Calcium and phosphorus excretion and retention in diets of laying hens fed soybean waste fermented by Trichoderma viride

J S Mandey, J R Leke¹ and C A Rahasia

Animal Nutrition Department, Animal Husbandry Faculty, Sam Ratulangi University,
Jl. Kampus Selatan, Unsrat. Manado 95115, Indonesia
jetsm_fapet@yahoo.co.id
¹ Animal Production Department, Animal Husbandry Faculty, Sam Ratulangi University, Jl. Kampus Selatan, Unsrat. Manado 95115,Indonesia

Abstract

A 19-day experiment was conducted to determine the effect of soybean waste (SBW) fermented at several duration time by Trichoderma viride in diets on calcium (Ca) and phosphorus (P) excretion and retention of laying hens. The experiment was designed to completely randomized design (CRD) with 5 treatments and 4 replications. Dietary treatments were SBW unincubated-based diet, fermented SBW 3 days, 5 days, 7 days and 9 days-based diet, as SBW-0, SBW-3, SBW-5, SBW-7 and SBW-9, respectively. The method of total excreta collection was applied to determine Ca and P excretion and retention.

The statistical evaluation of the chicks’ dietary response indicated no difference in Ca intake, in Ca excretion and in P excretion of the birds fed SBW fermented T. viride 3-9 days in diets compared to fermented SBW in the diet. There were differences in Ca retention, in P intake and in P retention. It could be concluded that SBW fermented T. viride 9 days was the best. Feed produced by microbial fermentation of SBW could be successfully used as poultry feed.

Key words: by-product, digestibility, fungi, poultry

Introduction

Tempe (or tempeh) is a solid fermented soybean product that is consumed widely in Indonesia, and in recent years, there has been considerable interest in the West, especially the USA. In the processing of tempeh, the seed coat or hull of soybean will be discarded after soaking and dehulling of tempeh processing as industrial waste (Nelwida 2011). Iriyani (2001) reported that soybean hulls contained 18.0% crude protein, 5.5% crude fat, 24.8% crude fibre and 2989 Kcal/kg metabolizable energy, and were considered to be exellent source of calcium (Ca) and phosphorus (P). And, although the Ca and P levels are variable, most products contain between 7 to 10% Ca and 3.5 to 5.0% P.

Soybean waste (SBW) can be an alternative feed ingredient for poultry, because it was available and do not compete with human needs. However, feed ingredients derived from agro waste are usually very limited use for poultry diet, because these materials generally contain high crude fiber. According to Ward and Reichert (1986) the dietary fiber in canola and soybean seeds are in the outter hull and the cell wall material. These fibers may affect nutrients and especially mineral digestion in animals (Van der Aar et al 1983). The absorption of certain dietary minerals depends on their bioavailability from digestible plants, interaction of the minerals with other dietary nutrients and physiological state of the animal. Higher fiber in feed ingredients has been shown to have negative effects on digestion and absorption of nutrients in chickens (Krogdahl 1986). Therefore, the necessary efforts to overcome the weaknesses of the materials through bioconversions process which can be made with fermentation technology, that is one alternative and inexpensive method to improve the nutritional value of a waste. Fermentation with the fungus Trichoderma viride has been used in various substrates, especially the high crude fiber such as SBW. The fungus Trichoderma viride have potency to produce cellulase in relatively large quantities in order to degrade cellulose (Sukaryana et al 2010), and also have potency to produce pentonase and xylanase (Hardjo et al 1984).

The digestion of Ca and P in poultry have been generally measured over the total digestive tract (Common et al 1948) and the inter-relationship between dietary Ca and P concentrations in poultry nutrition and metabolism has been recognized for many years. The importance of the identification of the Ca bioavailability in laying hens has been of great interest to researchers for a long time (De Witt et al 2008). Calcium is an important feedstuff for shell strength (William et al 2006), and is the main mineral component of the egg shells which responsible of the internal egg quality (Roudybush and Grau 1987). Roland et al (1996) reported that Ca deficiency lead to decreased egg production, feed consumption and bone density and strength. While excess calcium significantly reduced egg production, and feed consumption (Harms and Waldroup 1971), and high dietary Ca have negative effect on P absorption in different species (Liu et al 2000).

