Gupta, S.L., Tyagi, Pramod K., Tyagi, Praveen K., Mandal, A.B., Kolluri, G., Mir, N.A. and Khan, A. 2017. The response of rice based dry distiller’s grains with soluble (DDGS) feeding on gastrointestinal microbiota and immunity in layer’s diet. Indian Journal of Poultry Science, 52(2): 133-137.
A biological experiment was undertaken in a 4x2 factorial completely randomized design (CRD) with 45 wk old CARI Sonali layers (n=120) which was assigned to eight treatments (i.e. twocontrols + sixtest diets) in such a way that each treatment has 15 replicates with one layer in each replicate for ten weeks (45th-54th week of age). Theexperimental diets were prepared by incorporating rice DDGS at 0, 5, 7.5 and 10% levels (D1, D2, D3, D4); proteaseenzyme supplemented in rice DDGS at the above levels (D5, D6, D7, D8). All the diets wereis-caloric and is- nitrogenousin nature.Dietary inclusion of rice DDGS with and without proteaseenzyme improved the immune response in layers measured in terms of cell mediated immune response to phytohaemag glutinin-P and humoral immune response to sheep red blood cells.Rice DDGS with and without proteaseenzyme has beneficial effect on gut health in terms of bacterial population. Inclusion of 7.5 and 10% level of DDGS was found to lower (P<0.05) the total viable count TVC in crop. However, lactobacillus count in jejunum was significantly higher (P<0.01) at 5, 7.5 and 10% levels of DDGS with maximum effect at 10% level of DDGS but no effect was observed incrop region. Significantly (P<0.01) higher lactobacillus count was also observed in jejunum whenprotease enzyme was incorporated.
Key words: Crop, Protease, Imune response, Jejunum, Lactobacillus, Layer, Rice DDGS
High feed ingredient price around the world has forced to search for lower cost alternative feed ingredients to minimize the cost of feed. Dried distillers grain with soluble (DDGS) is a by-product of the ethanol production process and it has been documented as a promising feed ingredient in the poultry and livestock industries (Cheng and Hardy, 2004; Jacob et al., 2008) because it is highly nutritious and economical. It is a cereal by-product which is fermented and distilled to obtain alcoholic beverage (Hertrampf and Piedad-Pascual, 2000). The fermentation process concentrates the remaining nutrients approximately to three-fold. The sugars from starch in cereal grain are fermented by yeast to produce ethanol, releasing carbon dioxide. The remaining “wetcake” is distillers’ grains, to which the condensed solubles are added back at drying ending up with DDGS for feeding to livestock and poultry. The cereal grain yields about one-third ethanol, one-third CO2, and one third DDGS (Saunders and Rosentrater, 2009). After starch extraction, the by product contains protein, fibre, oil and essential amino acids which are suitable for livestock and poultry feed called DDGS.The high energy and protein content in rice DDGS can make it an important substitute forexpensive source of energy (corn) and protein (soybean meal) ingredients inpoultry feed. Exogenous enzyme supplementation in the diet improves production efficiency of poultry by increasing digestion of low quality products and reducing nutrient loss through excreta (Buchanan et al 2007; Costa et al 2008). This leads not only to more dietary energy being utilized by the bird, but also gut viscosity is decreased. Recent studies (Shim et al. 2011; Loaret al., 2010; Masa’dehet al. 2011) have shown that corn DDGS can be added to poultry diets to achieve excellent performance and improved gut health provided that accurate nutrient profiles specific to the DDGS source are used. However, no information is available on the potential use of rice DDGS in poultry feed. Therefore the present experiment was carried out to evaluatethe immune status and gut health of layers fed rice based DDGS with or without proteaseenzyme.
A biological experiment was undertaken in a 4x2 factorial completely randomized design (CRD) with twenty six weeks old CARI Sonali(RIR Male x WLH Female) layer birds (n=120) from a single hatch andwhich were randomly assigned into eight groups (D1 to D8) in such a way that each treatment had15 replicateseach. All the layer birds were reared in battery cages fitted with individual feeder, waterers and dropping trays and reared under standard management conditions. The allocation of birds in each treatment was based on egg production at the start of the experiment. The experiment was conducted for 10 weeks (45th to 54th weeks of age).
