Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

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Determination of In vitro Nutrient Digestibility of Silage based Diets with Varying Levels of Crude Fibre using Pig Faecal Inoculum

K. C. Barman1, K. Barman2, A. K. Verma1,*, S. E. Jadhav1, N. Dutta1, P. Singh3, P.Chaudhary1, S. L. Jat4
1Division of Animal Nutrition, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India.
2ICAR-National Research Centre on Pig, Guwahati-781 015, Assam, India.
3Division of Biochemistry, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India.
4ICAR-Indian Institute of Maize Research, Unit Office, New Delhi-110 012, India.
Background: The in-vitro nutrient digestibility and fermentation mimicking gastrointestinal process of pigs is a convenient method for rapid and cheaper measurement of degradation of feed ingredients. In the present study silage based diets containing varying levels of crude fibre were subjected to in vitro digestibility evaluation using pig faecal inoculum to determine the best crude fibre level for feeding pigs.  

Methods: Eight diets were prepared namely, B1, B2, V1, V2, V3, M1, M2 and M3 having 6.28, 8.43, 8.03, 10.0, 11.96, 8.17, 10.31 and 12.06 per cent crude fibre (on dry matter basis) respectively, where diets B1 and B2 were basal diets, V1, V2 and V3 contained vegetable waste silage; M1, M2 and M3 contained maize silage -as additional fibre source. An in-vitro study was carried out to investigate the nutrient digestibility of diets involving enzymatic hydrolysis and fermentation using pig faecal inoculum.