Reduction of P excretion into environment has become one of the most important tasks in modern poultry production. Poultry diets are usually based on cereals in which the mineral P is bound in the form of phytate, and also phytate can makes complexes with mineral Ca (Pintar et al 2005). Utilization of plant P by chickens is poor because a significant proportion of the total P in the vegetable feed ingredients is bound to phytate (Eeckhout and Paepe 1994). Several studies have shown that dietary P concentration is an important factor that affects apparent P digestibility by poultry (Qian et al 1996; Rodehutscord and Dieckmann 2005).

A high ratio of dietary Ca to P reduces the digestibility and absorption of Ca and P due to increased precipitation of Ca-P complexes (Plumstead et al 2008; Selle et al 2009). However, because Ca is the mineral added in highest concentrations in poultry diets, it has a greater impact in forming mineral-phytate chelates than other dietary minerals, making both Ca and P unavailable for absorption (Tamim et al 2004).

The research was conducted to investigate the chemical composition and mineral Ca and P retention of broiler chickens fed diets containing varying duration time of fermentation of soybean waste.

Materials and methods

Collecting and processing of soybean waste

The tempeh was prepared according to traditional methods such as soaking, boiling/heating, drying and fermentation (Hedger 1982; Wei 1991; Egounlety and Aworh 2003). Soaking is necessary to remove inhibitors from the soybeans and to loosen the hulls, and dehulling is necessary for successful fermentation (Hedger 1982). Clean soybean grains were dehulled, and the separated cotyledons were cooked in boiling water for 10 min. After the phase of soaking and dehulling soybean waste were collected and processing for treatments.

Soybean wastes were dried and ground and then covered with alumunium foil. The substrates then were autoclaved at 121 ⁰C for 15 minutes. The autoclaved samples were allowed to cool to ambient temperature before inoculation. All experiments were performed in triplicates.

Inoculum was made by the suspension of filamentous fungi Trichoderma viride (contain of T. viride and destilation water in 1:9 ratio) and nutrients (31.25 g (NH4)2PO4; 6.25 g NaH2PO4; 0.3 g KCl; 2.08 g MgSO4; 0.31 g FeSO4 and 0.35 g urea in 1 L destilation water) in 1:4 ratio. Soybean waste meal then was inoculated with inoculum of T. viride on 5L per 50 kg SBW for 3 days, 5 days, 7 days and 9 days of incubation. Then, the fermented soybean waste was dried and ground for treatments.

Diets

The experiment was designed to completely randomized design (CRD) with 5 treatments and 4 replications. Dietary treatments were unfermented SBW-based diet, fermented SBW 3 days, 5 days, 7 days and 9 days-based diet, as SBW-0, SBW-1, SBW-2, SBW-3 and SBW-4, respectively. The feed ingredients and composition of based diet were shown in Table 2.

Chicks were raised in 20 metabolic batteries constitutes of cages, which were equipped with feeder and drinker. Feed in collection period was made 80% restriction and water was made available for ad libitum consumption. The Chemical Composition of SBW is shown in Table 1 and compositions and calculated analysis of diets are shown in Table 2.

Retention and Excretion

The in vivo digestibility trial was conducted by using 7 months old Strain Brown laying hens. Twenty layers having similar body weight (1700 – 1725 g) were obtained from a commercial layer farm and were reared in individual metabolic cages.

The experiment was conducted using total collection method, period lasted 19 days: a 12-day precollection period and a 7-day collection period, and during experiment birds were fed same amount of feed. The collected excreta were sprayed by 2% boric acid solution to prevent any loss in ammonia. The excreta of all experimental units were collected daily on trays covered with plastic and then dried in an oven at 60 ⁰C for 24 hours.

At the end of the experimental period, the twenty samples of excreta were weighed and ground prior to analysis for calcium and phosphorus as described by AOAC (1990).

Retention was calculated as follows:

% Retention = (consumed – excreted) x 100/consumed (Pintar et al 2005)

Statistical Analysis

The data were subjected to analysis of variance technique using Completely Randomized Design (CRD) whereas, Duncan’s multiple range test (Snedecor and Cochran, 1967) was used for comparison of the treatment means. Software package Genstat 12.2 was used for statistical calculation.

