Effects of different physical forms of starter on digestibility, growth, health, selected rumen parameters and blood metabolites in Holstein calves

Effects of different physical forms of starter on digestibility, growth, health, selected rumen parameters and blood metabolites in Holstein calves

Animal Feed Science and Technology 271 (2021) 114759 Contents lists available at ScienceDirect Animal Feed Science and Technology journal homepage: ...

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Animal Feed Science and Technology 271 (2021) 114759

Contents lists available at ScienceDirect

Animal Feed Science and Technology journal homepage: www.elsevier.com/locate/anifeedsci

Effects of different physical forms of starter on digestibility, growth, health, selected rumen parameters and blood metabolites in Holstein calves C. Du a, b, c, 1, L. Ma a, 1, Y.G. Zhen b, A.F. Kertz d, W.J. Zhang c, D.P. Bu a, e, f, * a

State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China c College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China d ANDHIL LLC, St. Louis, MO 63122, USA e CAAS-ICRAF Joint Lab on Agroforestry and Sustainable Animal Husbandry, World Agroforestry Centre, East and Central Asia, Beijing 100193, China f Hunan Co-Innovation Center of Animal Production Safety, CICAPS, Changsha, Hunan 410128, China b

A R T I C L E I N F O

A B S T R A C T

Keywords: Calves Texturized starter Pelleted starter Diarrhea Digestibility

This study was designed to determine whether physical forms of starter feed could influence digestion, growth, rumen fermentation parameters, and blood metabolites in Holstein dairy calves. Sixty-three female calves (averaging 41 kg body weight (BW)) were randomly assigned to 1 of the following 3 starter diets differing in physical form or formulation: texturized starter (TS; n = 21), pelleted starter with the same formula as TS (PSA; n = 21), and a commercial pelleted starter (PSB; n = 21) with a different formulation. Twelve male calves were selected and randomly assigned to each treatment group to determine feed digestibility using indigestible neutral detergent fiber (iNDF) and acid insoluble ash (AIA) as the internal markers. The dry matter intake (DMI) of calves in three treatments was not different within the first 6 wk (P = 0.19), and there were also similar gain-to-feed ratios (P = 0.91) and Average daily gain (ADG) (P = 0.26). During wk 7, starter DMI of the calves in TS and PSA groups was greater than PSB animals (P < 0.01). The ADG of TS and PSA-fed groups were greater than the animals fed with PSB during wk 7. Offering TS resulted in tendentionally greater starter DMI and ADG than PSA during the following 2 wk after weaning (P = 0.08, P = 0.09). During the 9-wk study, calves receiving PSB had lower ADG than the other two treatments (P = 0.04), but there were no dif­ ferences in gain-to-feed ratio (P = 0.86). Serum biochemical parameters were not different among the 3 treatments. Calves began exhibiting ruminating behavior for TS about 2 d numerically earlier than that for PSA and PSB (P = 0.54). Butyrate in the rumen of PSB-fed animals was greater than in PSA and TS calves (P < 0.01) on day 63. Also, PSB-fed calves had a greater ruminal acetate: propionate ratio than calves receiving PSA and TS (P < 0.01) on day 49. The TS starter had the lowest digestibility of CP, DM, and ADF (P < 0.01) by both digestibility methods.

Abbreviations: ADF, acid detergent fiber; ADG, average daily gain; AIA, acid insoluble ash; BW, body weight; CP, crude protein; DM, dry matter; DMI, dry matter intake; EE, ether extract; iNDF, Indigestible neutral detergent fibre; Immunoglobulins G, IgG; NDF, neutral detergent fibre; TS, texturized starter; PSA, pelleted starter A; PSB, pelleted starter B; VFA, volatile fatty acid. * Corresponding author at: State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Science, No. 2 Yuanmingyuan Road, Haidian District, Beijing, 100193, China. E-mail address: [email protected] (D.P. Bu). 1 These authors have contributed equally. https://doi.org/10.1016/j.anifeedsci.2020.114759 Received 29 April 2020; Received in revised form 10 November 2020; Accepted 12 November 2020 Available online 21 November 2020 0377-8401/© 2020 Elsevier B.V. All rights reserved.

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Overall, the pelleted starter with soybeans and distillers dried grains had lower DMI and ADG than the other pelleted starter and texturized starter which had the same formulation.

