actual values found (Table 4), at any of the passion fruit by-product inclusion levels.

Dietary inclusion of passion fruit by-product induced a reduction in NDF and an increase in ADF contents when compared with the diet without the ingredient. Lignin levels also increased, likely due to the presence of seeds in the passion fruit by-product. The average DM digestibility of the diets was 52% and varied according to the estimate provided by each evaluated marker.

Results obtained with CO are abundant and varied in the literature because it is the most commonly used external marker in trials to estimate intake and digestibility in several categories of animals and diets. According to Owens and Hanson (1992Owens, F. N., & Hanson, C. F. (1992). External and internal markers for appraising site and extent of digestion in ruminants. Journal Dairy Sciense, 75(9), 2605-2617. doi: 10.3168/jds.S0022-0302(92)78023-0
https://doi.org/10.3168/jds.S0022-0302(9...
), variations in fecal CO excretion throughout the day are a relatively known limitation of this marker. Unlike the present results, Silva et al. (2010Silva, J. J., Saliba, E. O. S., Borges, I., Gonçalves, L. C., Rodriguéz, N. M., Aroeira, L. J. M., & Costa, F. J. N. (2010). Indicadores para estimativa de consumo total por novilhas holandês x zebu mantidas em confinamento. Revista Brasileira de Saúde e Produção Animal, 11(3), 838-848.) found that CO underestimated the intake of feedlot heifers fed diets based on of elephant grass silage or sugarcane, regardless of the diet, with results differing from those measured at the trough and obtained with the iNDF, iADF, Klason lignin and LIPE® markers. Ribeiro Filho, Zimermann, and Kozloski (2008Ribeiro Filho, H. M. N., Zimermann, F. C., & Kozloski, G. V. (2008). Baixa dosagem de óxido de cromo para estimativa da produção fecal em bovinos. Ciência Rural, 38(9), 2567-2573. doi: 10.1590/S0103-84782008005000014
https://doi.org/10.1590/S0103-8478200800...
) reported that the different CO recovery rates described in the literature can be attributed to different protocols used and to differences in the type and quality of the diet.