Results and discussion

Data on calcium and phosphorus excretion and retention of laying hens affected by soybean waste (SBW) in diet is shown in Table 3.

There were no difference among treatments in Ca intake and Ca excretion, however, the diets containing fermented SBW 7 and 9 days showed increased Ca retention when compared to those fed unfermented (SBW-0) and fermented 3 and 5 days diets. The diets containing SBW-9 were increased in mineral P intake when compared to those fed SBW-0 and fermented SBW-3, SBW-5 and SBW-7 diets. And the diets containing fermented SBW-7 and SBW-9 were increased in mineral P retention when compared to those fed SBW-0 and fermented SBW 3 and 5 days diets. However, there was no difference among treatments in P excretion.

The lowest retention of these minerals was exhibited by the birds fed diet containing SBW-0 followed by the diets containing fermented SBW 3 and 5 days. It means that the lower the fiber the higher the retention of Ca and P. The result of the study revealed that the digestibility of minerals Ca and P were affected in the birds fed different term of fermentation of SBW. There was a corresponding increase in the mean digestibility values with increase the term of fermentation of SBW in the diets.

Cellulose in fiber produced the bulkiest fecal material and had the fastest transit rate. This may indicate that cellulose affects absorption of minerals mainly through increased bulkiness and decreased transit time will decrease the frequency of mucosal-mineral interactions necessary for absorption. Harmuth-Hoene and Schelenz (1980) reported that the inclusion of various fibers into the rat diets produced lower apparent availability of copper, iron, zinc, magnesium, calcium and phosphorus. These decreases were depending on the type of fiber added to the diet and the particular mineral measured, that were intestinal transit time was shortened and fecal bulk was greater in comparison to the control diet, chelation of the minerals to the fiber matrix. Minerals are found in highest concentrations in the germ and outer layers of the grains. With fermentation, the dietary fibre matrix is often altered or destroyed and more minerals would remove (Harland, 1989), so it successfully yielded Ca and P and it is easily digestible.

Conclusion

• From the analysis of Ca and P retention in soybean waste it is concluded that soybean waste fermented T. viride has a potential in laying hens production. Because, when used as a feed ingredient, it successfully yielded Ca and P and it is easily digestible.

References

Association of Official Analytical Chemists 1990 Official Methods of Analysis; 15th ed. Washington, DC.

Common R H, Rutledge N A and Hale R W 1948 Observations on the mineral metabolism of pullets. VIII. The influence of gonadal hormones on retention of calcium and phosphorus. Journal of Agricultural Science, 38: 64-80.

De Witt F H, Kuleile N P, Van der Merwe H J and Fair M D 2008 Influence of limestone particle size on egg production and eggshell quality of layers. World’s Poultry Science Journal 64 (Suppl. 2): 411-416.

Egounlety M and Aworh O C 2003 Effect of soaking, dehulling, boiling and fermentation with Rhizopus oligosporus on the oligosaccharides, trypsin inhibitor, phytic acid and tannis of soya bean (Glycine max Merr.), cowpea (Vigna unguiculata L. Walp) and groundbean (Macrotyloma geocarpa Harms). Journal of Food Engineering 56:249-254.

Eeckhout W and Paepe M D 1994 Total phosphorus, phytate phosphorus and phytase activity in plant feedstuffs. Animal Feed Science Technology 47:19–29.

Hardjo S. Indrastuti N S dan Bantacut T 1989 Biokonversi: Pemanfaatan Limbah Industri Pertanian. PAU-Gizi, IPB, Bogor.

Harland B F 1989 Dietary Fiber and Mineral Bioavailability. Nutrition Research Review 2:133-147.

Harms R H and Waldroup P W 1971 The effect of high dietary calcium on the performance of laying hens. Poultry Science 50: 967-969.

Hedger J N 1982 Production of Tempe, an Indonesian Fermented Food. The Society for General Microbiology. Depart. of Botany and Microbiology, Univ. College of Wales.