1. Introduction Pelleting feed has been shown to increase consumption when eating time was limited (Kertz et al., 1981). Many studies have evaluated approaches for optimizing calf feeding programs. Most of those studies evaluated the effect of nutrient composition on calf performance (NRC, 2001; Kertz et al., 2017). Only a few studies focused on how physical forms of starter could affect the growth of calves (Franklin et al., 2003; Lesmeister and Heinrichs, 2004; Bach et al., 2007; Porter et al., 2007). Reported effects of physical forms of starter on calf performance are mixed (Terr´ e et al., 2015). Pelleted starter resulted in reduced feed consumption as compared to texturized starter, especially after calves were weaned (Bach et al., 2007). Franklin et al. (2003) reported reduced starter intake and ADG in calves fed a pelleted versus texturized starter, but ingredients differed in these formulations. Khan et al. (2011) found lower rumen pH when calves were fed a texturized starter containing 37 % processed grains than calves fed a texturized starter with libitum access to chopped hay, and calves fed hay resulted in 4.7 kg more gut fill and less true growth because ADG did not differ by treatment. Thus, the amount of texturization was inadequate in this study without also feeding hay. Suarez-Mena et al. (2015) reported whole oats in texturized starter had similar digestibility with pelleted starter containing ground oats. While DM digestibility tends to decrease with increasing ration particle size (Maulfair et al., 2011) in lactating dairy cow rations, in calf starters texturization increased rumen pH and digestibility (Porter et al;, 2007). We hypothesized that texturized starter would improve digestibility, DMI, and rumen fermentation of calves. We further hypothesized that texturized starter fed calves would have greater DMI and ADG and greater BW when weaned. The objective of this study was to evaluate effects of two physical forms and two formulations of starters on digestibility, DMI, ADG, rumen and blood parameters in dairy calves. 2. Material and methods 2.1. Calves, treatments, and management The study was conducted (April 18 to August 11, 2015) at the ZhongDi-Dairy-Farm (Beijing, China) using 63 female Holstein calves and 12 male calves. All calves were separated from the cow immediately after birth and each received 4 l of colostrum via esophageal tube only at this feeding. Colostrum Immunoglobulins G (IgG) was tested using an Brix refractometer (HT113ATC; Httechltd Tianyuan Optical Instruments Co. Ltd., China). Average Brix value of colostrum was 25.8 during this experiment. Only colostrum with Brix values above 22 were fed to calves (Chigerwe et al., 2008). Navels were disinfected with diluted iodine solution, and calves were transferred to individual outside calf hutches (1.5m × 2m) 24 h after birth. Calves were bedded with sand. All calves were fed pasteurized whole milk (average composition: 3.41 % protein, 3.81 % fat, 12.01 % total solids) collected from the herd and using a teat Table 1 Ingredient and chemical composition of starters fed to calves. Treatment1 Ingredient composition, g/kg DM Corn (cracked in TS) Oats (whole in TS) Molasses Corn (ground) Soybean meal Cottonseed meal Wheat bran Full fat soybeans Distillers dried grains Premix Chemical composition, g/kg DM DM CP NFC2 NDF ADF Ether extract Ash

PSA

PSB

TS

300 100 50 100 250 50 100 0 0 50

0 0 40 500 100 120 50 50 9 50

300 100 50 100 250 50 100 0 0 50

889 210 476 299 54.7 14.5 72.9

882 211 485 291 61.2 13.6 71.1

890 197 479 311 55.4 13.8 71.8

1 Treatments: TS = texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface; PSA = pelleted starter with the same ingredient formula as TS; PSB = commercially available pelleted starter with a different formulation than TS or PSA. 2 Non- fibre -carbohydrate was calculated as [DM - (NDF + CP + ether extract + ash)] (NRC, 2001).

