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Broccoli byproduct-wheat straw silage as a feed resource for fattening lambs

Broccoli byproduct-wheat straw silage as a feed resource for fattening lambs Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 † †,2 ‡ †, Edris Partovi, Yousef Rouzbehan, Hasan Fazaeli, and Javad Rezaei † ‡ Department of Animal Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran; and Animal Science Research Institute of Iran, Agricultural Research, Education and Extension Organization, Karaj, Iran ABSTRACT:  The effect of feeding broccoli retention, rumen, and blood parameters were byproduct-wheat straw silage [BBWS; 69:31 measured. The BBWS diets had no influence ratio, dry matter (DM) basis] on performance, on DMI, ADG, feed conversion efficiency, in microbial N synthesis (MNS), rumen, and blood vivo apparent digestibility coefficients of DM, parameters in Fashandy lambs were evaluated. organic matter, CP, and ash-free neutral deter- Three diets, with equal metabolizable energy gent fiber. Neither MNS and N retention nor and crude protein (CP) with a forage to con- serum concentrations of glucose, triglycerides, centrate ratio of 27:73 (DM basis), were formu- creatinine, cholesterol, urea N, triiodothyronine, lated in which forage (lucerne and wheat straw) thyroxine, total protein, albumin, and globulin was replaced by BBWS (0, 100, or 200  g/kg of were affected. Rumen pH, NH -N, short-chain diet DM). These were assigned to three groups fatty acid concentrations, the ratio of acetic to (n = 15/group) in a completely randomized block propionic acid, and protozoa numbers were, design for a 70-d period in which diets were also, not influenced. In summary, BBWS may be offered as a total mixed ration. For each animal, fed to Fashandy lambs up to 200  g/kg of diet dry matter intake (DMI), average daily gain DM without any adverse impacts on growth (ADG), in vivo apparent digestibility, MNS, N performance. Key words: broccoli byproduct, lamb, lamb growth, protozoa, rumen © The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribu- tion, and reproduction in any medium, provided the original work is properly cited. Transl. Anim. Sci. 2020.4:1-11 doi: 10.1093/tas/txaa078 Broccoli (Brassica oleracea L. var. italica) byprod- INTRODUCTION uct comprised of stem and leaves could be an The increasing frequency and intensity of alternative. droughts in the world have led to shortages The broccoli flower bud is harvested for of common livestock feedstuffs (Alipour and human consumption and more than two-thirds Rouzbehan, 2010). Alternative feed sources, of the plant (outer leaves and stems) is left in the including agricultural waste byproducts, need to field. The use of this discarded crop waste is a pos- be investigated as ingredients for livestock diets. sible way to supply animal feedstuffs and decrease the environmental pollution. Globally, 20 million The authors wish to gratefully acknowledge Dr. Ali tonnes of cauliflower and broccoli crops are pro- Mokhtassi-Bidgoli, from Tarbiat Modares University, for his duced yearly and 15 million tonnes of byproduct assistance with statistical analysis of data and Mr. Gary Eas- are left in the field (Jian et al., 2017). In Iran, the ton for his English language correction of the manuscript. estimated production of cauliflower and broccoli Corresponding author: rozbeh_y@modares.ac.ir for the year 2016 was 60,000 tonnes with 45,000 Received March 28, 2020. tonnes of byproduct available for ruminant feed Accepted June 5, 2020. 1 Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. (FAO, 2018). Vegetable growers and ruminant (Halkier and Gershenzon, 2006). There is a concern farmers exist side by side in many parts of the over the adverse effects of the profile of glucosi- world, and this could facilitate the use of these nolates when feeding Brassica oleracea, as glucosi- waste products as ruminant feed. However, the nolates and subsequent metabolites may negatively fast rate of spoilage due to high moisture (>70%) affect growth performance [European Food Safety make the use of these waste products not practical, Authority (EFSA, 2008)]. Hence, it was hypothe- so ensiling is potentially a good preservation tech- sized that BBWS may be fed to Fashandy lambs nique for broccoli byproduct (BB) to reach the ideal up to medium concentration (i.e., 200 g/kg of diet moisture content for ensiling is between 60% and DM) without any adverse impacts on health and 70% (McDonald et al., 1991). The preservation of growth performance. the forage crops as silage is based on a fermenta- Therefore, this study was carried out to evaluate tion process that lowers the pH and preserves the the effects of a mixed BB-wheat straw silage nutritive value of the fresh crop. The main principle (BBWS) substituted only up to 20% of the forage is the production of lactic acid by the lactic acid ration (DM basis; i.e., lucerne and wheat straw) bacteria from the metabolism of the water-soluble in finishing diets for lambs by measuring in vivo carbohydrates in the fresh crop. Since fresh BB apparent digestibility of nutrients, rumen metab- contains about 186  g/kg dry matter (DM) soluble olites, N retention, microbial N synthesis (MNS), sugars (Wadhwa et al., 2006), mixing BB with dry blood biochemistry parameters, and performance feeds, such as straw, before ensiling probably re- in Fashandy lambs. sults in good preservation of this forage (Jian et al., 2017). Ensilage of BB also leads to a reduction in MATERIALS AND METHODS the antinutritive compound (i.e., glucosinolate), The Guide for the Care and Use of Agricultural improving the nutritional value of the feed (Vipond Animals in Research and Teaching (FASS, 2010) et  al., 1998). Fresh BB is a good source of crude was followed for housing, feeding, transport, protein (CP; 270 g/kg DM) with relatively low neu- proper and humane care and use of animals, vet- tral detergent fiber (NDF; 280  g/kg DM) and a erinary care, occupational health and safety, pro- metabolizable energy (ME) content of 9.87 MJ/kg gram management and procedures. The Committee DM (Hu et al. 2011) and an in vivo organic matter of Animal Science of Tarbiat Modares University (OM) digestibility of 86.9% (Wadhwa et al. 2006). (Iran) approved the experimental protocols. In an in vitro work, the chemical composition, fermentation characteristics, and aerobic stability of cabbage silages either mixed with 40 g ground corn Silage Preparation or treated with bacterial inoculant in an attempt to increase DM have been evaluated by Rezende Mature BB was harvested according to com- et al. (2015), and it was observed that the inoculant mercial production practices in Iran (i.e., when the was unnecessary, and the inclusion of ground corn buds of the head are firm and tight before flowering) would increase the cost of the silage and, there- and chopped into pieces of approximately 5 × 5 cm fore, was impracticable. Previous work on feeding using an electric cutter (Hallde, RG-200, Sweden). fresh cabbage leaves or cauliflower leaves to goats Wheat straw was harvested at full maturity and showed no difference in performance [in terms of chopped into 3–5  cm lengths. Broccoli byproduct apparent digestibility of DM, OM, CP, acid deter- has a high moisture content (836 g/kg fresh weight), gent fiber (ADF), efficiency of nutrient utilization, so it was mixed with wheat straw at a ratio of 90:10 and dry matter intake (DMI)] compared to other on fresh basis (a ratio of 69:31 on a DM basis) to green forages (Wadhwa et al., 2006), but it has been give a fresh mixture containing above 260  g DM/ illustrated that dietary inclusion of fresh cabbage kg of fresh weight, as this DM content promotes (up to 200  g/kg diet in concentrate basal feed) to relatively well-preserved silage (McDonald et  al., lambs reduced their growth rate (232 vs. 271  g/d) 1991). The chopped mixture of BB and wheat straw probably due to the presence of S-methylcysteine was ensiled in three trench silos of approximately 1 sulfoxide and glucosinolates, which depress feed m height and 5 m width, which were each covered consumption (Nkosi et al., 2016). Moreover, lambs with a thick plastic sheet. The mixture was ensiled fed on Brassica fodder crops, such as broccoli, may and compacted using a tractor, with a compac- develop goiter (broccoli contain goitrogenic com- tion density of approximately 900  kg wet matter/ pounds, which interfere with the availability of m . After ensiling for 60 d, representative sam- iodine) and this can reduce the lamb performance ples (500 g) were taken (weekly from five different Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb locations within each silo, which were opened at lactic acid and volatile fatty acids (VFA) using a gas different time points and stored frozen at −20  °C chromatograph (UNICAM 4600; SB Analytical, until analysis) to determine chemical composition Cambridge, UK) equipped with a flame ioniza- and fermentation characteristics. tion detector (250 °C), split-injection port (1.0 μL injection), and a capillary column (Agilent J&W HP-FFAP, 10 m by 0.535 mm by 1.00 μm, 19095F- Chemical Analysis and Fermentation 121; Agilent, Santa Clara, CA). The carrier gas Characteristics of Silage was He (column head pressure of 68.9 kPa) and the working temperature of the injector and de- Samples of fresh materials and silage were tector were 250 and 300 °C, respectively. The initial oven-dried at 60  °C to a constant weight and column temperature was set at 80 °C for 1 min and ground to pass through a 1-mm sieve (Wiley mill, then increased with 20 °C/min to 120 °C followed Swedesboro, NJ), and the determined DM was cor- by 6.2 °C/min to 140 °C and thereafter with 20 °C/ rected for the loss of volatiles during drying. The min to 205 °C. samples were analyzed for ash (method 924.05), ether extract (EE; method 920.39), and CP (method Treatments, Animals, and Diets 988.05) according to AOAC (1998). Ash-free NDF (NDFom) was determined without sodium sulfite Forty-five male Fashandy lambs with an (Mertens, 2002). The determination of ADF was average BW of 29.9 ± 0.9 kg and 6 months of age performed (method 973.18; AOAC, 1998) and ex- were used in this study. Animals were housed in pressed exclusive of residual ash (ADFom). Acid individual wooden-slatted pens (1.5  × 1.5 m) and detergent lignin (ADL) was determined by solubil- given an adaptation period of 14 d followed by a ization of cellulose with 72% sulfuric acid (method data collection period of 56 d. Lambs were offered 973.18), and phenolic compounds using the Folin– Ciocalteu method. The water-soluble carbohy- drates (WSC) were measured by colorimetry after Table 1.  