The results reported here using the LIPE® marker corroborate other authors who have shown positive outcomes with the use of this marker in the estimation of intake, fecal output and nutrient digestibility in several animal species (Ferreira et al., 2009Ferreira, M. A., Valadares Filho, S. C., Marcondes, M. I., Paixão, M. L., Paulino, M. F., & Valadares, R. F. D. (2009). Avaliação de indicadores em estudos com ruminantes: digestibilidade. Revista Brasileira de Zootecnia, 38(8), 1568-1573. doi: 10.1590/S1516-35982009000800022
https://doi.org/10.1590/S1516-3598200900...
; Lanzetta et al., 2009Lanzetta, V. A. S., Rezende, A. S. C., Saliba, E. O. S., Lana, Â. M. Q., Rodriguez, N. M., & Moss, P. C. B. (2009). Validação do Lipe® como método para determinar a digestibilidade dos nutrientes em eqüinos. Revista Brasileira de Zootecnia, 38(1), 69-74. doi: 10.1590/S1516-35982009000100009
https://doi.org/10.1590/S1516-3598200900...
; Andrade et al., 2013Andrade, R. P. X., Ferreira, M. A., Urbano, S. A., Azevedo, M., Félix, S. C. R., Siqueira, M. C. B., ... Chagas, J. C. C. (2013). Castor beans hulls as a replacement for Tifton 85 hay in lamb diets. Tropical Animal Health and Production, 45(5), 1191-1196. doi: 10.1007/s11250-012-0345-4
https://doi.org/10.1007/s11250-012-0345-...
; Silva et al., 2014Silva, L. G., Torrecilhas, J. A., Ornaghi, M. G., Eiras, C. E., Prado, R. M., & Prado, I. N. (2014). Glycerin and essential oils in the diet of Nellore bulls finished in feedlot: animal performance and apparent digestibility. Acta Scientiarum. Animal Sciences, 36(2), 177-184. doi: 10.4025/actascianimsci.v36i2.23089
https://doi.org/10.4025/actascianimsci.v...
; Saliba et al., 2015Saliba, E. O. S., Faria, E. P., Rodriguez, N. M., Moreira, G. R., Sampaio, I. B. M., Saliba, J. S., & Borges, A. L. C. C. (2015). Use of infrared spectroscopy to estimate fecal output with marker Lipe. International Journal of Food Science, Nutrition and Dietetics, 4(001), 1-10. doi: 10.19070/2326-3350-SI04001
https://doi.org/10.19070/2326-3350-SI040...
). LIPE® has the advantage of being provided in only two days, which reduces animal stress when the marker is given orally. NANOLIPE®, in turn, offers the advantage of coming in the form of nanoparticles, which favors its incorporation into the digesta, with a better fecal recovery rate. The results obtained with this marker in the present study are promising, and the intake and digestibility trials that have used it are recent. A noteworthy advantage of NANOLIPE® is the shorter time of adaptation of the animals and shorter period of fecal collection compared with those of the other evaluated markers, which makes it possible to minimize the work, labor and animal stress, demonstrating its use potential, as shown in the studies of Moss et al. (2017Moss, P. C. B., Rezende, A. S. C., Saliba, E. O., Lana, Â. M. Q., Moura, R. S., Cassou, F., & Alves, G. E. S. (2017). Validação do Nanolipe ® como método para determinar a digestibilidade aparente dos nutrientes pelos equinos. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 69(3), 687-694. doi: 10.1590/1678-4162-8270
https://doi.org/10.1590/1678-4162-8270...
) and Figueiredo et al. (2019Figueiredo, M. R. P., Saliba, E. O. S., Barbosa, G. S. S. C., Silva, F. A., Nunes, A. N., Silva, C. R. M., & Moreira, G. R. (2019). Use of indigestible markers to estimate the apparent dry matter digestibility of diets containing a cocoa by-product. Semina: Ciências Agrárias, 40(6), 2771-2782. doi: 10.5433/1679-0359.2019v40n6p2771
https://doi.org/10.5433/1679-0359.2019v4...
).

Table 4
. Mean values (%) of apparent digestibility of dry matter (DMd), crude protein (CPd) and neutral detergent fiber (NDFd), standard error of the mean (SEM) and coefficient of variation (CV, %) of the diets as estimated by the external and internal markers and compared with total collection.

As regards the internal markers iDM, iNDF and iADF, results presented in the literature indicate that their behavior may vary depending on the analysis methodology and type of diet provided to the animals-more specifically, the fiber used. This is because the constitution of the fibrous fraction of each roughage changes the rate and extent of degradation (Berchielli, Oliveira, Carrilho, Feitosa, & Lopes, 2005Berchielli, T. T., Oliveira, S. G., Carrilho, E. N. V. M., Feitosa, J. V., & Lopes, A. D. (2005). Comparação de marcadores para estimativas de produção fecal e de fluxo de digesta em bovinos. Revista Brasileira de Zootecnia, 34(3), 987-996. doi: 10.1590/S1516-35982005000300032
https://doi.org/10.1590/S1516-3598200500...
). Berchielli et al. (2006Berchielli, T. T., Garcia, A. V., & Oliveira, S. G. (2006). Principais técnicas de avaliação aplicadas em estudo de nutrição. In T. T. Berchielli, A. V. Pires, & S. G. Oliveira (Orgs.), Nutrição de Ruminantes (p. 397-421). Jaboticabal, SP: Funep.) also stated that errors in the methodology of analysis of these markers lead to variations in the results and that some markers are probably more adequate than others depending on the roughage used. In addition, other factors such as the form of incubation (in situ or in vitro), incubation time and loss of particles through the pores of the nylon bags may be the main causes of the observed variations in the fecal recovery of the internal markers. The internal marker iNDF came closest to the total collection method in the estimates of fecal output and digestibility (Tables 3 and 4).