2

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bucket twice daily (07:30 and 14:30). Calves were fed twice daily 2 l milk per feeding during the first wk, 3 l per feeding during wk 2–6, and then only 2 l at one feeding daily during wk 7, with full weaning at the end of that wk. Buckets were cleaned immediately after each feeding. Calves were fed ad libitum starter beginning at 5 d of age. Leftovers were weighed at 8 a.m. daily. Then new starter was fed. Calves had free access to water 2 h after each milk feeding, because this is standard practice to encourage calves to drink more water and this leads to more starter consumption. Calves were then fed only starter feed during wk 8 to wk 9 and had free access to water. Calves were randomly assigned to three treatment groups, with each fed starter diets with similar nutrient content, but differing in physical form or formulation: texturized starter (TS; a mixture of a pellet, cracked corn and whole oats, which was surface dressed with 5% molasses) (25 calves, 21 females with 4 males); pelleted starter produced specifically for this study with the same ingredient formula as TS (PSA; 25 calves, 21 females with 4 males); and a commercially available pelleted starter (PSB; with some different ingredients than TS and PSA, (25 calves, 21 females with 4 males). There are many differently formulated pelleted starters available, but no real evaluations of impact of differing formulations on calf performance. In this study all formulations were provided and manufactured by the same feed company (Apichat Wantanasak of Chia Tai Group, Beijing, China). The PSB starter was selected because it was a pelleted commercially available starter with a known formula. Ingredient and chemical composition of starter feeds are shown in Table 1. 2.2. Data collection and sampling Offered and refused amounts of starter were recorded daily on an individual basis for female calves. All female calves were measured for body weight, body length, heart girth, withers height, hip height, cannon circumference at birth and subsequently every wk until the end of the experiment (9 wk, 63 d). Average daily gain was calculated as the average amount of weight gain per day during each feeding period by subtracting initial and final BW divided by d, while gain to feed ratio was the amount of body weight gain coming from one unit of feed (feed efficiency = kg of BW gain/kg of total DMI). Samples of starters (100 g) were collected every wk during the experiment, composited and stored for analyses. Feces samples (approximately 30 g, wet weight) were collected through the rectum every 6 h from the 4 male calves of each treatment group at d 51, 52, 53, for a total of 12 feces samples per calf. Samples were pooled across sampling times for each calf. One portion of the feces samples was pooled from each calf, and a 10 % volume of 6 M hydrochloric acid was added to preserve the samples for nutrient analysis (Meyer et al., 2019). Starter and fecal samples were mixed thoroughly after drying and ground to pass through a 1-mm screen using a Wiley mill (Arthur H. Thomas Co) for nutrient analyses. The farm veterinarian scored whether the female calves had diarrhea based on the scoring system of 1 (firm), 2 (soft), 3 (runny) or 4 (watery) according to the standard advised by Kertz and Chester-Jones (2004), A score of 3 or greater was considered to be diarrhea. Blood samples (10 mL) were collected from female calves before morning feeding on d 1, 42, 49, and 63 of age, because sampling before morning feeding would reduce variation caused by milk feeding. Blood was drawn from the jugular vein of each calf into evacuated tubes without anticoagulant (Kangjian Medical Appartus CO.LTD, JiangSu, China), and serum was harvested by centri­ fugation at 3000 × g for 15 min at 4 ◦ C and stored at − 20 ◦ C until further analysis. Ruminating behaviours (age calves started rumination behaviours) were observed by walking around the pen area 1 h each morning with ruminating behaviour defined as repetitive movements of lower jaw in the lateral plane (Hepola et al., 2006). Rumen fluid samples were collected from all female calves using a stomach tube (K0021; Anscitech Ltd. China) before morning feeding on d 49 and 63 of age and strained through 4 layers of cheesecloth. Rumen fluid pH was immediately measured using a portable pH meter (pH Cube, TPS Pty Ltd., Springwood, QLD, Australia). Two mL of rumen fluid were acidified with 400 ul metaphosphoric acid and stored (-20℃) until analysis for volatile fatty acid (VFA), Another 10 mL sample was acidified with 0.1 mL of 6 M HCI for ammonia concentration determination. Ambient temperature (℃) and relative humidity were recorded every 15 min with a data logger (DSR-TH; ZOGLAB Microsystem Co., Ltd. China). Temperature-humidity index (THI) was calculated using the following equation: THI = (0.8 × ambient temperature (◦ C) +(% relative humidity/100) × (ambient temperature - 14.4) + 46.4. 2.3. Laboratory analyses Colostrum quality was determined by several drops of colostrum placed on the prism and the sample cover was lowered. The refractometer was then held up to a light source, and the Brix value was read at the line between the light and dark areas that appear on the scale. Starter and fecal samples were analysed for content of DM (method 934.01;AOAC, 1990), CP (method 988.05;AOAC, 1990), and Ash (method 942.05;AOAC, 1990). Neutral detergent fibre (NDF) and acid detergent fibre (ADF) were determined in sequential an­ alyses using an ANKOM fibre analyser (Fiber Analyzer A200; Ankom Technology, NY, USA) using the procedure of Van Soest et al. (1991). NDF was assayed with a heat stable amylase. Both NDF and ADF were expressed inclusive of residual ash. Ether extract (EE) content was determined by a Soxtec system HT6 apparatus (Tecator AB, Hillerød, Denmark) according to AOAC (1990) method 920.39. Rumen liquid samples treated with meta-phosphoric acid were then centrifuged at 10,000 × g for 30 min at 4 ◦ C, the clear su­ pernatant was injected for analyses by gas chromatography (Agilent 7890A GC, Column type: DB-FFAP) using the procedure of Filípek and Dvoˇr´ ak (2009). Ammonia-N concentrations were determined using the Berthelot reaction and an Infinite® F200 microplate reader (TECAN, Switzerland) using the alkaline phenol hypochlorite colorimetric procedure of Chaney and Marach (1962). Serum glucose concentrations were determined using AEROSET (CX5 pro, 143 Beckman), and IgG, growth hormone(GH), insulin concentrations were determined using enzyme-linked immunosorbent assay with commercially available kits H106, H091, H203 3