Result: The in-vitro total tract digestibility (IVTTD) of dry matter (DM), crude protein (CP) and organic matter (OM) was significantly higher (P<0.001) in B1 diet than the other diets. The digestibility of DM, CP and OM of B2 diet was also significantly higher (p<0.001) than the M2, M3, V2 and V3 diets but comparable to M1 and V1 diets. The IVTTD of crude fibre (CF) was higher in V3 diet (25.85%) and lowest (P<0.001) in M1 diet (11.65%) and both showed a highly significant (P<0.001) difference (to other diets. The DM, CP and OM digestibility were significantly higher (P<0.001) in the diets containing normal fibre sources than silage containing diets. Cumulative gas production (ml/0.5g feed) at 24 h of in-vitro fermentation were significantly higher (P<0.001) in B2 (34.95) and B1 (31.00) diets than other diets. As the CF level in diets increased from 6 to 12%, the digestibility of nutrients linearly decreased except CF digestibility. It was revealed from the study that the level and sources of crude fibre in diets influenced the digestibility of nutrients.
The use of fibre rich feed ingredients in pig diets especially in small scale farming is rising in recent times due to their wide availability and relatively low cost as compared to commercial feeds. Similarly, offering silages of forages and vegetables in the diets of pigs is becoming an efficient practice in organic pig production system (Edwards, 2002; Bikker et al., 2014; Wüstholz et al., 2017 and Friman et al., 2021). High fibre diets increase cellulolytic bacteria in the large intestines of pigs (Anugwa et al., 1989) and have prebiotic effects and improve gut-associated immune system (Lindberg, 2014). The fermentation products of crude fibre at large intestine such as volatile fatty acids contribute 7 to 25% of maintenance energy needs of pigs (Yen et al., 1991) and butyric acid acts as instant energy source for the colonocytes and regulates cell growth and differentiation (Roediger, 1982). As a part of plant cell wall, crude fibre is generally less digestible in pigs and it adversely effects nutrient utilization (Zhang et al., 2013; Wilfart et al., 2007). The impact of fibre level on nutrients digestibility may vary with fibre source, properties and composition of fibre (Hogberg and Lindberg, 2004; Van Soest, 1978).
Hence, it is imperative to understand the effect of fibre level on nutrient digestibility and voluntary feed intake for optimum swine production. Numerous studies by different scientists regarding digestibility of fibre and fibrous feeds in pigs have been executed. However, information about nutrient digestibility and fermentation characteristics of fibre from vegetable waste silages and maize silages are meagre.
Determination of digestibility of nutrients is an important criterion for quality evaluation and feeding value. Digestibility evaluation using animal is a time-consuming and tedious process and requires large quantities of feeds and animals. In pigs, digestibility of nutrients starts in the stomach (pepsin and hydrochloric acid), continues in small intestine (enzymatic hydrolysis) and finally in caecum and colon fermentation occurs and help in the breakdown of fibre and other nutrients that had escaped enzymatic hydrolysis. The in vitro nutrient digestibility and fermentation that mimics gastrointestinal process of pigs are convenient methods for rapid and inexpensive measurement of the degradation of feed ingredients (Boisen and Fernandez, 1997; Guo et al., 2004). There may be numerous modifications, where some may use commercially available multi-enzymes (Boisen and Fernandez, 1997) or microbial inoculum either rumen or faecal inoculum (Dung et al., 2002; Bindelle et al., 2007; Youssef and Kamphues, 2020) for simulation of large intestinal fermentation processes. Therefore, vegetable waste silage and maize silage were incorporated as a source of fibre to increase the CF level in experimental diets and were subjected to in-vitro digestion comprising two steps enzymatic hydrolysis and fermentation to investigate the higher level of incorporation of CF in pig feeds.
The study was conducted at Indian Council of Agricultural Research-National Research Centre on Pig, Rani, Guwahati, Assam, India (91°17' 49" E longitude, 25°48' 30" N latitude and 467m altitude) during the month of April-September, 2021. The experiment was carried out taking prior approval of the Institutional Animal Ethics Committee (IAEC) of Indian Council of Agricultural Research-National Research Centre on Pig, Rani, Guwahati and ensured that no potential harm toward animal welfare would be done.
Silages were prepared from maize fodder (Zea mays) and vegetable waste (cabbage and cauliflower at the ratio 60:40 in raw basis) individually in silage bags following standard procedure (3 kg jaggery and 250 g salt per 100 kg raw materials) and opened for sampling after 4 weeks. Feed ingredients were analysed for proximate principles (AOAC, 2012) and neutral detergent soluble and fibre fractions as per the procedure of Van Soest et al., (1991). Energy content of feeds was calculated from the proximate composition using the formula as mentioned by NRC (2012). The pH of silages was estimated following the method described by Bernardes et al., (2019) using digital pH meter (pHTestr 10, Eutech Instruments). The silage quality was determined by using Flieg’s score formula (Kilic, 2010). All feed ingredients were dried in oven following standard procedure and ground through a 1mm mesh screen with Wiley No. 4 Laboratory Mill. Low temperature (60°C) oven drying method was followed for estimation of dry matter (DM) in silages. 
Eight types of diets were formulated as per the nutrient requirement of swine (ICAR, 2013) for in-vitro studies with commonly available feed ingredients and silages (vegetable waste and maize silage) as fibre source as mentioned in Table 1.

Table 1: Ingredient and nutrient composition of diets used for in-vitro digestibility experiments.

Representative samples from formulated diets were subjected to a 2-step in-vitro enzymatic hydrolysis to mimic the gastro-intestinal digestion using pepsin and pancreatin solutions (Boisen and Fernández, 1997) followed by in-vitro fermentation as per the procedure described by Youssef and Kamphues (2020) using faecal materials from pigs slaughtered in the institute slaughter house of ICAR-NRC on Pig, Rani, Guwahati.
In-vitro total tract digestibility of DM, CP, OM and CF were obtained after enzymatic hydrolysis and in-vitro fermentation. Cumulative gas production (ml/0.5 g feed) during in-vitro fermentation of experimental diets was also estimated. The data generated were subjected to one way ANOVA as per the methods described by Snedecor and Cochran (1989).
Silage parameters
The prepared silages were evaluated for moisture, pH, fleig point, colour, odour and mould growth (Table 2). The average moisture content and pH level of vegetable waste silage (VS) was more (75.35%; 4.9) compared to maize silage (MS, 69.35%; 4.0). The moisture content in silage greatly influenced the pH level.  Yahaya et al., (2002) opined that at 65-75% moisture level in silage the pH remained below 5; whereas moisture content below or above this value caused increase the pH of orchard grass silages. The average fleig point for maize silage (MS) and vegetable waste silage (VS) was 106 and 59.3, respectively and as per fleig score maize silage was excellent and vegetable silage was fair in quality (Kilic, 2010). VS contained comparatively more CP and energy (18.25%; ME 2536 kcal/kg) than MS (CP 8.3%, ME 2000.8 kcal/kg). Inversely, MS was richer in CF (24.35 vs18.5) and NDF (45.5 vs 28.5) content than VS. Literature pertaining to the chemical composition of vegetable waste silage is scanty to relate the observed findings and it differs as per the content and type of vegetable waste used for silage preparation. The chemical composition of MS was comparable to the report of NDDB (2012) and Htet et al., (2016).