Iriyani N 2001 Pengaruh penggunaan kulit biji kedelai sebagai pengganti jagung dalam ransum terhadap kecernaan energi, protein dan kinerja domba. Jurnal Produksi Ternak Vol. 2, Nov. 2001.

Krogdahl A 1986 Antinutrients affecting digestive function and performance in poultry. Proceedings of the 7th European Poultry Conference. Paris. Vol. 1, pp 239-248.

Liu J, Bollinger D W, Ledoux D R and Veum T L 2000 Effects of dietary calcium:phosphorus ratios on apparent absorption of calcium and phosphorus in the small intestine, cecum, and colon of pigs. Journal of Animal Science 78: 106-109.

Nelwida 2011 Pengaruh pemberian kulit ari biji kedelai hasil fermentasi dengan Aspergillus niger dalam ransum terhadap bobot karkas ayam pedaging. Jurnal Ilmiah Ilmu-Ilmu Peternakan, Vol. XIV, No. 1:23-29.

Pintar J, Homen B, Gazic K, Janjecic Z, Sikiric M and Cerny T 2005 Effect of supplemental phytase on nutrient excretion and retention in broilers fed different cereal based diets. Czech Journal Animal Science 50 (1):40-46.

Plumstead P W, Leytem A B, Maguire R O, Spears J W, Kwanyuen P and Brake J 2008 Interaction of calcium and phytate in broiler diets.1. Effects on apparent prececal digestibility and retention of phosphorus. Poultry Science 87:449–458.

Qian H, Kornegay E T and Denbow D M 1996 Phosphorus equivalence of microbial phytase in turkey diets as influenced by calcium to phosphorus ratios and phosphorus levels. Poultry Science 75:69–81.

Roland D A, Sr, Bryant M M and Rabon H W 1996 Influence of calcium and environmental temperature on performance of first –cycle (phase 1) commercial leghorn. Poultry Science 75: 62-68.

Roudybush T E and Grau C R 1987 Calcium need and danger. Exotic bird report. Avian Science Department, University of California, Davis, California, 95616,7:1.

Rodehutscord M and Dieckmann A 2005 Comparative studies with three-week-old chickens, turkeys, ducks, and quails on the response in phosphorus utilization to a supplementation of monobasic calcium phosphate. Poultry Science 84:1252–1260.

Harmuth-Hoene A E and Schelenz R 1980 Effect of dietary fiber on mineral absorption in growing rats. Journal Nutrition 110: 1774-1784.

Selle P H, Cowieson A J and Ravindran V 2009 Consequences of calcium interaction with phytate and phytase for poultry and pigs. Livestock Science 124:126–141.

Snedecor G W and Cochran W G 1967 Statistical Methods, 6^th Ed. Oxford and IBH Publication, Calcutta, India.

Sukaryana Y, Atmomarsono U, Yunianto V D dan Supriyatna E 2010 Bioconversions of Palm Kernel Cake and Rice Bran Mixtures by Trichoderma viride Toward Nutritional Contents. International Journal of Science and Engineering Vol. 1(2):27-32, Dec.

Tamim N M, Angel R and Christman M 2004 Influence of dietary calcium and phytase on phytate phosphorus hydrolysis in broiler chickens. Poultry Science 83:1358–1367.

Van der Aar P J, Fahey G C, Jr. Ricke, S C, Allen S E and Berger L L 1983 Effects of dietary fibers on mineral status of chicks. Journal Nutrition 113:653-661.

Wei I S 1991 Domestic soybean consumption in Asia. In: Tanteeratarm, K. (Ed.). Soybean processing for food uses. Urbana-Champaign: INTSOY: University of Illinois. p. 162-183.

Ward A T and Reichert R D 1986 Comparison of the effect of cell wall and hull fiber from canola and soybean on the bioavailability for rats of minerals, protein and lipid. Journal Nutrition 116: 233-241.

William N S, Horracio S R, Paulo R S, Luis F U and Marcelo A S 2006 Nutritional requirements of Calcium in white laying hens from 46 to 62 week of age. International Journal of Poultry Science, 2;181-184.

Received 17 June 2015; Accepted 11 October 2015; Published 1 November 2015

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