Feed ingredients and supplements for formulation of control and test diets were procured from the feed storage and processing unit of the institute. All the feed ingredients were analyzed for proximate constituents (AOAC, 2000) prior to formulation of experimental diets. Eight test diets containing dry distiller grains with soluble at 0, 5, 7.5, 10% levels with and without protease enzyme was prepared for layer ration. All the diets contained uniform package of essential nutrients. Weighed amount of respective diet was offered to individual birds daily in the morning and fresh wholesome potable water was always made available to the birds throughout the experimental period. Every attempt was made to minimize feed spillage/wastage. From the basal diet as maintained above, eight experimental diets (designated as D1 to D8) was formulated by adding desired levels of DDGS with and withoutprotease enzyme.
After 4th week of experimental peried, ten layers per treatment were taken to evaluate humoral immune response estimated by method of Siegel and Gross (1980) with slight modification by Saxenaet al. (1997) was followed for assaying the immune response to sheep red blood cells (SRBCs).The in vivo cell mediated immune response to PHA-P was evaluated after 8th week of experimental period by the method of Cheng and Lamont (1988).At the end of trail, four birds from each dietary treatment were sacrificed by cervical dislocation; crop and jejunum scraping were collected in sterile vials for evaluation of total microbial load colonization. Microbial populations were determined by serial dilution (104 to 106) of crop and jejunum samples in anaerobic diluents before inoculation onto petri dishes of sterile agar as described by Bryant and Burkey (1953). Total bacterial count and Lactobacilli was grown on nutrient agar and Rogosa SL agar respectively (Demanet al, 1960). Plates were counted between 24 and 48 h after inoculation. Colony forming units were defined as distinct colonies measuring at least 1 mm in diameter.
cfu/g= Total No. of colony counted x Dilution factor Volume of aliquot taken The data obtained for these parameters was subjected to statistical analysis as per factorial design (Snedecor and Cochran, 1989). The significant mean differences were sort out as per Duncan’s multiple range (Duncan, 1955) test.
|Nutrient composition (%)||0.04||0.04||0.0||0.0||0.0||0.0||0.0||0.0|
Constant includes salt 0.25%, trace mineral premix 0.11%, vitamin premix 0.15%, vit. B complex & E 0.015%, choline chloride 0.05% and Toxin binder 0.05%. Trace mineral premix supplied mg / kg diet: Mg, 300; Mn, 55; I, 0.4; Fe, 56; Zn, 30; Cu, 4. The vitamin premix supplied per kg diet: Vit.A, 8250 IU; Vit. B2 5mg; Vit.D3, 1200 IU; Vit.K, 1mg. Vitamin B complex & E supplied per kg diet: Vit. B1, 4 mg; Vit. B6 8 mg; Vit.B12, 40mcg; niacin, 60mg; pantothenic acid, 40mg; Vit.E, 80 IU. Choline chloride supplied per kg diet: choline, 500mg.
DDGS –Dry distiller grains with soluble, DORB- De-oiled rice bran, DCP-Di-calcium phosphate and E-Enzyme. 1Analyzed values, 2Calculated values.