Table 2: Chemical and physical qualities of silages.

In-vitro dry matter digestibility in enzymatic hydrolysis (IVDMDh)
Diet B1 (62.57%) had significantly higher (P<0.001) IVDMDh; whereas M3 and V3 diets had significantly lower (P<0.001) IVDMDh as compared to other diets (Table 3). The IVDMDh of diets with normal fibre source was found to be significantly higher (50.80%; P<0.001) as compared to MS or VS containing diets. The CF levels irrespective of fibre sources had significant influence on IVDMDh as shown in Table 4 and digestibility was significantly (P<0.001) reduced as CF level increased in the diet. It may be due to negligible action of pepsin and pancreatin enzymes on fibre and fibre bound nutrients in upper digestive tract.

Table 3: In-vitro dry matter digestibility (IVDMDh) and total tract digestibility of nutrients (IVTTD) of experimental diets.

In-vitro total tract digestibility (IVTTD) of different nutrients
The IVTTD of DM of different diets ranged from 60.80 to 76.54%, where B1 diet showed a significantly higher value (P<0.001) than the other diets. The digestibility of B2 diet was significantly higher (P<0.001) than the M2, M3, V2 and V3 but it was non-significant to M1 and V1 diets. The IVTTD of CP and OM were also significantly higher (P<0.001) for B1 diet compared to other diets, which showed similar trend as the digestibility of DM. The IVTTD of CF was higher in V3 diet (25.85%) and lowest (P<0.001) in M1 diet (11.65%) and showed a highly significant difference to B1, M1, M2, M3, V1 and V2 diets. The higher CF digestibility of high fibre diets may be due to higher concentration of CF as substrate for microbial fermentation (Anguita et al., 2006). The DM, CP and OM digestibility were significantly higher (P<0.001) in the diets that contained normal fibre sources. The CF digestibility of diet containing MS was significantly lower (P<0.01) than the diet with normal feed or vegetable silage as fibre sources. This could be due the quality of fibre and degree of lignification. The diet containing 6% CF level showed a significantly higher (P<0.001) IVTTD of DM and OM followed by 8% CF level; while comparable in diets with 10 and 12% CF level (Table 3). A highly significant difference was recorded for CP digestibility (P<0.001) among the diets with different CF level and diet with 6% CF level showed higher CP digestibility. As the level of CF increased in the diets the IVTTD of CP significantly decreased. On the other hand, the IVTTD of CF was comparable among the diets with different levels of CF (P>0.05). The result obtained is comparable with the outcomes from previous experiment (Gürsoy et al., 2021). Youssef and Kamphues (2020) also reported similar findings when fibre rich ingredients were exposed to in vitro fermentation with faecal inoculum of swine and turkey. The IVTTD of CF was significantly lower (P<0.01) in diets containing MS than normal diets or VS containing diets. The overall IVTTD of nutrients in all the formulated diets were found satisfactory because the diets were properly balanced as per the requirement of growing-finishing pigs.
Cumulative gas production during fermentation
Cumulative gas production at 24 h of in-vitro fermentation ranged from 16.00 to 34.95 (ml/0.5 g feed) and gas production in B2 diet was significantly higher (P<0.001) followed by B1 diet (Table 4).  Cumulative gas production of diets with normal fibre source was significantly higher (P<0.001) than MS or VS added diets. It was found that increase fibre level in diet reduces the gas production (Table 4). The higher production of gas in B1 and B2 diets may be due to higher content of fermentable nutrients and easily digestible fibre. However, cumulative gas production (ml/0.5g feed) in in-vitro fermentation was less compared to experiments of Youssef and Kamphues, (2020) who used substrate without enzymatic hydrolysis; but results were comparable to that reported by Bachmann et al., (2021) for fibre rich ingredients like sugar beet pulp, soybean shell, lucerne and wheat bran after pre-digestion with enzymes. This may be due to the fact that pre-digested substrate mostly produces less gas, NH3 and short chain fatty acids as compared to substrate without pre-digestion (Bachmann et al., 2021).