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(Nanjing Jiancheng Bioengineering Institute, Jiangsu, China). Milk samples were analysed for total solids using an infrared analyzer (Milkoscan TM FT 120; FOSS Analytical). Apparent total tract nutrient digestibility was determined using both 4 N hydrochloric acid insoluble ash (Van Keulen and Young, 1977) and indigestible NDF as internal markers. Indigestible NDF of starter and fecal samples was determined by placing samples using a nylon bag placed into a dairy cow rumen through a fistula for 12 d (Wang et al., 2014). 2.4. Statistical analyses Completely randomized design was used in this study. Normality of data was tested first. Then all data were statistically analyzed using the Proc Mixed method of SAS 9.4 (SAS Institute, Cary, NC). The frequencies of diarrhea of different treatments were analyzed using the Chi-squared 193 test (Proc Freq along with the option CHISQ). The statistical model used for analysis of DMI data was: Yijk = μ+ Ti + Dj(i)+ (TW)ik + εijk, where i = treatment, j = calf, k = week (age); μ = the overall mean; Ti = effect of ith treatment, Dj(i) = random effect of jth calf within treatment, (TW)ik = interaction effect between ith treatment and time, εijk = random error associated with the jth calf assigned to the ith treatment at kth week. The model included treatment, week(age), and interaction between treatment and week (age) as fixed effects, calf within treatment as random effect, and week(age) as repeated effect with autoregressive as the covariance structures. The MIXED statistical model was used to analyze gain to feed ratio, ADG, body measurements, rumen fermentation, blood pa­ rameters, and nutrient digestibility as: Yijk = μ+ Ti + Dj(i) + εijk, where i = treatment, j = calf; μ = the overall mean; Ti = effect of ith treatment, Dj(i) = random effect of jth calf within treatment, εijk = random error associated with the jth calf assigned to the ith treatment. The model included treatment as fixed effect, calf with treatment as random effect. Significance was declared at P ≤ 0.05 and trends at 0.05


SEM

P-value

0.52

0.019

0.26

0.83 0.13 0.97 0.58

0.83 0.17 1.00 0.57

— 0.024 0.024 0.019

— 0.19 0.19 0.91

0.64

0.55

0.72

0.074

0.26

0.50 0.80a 1.24a 0.59

0.50 0.50b 1.00b 0.67

0.50 0.74a 1.23a 0.71

— 0.091 0.091 0.068

— <0.01 <0.01 0.45

0.74ab

0.63b

0.81a

0.058

0.09

1.48ab 0.49 0.57 0.60a

1.30b 0.50 0.58 0.53b

1.57a 0.52 0.58 0.60a

0.085 0.035 0.016 0.021

0.08 0.81 0.86 0.04

PSA1

PSB2

TS3

0.55

0.50

0.83 0.20 1.03 0.58

Different superscripts within a row indicate significant difference (P < 0.05). 1 PSA: pelleted starter with same ingredient formula as TS (n = 21). 2 PSB: commercially available pelleted starter with a different formulation than TS or PSA (n = 21). 3 TS: texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface (n = 21). 4 Milk: milk feed to the calves have average 12.01 % total solid. 5 Total DMI: DM from milk and starter. 4