Dias et al. (2008Dias, M., Leão, M. I., Detmann, E., Valadares Filho, S. C., Vasconcelos, A. M., Souza, S. M., ... Murça, T. B. (2008). Técnicas para estimativa da digestibilidade e produção microbiana em bovinos. Revista Brasileira de Zootecnia, 37 (3), 504-512. doi: 10.1590/S1516-35982008000300016
https://doi.org/10.1590/S1516-3598200800...
) found that iADF accurately estimated DM digestibility and fecal recovery like total fecal collection, in a trial with cattle fed a diet based on Tifton grass hay and concentrate. In an experiment with cattle fed diets based on ground corn, soybean hulls or corn germ meal, Watanabe, Ezequiel, Galati, Biagioli, and Silva (2010Watanabe, P. H., Ezequiel, J. M. B., Galati, R. L., Biagioli, B., & Silva, O. G. C. (2010). Indicadores internos indigestíveis para a estimativa das digestibilidades de dietas à base de coprodutos. Revista Brasileira de Saúde e Produção Animal, 11(3), 849-857.) recommended using iNDF and iADF together, as they provided accurate digestibility estimates of the evaluated diets. Maeda et al. (2011Maeda, E. M., Zeoula, L. M., Gomes, H. C. C., Jacobi, G., Simioni, F. L., & Oliviera, R. A. (2011). Avaliação de indicadores usados nos estudos de ingestão e digestibilidade de bovinos e bubalinos. Archivos de Zootecnia, 60(229), 123-131. doi: 10.4321/S0004-05922011000100014
https://doi.org/10.4321/S0004-0592201100...
) found that iDM accurately estimated omasal flow in cattle and buffalo. As for fecal flow, CO showed fecal recovery close to 100%, with iNDF overestimating and iADF underestimating fecal output. Sampaio et al. (2011Sampaio, C. B., Detmann, E., Valente, T. N. P., Costa, V. A. C., Valadares Filho, S. C., & Queiroz, A. C. (2011). Fecal excretion patterns and short term bias of internal and external markers in a digestion assay with cattle. Revista Brasileira de Zootecnia, 40(3), 657-665. doi: 10.1590/S1516-35982011000300026
https://doi.org/10.1590/S1516-3598201100...
) evaluated the fecal recovery of the iDM, iNDF, iADF, CO and TD markers in cattle fed elephant grass silage, corn silage or Brachiaria grass hay, supplemented or unsupplemented with a 20% concentrate mixture, and found satisfactory results for all evaluated markers, with fecal recovery not differing from 100%.

Pombo, Valle, Bradi, and Bueno (2016Pombo, G. V., Valle, T. D., Bradi, R. A., & Bueno, I. C. S. (2016). Acurácia, precisão e robustez de indicadores internos para predição da digestibilidade aparente total de matéria seca em equinos. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 68(3), 769-775. doi: 10.1590/1678-4162-8441
https://doi.org/10.1590/1678-4162-8441...
) evaluated the accuracy, precision and robustness of the internal markers iNDF, iADF, indigestible cellulose and acid detergent indigestible lignin in the prediction of apparent DM digestibility in horses and also recommended iADF. Magalhães et al. (2018Magalhães, A. L. R., Teodoro, A. L., Souza, M. F. S., Nascimento Júnior, J. R. S., Souza, D. R., Santos, K. C., ... Silva, S. M. C. (2018). Dietas para ovinos contendo capim elefante maduro e raspa de mandioca: estimativa de digestibilidade aparente por meio de indicadores. Archivos de Zootecnia, 67(258), 244-252. doi: 10.21071/az.v67i258.3660
https://doi.org/10.21071/az.v67i258.3660...
) conducted a trial evaluating the digestibility of diets based on cassava chips and mature elephant grass in sheep and recommended the internal markers iNDF and iADF. Additionally, Sousa et al. (2018Sousa, S. V., Araújo, M. J., Marques, C. A. T., Torreão, J. N. C., Lima, L. A., Gottardi, F. P., & Bezerra, L. R. (2018). Internal markers for predicting dry matter intake and digestibility in Santa Inês ewes on a pasture. Grassland Science, 18(1), 1-11. doi: 10.1111/grs.12223
https://doi.org/10.1111/grs.12223...
) reported that iDM showed satisfactory results in estimating nutrient intake and digestibility compared with iNDF and iADF, in a trial with grazing sheep.