Table 4: Cumulative gas production (ml/0.5g feed) during in-vitro fermentation of experimental diets.

The present study revealed that the level and sources of crude fibre in diets influence the digestibility of nutrients. The results obtained from this experiment confirmed the possibilities to use maize silage and vegetable waste silage as CF source in diets of growing-finishing pigs. It is suggested that for better understanding of the role of vegetable waste silage and maize silage as fibre source and their level of incorporation in diets for better health and production needs extensive in-vivo studies in pigs.
The authors are grateful to the Director of ICAR-NRC on Pig, Rani, Guwahati for the permission to conduct feeding trial and avail the laboratory facilities and ICAR-Indian Institute of Maize Research, Ludhiana for financial support to do the experiment successfully.

  1. Anguita, M., Canibe, N., Perez, J.F. and Jensen, B.B. (2006). Influence of amount of dietary fibre on the available energy from hindgut fermentation in growing pigs: Use of cannulated pigs and in vitro fermentation. Journal of Animal Science. 84: 2766-2778

  2. Anugwa F.O., Varel, V.H., Dickson, J.S., Pond, W.G. and Krook, L.P. (1989). Effects of dietary fiber and protein concentration on growth, feed efficiency, visceral organ weights and large intestine microbial populations of swine. The Journal of Nutrition. 119: 879-886.

  3. AOAC. (2012). Official Method of Analysis, 19th ed. Association of Analytical Chemists, Washington DC, USA. 

  4. Bachmann, M., Michel, S., Greef, J.M. and Zeyner, A. (2021). Fermentation characteristics and in vitro digestibility of fibers and fiber-rich by-products used for the feeding of pigs. Animals (Basel). 11(2): 341. 

  5. Bernardes, T.F., Gervásio, J.R.S., De Morais, G. and Casagrande, D.R. (2019). Technical note: A comparison of methods to determine pH in silages. Journal of Dairy Science. 102(10): 9039-9042. 

  6. Bikker, P., Binnendijk, G., Vermeer, H. and Peet-Schwering Carola Van Der (2014). Grass Silage in Diets for Organic Growing -finishing Pigs. In: Proceedings of the 4th ISOFAR Scientific Conference. [Rahmann, G. and Aksoy, U. (Eds.)], ‘Building Organic Bridges’, at the Organic World Congress 2014, 13-15 Oct., Istanbul, Turkey (eprint ID 24257).

  7. Bindelle, J., Buldgen, A., Lambotte, D., Wavreille, J. and Leterme, P. (2007). Effect of pig faecal donor and of pig diet composition on in vitro fermentation of sugar beet pulp. Animal Feed Science and Technology. 132(3): 212-226.

  8. Boisen, S. and Fernández, J.A. (1997). Prediction of the total tract digestibility of energy in feedstuffs and pig diets by in vitro analyses. Animal Feed Science and Technology. 68(3): 277-286.

  9. Dung, N.N.X., Manh, L.H. and Uden, P. (2002). Tropical fibre sources of pigs-digestibility, digesta retention and estimation of fibre digestibility in vitro. Animal Feed Science and Technology. 102:109-124. 

  10. Edwards, S. (2002). Feeding Organic Pigs: A handbook of Raw Materials and Recommendations for Feeding Practice. Published by University of Newcastle, Newcastle upon Tyne, UK.