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among the three treatment groups during the first 6 wk (Table 2, Fig. 1). At weaning, PSB starter consumption was lower than PSA and TS starter consumption (P < 0.01). Following weaning, TS calves had greater starter DMI than PSB calves while the DMI of PSA calves was intermediate (P = 0.09). During the 9 wk trial, PSB calves had a lower ADG than the other two treatment calves (P < 0.05). However, there were no differences in gain:feed ratios among the three groups (Table 2). The TS calves achieved a tendentionally greater (P = 0.06) final BW than the PSB animals, but TS group had a similar final BW to the PSA calves (Table 3). The different physical forms of starter did not have significant effects on body length, heart girth, wither height, hip height and cannon circumference. 3.2. Ruminal fermentation parameters Age of calves initiating rumination was about 1.5–1.7 d sooner for TS (Table 4) than for PSA and PSB calves. Calves from each group started ruminating after d 20 of age. Ruminal pH, ammonia nitrogen, and total VFA concentrations were unaffected by the physical forms of starters. Concentration of butyrate in the rumen of PSB calves was greater than in calves receiving PSA and TS (P < 0.01) on day 63. Also, PSB group had a greater acetate:propionate ratio than the animals fed with PSA and TS (P < 0.01) on day 49. 3.3. Blood serum metabolites Glucose concentration was not found to be different among treatments (Table 5). The PSA calves had similar blood insulin con­ centration as for PSB and TS calves at d 42, but PSB tended to have lower blood insulin concentration than PSB and TS calves at d 49 and d 63. No differences were observed for blood IgG and GH content among the three groups. 3.4. Digestibility Digestibility of DM, CP, and ADF was greater by iNDF method for PSB (P < 0.05) treatment than for TS treatment; but PSA treatment was found to be similar to PSB treatment in these digestibilities (Table 6), while by acid insoluble ash PSA was also found to have greater NDF digestibility than for PSB. 3.5. Diarrhea No differences were observed among groups for diarrhea occurrence (P = 0.645) (Fig. 2). The most diarrhea was observed on day 12 of the experiment. Calves receiving PSA treatment exhibited diarrhea later than those of TS and PSB treatments, and TS calves had diarrhea that lasted longer, but only 1 d longer for 1 calf. Immediately after diarrhea ceased, calves showed rumination behaviour. 4. Discussion 4.1. Effect of the physical form of starter on intake and performance Lower DMI and ADG of PSB calves were likely due to inclusion of roasted soybeans and distillers dried grains, both of which have been shown to reduce DMI and ADG (Kuehn et al., 1994; Suarez-Mena et al., 2011). Addition of fat (Hill et al., 2015) to calf starters

Fig. 1. Effects of different physical forms of starter on starter intake of dairy calves. TS = texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface (▴), n = 21; PSA = pelleted starter with the same ingredient formula as TS (◼), n = 21; (3) PSB: available pelleted starter with a different formulation than TS or PSA (×), n = 21. 5

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Table 3 Different physical forms of starter effects on female calf body parameters. Items Body weight, kg d 1(Initial) d 42 d 49(At weaning) d 63(Final) Body length, cm d 1(Initial) d 42 d 49(At weaning) d 63(Final) Heart girth, cm d 1(Initial) d 42 d 49(At weaning) d 63(Final) Withers height, cm d 1(Initial) d 42 d 49(At weaning) d 63(Final) Hip height, cm d 1(Initial) d 42 d 49(At weaning) d 63(Final) Cannon circumference, cm d 1(Initial) d 42 d 49(At weaning) d 63(Final)

Treatment

SEM

P-value

41.4 63.2 68.2 79.5a

0.62 1.06 1.17 1.52

0.92 0.33 0.19 0.06

70.7 80.9 82.8 84.5

71.8 81.5 83.1 85.1

0.63 0.65 0.60 0.82

0.34 0.53 0.82 0.87

80.0 92.6 93.7 99.4

80.3 91.8 93.7 97.4

79.8 92.9 94.9 99.3

0.48 0.50 0.71 0.74

0.81 0.25 0.37 0.12

75.2 84.9 86.3 88.4

75.6 84.4 86.1 87.1

75.4 84.7 86.3 88.3

0.65 0.46 0.50 0.51

0.91 0.79 0.96 0.16

80.0 88.8 91.0 93.8

79.7 89.8 90.9 92.6

80.8 89.5 90.9 93.8

0.61 0.59 0.80 0.56

0.41 0.43 0.99 0.21

10.9 11.2 11.5 11.7

10.9 11.3 11.4 11.5

10.9 11.5 11.5 11.8

0.10 0.10 0.09 0.10

0.91 0.22 0.58 0.10

PSA1

PSB2

TS3

41.4 64.4 68.9 79.2ab

41.1 62.2 66.0 74.8b

71.9 82.0 83.3 84.8

Different superscripts within a row indicate significant difference (P < 0.05). 1 PSA: pelleted starter with same ingredient formula as TS (n = 21). 2 TS: texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface (n = 21). 3 PSB: commercially available pelleted starter with a different formulation than TS or PSA (n = 21).