Ingredients and chemical composition reaction with anthrone reagent (MAFF, 1982). The (%  of DM) of the diets containing different con- ME contents of pre-ensiled and BBWS were calcu- centrations of BBWS lated according to Menke et al. (1979) as: Concentration of BBWS in diet (% of DM) ME (MJ/kg DM)= 2.20 + 0.136 × GP Item 0 10 20 + 0.057 × CP + 0.0029 × EE Ingredient Alfalfa hay 15 12 7.5 where GP is 24-h net gas production (mL/200 mg Wheat straw 12.5 5.5 — of DM), and CP is measured in g/kg of DM. Wheat bran 19.6 19.6 19.6 For measuring silage pH, 50  g of fresh silage BBWS — 10.0 20.0 was blended with 125  mL of distilled water and Corn grain 44.3 44.3 44.3 allowed to stand at room temperature for 1  h Soybean meal 5.9 5.9 5.9 (Faithfull, 2002). After decanting the silage ex- Urea 0.5 0.5 0.5 tract into a small beaker, the pH was measured Sodium bicarbonate 0.7 0.7 0.7 Calcium carbonate 0.7 0.7 0.7 using a portable digital pH meter (Sartorius Vitamin–mineral premix* 0.5 0.5 0.5 PT-10; Sartorius AG, Göttingen, Germany). To Salt 0.3 0.3 0.3 determine ammonia-N (NH -N), an extract was Chemical composition† obtained by squeezing the silage material, filtered DM using Whatman 54 filter paper, then a 9 mL of ali- OM 94.7 95.1 93.3 quot was taken, mixed with 1 mL of 7.2 N H SO , 2 4 CP 14.6 14.8 14.5 and stored at −20 °C. After thawing, the silage ex- NDFom 28.3 27.1 25.8 tracts were analyzed for NH -N using a phenol-hy- 3 EE 7.0 7.9 7.9 pochlorite assay (Galyean, 1997). Two milliliters of ME, Mcal/kg of DM 2.69 2.69 2.67 the silage juice was pipetted into a microcentrifuge *Contained (per kilogram) 190 g of Ca, 50 g of Na, 19 g of Mg, 3 g tube with 0.5  mL of an acid solution (containing of Zn, 3 g of Fe, 2 g of Mn, 300 mg of Cu, 100 mg of Co, 1 mg of Se, 20% orthophosphoric acid and 20  mM 2-ethyl 100 mg of I, 100 mg of vitamin E, 500,000 IU of vitamin A, 100,000 butyric acid, as the internal standard) and centri- IU of vitamin D3, 300 mg of antioxidant. fuged at 15,000 × g for 15 min at 4 °C (Rezaei et al., †Calculated from analysis of each feed composition, which were 2014). The supernatant was used to quantify the analyzed. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. isonitrogenous and isocaloric diets, which were and at 15-d intervals before the morning feeding, daily (Table  1) formulated according to National after 16  h fasting, to calculate average daily gain Research Council (NRC, 1985) guidelines, in which (ADG) and feed efficiency (FE). The ADG was forage (lucerne + wheat straw) was replaced with calculated for each lamb by using the following different concentrations (0, 100, or 200 g/kg of diet equation: DM) of BBWS (Table 2). Diets were randomly as- ADG =(f inal BW − initial BW) / signed by BW into three groups of 15 lambs each in number days on feeding. a completely randomized design. Animals received the diets as a total mixed ration, at 0800 and 1700 h, to ensure 10% feed refusal and had constant access In Vivo Apparent Digestibility of Nutrients, to freshwater. Microbial N Supply, and N Retention Feed Consumption and Growth Performance On day 56 of the data collection period, ran- domly four animals per treatment were placed into The feed eaten by each animal and corres- individual metabolism crates (0.6 × 1.3 m), allowing ponding feed refusals were recorded daily to esti- feces and urine to be collected. Apparent digest- mate the consumption of DM and nutrients, and ibility was determined by the total fecal collection representative samples were kept frozen at –20  °C method, with 6 d for sample collection. During the for subsequent analyses. Samples of feed offered collection period, the amounts of feed offered, feed and refusals were oven-dried at 60 °C to a constant refusals, and feces from each animal were recorded weight, ground to pass through a 1-mm sieve, then daily and samples were stored (at –20 °C) for later analyzed for OM, N, and EE as described above. analysis of DM, OM, CP, NDFom, and EE as The silos of BB-wheat straw, which were opened at described above. different time points (i.e., on days 1, 21, 41, and 61 Total urine produced daily was collected in of the trial), were fed to animals. The body weight plastic vessels containing 100  mL of 10% sulfuric (BW) of animals was individually recorded at the acid. From the daily urine collected, 10% was sam- beginning and the end of the experiment period (on pled, diluted fivefold with distilled water, and then two consecutive days to provide baseline weights) stored at –20 °C to estimate purine derivatives and nitrogen (Chen and Gomes, 1992). Urine allantoin Table 2.  Chemical composition (%  of DM), fer- was measured by a colorimetric method at 522 nm mentation characteristics, and ME values (Mcal/kg after its conversion to phenylhydrazine. The sum of DM) of pre-ensiled and ensiled BBWS of xanthine and hypoxanthine was calculated by Item Pre-ensiled Ensiled SEM P-value their conversion to uric acid with xanthine oxidase Chemical composition (X-1875; Sigma-Aldrich, St. Louis, MO) and meas- DM, % of fresh weight 26 30 1.4 0.003 uring the subsequent optical density at 293  nm. CP 9.0 8.0 0.46 0.05 Uric acid was measured from the reduction in op- NDFom 39 40 0.97 0.16 tical density at 293  nm after the degradation of ADFom 32.6 36.4 1.3 0.03 uric acid to allantoin with uricase (U-9375; Sigma- Acid detergent lignin 6.5 11.2 1.7 0.001 Aldrich). Total purine derivative excretion per day Ash 18 27 0.36 0.001 was calculated, the daily absorbed exogenous pur- EE 5.9 7.4 0.16 0.02 ines were estimated, and the MNS was predicted. Water-soluble carbohydrate 7.5 0.5 0.08 0.0001 The concentration of urinary N was estimated by ME, Mcal/kg of DM 1.91 1.72 0.02 0.015 TP* 0.6 0.55 0.08 0.16 the Kjeldahl method (AOAC 1998), and N reten- Fermentation characteristics tion was calculated as daily N excretion (urinary N pH value — 4.4 — plus fecal N) subtracted from daily N intake. Ammonia-N, % of total N — 6.1 — Fermentative acids, % of DM Lactic acid — 4.5 — Rumen Metabolites Acetic acid — 1.7 — Propionic acid — 0.2 — Just before the morning feeding and 2, 4, and Butyric acid — 0.5 — 6 h afterward, rumen fluid from four animals ran- Valeric acid — 0.04 — domly allocated to each treatment was sampled Isovaleric acid — 0.05 — (using an esophageal tube) on day 56. After dis- carding the first 15  mL of rumen fluid, pH was *Total phenolic compounds. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb determined. For NH -N determination, a 25-mL (treatment) of the ith concentration of BBWS sample of strained rumen fluid was preserved with (i  =  three concentrations of BBWS), e represents ij 5  mL of 0.2  N HCl at –20  °C and then analyzed the experimental error, and e represents the sam- ijk using a phenol-hypochlorite assay (Broderick and pling error. The random effect included treatment Kang, 1980). To measure ruminal VFA, 8  mL of × replication. The model included the fixed effect strained rumen fluid was preserved (at –20 °C) with of dietary treatment. Moreover, data on digest- 2  mL of an acid solution (containing 20% ortho- ibility, MNS, and N balance were measured once phosphoric acid and 20 mM 2-ethyl-butyric acid as and examined using the same model with four rep- internal standard). After thawing, rumen fluid was licates for each diet. centrifuged (15,000 × g for 15 min at 4 °C) and the Repeated measures were used to statistically supernatant was used to quantify VFA as described examine the data of rumen metabolites and blood previously. biochemistry parameters. The model used was Rumen protozoa were enumerated using y = μ + T + σ + S + T S + ε ijk i ij k i k ijk 5 mL of rumen fluid pipetted into a test tube con- where y represents the value of each individual taining 20  mL of formalinized physiological sa- ijk observation, µ is the overall mean, T is the fixed ef- line (20 mL of formaldehyde in 100 mL of saline fect (treatment) of the ith concentration of BBWS containing 0.85 g of NaCl in 100 mL of distilled (i  =  three concentrations of BBWS), σ is the main water). Total and subfamily counts of protozoa ij random error, S is the sampling time, T S is detecting were determined using a light microscope k i k interaction between treatment and sampling time, and and a Hemocytometer (Neubauer-improved; Ɛ is the subrandom error. The influence of diet (treat- MarienfeldSuperior, Lauda-Königshofen, ijk ment), sampling time, and their interactions were con- Germany; Dehority 2003). sidered fixed. Compound symmetry was used based on the lowest Akaike information criterion (AIC) as Blood Biochemistry Parameters a covariance structure in the model. Also, repeated measure was used to test the data on feed consump- Just before the morning feeding and 4  h after tion, whereas the influence of dietary treatment, and feeding, blood samples (15  mL) were randomly their interactions were considered fixed. collected from four animals assigned to each treat- Additionally, a polynomial contrast was used ment on days 56 and 70 (the same animals were to test the linear or quadratic effects of the diet (i.e., used on both times). The samples were transferred BB inclusion concentration) on parameters meas- to the laboratory on ice, and the harvested serum ured. The result is then illustrated as statistically was stored at –20 °C until analysis. Glucose, trigly- significant at P ≤ 0.05. ceride, cholesterol, urea N, total protein, albumin, The SAS program was used to calculate the and creatinine concentrations were measured using number of replications needed to obtain the desired kits from Pars Azmun (Tehran, Iran) using a spec- power (>0.8). The power of the test is given by: trophotometer (Genova; Jenway, Essex, UK). Blood thyroxine and triiodothyronine concentra- Power = P F > F = F α,(a−1),(N−a) β tions were measured using kits from Pars Azmun (Tehran, Iran) using an ELISA reader. using a noncentral F distribution for H with a non- centrality parameter λ  =  SS /MS , and de- treatment error grees of freedom (a − 1)  and (N − a). Here, N is Statistical Analysis the number of replications per treatment, a is the Using a t-test, the chemical composition of number of treatments, and F is the critical α,(a − 1),(N − a) pre-ensiled and BBWS were compared to quan- value (Kaps and Lamberson 2009). tify changes made during the process of silage fer- mentation (Table  2). Data on ADG, digestibility, RESULTS MNS, and N retention were statistically examined using the PROC MIXED of SAS (Version 9.1; SAS Chemical Analysis and Fermentation Inst. Inc.) in a completely randomized design. The Characteristics of Silage model used was Compared to the fresh product (Table  2), y = μ + T + e + e , ijk i ij ijk BBWS contained higher concentrations of ADFom where y represents the value of each individual ob- (P  <  0.03), ADL and ash (P  <  0.001), and lower ij servation, µ is the overall mean, T is the fixed effect WSC concentration (P < 0.0001). Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. In Vivo Apparent Digestibility of Nutrients, Feed propionic (P < 0.63), or butyric acid (P < 0.41) con- Intake, and Growth Performance centrations. Ruminal enumeration of total protozoa, Entodiniinae, Diplodiniinae, Isotrichidae, Epidiniinae, The apparent digestibility of DM, OM, CP, and Ophrioscolecinae subfamilies were not affected by NDFom, and EE was not influenced (P   <  0.34, feeding lambs with different concentration of BBWS P < 0.39, P < 0.89, P < 0.69, and P < 0.35, respect- (P < 0.55, P < 0.78, P < 0.57, P < 0.17, P < 0.65, and ively) by feeding BBWS (Table 3). The DMI, organic P < 0.29, respectively). matter intake, crude protein intake, ether extract in- take, neutral detergent fiber intake, metabolisable en- Blood Biochemistry Parameters ergy intake, ADG, and feed conversion ration (FCR) were not affected (P  <  0.67, P  <  0.67, P  <  0.66, Table  6 illustrated that the blood concentra- P < 0.30, P < 0.63, P < 0.32, P < 0.55, and P < 0.55, tions of glucose, triglyceride, cholesterol, urea N, respectively) by feeding the experimental diets. creatinine, triiodothyronine, thyroxin, total protein, albumin, and globulin, did not change (P  <  0.59, P  <  0.85, P  <  0.75, P  <  0.52, P  <  0.63, P  <  0.59, Microbial N supply and N retention P  <  0.76, P  <  0.72, P  <  0.68, and P  <  0.75, respectively). Feeding lambs with BBWS had no effect (P  <  0.53) on total purine derivatives absorbed, DISCUSSION total purine derivative excretion (P  <  0.56), and MNS (P < 0.57; Table 4). Among the lambs, there Chemical Analysis and Fermentation were no significant changes in N consumption Characteristics of Silage (P < 0.72), fecal N (P < 0.88), urinary N (P < 0.61), and N retained (P < 0.81). Changes in the chemical composition of BBWS after ensilage occur because of microbial fermentation and losses from effluent throughout Rumen Metabolites and Protozoa Enumeration the ensiling process (Buxton and O’Kiely, 2003), Dietary inclusion of BBWS had no effect on but the DM of BBWS is within the range of me- ruminal pH (P  <  0.82) and NH -N concentration dium- to good-quality silage. Low-moisture silage (P < 0.61; Table 5). Feeding BBWS had no influence on depresses undesirable clostridial growth (Pahlow the content of total VFA (P < 0.83), acetic (P < 0.49), et al., 2003) because of the affinity of Clostridium Table 3. Effect of dietary inclusion of BBWS on in vivo apparent digestibility (n = 4), intake, and growth performance of fattening lambs Concentration of BBWS in diet (g/kg of DM) Item 0 100 200 SEM P-value Digestibility, % DM 76 75 73 1.50 0.34 OM 77.1 76 78.1 0.75 0.39 CP 72.1 72.5 72.4 2.4 0.89 NDFom 62 61 63 2.85 0.69 EE 77 77 79 1.34 0.35 Intake, g/d DM 1,432 1,435 1,420 14.4 0.67 OM 1,332 1,316 1,315 16.25 0.67 CP 204 200 202 2.59 0.66 EE 112 113 115 1.77 0.3 NDFom 420 414 416 4.67 0.63 ME, Mcal/d 2.94 2.91 2.96 0.03 0.32 Performance Initial BW 29.41 29.28 29.06 0.87 0.28 Finial BW 42.8 42.47 41.94 1.20 0.32 ADG, g/d 239 235 230 9.34 0.55 FCR* 5.99 6.12 6.13 0.4 0.55 *Feed conversion ratio (DMI:ADG). Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb Table 4. Effect of dietary inclusion of BBWS on total purine derivatives (TPD), MNS, and N retention in fattening lambs (n = 4) Concentration of BBWS in diet (g/kg of DM) Item 0 100 200 SEM P-value Urinary excretion, mmol/d Allantoin 8.58 8.04 8.10 0.56 0.41 Xanthine + hypoxanthine 0.73 0.73 0.74 0.51 0.51 Uric acid 3.61 4.10 4.37 0.55 0.52 TPD absorbed, mmol/d 14.69 14.67 15.11 1.07 0.53 TPD excretion, mmol/d 12.92 12.87 13.22 0.91 0.56 MNS, g/d 10.68 10.66 10.99 0.78 0.57 N retention, g/d N intake 29.7 29.02 29.12 1.10 0.72 Fecal N 8.17 8 8.01 0.69 0.88 Urinary N 11.27 11.07 11.10 0.50 0.61 Retained N 10.28 9.95 10.01 0.65 0.81 Retained N, g/kg of N intake 346 343 344 2.81 0.96 Table 5. Effect in vivo of dietary inclusion of BBWS on rumen metabolites and protozoa numeration in fattening lambs (n = 4) Concentration of BBWS in diet (g/kg of DM) Item 0 100 200 SEM P-value pH 6.38 6.37 6.43 0.14 0.82 Ammonia-N, mg/dL 10.12 10.33 10.50 0.86 0.61 Total VFA, mmol/L 82.3 79.63 81.95 1.7 0.83 VFA, mmol/100 mmol Acetic acid 56.29 54.53 54 1.66 0.49 Propionic acid 27.11 28.5 28.58 1.60 0.63 Butyric acid 15.9 16.3 16.8 0.44 0.41 Valeric acid 0.56 0.55 0.50 0.08 0.76 Isovaleric acid 0.14 0.12 0.12 0.02 0.63 Acetate:propionate 2.08 2.08 2.06 0.22 0.94 Protozoa, log /g of digesta Total protozoa 6.09 6.06 6.06 0.032 0.55 Entodiinina 5.86 5.83 5.86 0.039 0.78 Diplodiinina 5.45 5.40 5.35 0.086 0.57 Isotrichidea 4.87 4.86 4.98 0.042 0.17 Epidiniina 4.79 4.78 4.79 0.013 0.65 Ophrioscolecinae 4.89 5.01 5.04 0.06 0.29 No significant interaction between treatment and sampling time were observed. spp. for moisture. The production of butyric acid concentration (Megías et  al., 2014). The preserva- was low in the present study, indicating low growth tion of BBWS was suitable in terms of pH value of Clostridium spp., which derive energy for growth (i.e., 4.8), low NH -N content (<100 g/kg total N), by fermenting sugars and lactate into butyric acid and low concentrations of acetic, propionic, and bu- (Pahlow et al., 2003). In agreement with our results, tyric acids (McDonald et al., 2011). A good-quality Shao et  al. (2007) documented improved fermen- silage is characterized by a lactic acid concentra- tation of perennial ryegrass with decreased con- tion between 30 and 140 g/kg of DM (McDonald centrations of butyric. The inverse relationship et  al., 2011), and BBWS lactic acid concentration between ADFom, ADL, and WSC show that a re- was 45 g/kg of DM. The pH values of our BBWS duction in WSC increases the content of ADFom were the same as those (pH = 4.76) noted by Jian and ADL resulting in a proportional rise in ash et al. (2017) who ensiled rice straw, broccoli residue, Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. Table 6. Effect of dietary inclusion of BBWS on blood biochemistry parameters in fattening lambs Concentration of BBWS in diet (g/kg of DM) Item* 0 100 200 SEM P-value Metabolites, mg/dL Glucose 67.22 69.41 70.76 1.52 0.59 Triglyceride 21.98 21.23 21.90 2.10 0.85 Cholesterol 59.60 61.70 63.99 5.38 0.75 Blood urea N 19.45 22.44 19.58 1.27 0.52 Creatinine 1.40 1.42 1.45 0.07 0.63 Thyroid hormones, ng/mL Triiodothyronine 5.13 5.25 5.33 0.28 0.59 Thyroxin 69.00 70.76 71.80 3.30 0.76 Protein, g/dL Total protein 6.77 6.67 6.76 0.18 0.72 Albumin 3.59 3.51 3.55 0.11 0.68 Globulin 3.18 3.16 3.20 0.15 0.75 Albumin:globulin 1.14 1.10 1.11 0.06 0.68 *Average of repeated sampling of blood collected from six animals assigned to each treatment on days 56 and 70 of data collection period, just before the morning feeding and 4 h after the morning feeding. and lucerne at a ratio of 5:4:1 for 30 d, whereas, for The similar ADG among the lambs reflected other silage characteristics, such as NH -N, lactic their comparable ruminal VFA concentration and acid, and VFA content, Jian et al. (2017) reported a MNS (Ben Salem et  al., 2002). The FCR showed lower silage quality than those observed in BBWS. no difference among the lambs due to their similar DMI and ADG. The experimental diets, in the cur- rent study, were formulated to meet the require- In Vivo Apparent Digestibility of Nutrients, Feed ments of a growth rate of 300 g/d for fattening male Consumption, and Growth Performance lambs at 8.5 mo of age according toNRC (1985) The inclusion of BBWS in the diets had no recommendations. However, the ADG of our effect on nutrient digestibility and this relates to lambs was not as predicted in NRC (1985) tables the similar chemical composition of the diets and because the equations used to predict lamb growth the comparable ruminal pH in the lambs, factors were not obtained in Iranian sheep. Iranian sheep which influence rumen microbial activity (Petit and breeds (except the Zel breed) are fat tailed and their Castonguay, 1994). The low concentration of phen- physiological characteristics and genetic potential olic compounds (1.0 g/kg of DM) had no adverse for growth and, to some extent, the rumen micro- effect on diet apparent digestibility. Previous work biota of these breeds differ from sheep in other on fresh cauliflower (Brassica oleracea var. botrytis) countries, which can lead to a difference in overall reported good nutrient digestibility of DM, OM, performance. CP, and NDF (80.9%, 86.9%, 84.9%, and 71.8%, re- spectively) when fed to goats (Wadhwa et al., 2006). Microbial N Supply and N Retention It has been stated that increased concentrations of fresh cabbage (Brassica oleracea var. capitata) in Diets that maximize MNS often increase the diet of lambs reduced OM and NDF digest- animal performance (Chen and Gomes, 1992). ibility (73% vs. 65%; 56% vs. 47%, respectively; In the present study, the MNS reached its high- Nkosi et al., 2016). est value (10.99  g/d) with 200  g of BBWS per Feeding BBWS had no influence on feed con- kilogram of diet DM. Similar concentrations of sumption and this may be due to the similar diet digest- urinary purine derivatives (PD) and MNS among ibility, particularly NDFom digestibility. It was noted treatments were due