  11. Friman, J., Lundh, T. and Åkerfeldt, M.P. (2021). Grass/clover silage for growing/finishing pigs-effect of silage pre-treatment and feeding strategy on growth performance and carcass traits. Acta Agriculturae Scandinavica, Section A-Animal Science 70(3-4): 151-160. 

  12. Guo, L., Piao, X., Li, D. and Li, S. (2004). The apparent digestibility of corn by-products for growing-finishing pigs in vivo and in vitro. Asian-Australian Journal of Animal Science. 17(3):  379-385.

  13. Gürsoy, E., Kaya, A. and Gül, M. (2021). Determining the nutrient content, energy and in vitro true digestibility of some grass forage plants. Emirates Journal of Food and Agriculture. 33(5): 417-422.

  14. Hogberg, A. and Lindberg, J.E. (2004). Influence of cereal non- starch polysaccharides and enzyme supplementation on digestion site and gut environment in weaned piglets. Animal Feed Science and Technology. 116: 113-128.

  15. Htet, M.N.S., Than, N.N., Soomro, R.N., Ya-Dong, X., Jiang-Bo, H. (2016). Comparison of nutrients composition, forage and silage yields of maize (Zea mays L). Scholars Journal of Agriculture and Veterinary Science. 3(7): 474-479.

  16. ICAR, (2013). Nutrient Requirement of Swine. Indian Council of Agricultural Research.

  17. Kilic, A. (2010). Silo Feed (Teaching, learning and practice suggestions). Manuel. Hasad Publ. Istanbul - Turkey.

  18. Lindberg, J.E. (2014). Fiber effects in nutrition and gut health in pigs. Journal of Animal Science and Biotechnology. 5: 15.

  19. NDDB, (2012). Nutritive value of commonly available feeds and fodders in India. Animal Nutrition Group, National Dairy Development Board, Anand.

  20. NRC. (2012). Nutrient Requirements of Swine, 11th Revised edition, National Academies Press, Washington DC.

  21. Roediger, W.E. (1982). Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology. 83(2): 424-9. 

  22. Snedecor, G.W. and Cochran, W.G. (1989). Statistical Methods, 8th Ed., Iowa University Press, Ames, Iowa (USA).

  23. Van Soest, P.J. (1978). Dietary fibre: their definition and nutritional properties. American Journal of Clinical Nutrition. 31: 512-520.

  24. Van Soest, P.J., Robertson, J. B. and Lewis, B.A. (1991). Methods for dietary fibre, neutral detergent fibre and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science. 74 (10): 3583-3597.

  25. Wilfart, A., Montegne, L., Simmins, H., Noblet, J. and Van Miligan, J. (2007). Effect of fibre content in the diet on the mean retention time in different segments of the digestive tract in growing pigs. Livestock Science. 109: 27-29.

  26. Wüstholz, J., Carrasco, S., Berger, U., Sundrum, A. and Bellof, G.(2017). Fattening and slaughtering performance of growing pigs consuming high levels of alfalfa silage (Medicago sativa) in organic pig production. Livestock Science. 200:46-52.

  27. Yahaya, M.S., Kawai. M., Takahashi, J. and Matsuoka, S. (2002). The Effects of different moisture content and ensiling time on silo degradation of structural carbohydrate of orchard grass. Asian-Australian Journal of Animal Science. 15(2): 213-217.

  28. Yen, J.T., Nienaber, J.A., Hill, D.A. and Pond, W.G. (1991). Potential contribution of absorbed volatile fatty acids to whole- animal energy requirement in conscious swine. Journal of Animal Science. 69: 2001-2012.

  29. Youssef, I.M.I and Kamphues, J. (2020). Fermentation of fibre rich ingredients exposed in vitro to the faecal inoculums of swine and turkeys. Veterinary Medicine and Science. 6: 511-517.

  30. Zhang, W., Li, D., Liu, L., Zang, J., Duan, Q., Yang, W. and Zhang, L. (2013). The effect of dietary fiber level on nutrient digestibility in growing pigs. Journal of Animal Science and Biotechnology. 4: 17. 

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