reduced intake and ADG. Ingredients containing fat such as distillers dried grains had lower digestibility, intake, and ADG (Suar­ ez-Mena et al., 2011) as young calves are sensitive to dietary fat perhaps because they do not yet have a resident protozoa population, and because rumen function is not yet fully developed. Based on data of previous studies (Porter et al., 2007; Shiasi et al., 2015), greater intake of TS was expected. This might have been due to surface dressing of TS starter with molasses which was observed to attract flies which calves’ behaviour indicated they did not like. Lesmeister and Heinrichs (2005) found that increasing molasses from 5 to 12 % in a texturized starter reduced intake and ADG during the first 6 wk when calves had been abruptly weaned at the end of 4 wk. Bateman et al. (2009) found (in Trial 2) that when a texturized starter was mixed with a pelleted starter of the same formulation at equal ratio, starter intake and ADG decreased more in the second 28 d than in the first 28 d. In both trials (Bateman et al., 2009), they found no differences in intake and ADG when corn in the starter was whole, flaked, or rolled. Likewise, Lesmeister and Heinrichs (2004) found no differences in intake and ADG when corn in the starter was steam-flaked, rolled, roasted and rolled, or whole before abrupt weaning after 28 d; however, in the following 2 wk, intake and ADG were reduced by steam-flaked and roasted rolled versus whole and rolled in the starter. In our study, no differences were found in ADG among the 3 treatments from 0 to 49 d, which is consistent with the results of Franklin et al. (2003). It was reported by Yavuz et al. (2015) that starter containing whole maize grain and pelleted starter had similar intakes, performance and health status as compared to those of calves fed starters with coarsely ground maize. When starters contained similar ingredient and nutrient contents, manufacturing processes did not affect calf performance unless the diet contained a significant amount of fine particles, which reduced intake and ADG (Bateman et al., 2009). Differences in starch granule size, shape, and interactions between amylose and surface compounds can also alter the rate of enzymatic digestion of corn and wheat starches (Nocek and Tamminga, 1991; Kotarski et al., 1992). The PSA starter may have been expected to result in lower dry feed consumption compared with TS starter, but because final BW was similar in PSA and TS calves in this study, feed efficiency of calves consuming PSA was numerically lower. Dry matter intake is related with rumen development, but physical form of starter can affect rumen development because of its effects on fermentation. Rumen passage rate of solid phase may be different because of physical form (Pazoki et al., 2017). Physical form of starter may affect taste, dustiness and palatability, all of which can affect DMI and digestion. Calves generally do not like finely ground (meal) feeds, which usually decrease palatability and intake (Bateman et al., 2009). During the 63-d trial, the PSB-fed calves had the lowest ADG, mainly because of reduced DMI.

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Table 4 Effects of different physical forms of starter on rumen fermentation parameters. Item Age calves starting Rumination, d Ruminal pH d 49 d 63 NH3-N, mg/dl d 49 d 63 Total VFA, mmol/L d 49 d 63 Individual VFA, mmol/L Acetate d 49 d 63 Propionate, mmol/L d 49 d 63 Butyrate, mmol/L d 49 d 63 Isobutyrate, mmol/L d 49 d 63 Valerate, mmol/L d 49 d 63 Isovalerate, mmol/L d 49 d 63 Acetate:propionate d 49 d 63