to the similar intakes of di- that NDF digestibility provides an accurate estimation gestible OM and N (Ben Salem et al., 2002) and of intake (Harper and McNeill, 2015). A correlation the equal ruminal synchrony of ME and N re- between NDF digestibility and intake was stated by sulting from a similar daily ME intake and di- Nkosi et al. (2016) who noted a lower DMI for lambs gestible CP consumption among the treatments. fed fresh Brassica (oleracea var. capitata) compared to Urinary PD excretions are indicators of MNS a Brassica-free diet (1,420 vs. 1,600 g). arriving at the beginning of the small intestine Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb (Chen and Gomes, 1992). It was reported by was not affected by the diets. The similar daily OM Wadhwa et  al. (2006) that higher MNS values intake and OM digestibility, which resulted in equal in goats offered fresh cauliflower (Brassica oler - fermented OM per day, reflects the similar ruminal acea var. botrytis; 25.40  g/d) compared to those concentrations of total and proportions of indi- in the current study. This difference could be due vidual VFA among the dietary treatments (Morvay to the different experimental animals and diets et al., 2011). (i.e., lambs vs. goats; BBWS vs. fresh Brassica Rumen protozoa, exclusively ciliates, rank oleracea). second only to bacteria in cellular biomass of Nitrogen retention among the animals did not the ruminal microbiota (Dehority, 2003). They differ because digestible OM and N (Ben Salem are found in the rumen where they play im- et  al., 2002), N excreted in feces and urine (Zuo, portant roles in feed digestion and homeostasis 2011), and ADG were not influenced by feeding of the rumen ecosystem (Dehority, 2003). Feeding BBWS. A  lower N consumption and N retention BBWS had no effect on rumen protozoa enumer- were noted in animals offered diets with cabbage ation and this was due to the normal ruminal pH, compared to those fed a Brassica-free diet, sug- which plays an important role in rumen protozoa gesting that the lower N intake and retention could activity (Dehority, 2003). be related to the decrease in the digestibility of DM and OM in this diet (Nkosi et al., 2016). Blood Biochemistry Parameters Concentrations of blood parameters represent Rumen Metabolites and Protozoa Enumeration an integrated index of the adequacy of nutrient For all lambs, ruminal pH values varied from supply in relation to the utilization of nutrients, 6.37 to 6.43, which were within the optimum range which is a self-governing aspect of the physiological (i.e., 6.1–6.6) for maintaining normal cellulolytic condition of the animal and provides an immediate organism populations (Van Soest, 1994). Feeding indication of nutritional status at a specific point BBWS had no effect on rumen pH and this was re- in time (Pambu-Gollah et al., 2000). In the present lated to the similar DMI and nutrient digestibility study, the lack of differences in blood biochemistry among lambs in different treatments. Dietary add- parameters among the treatments was probably due ition of BBWS had no adverse effects on ruminal to similar intakes and nutrient digestibilities. The metabolism. It was noted by Keogh et  al. (2009) serum concentrations of glucose, cholesterol, urea that replacing grass silage with fresh kale (Brassica N, total protein, albumin, and globulin and the al- oleracea) had no effect on the ruminal pH of dry bumin to globulin ratio in all treatments were within dairy cows (mean pH  =  6.2). In contrast, feeding the normal ranges for sheep (Radostits et al., 2007). fresh brassica forage (Brassica spp.) to lambs re- However, the concentration of triglyceride was sulted in a lower ruminal pH (<6.0) as a result of greater than the range for sheep (Radostits et  al., the greater ratio of readily fermentable to struc- 2007) but was in the usual range reported for fat- tural carbohydrates in Brassica forage (Sun et al., tailed Iranian sheep (18.03 to 50.93 mg/dL; Mojabi, 2015). Ruminal NH -N content in all the animals 2011). The similar concentrations of blood albumin was higher than 5  mg/dL, which is the minimum among the lambs suggested normal liver function concentration required by rumen microorganisms as the liver is the main organ of albumin synthesis. to support their optimum growth (McDonald The values of albumin and globulin in the serum of et  al., 2011). The lack of difference in rumen the lambs in the current study show that these ani- NH -N concentration among the lambs suggests mals did not suffer from any health problems that that the release of N in the rumen and its uptake might have affected their performance (Radostits by microbes to synthesize microbial CP were not et al., 2007). affected by the diets. In Brassicas, the antinutritional S-methylcysteine Ruminal VFA is the main energy source for sulfoxide and glucosinolates were reported to be goi- ruminants (Abarghuei et  al., 2013); consequently, trogenic and could affect triiodothyronine and thy- diets that increase the ruminal production of VFA roxin production (Chorfi et  al., 2015). The lambs lead to higher animal performance (Penner, 2014). offered BBWS had no clinical indicators of gluco- In the current study, dietary BBWS had no influence sinolate toxicity because the concentration was low on the total and proportions of individual VFA and there were no symptoms of toxicity (EFSA, content; thus, the performance of fattening lambs 2008). This may be due to the ensiling process Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. European Food Safety Authority. 2008. Glucosinolates as un- causing a reduction in the concentration of gluco- desirable substances in animal feed—Scientific Panel on sinolates and S-methylcysteine sulfoxide improving Contaminants in the Food Chain. EFSA J. 590:1–76. the feed quality (Vipond et al., 1998). Similarly, feed- Faithfull,  N.  T. 2002. Methods in agricultural chemical ana- ing different concentrations of ensiled kale (Brassica lysis: a practical handbook. CABI Int., Wallingford, UK. oleracea) in lambs did not affect their performance FAO. 2018. FAO statistics division. Crop production. Food and Agriculture Organization of the United Nations, or blood triiodothyronine and thyroxin concentra- Rome, Italy. tions probably because of the reduction in glucosi- FASS. 2010. Guide for the care and use of agricultural animals nolate during ensilage (Vipond et al., 1998). in research and teaching. 3rd ed. Federation of Animal Science Societies, Champaign, IL. CONCLUSIONS Galyean, M. L. 1997. Laboratory procedures in animal nutri- tion research. Department of Animal and Food Sciences, The chemical composition and fermentation Texas Tech University, Lubbock, TX. characteristics of BBWS indicates its potential as a Halkier, B. A., and J. Gershenzon. 2006. Biology and biochem- ruminant feedstuff. BBWS inclusion at up to 200 g/ istry of glucosinolates. Annu. Rev. Plant Biol. 57:303–333. kg of DM in the diet of fattening Fashandy lambs doi:10.1146/annurev.arplant.57.032905.105228. Harper, K. J., and D. M. McNeill. 2015. The role iNDF in the is equivalent to an Alfalfa-wheat straw diet in terms regulation of feed intake and the importance of its as- of effects on nutrient digestibility, feed intake, sessment in subtropical ruminant systems. Agriculture. growth performance, FE, rumen parameters and 5:778–790. blood parameters. BBWS at this level is a safe feed- Hu,  C.  H., A.  Y.  Zuo, D.  G.  Wang, H.  Y.  Pan, W.  B.  Zheng, stuff for sheep having no adverse effects on their Z.  C.  Qian, and X.  T.  Zou. 2011. Effects of broccoli stems and leaves meal on production performance and health status or growth performance. egg quality of laying hens. Anim. Feed Sci. Technol. Conflict of interest statement. None declared. 170:117–121. Jian,  W., C.  Lei, X-j.  Yuan, G.  Gang, J-f.  Li, Y-f.  Bai, and LITERATURE CITED S. Tao. 2017. Effects of molasses on the fermentation char- acteristics of mixed silage prepared with rice straw, local Abarghuei,  M.  J., Y.  Rouzbehan, A.  Z.  M.  Salem, and vegetable by-products and alfalfa in Southeast China. J. M. J. Zamiri. 2013. Nutrient digestion, ruminal fermenta- Integr. Agr. 16:664–670. tion and performance of dairy cows fed pomegranate peel Kaps, M., and W. Lamberson. 2009. Biostatistics for animal sci- extract. Livest. Sci. 157:452–461. ence: an introductory text. 2nd ed. CAB Int., Wallingford, Alipour, D., and Y. Rouzbehan. 2010. Effects of several levels UK. of extracted tannin from grape pomace on intestinal di- Keogh,  B., P.  French, J.  J.  Murphy, J.  F.  Mee, T.  McGrath, gestibility of soybean meal. Livest. Sci. 128:87–91. T.  Storey, J.  Grant, and F.  J.  Mulligan. 2009. A note AOAC. 1998. Official methods of analysis. 16th ed. 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McDonald, P., A. R. Henderson, and S. J. E. Heron. 1991. The doi:10.3168/jds.S0022-0302(80)82888-8. biochemistry of silage. 2nd ed. Chalcombe Publication, Buxton, D. R., and P. O’Kiely. 2003. Preharvest plant factors af- Marlow, UK. fecting ensiling. In: Buxton DR, Muck RE, Harrison JH, Megías,  M.  D., M.  Meneses, J.  Madrid, F.  Hernández, editors. Silage science and technology. ASA-CSSA-SSSA, A. Martínez-Teruel, and J. A. Cano. 2014. Nutritive, fer- Madison, WI. p. 199–250. mentative and environmental characteristics of silage of Chen,  X.  B., and J.  J.  Gomes 1992. Estimation of microbial two industrial broccoli (Brassica oleracea var. Italica) protein supply to sheep and cattle based on urinary excre- by-products for ruminant feed. Int. J.  Agric. Biol. tion of purine derivates—an overview of the technical de- 16:307–313. tails. International Feed Resources Unit, Rowett Research Menke, K., L. Raab, A. Salewski, H. Steingass, D. Fritz, and Institute, Aberdeen, UK. W. Schneider. 1979. The estimation of the digestibility and Chorfi,  Y., Y.  Couture, G.  F.  Tremblay, R.  Berthiaume, and metabolizable energy content of ruminant feedingstuffs D.  Cinq-Mars. 2015. Growth and blood parameters of from the gas production when they are incubated with weaned crossbred beef calves fed forage kale (Brassica rumen liquor in vitro. J. Agric. Sci. 93:217–222. oleracea spp. acephala). Adv. Agric. 