Fecal output as estimated by TD in this study does not agree with values reported in the literature (Ferreira et al., 2009Ferreira, M. A., Valadares Filho, S. C., Marcondes, M. I., Paixão, M. L., Paulino, M. F., & Valadares, R. F. D. (2009). Avaliação de indicadores em estudos com ruminantes: digestibilidade. Revista Brasileira de Zootecnia, 38(8), 1568-1573. doi: 10.1590/S1516-35982009000800022
https://doi.org/10.1590/S1516-3598200900...
; Glindemann, Tas, Wang, Alvers, & Susenbeth, 2009Glindemann, T., Tas, B. M., Wang, C., Alvers, S., & Susenbeth, A. (2009). Evaluation of titanium dioxide as an inert marker for estimating faecal excretion in grazing sheep. Animal Feed Science and Technology, 152(3-4), 186-197. doi: 10.1016/j.anifeedsci.2009.04.010
https://doi.org/10.1016/j.anifeedsci.200...
). Wang, Ragland, and Adeola (2017Wang, T., Ragland, D., & Adeola, O. (2017). Combination of digestibility marker and fiber affect energy and nitrogen digestibility in growing pigs. Animal Feed Science and Technology, 230, 23-29. doi: 10.1016/j.anifeedsci.2017.05.012
https://doi.org/10.1016/j.anifeedsci.201...
) reported a low fecal recovery rate of the TD marker when compared with total fecal collection in pigs fed diets with corn starch, corn meal or oatmeal. Sampaio et al. (2011Sampaio, C. B., Detmann, E., Valente, T. N. P., Costa, V. A. C., Valadares Filho, S. C., & Queiroz, A. C. (2011). Fecal excretion patterns and short term bias of internal and external markers in a digestion assay with cattle. Revista Brasileira de Zootecnia, 40(3), 657-665. doi: 10.1590/S1516-35982011000300026
https://doi.org/10.1590/S1516-3598201100...
) investigated the excretion profile of the CO, TD, iDM, iNDF and iADF markers, as well as different fecal sampling designs, to obtain representative point samples. According to these authors, there must be at least four fecal collections, which can be distributed throughout the day, for a significant representativeness of the total excreted feces; and four days of collection, so that there the marker is accurately represented in the fecal sample. This protocol would reduce sampling errors that could interfere with the determination of fecal output and, consequently, nutrient digestibility by the marker. In the present study, the external markers were supplied once daily and the feces were also collected once, in the morning.

In view of the foregoing, it is imperative to standardize the methodologies of use and analysis of markers in digestibility and intake trials so that better responses can be obtained with the most varied diets and animal categories evaluated.

Conclusion

The chromium oxide, LIPE®, NANOLIPE® and indigestible neutral detergent fiber markers accurately estimated fecal output and nutrient digestibility.

The titanium dioxide, indigestible dry matter and indigestible acid detergent fiber markers were inefficient in estimating fecal output and nutrient digestibility.

Acknowledgments

The authors thank the Federal University of Minas Gerais; the Institute of Research, Technical Assistance and Rural Extension of Espírito Santo State; the Center for Agrarian Education and Development of Florestal - MG; and Trop Frutas (Leão Alimentos) - ES, for their support in the development of this study

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Publication Dates

  • Publication in this collection
    25 Sept 2020
  • Date of issue
    2021

History

  • Received
    10 Jan 2019
  • Accepted
    20 Mar 2020
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