The rice DDGS contains moisture 8.28%, DM 91.72%, CP 45.00%, EE 4.49%, CF 4.89%, TA 10.22%, NFE 27.12%, AIA 4.28%, Ca 0.73%, P 0.77% and GE 4097kcal/kg. The data pertaining to influence of different levels of DDGS with and without enzyme supplementation on total viable count (TVC) and Lactobacillus count in crop and jejunum have been presented in table 2, and figure 1 & 2. Lactobacillus is the major component of the microbial barrier to infection (Ewing andCole, 1994). The earlier reports have also shownthe beneficial effect of DDGS on gut health.The corn DDGS diet had a greater (P < 0.05) lactic acid concentration than the wheat DDGSdiet and lactic acid is implicated aspotentially inhibitorof the growth of pathogens (Axelsson, 1998). In present experiment, inclusion of different levels of rice DDGS with andwithout proteaseenzyme hadsignificant (P<0.01) impact on TVC and Lactobacilluscount in crop and Jejunum. Inclusion of 7.5 and 10% levelof DDGS significantly (P<0.05) lower the TVC in crop as compare to other inclusionlevel of DDGS, while TVC significantly (P<0.01) lowered in jejunum at all three levels ofDDGS as compare to 0% inclusion level (reference diet). However, Lactobacillus have showed a significant (P<0.01) effect at 5, 7.5 and
Table 2: Effect of feeding rice DDGS with and without protease enzyme supplementation on TVC and Lactobacillus count (cfu/g)
|0||-||6.73 6.62||3.71 3.29|
|D2||0||+||6.64 6.56||3.89 3.38
|D3||5||-||6.31 6.22||3.82 3.49|
10% levels of DDGS with maximum effect at 10% level ofDDGS in jejunum region.No significant (P>0.05) effect was observed in all DDGS level in crop region. Addition of proteaseenzymehas no significant (P>0.05) effect on TVC in crop and jejunum and also on Lactobacillus incrop region whereas, significantly (P<0.01) higher Lactobacillus count was observed in jejunumin presence of proteaseenzyme. Interaction between different levels of DDGS and proteaseenzyme has noeffect on TVC and Lactobacillus in crop or jejunum region.In this respect, Hahn (2010) observed that the diets containing corn DDGS had asignificantly higher count of 1.998 x 10-6 CFU/ml as compared to control diet. This relationshiphas been previously seen, as Yang et al. (2010) reported that diets containing corn DDGS hadgreater lactic acid containing bacteria than diets containing wheat DDGS. This could be becauseof the composition of DDGS in that it is more fibrous offers more nutrients to Lactobacillus.Our results are coincide with (Yan et al., 2013) who suggested that addition of NSP degrading enzymes improved growth performance and gut health in broilers.
Table 3: Effect of feeding rice DDGS with and without proteaseenzyme supplementation on immunological parameters
|Diet||DDGS(%)||Enz.||CMI response (mm)||HA titre(log)2|
|NS- Non-significant (P>0.05)|
Effect of feeding rice DDGS with and without enzyme supplementation on hemato-immunological parameters have been shown in tables 3. Heamagglutin in antigen titre (HA) was significantly (P<0.01) increased at 7.5 and 10% inclusion levels with highest value (7.65) at 10% level of inclusion of rice DDGS in layer diet. Dietary addition ofprotease enzyme hadsignificant (P<0.05) impact on HA titre, where thevalues were improved, however, CMI response did not show any significant (P>0.05) change due toprotease enzyme. These results are similar to Barekatain et al. (2013) who investigated the effect of a high level of sorghum distillers dried grains with solubles (DDGS; 20%), with or without a combination of protease and xylanase in broiler chickens, under a necrotic enteritis disease challenge and found that incorporation of DDGS in the diets improved (P<0.01) the IgA and IgG titer at d 13 but interacted with the disease challenge, reducing the concentration of IgA at d 21 and IgM at d 35 in the infected birds.
Inclusion of 7.5 and 10% level of DDGS was found tolower (P<0.05) the TVC in crop andjejunum. However, lactobacilluscount lowered at 10% level of DDGS only in jejunum.Addition of protease enzyme had no significant (P>0.05) effect on TVC in crop and jejunum and lactobacillus in crop region whereas, significantly (P<0.01) higher lactobacillus count was observed in jejunum in presence of protease enzyme.Interaction between different levels of DDGS and proteaseenzyme had no effect on TVC and Lactobacillus in crop or jejunum region.Interaction between different levels of DDGS and proteaseenzyme hadno effect on TVC and Lactobacillus in crop or jejunum region.Cell mediated immune response to PHA-P did not have any significant (P>0.05) effect of inclusion of different levels of rice DDGS either sole or in combination with proteaseenzyme. However, heamagglutin in antigen titre (HA) was significantly (P<0.01) increased at 7.5 and 10% inclusion level with highest value (7.65) at 10% level of rice DDGS, which was significantly increased by protease enzyme addition in diets.
The study envisaged that incorporation of 10% rice based dried distillers grain with soluble (DDGS)proved to be beneficial for immune response & gut health. Further, supplementation of protease enzyme in layer diet containing DDGS improved the immune response.