2015:1–7. doi: Mertens, D. R. 2002. Gravimetric determination of amylase-treated 10.1155/2015/410497. neutral detergent fiber in feeds with refluxing in beakers or Dehority, B. A. 2003. Rumen microbiology. 1st ed. Nottingham crucibles: collaborative study. J. AOAC Int. 85:1217–1240. University Press, Nottingham, UK. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb Mojabi,  A. 2011. Veterinary clinical biochemistry. 2nd ed. London, UK. Noorbakhsh Publ., Tehran, Iran. Rezaei, J., Y. Rouzbehan, H. Fazaeli, and M. Zahedifar. 2014. Morvay, Y., A. Bannink, J. France, E. Kebreab, and J. Dijkstra. Effects of substituting amaranth silage for corn silage on 2011. Evaluation of models to predict the stoichiometry intake, growth performance, diet digestibility, microbial of volatile fatty acid profiles in rumen fluid of lactating protein, nitrogen retention and ruminal fermentation in Holstein cows. J. Dairy Sci. 94:3063–3080. doi:10.3168/ fattening lambs. Anim. Feed Sci. Technol. 192:29–38. jds.2010-3995. Rezende,  A.  V., C.  H.  S.  Rabelo, and M.  R.  M.  Silva. 2015. Nkosi,  B.  D., R.  Meeske, M.  M.  Ratsaka, T.  Langa, Wasted cabbage (Brassica oleracea) silages treated with M.  D.  Motiang, and I.  B.  Groenewald. 2016. Effects of different levels of ground corn and silage inoculant. Rev. dietary inclusion of discarded cabbage (Brassica oleracea Bras. 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American Society of Agronomy, fresh winter forage rape (Brassica napus L.) emit less me- Madison, WI. p. 31–93. thane than those fed perennial ryegrass (Lolium perenne Pambu-Gollah,  R., P.  Cronje, and N.  Casey. 2000. An evalu- L.), and possible mechanisms behind the difference. Plos ation of the use of blood metabolite concentrations as in- One 10:e0119697. doi:10.1371/journal.pone.0119697. dicators of nutritional status in free-ranging indigenous Van Soest, P. J. 1994. Nutritional ecology of the ruminant. 2nd goats. S. Afr. J. Anim. Sci. 30:115–120. ed. Cornell University Press, Ithaca, NY. Penner, G. B. 2014. Mechanisms of volatile fatty acid absorption Vipond,  J.  E., A.J.  Duncan, D.  Turner, L.  Goddyn, and and metabolism and maintenance of a stable rumen environ- G.  W.  Horgan. 1998. Effects of feeding ensiled kale ment. In: Proceedings of the 25th Annual Florida Ruminant (Brassica oleracea) on the performance of finishing lambs. 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Broccoli byproduct-wheat straw silage as a feed resource for fattening lambs

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Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 † †,2 ‡ †, Edris Partovi, Yousef Rouzbehan, Hasan Fazaeli, and Javad Rezaei † ‡ Department of Animal Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran; and Animal Science Research Institute of Iran, Agricultural Research, Education and Extension Organization, Karaj, Iran ABSTRACT:  The effect of feeding broccoli retention, rumen, and blood parameters were byproduct-wheat straw silage [BBWS; 69:31 measured. The BBWS diets had no influence ratio, dry matter (DM) basis] on performance, on DMI, ADG, feed conversion efficiency, in microbial N synthesis (MNS), rumen, and blood vivo apparent digestibility coefficients of DM, parameters in Fashandy lambs were evaluated. organic matter, CP, and ash-free neutral deter- Three diets, with equal metabolizable energy gent fiber. Neither MNS and N retention nor and crude protein (CP) with a forage to con- serum concentrations of glucose, triglycerides, centrate ratio of 27:73 (DM basis), were formu- creatinine, cholesterol, urea N, triiodothyronine, lated in which forage (lucerne and wheat straw) thyroxine, total protein, albumin, and globulin was replaced by BBWS (0, 100, or 200  g/kg of were affected. Rumen pH, NH -N, short-chain diet DM). These were assigned to three groups fatty acid concentrations, the ratio of acetic to (n = 15/group) in a completely randomized block propionic acid, and protozoa numbers were, design for a 70-d period in which diets were also, not influenced. In summary, BBWS may be offered as a total mixed ration. For each animal, fed to Fashandy lambs up to 200  g/kg of diet dry matter intake (DMI), average daily gain DM without any adverse impacts on growth (ADG), in vivo apparent digestibility, MNS, N performance. Key words: broccoli byproduct, lamb, lamb growth, protozoa, rumen © The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribu- tion, and reproduction in any medium, provided the original work is properly cited. Transl. Anim. Sci. 2020.4:1-11 doi: 10.1093/tas/txaa078 Broccoli (Brassica oleracea L. var. italica) byprod- INTRODUCTION uct comprised of stem and leaves could be an The increasing frequency and intensity of alternative. droughts in the world have led to shortages The broccoli flower bud is harvested for of common livestock feedstuffs (Alipour and human consumption and more than two-thirds Rouzbehan, 2010). Alternative feed sources, of the plant (outer leaves and stems) is left in the including agricultural waste byproducts, need to field. The use of this discarded crop waste is a pos- be investigated as ingredients for livestock diets. sible way to supply animal feedstuffs and decrease the environmental pollution. Globally, 20 million The authors wish to gratefully acknowledge Dr. Ali tonnes of cauliflower and broccoli crops are pro- Mokhtassi-Bidgoli, from Tarbiat Modares University, for his duced yearly and 15 million tonnes of byproduct assistance with statistical analysis of data and Mr. Gary Eas- are left in the field (Jian et al., 2017). In Iran, the ton for his English language correction of the manuscript. estimated production of cauliflower and broccoli Corresponding author: rozbeh_y@modares.ac.ir for the year 2016 was 60,000 tonnes with 45,000 Received March 28, 2020. tonnes of byproduct available for ruminant feed Accepted June 5, 2020. 1 Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. (FAO, 2018). Vegetable growers and ruminant (Halkier and Gershenzon, 2006). There is a concern farmers exist side by side in many parts of the over the adverse effects of the profile of glucosi- world, and this could facilitate the use of these nolates when feeding Brassica oleracea, as glucosi- waste products as ruminant feed. However, the nolates and subsequent metabolites may negatively fast rate of spoilage due to high moisture (>70%) affect growth performance [European Food Safety make the use of these waste products not practical, Authority (EFSA, 2008)]. Hence, it was hypothe- so ensiling is potentially a good preservation tech- sized that BBWS may be fed to Fashandy lambs nique for broccoli byproduct (BB) to reach the ideal up to medium concentration (i.e., 200 g/kg of diet moisture content for ensiling is between 60% and DM) without any adverse impacts on health and 70% (McDonald et al., 1991). The preservation of growth performance. the forage crops as silage is based on a fermenta- Therefore, this study was carried out to evaluate tion process that lowers the pH and preserves the the effects of a mixed BB-wheat straw silage nutritive value of the fresh crop. The main principle (BBWS) substituted only up to 20% of the forage is the production of lactic acid by the lactic acid ration (DM basis; i.e., lucerne and wheat straw) bacteria from the metabolism of the water-soluble in finishing diets for lambs by measuring in vivo carbohydrates in the fresh crop. Since fresh BB apparent digestibility of nutrients, rumen metab- contains about 186  g/kg dry matter (DM) soluble olites, N retention, microbial N synthesis (MNS), sugars (Wadhwa et al., 2006), mixing BB with dry blood biochemistry parameters, and performance feeds, such as straw, before ensiling probably re- in Fashandy lambs. sults in good preservation of this forage (Jian et al., 2017). Ensilage of BB also leads to a reduction in MATERIALS AND METHODS the antinutritive compound (i.e., glucosinolate), The Guide for the Care and Use of Agricultural improving the nutritional value of the feed (Vipond Animals in Research and Teaching (FASS, 2010) et  al., 1998). Fresh BB is a good source of crude was followed for housing, feeding, transport, protein (CP; 270 g/kg DM) with relatively low neu- proper and humane care and use of animals, vet- tral detergent fiber (NDF; 280  g/kg DM) and a erinary care, occupational health and safety, pro- metabolizable energy (ME) content of 9.87 MJ/kg gram management and procedures. The Committee DM (Hu et al. 2011) and an in vivo organic matter of Animal Science of Tarbiat Modares University (OM) digestibility of 86.9% (Wadhwa et al. 2006). (Iran) approved the experimental protocols. In an in vitro work, the chemical composition, fermentation characteristics, and aerobic stability of cabbage silages either mixed with 40 g ground corn Silage Preparation or treated with bacterial inoculant in an attempt to increase DM have been evaluated by Rezende Mature BB was harvested according to com- et al. (2015), and it was observed that the inoculant mercial production practices in Iran (i.e., when the was unnecessary, and the inclusion of ground corn buds of the head are firm and tight before flowering) would increase the cost of the silage and, there- and chopped into pieces of approximately 5 × 5 cm fore, was impracticable. Previous work on feeding using an electric cutter (Hallde, RG-200, Sweden). fresh cabbage leaves or cauliflower leaves to goats Wheat straw was harvested at full maturity and showed no difference in performance [in terms of chopped into 3–5  cm lengths. Broccoli byproduct apparent digestibility of DM, OM, CP, acid deter- has a high moisture content (836 g/kg fresh weight), gent fiber (ADF), efficiency of nutrient utilization, so it was mixed with wheat straw at a ratio of 90:10 and dry matter intake (DMI)] compared to other on fresh basis (a ratio of 69:31 on a DM basis) to green forages (Wadhwa et al., 2006), but it has been give a fresh mixture containing above 260  g DM/ illustrated that dietary inclusion of fresh cabbage kg of fresh weight, as this DM content promotes (up to 200  g/kg diet in concentrate basal feed) to relatively well-preserved silage (McDonald et  al., lambs reduced their growth rate (232 vs. 271  g/d) 1991). The chopped mixture of BB and wheat straw probably due to the presence of S-methylcysteine was ensiled in three trench silos of approximately 1 sulfoxide and glucosinolates, which depress feed m height and 5 m width, which were each covered consumption (Nkosi et al., 2016). Moreover, lambs with a thick plastic sheet. The mixture was ensiled fed on Brassica fodder crops, such as broccoli, may and compacted using a tractor, with a compac- develop goiter (broccoli contain goitrogenic com- tion density of approximately 900  kg wet matter/ pounds, which interfere with the availability of m . After ensiling for 60 d, representative sam- iodine) and this can reduce the lamb performance ples (500 g) were taken (weekly from five different Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb locations within each silo, which were opened at lactic acid and volatile fatty acids (VFA) using a gas different time points and stored frozen at −20  °C chromatograph (UNICAM 4600; SB Analytical, until analysis) to determine chemical composition Cambridge, UK) equipped with a flame ioniza- and fermentation characteristics. tion detector (250 °C), split-injection port (1.0 μL injection), and a capillary column (Agilent J&W HP-FFAP, 10 m by 0.535 mm by 1.00 μm, 19095F- Chemical Analysis and Fermentation 121; Agilent, Santa Clara, CA). The carrier gas Characteristics of Silage was He (column head pressure of 68.9 kPa) and the working temperature of the injector and de- Samples of fresh materials and silage were tector were 250 and 300 °C, respectively. The initial oven-dried at 60  °C to a constant weight and column temperature was set at 80 °C for 1 min and ground to pass through a 1-mm sieve (Wiley mill, then increased with 20 °C/min to 120 °C followed Swedesboro, NJ), and the determined DM was cor- by 6.2 °C/min to 140 °C and thereafter with 20 °C/ rected for the loss of volatiles during drying. The min to 205 °C. samples were analyzed for ash (method 924.05), ether extract (EE; method 920.39), and CP (method Treatments, Animals, and Diets 988.05) according to AOAC (1998). Ash-free NDF (NDFom) was determined without sodium sulfite Forty-five male Fashandy lambs with an (Mertens, 2002). The determination of ADF was average BW of 29.9 ± 0.9 kg and 6 months of age performed (method 973.18; AOAC, 1998) and ex- were used in this study. Animals were housed in pressed exclusive of residual ash (ADFom). Acid individual wooden-slatted pens (1.5  × 1.5 m) and detergent lignin (ADL) was determined by solubil- given an adaptation period of 14 d followed by a ization of cellulose with 72% sulfuric acid (method data collection period of 56 d. Lambs were offered 973.18), and phenolic compounds using the Folin– Ciocalteu method. The water-soluble carbohy- drates (WSC) were measured by colorimetry after Table 1.  Ingredients and chemical composition reaction with anthrone reagent (MAFF, 1982). The (%  of DM) of the diets containing different con- ME contents of pre-ensiled and BBWS were calcu- centrations of BBWS lated according to Menke et al. (1979) as: Concentration of BBWS in diet (% of DM) ME (MJ/kg DM)= 2.20 + 0.136 × GP Item 0 10 20 + 0.057 × CP + 0.0029 × EE Ingredient Alfalfa hay 15 12 7.5 where GP is 24-h net gas production (mL/200 mg Wheat straw 12.5 5.5 — of DM), and CP is measured in g/kg of DM. Wheat bran 19.6 19.6 19.6 For measuring silage pH, 50  g of fresh silage BBWS — 10.0 20.0 was blended with 125  mL of distilled water and Corn grain 44.3 44.3 44.3 allowed to stand at room temperature for 1  h Soybean meal 5.9 5.9 5.9 (Faithfull, 2002). After decanting the silage ex- Urea 0.5 0.5 0.5 tract into a small beaker, the pH was measured Sodium bicarbonate 0.7 0.7 0.7 Calcium carbonate 0.7 0.7 0.7 using a portable digital pH meter (Sartorius Vitamin–mineral premix* 0.5 0.5 0.5 PT-10; Sartorius AG, Göttingen, Germany). To Salt 0.3 0.3 0.3 determine ammonia-N (NH -N), an extract was Chemical composition† obtained by squeezing the silage material, filtered DM using Whatman 54 filter paper, then a 9 mL of ali- OM 94.7 95.1 93.3 quot was taken, mixed with 1 mL of 7.2 N H SO , 2 4 CP 14.6 14.8 14.5 and stored at −20 °C. After thawing, the silage ex- NDFom 28.3 27.1 25.8 tracts were analyzed for NH -N using a phenol-hy- 3 EE 7.0 7.9 7.9 pochlorite assay (Galyean, 1997). Two milliliters of ME, Mcal/kg of DM 2.69 2.69 2.67 the silage juice was pipetted into a microcentrifuge *Contained (per kilogram) 190 g of Ca, 50 g of Na, 19 g of Mg, 3 g tube with 0.5  mL of an acid solution (containing of Zn, 3 g of Fe, 2 g of Mn, 300 mg of Cu, 100 mg of Co, 1 mg of Se, 20% orthophosphoric acid and 20  mM 2-ethyl 100 mg of I, 100 mg of vitamin E, 500,000 IU of vitamin A, 100,000 butyric acid, as the internal standard) and centri- IU of vitamin D3, 300 mg of antioxidant. fuged at 15,000 × g for 15 min at 4 °C (Rezaei et al., †Calculated from analysis of each feed composition, which were 2014). The supernatant was used to quantify the analyzed. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. isonitrogenous and isocaloric diets, which were and at 15-d intervals before the morning feeding, daily (Table  1) formulated according to National after 16  h fasting, to calculate average daily gain Research Council (NRC, 1985) guidelines, in which (ADG) and feed efficiency (FE). The ADG was forage (lucerne + wheat straw) was replaced with calculated for each lamb by using the following different concentrations (0, 100, or 200 g/kg of diet equation: DM) of BBWS (Table 2). Diets were randomly as- ADG =(f inal BW − initial BW) / signed by BW into three groups of 15 lambs each in number days on feeding. a completely randomized design. Animals received the diets as a total mixed ration, at 0800 and 1700 h, to ensure 10% feed refusal and had constant access In Vivo Apparent Digestibility of Nutrients, to freshwater. Microbial N Supply, and N Retention Feed Consumption and Growth Performance On day 56 of the data collection period, ran- domly four animals per treatment were placed into The feed eaten by each animal and corres- individual metabolism crates (0.6 × 1.3 m), allowing ponding feed refusals were recorded daily to esti- feces and urine to be collected. Apparent digest- mate the consumption of DM and nutrients, and ibility was determined by the total fecal collection representative samples were kept frozen at –20  °C method, with 6 d for sample collection. During the for subsequent analyses. Samples of feed offered collection period, the amounts of feed offered, feed and refusals were oven-dried at 60 °C to a constant refusals, and feces from each animal were recorded weight, ground to pass through a 1-mm sieve, then daily and samples were stored (at –20 °C) for later analyzed for OM, N, and EE as described above. analysis of DM, OM, CP, NDFom, and EE as The silos of BB-wheat straw, which were opened at described above. different time points (i.e., on days 1, 21, 41, and 61 Total urine produced daily was collected in of the trial), were fed to animals. The body weight plastic vessels containing 100  mL of 10% sulfuric (BW) of animals was individually recorded at the acid. From the daily urine collected, 10% was sam- beginning and the end of the experiment period (on pled, diluted fivefold with distilled water, and then two consecutive days to provide baseline weights) stored at –20 °C to estimate purine derivatives and nitrogen (Chen and Gomes, 1992). Urine allantoin Table 2.  Chemical composition (%  of DM), fer- was measured by a colorimetric method at 522 nm mentation characteristics, and ME values (Mcal/kg after its conversion to phenylhydrazine. The sum of DM) of pre-ensiled and ensiled BBWS of xanthine and hypoxanthine was calculated by Item Pre-ensiled Ensiled SEM P-value their conversion to uric acid with xanthine oxidase Chemical composition (X-1875; Sigma-Aldrich, St. Louis, MO) and meas- DM, % of fresh weight 26 30 1.4 0.003 uring the subsequent optical density at 293  nm. CP 9.0 8.0 0.46 0.05 Uric acid was measured from the reduction in op- NDFom 39 40 0.97 0.16 tical density at 293  nm after the degradation of ADFom 32.6 36.4 1.3 0.03 uric acid to allantoin with uricase (U-9375; Sigma- Acid detergent lignin 6.5 11.2 1.7 0.001 Aldrich). Total purine derivative excretion per day Ash 18 27 0.36 0.001 was calculated, the daily absorbed exogenous pur- EE 5.9 7.4 0.16 0.02 ines were estimated, and the MNS was predicted. Water-soluble carbohydrate 7.5 0.5 0.08 0.0001 The concentration of urinary N was estimated by ME, Mcal/kg of DM 1.91 1.72 0.02 0.015 TP* 0.6 0.55 0.08 0.16 the Kjeldahl method (AOAC 1998), and N reten- Fermentation characteristics tion was calculated as daily N excretion (urinary N pH value — 4.4 — plus fecal N) subtracted from daily N intake. Ammonia-N, % of total N — 6.1 — Fermentative acids, % of DM Lactic acid — 4.5 — Rumen Metabolites Acetic acid — 1.7 — Propionic acid — 0.2 — Just before the morning feeding and 2, 4, and Butyric acid — 0.5 — 6 h afterward, rumen fluid from four animals ran- Valeric acid — 0.04 — domly allocated to each treatment was sampled Isovaleric acid — 0.05 — (using an esophageal tube) on day 56. After dis- carding the first 15  mL of rumen fluid, pH was *Total phenolic compounds. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb determined. For NH -N determination, a 25-mL (treatment) of the ith concentration of BBWS sample of strained rumen fluid was preserved with (i  =  three concentrations of BBWS), e represents ij 5  mL of 0.2  N HCl at –20  °C and then analyzed the experimental error, and e represents the sam- ijk using a phenol-hypochlorite assay (Broderick and pling error. The random effect included treatment Kang, 1980). To measure ruminal VFA, 8  mL of × replication. The model included the fixed effect strained rumen fluid was preserved (at –20 °C) with of dietary treatment. Moreover, data on digest- 2  mL of an acid solution (containing 20% ortho- ibility, MNS, and N balance were measured once phosphoric acid and 20 mM 2-ethyl-butyric acid as and examined using the same model with four rep- internal standard). After thawing, rumen fluid was licates for each diet. centrifuged (15,000 × g for 15 min at 4 °C) and the Repeated measures were used to statistically supernatant was used to quantify VFA as described examine the data of rumen metabolites and blood previously. biochemistry parameters. The model used was Rumen protozoa were enumerated using y = μ + T + σ + S + T S + ε ijk i ij k i k ijk 5 mL of rumen fluid pipetted into a test tube con- where y represents the value of each individual taining 20  mL of formalinized physiological sa- ijk observation, µ is the overall mean, T is the fixed ef- line (20 mL of formaldehyde in 100 mL of saline fect (treatment) of the ith concentration of BBWS containing 0.85 g of NaCl in 100 mL of distilled (i  =  three concentrations of BBWS), σ is the main water). Total and subfamily counts of protozoa ij random error, S is the sampling time, T S is detecting were determined using a light microscope k i k interaction between treatment and sampling time, and and a Hemocytometer (Neubauer-improved; Ɛ is the subrandom error. The influence of diet (treat- MarienfeldSuperior, Lauda-Königshofen, ijk ment), sampling time, and their interactions were con- Germany; Dehority 2003). sidered fixed. Compound symmetry was used based on the lowest Akaike information criterion (AIC) as Blood Biochemistry Parameters a covariance structure in the model. Also, repeated measure was used to test the data on feed consump- Just before the morning feeding and 4  h after tion, whereas the influence of dietary treatment, and feeding, blood samples (15  mL) were randomly their interactions were considered fixed. collected from four animals assigned to each treat- Additionally, a polynomial contrast was used ment on days 56 and 70 (the same animals were to test the linear or quadratic effects of the diet (i.e., used on both times). The samples were transferred BB inclusion concentration) on parameters meas- to the laboratory on ice, and the harvested serum ured. The result is then illustrated as statistically was stored at –20 °C until analysis. Glucose, trigly- significant at P ≤ 0.05. ceride, cholesterol, urea N, total protein, albumin, The SAS program was used to calculate the and creatinine concentrations were measured using number of replications needed to obtain the desired kits from Pars Azmun (Tehran, Iran) using a spec- power (>0.8). The power of the test is given by: trophotometer (Genova; Jenway, Essex, UK). Blood thyroxine and triiodothyronine concentra- Power = P F > F = F α,(a−1),(N−a) β tions were measured using kits from Pars Azmun (Tehran, Iran) using an ELISA reader. using a noncentral F distribution for H with a non- centrality parameter λ  =  SS /MS , and de- treatment error grees of freedom (a − 1)  and (N − a). Here, N is Statistical Analysis the number of replications per treatment, a is the Using a t-test, the chemical composition of number of treatments, and F is the critical α,(a − 1),(N − a) pre-ensiled and BBWS were compared to quan- value (Kaps and Lamberson 2009). tify changes made during the process of silage fer- mentation (Table  2). Data on ADG, digestibility, RESULTS MNS, and N retention were statistically examined using the PROC MIXED of SAS (Version 9.1; SAS Chemical Analysis and Fermentation Inst. Inc.) in a completely randomized design. The Characteristics of Silage model used was Compared to the fresh product (Table  2), y = μ + T + e + e , ijk i ij ijk BBWS contained higher concentrations of ADFom where y represents the value of each individual ob- (P  <  0.03), ADL and ash (P  <  0.001), and lower ij servation, µ is the overall mean, T is the fixed effect WSC concentration (P < 0.0001). Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. In Vivo Apparent Digestibility of Nutrients, Feed propionic (P < 0.63), or butyric acid (P < 0.41) con- Intake, and Growth Performance centrations. Ruminal enumeration of total protozoa, Entodiniinae, Diplodiniinae, Isotrichidae, Epidiniinae, The apparent digestibility of DM, OM, CP, and Ophrioscolecinae subfamilies were not affected by NDFom, and EE was not influenced (P   <  0.34, feeding lambs with different concentration of BBWS P < 0.39, P < 0.89, P < 0.69, and P < 0.35, respect- (P < 0.55, P < 0.78, P < 0.57, P < 0.17, P < 0.65, and ively) by feeding BBWS (Table 3). The DMI, organic P < 0.29, respectively). matter intake, crude protein intake, ether extract in- take, neutral detergent fiber intake, metabolisable en- Blood Biochemistry Parameters ergy intake, ADG, and feed conversion ration (FCR) were not affected (P  <  0.67, P  <  0.67, P  <  0.66, Table  6 illustrated that the blood concentra- P < 0.30, P < 0.63, P < 0.32, P < 0.55, and P < 0.55, tions of glucose, triglyceride, cholesterol, urea N, respectively) by feeding the experimental diets. creatinine, triiodothyronine, thyroxin, total protein, albumin, and globulin, did not change (P  <  0.59, P  <  0.85, P  <  0.75, P  <  0.52, P  <  0.63, P  <  0.59, Microbial N supply and N retention P  <  0.76, P  <  0.72, P  <  0.68, and P  <  0.75, respectively). Feeding lambs with BBWS had no effect (P  <  0.53) on total purine derivatives absorbed, DISCUSSION total purine derivative excretion (P  <  0.56), and MNS (P < 0.57; Table 4). Among the lambs, there Chemical Analysis and Fermentation were no significant changes in N consumption Characteristics of Silage (P < 0.72), fecal N (P < 0.88), urinary N (P < 0.61), and N retained (P < 0.81). Changes in the chemical composition of BBWS after ensilage occur because of microbial fermentation and losses from effluent throughout Rumen Metabolites and Protozoa Enumeration the ensiling process (Buxton and O’Kiely, 2003), Dietary inclusion of BBWS had no effect on but the DM of BBWS is within the range of me- ruminal pH (P  <  0.82) and NH -N concentration dium- to good-quality silage. Low-moisture silage (P < 0.61; Table 5). Feeding BBWS had no influence on depresses undesirable clostridial growth (Pahlow the content of total VFA (P < 0.83), acetic (P < 0.49), et al., 2003) because of the affinity of Clostridium Table 3. Effect of dietary inclusion of BBWS on in vivo apparent digestibility (n = 4), intake, and growth performance of fattening lambs Concentration of BBWS in diet (g/kg of DM) Item 0 100 200 SEM P-value Digestibility, % DM 76 75 73 1.50 0.34 OM 77.1 76 78.1 0.75 0.39 CP 72.1 72.5 72.4 2.4 0.89 NDFom 62 61 63 2.85 0.69 EE 77 77 79 1.34 0.35 Intake, g/d DM 1,432 1,435 1,420 14.4 0.67 OM 1,332 1,316 1,315 16.25 0.67 CP 204 200 202 2.59 0.66 EE 112 113 115 1.77 0.3 NDFom 420 414 416 4.67 0.63 ME, Mcal/d 2.94 2.91 2.96 0.03 0.32 Performance Initial BW 29.41 29.28 29.06 0.87 0.28 Finial BW 42.8 42.47 41.94 1.20 0.32 ADG, g/d 239 235 230 9.34 0.55 FCR* 5.99 6.12 6.13 0.4 0.55 *Feed conversion ratio (DMI:ADG). Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb Table 4. Effect of dietary inclusion of BBWS on total purine derivatives (TPD), MNS, and N retention in fattening lambs (n = 4) Concentration of BBWS in diet (g/kg of DM) Item 0 100 200 SEM P-value Urinary excretion, mmol/d Allantoin 8.58 8.04 8.10 0.56 0.41 Xanthine + hypoxanthine 0.73 0.73 0.74 0.51 0.51 Uric acid 3.61 4.10 4.37 0.55 0.52 TPD absorbed, mmol/d 14.69 14.67 15.11 1.07 0.53 TPD excretion, mmol/d 12.92 12.87 13.22 0.91 0.56 MNS, g/d 10.68 10.66 10.99 0.78 0.57 N retention, g/d N intake 29.7 29.02 29.12 1.10 0.72 Fecal N 8.17 8 8.01 0.69 0.88 Urinary N 11.27 11.07 11.10 0.50 0.61 Retained N 10.28 9.95 10.01 0.65 0.81 Retained N, g/kg of N intake 346 343 344 2.81 0.96 Table 5. Effect in vivo of dietary inclusion of BBWS on rumen metabolites and protozoa numeration in fattening lambs (n = 4) Concentration of BBWS in diet (g/kg of DM) Item 0 100 200 SEM P-value pH 6.38 6.37 6.43 0.14 0.82 Ammonia-N, mg/dL 10.12 10.33 10.50 0.86 0.61 Total VFA, mmol/L 82.3 79.63 81.95 1.7 0.83 VFA, mmol/100 mmol Acetic acid 56.29 54.53 54 1.66 0.49 Propionic acid 27.11 28.5 28.58 1.60 0.63 Butyric acid 15.9 16.3 16.8 0.44 0.41 Valeric acid 0.56 0.55 0.50 0.08 0.76 Isovaleric acid 0.14 0.12 0.12 0.02 0.63 Acetate:propionate 2.08 2.08 2.06 0.22 0.94 Protozoa, log /g of digesta Total protozoa 6.09 6.06 6.06 0.032 0.55 Entodiinina 5.86 5.83 5.86 0.039 0.78 Diplodiinina 5.45 5.40 5.35 0.086 0.57 Isotrichidea 4.87 4.86 4.98 0.042 0.17 Epidiniina 4.79 4.78 4.79 0.013 0.65 Ophrioscolecinae 4.89 5.01 5.04 0.06 0.29 No significant interaction between treatment and sampling time were observed. spp. for moisture. The production of butyric acid concentration (Megías et  al., 2014). The preserva- was low in the present study, indicating low growth tion of BBWS was suitable in terms of pH value of Clostridium spp., which derive energy for growth (i.e., 4.8), low NH -N content (<100 g/kg total N), by fermenting sugars and lactate into butyric acid and low concentrations of acetic, propionic, and bu- (Pahlow et al., 2003). In agreement with our results, tyric acids (McDonald et al., 2011). A good-quality Shao et  al. (2007) documented improved fermen- silage is characterized by a lactic acid concentra- tation of perennial ryegrass with decreased con- tion between 30 and 140 g/kg of DM (McDonald centrations of butyric. The inverse relationship et  al., 2011), and BBWS lactic acid concentration between ADFom, ADL, and WSC show that a re- was 45 g/kg of DM. The pH values of our BBWS duction in WSC increases the content of ADFom were the same as those (pH = 4.76) noted by Jian and ADL resulting in a proportional rise in ash et al. (2017) who ensiled rice straw, broccoli residue, Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. Table 6. Effect of dietary inclusion of BBWS on blood biochemistry parameters in fattening lambs Concentration of BBWS in diet (g/kg of DM) Item* 0 100 200 SEM P-value Metabolites, mg/dL Glucose 67.22 69.41 70.76 1.52 0.59 Triglyceride 21.98 21.23 21.90 2.10 0.85 Cholesterol 59.60 61.70 63.99 5.38 0.75 Blood urea N 19.45 22.44 19.58 1.27 0.52 Creatinine 1.40 1.42 1.45 0.07 0.63 Thyroid hormones, ng/mL Triiodothyronine 5.13 5.25 5.33 0.28 0.59 Thyroxin 69.00 70.76 71.80 3.30 0.76 Protein, g/dL Total protein 6.77 6.67 6.76 0.18 0.72 Albumin 3.59 3.51 3.55 0.11 0.68 Globulin 3.18 3.16 3.20 0.15 0.75 Albumin:globulin 1.14 1.10 1.11 0.06 0.68 *Average of repeated sampling of blood collected from six animals assigned to each treatment on days 56 and 70 of data collection period, just before the morning feeding and 4 h after the morning feeding. and lucerne at a ratio of 5:4:1 for 30 d, whereas, for The similar ADG among the lambs reflected other silage characteristics, such as NH -N, lactic their comparable ruminal VFA concentration and acid, and VFA content, Jian et al. (2017) reported a MNS (Ben Salem et  al., 2002). The FCR showed lower silage quality than those observed in BBWS. no difference among the lambs due to their similar DMI and ADG. The experimental diets, in the cur- rent study, were formulated to meet the require- In Vivo Apparent Digestibility of Nutrients, Feed ments of a growth rate of 300 g/d for fattening male Consumption, and Growth Performance lambs at 8.5 mo of age according toNRC (1985) The inclusion of BBWS in the diets had no recommendations. However, the ADG of our effect on nutrient digestibility and this relates to lambs was not as predicted in NRC (1985) tables the similar chemical composition of the diets and because the equations used to predict lamb growth the comparable ruminal pH in the lambs, factors were not obtained in Iranian sheep. Iranian sheep which influence rumen microbial activity (Petit and breeds (except the Zel breed) are fat tailed and their Castonguay, 1994). The low concentration of phen- physiological characteristics and genetic potential olic compounds (1.0 g/kg of DM) had no adverse for growth and, to some extent, the rumen micro- effect on diet apparent digestibility. Previous work biota of these breeds differ from sheep in other on fresh cauliflower (Brassica oleracea var. botrytis) countries, which can lead to a difference in overall reported good nutrient digestibility of DM, OM, performance. CP, and NDF (80.9%, 86.9%, 84.9%, and 71.8%, re- spectively) when fed to goats (Wadhwa et al., 2006). Microbial N Supply and N Retention It has been stated that increased concentrations of fresh cabbage (Brassica oleracea var. capitata) in Diets that maximize MNS often increase the diet of lambs reduced OM and NDF digest- animal performance (Chen and Gomes, 1992). ibility (73% vs. 65%; 56% vs. 47%, respectively; In the present study, the MNS reached its high- Nkosi et al., 2016). est value (10.99  g/d) with 200  g of BBWS per Feeding BBWS had no influence on feed con- kilogram of diet DM. Similar concentrations of sumption and this may be due to the similar diet digest- urinary purine derivatives (PD) and MNS among ibility, particularly NDFom digestibility. It was noted treatments were due to the similar intakes of di- that NDF digestibility provides an accurate estimation gestible OM and N (Ben Salem et al., 2002) and of intake (Harper and McNeill, 2015). A correlation the equal ruminal synchrony of ME and N re- between NDF digestibility and intake was stated by sulting from a similar daily ME intake and di- Nkosi et al. (2016) who noted a lower DMI for lambs gestible CP consumption among the treatments. fed fresh Brassica (oleracea var. capitata) compared to Urinary PD excretions are indicators of MNS a Brassica-free diet (1,420 vs. 1,600 g). arriving at the beginning of the small intestine Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Broccoli byproduct-wheat straw silage to lamb (Chen and Gomes, 1992). It was reported by was not affected by the diets. The similar daily OM Wadhwa et  al. (2006) that higher MNS values intake and OM digestibility, which resulted in equal in goats offered fresh cauliflower (Brassica oler - fermented OM per day, reflects the similar ruminal acea var. botrytis; 25.40  g/d) compared to those concentrations of total and proportions of indi- in the current study. This difference could be due vidual VFA among the dietary treatments (Morvay to the different experimental animals and diets et al., 2011). (i.e., lambs vs. goats; BBWS vs. fresh Brassica Rumen protozoa, exclusively ciliates, rank oleracea). second only to bacteria in cellular biomass of Nitrogen retention among the animals did not the ruminal microbiota (Dehority, 2003). They differ because digestible OM and N (Ben Salem are found in the rumen where they play im- et  al., 2002), N excreted in feces and urine (Zuo, portant roles in feed digestion and homeostasis 2011), and ADG were not influenced by feeding of the rumen ecosystem (Dehority, 2003). Feeding BBWS. A  lower N consumption and N retention BBWS had no effect on rumen protozoa enumer- were noted in animals offered diets with cabbage ation and this was due to the normal ruminal pH, compared to those fed a Brassica-free diet, sug- which plays an important role in rumen protozoa gesting that the lower N intake and retention could activity (Dehority, 2003). be related to the decrease in the digestibility of DM and OM in this diet (Nkosi et al., 2016). Blood Biochemistry Parameters Concentrations of blood parameters represent Rumen Metabolites and Protozoa Enumeration an integrated index of the adequacy of nutrient For all lambs, ruminal pH values varied from supply in relation to the utilization of nutrients, 6.37 to 6.43, which were within the optimum range which is a self-governing aspect of the physiological (i.e., 6.1–6.6) for maintaining normal cellulolytic condition of the animal and provides an immediate organism populations (Van Soest, 1994). Feeding indication of nutritional status at a specific point BBWS had no effect on rumen pH and this was re- in time (Pambu-Gollah et al., 2000). In the present lated to the similar DMI and nutrient digestibility study, the lack of differences in blood biochemistry among lambs in different treatments. Dietary add- parameters among the treatments was probably due ition of BBWS had no adverse effects on ruminal to similar intakes and nutrient digestibilities. The metabolism. It was noted by Keogh et  al. (2009) serum concentrations of glucose, cholesterol, urea that replacing grass silage with fresh kale (Brassica N, total protein, albumin, and globulin and the al- oleracea) had no effect on the ruminal pH of dry bumin to globulin ratio in all treatments were within dairy cows (mean pH  =  6.2). In contrast, feeding the normal ranges for sheep (Radostits et al., 2007). fresh brassica forage (Brassica spp.) to lambs re- However, the concentration of triglyceride was sulted in a lower ruminal pH (<6.0) as a result of greater than the range for sheep (Radostits et  al., the greater ratio of readily fermentable to struc- 2007) but was in the usual range reported for fat- tural carbohydrates in Brassica forage (Sun et al., tailed Iranian sheep (18.03 to 50.93 mg/dL; Mojabi, 2015). Ruminal NH -N content in all the animals 2011). The similar concentrations of blood albumin was higher than 5  mg/dL, which is the minimum among the lambs suggested normal liver function concentration required by rumen microorganisms as the liver is the main organ of albumin synthesis. to support their optimum growth (McDonald The values of albumin and globulin in the serum of et  al., 2011). The lack of difference in rumen the lambs in the current study show that these ani- NH -N concentration among the lambs suggests mals did not suffer from any health problems that that the release of N in the rumen and its uptake might have affected their performance (Radostits by microbes to synthesize microbial CP were not et al., 2007). affected by the diets. In Brassicas, the antinutritional S-methylcysteine Ruminal VFA is the main energy source for sulfoxide and glucosinolates were reported to be goi- ruminants (Abarghuei et  al., 2013); consequently, trogenic and could affect triiodothyronine and thy- diets that increase the ruminal production of VFA roxin production (Chorfi et  al., 2015). The lambs lead to higher animal performance (Penner, 2014). offered BBWS had no clinical indicators of gluco- In the current study, dietary BBWS had no influence sinolate toxicity because the concentration was low on the total and proportions of individual VFA and there were no symptoms of toxicity (EFSA, content; thus, the performance of fattening lambs 2008). This may be due to the ensiling process Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/article/4/3/txaa078/5873894 by DeepDyve user on 03 September 2020 Partovi et al. European Food Safety Authority. 2008. Glucosinolates as un- causing a reduction in the concentration of gluco- desirable substances in animal feed—Scientific Panel on sinolates and S-methylcysteine sulfoxide improving Contaminants in the Food Chain. EFSA J. 590:1–76. the feed quality (Vipond et al., 1998). Similarly, feed- Faithfull,  N.  T. 2002. Methods in agricultural chemical ana- ing different concentrations of ensiled kale (Brassica lysis: a practical handbook. CABI Int., Wallingford, UK. oleracea) in lambs did not affect their performance FAO. 2018. FAO statistics division. Crop production. Food and Agriculture Organization of the United Nations, or blood triiodothyronine and thyroxin concentra- Rome, Italy. tions probably because of the reduction in glucosi- FASS. 2010. Guide for the care and use of agricultural animals nolate during ensilage (Vipond et al., 1998). in research and teaching. 3rd ed. Federation of Animal Science Societies, Champaign, IL. CONCLUSIONS Galyean, M. L. 1997. Laboratory procedures in animal nutri- tion research. Department of Animal and Food Sciences, The chemical composition and fermentation Texas Tech University, Lubbock, TX. characteristics of BBWS indicates its potential as a Halkier, B. A., and J. Gershenzon. 2006. Biology and biochem- ruminant feedstuff. BBWS inclusion at up to 200 g/ istry of glucosinolates. Annu. Rev. Plant Biol. 57:303–333. kg of DM in the diet of fattening Fashandy lambs doi:10.1146/annurev.arplant.57.032905.105228. Harper, K. J., and D. M. McNeill. 2015. The role iNDF in the is equivalent to an Alfalfa-wheat straw diet in terms regulation of feed intake and the importance of its as- of effects on nutrient digestibility, feed intake, sessment in subtropical ruminant systems. Agriculture. growth performance, FE, rumen parameters and 5:778–790. blood parameters. BBWS at this level is a safe feed- Hu,  C.  H., A.  Y.  Zuo, D.  G.  Wang, H.  Y.  Pan, W.  B.  Zheng, stuff for sheep having no adverse effects on their Z.  C.  Qian, and X.  T.  Zou. 2011. Effects of broccoli stems and leaves meal on production performance and health status or growth performance. egg quality of laying hens. Anim. Feed Sci. Technol. Conflict of interest statement. None declared. 170:117–121. 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