Treatment

SEM

P-value

21.9

1.20

0.54

6.09 5.65

6.06 5.65

0.126 0.085

0.83 0.99

19.5 26.4

20.6 24.6

20.4 22.5

1.90 1.64

0.91 0.24

78.1 88.8

79.5 87.3

80.9 82.9

4.92 5.10

0.92 0.70

38.0 41.1

41.4 39.4

40.0 39.3

2.21 2.53

0.55 0.86

30.4 37.0

27.5 34.2

31.4 33.4

2.29 2.25

0.45 0.55

5.79 6.36b

7.16 9.22a

5.66 6.08b

0.679 0.664

0.23 <0.01

0.61 0.81

0.72 0.88

0.66 0.74

0.053 0.060

0.34 0.28

2.54 2.50

1.87 2.39

2.36 2.14

0.236 0.159

0.12 0.25

0.76 1.02

0.89 1.14

0.79 0.93

0.081 0.077

0.49 0.17

1.30b 1.11

1.63a 1.16

1.32b 1.13

0.073 0.032

<0.01 0.61

PSA1

PSB2

TS3

23.6

23.4

5.99 5.64

Different superscripts within a row indicate significant difference (P < 0.05). 1 PSA: pelleted starter with same ingredient formula as TS (n = 21). 2 PSB: commercially available pelleted starter with a different formulation than TS or PSA (n = 21). 3 TS: texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface (n = 21). Table 5 Effects of different physical forms of starter on blood parameters. Items Glucose, mg/dL d 42 d 49 d 63 IgG, ug/mL d 42 d 49 d 63 Growth hormone, ug/L d 42 d 49 d 63 Insulin, mIU/L d 42 d 49 d 63 1 2 3

Treatment

SEM

P-value

97.2 93.6 90.0

2.52 2.16 2.52

0.82 0.77 0.35

84.0 82.4 88.5

80.1 80.7 84.5

3.75 3.88 4.13

0.47 0.43 0.22

40.4 38.2 38.2

40.9 38.8 36.9

40.5 39.9 37.1

1.89 1.97 1.85

0.98 0.83 0.86

47.9 50.0 54.3

45.3 43.2 47.1

44.5 48.1 50.8

2.36 2.19 2.03

0.58 0.09 0.06

PSA1

PSB2

TS3

97.2 93.6 93.6

99.0 91.8 90.0

86.6 75.6 78.3

PSA: pelleted starter with same ingredient formula as TS (n = 21). PSB: commercially available pelleted starter with a different formulation than TS or PSA (n = 21). TS: texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface (n = 21).

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Table 6 Effects of different physical forms of starters on nutrient digestibility of male calves determined by indigestible NDF(iNDF) and acid insoluble ash (AIA). Treatment Digestibility iNDF, g/kg DM CP Fat NDF ADF Digestibility AIA, g/kg DM CP Fat NDF ADF

SEM

P-value

656b 639b 640 497 657b

16.7 24.0 59.5 21.5 16.7

<0.01 <0.01 0.85 0.42 <0.01

502b 478b 485 274 512b

37.3 44.4 80.2 46.9 22.3

0.04 0.02 0.50 0.29 <0.01

PSA1

PSB2

TS3

718ab 736a 622 521 764a

762a 784a 592 539 721ab

636.9a 659.5a 521.5 383.2 696a

645a 678a 388 312 545b

Different superscripts within a row indicate significant difference (P < 0.05). 1 PSA: pelleted starter with same ingredient formula as TS (n = 4). 2 PSB: commercially available pelleted starter with a different formulation than TS or PSA (n = 4). 3 TS: texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface (n = 4).

Fig. 2. Prevalence of diarrhea within the first 3 wk of age. TS = texturized starter, mixture of a pellet, cracked corn and whole oats, and with 5% molasses on the surface, n = 21; PSA = pelleted starter with the same ingredient formula as TS, n = 21; (3) PSB: commercially available pelleted starter with a different formulation than TS or PSA, n = 21. Percentage of calves with diarrhea.

4.2. Effect of physical forms of starter on ruminal fermentation In our experiment, calves fed TS had earlier initiating rumination time than other calves, this indicates that their rumen function may have been more developed than calves from other starter treatments. Each group started ruminating after d 20 of age, which is similar to previous studies (Ghassemi Nejad et al., 2013; Porter et al., 2007). In contrast to the rumen variables observed in the present study, Porter et al. (2007) found numerically higher rumen pH, papillae length, acetate: propionate ratio, and molar percentage of butyric acid at 8 wk of age for texturized than for pelleted starters, but only 4 calves per treatment were evaluated. Another experiment conducted by Lesmeister and Heinrichs (2004) found that the whole corn treatment had higher ruminal pH at 6 wk than dry-rolled, roasted-rolled, and steam-flaked corn treatments. Greater molar proportion of butyrate in the rumen of PSB calves on day 63 may have been due to negative effects on DMI as noted above. Also, as noted by Hill et al. (2015), unsaturated fat in roasted soybeans of PSB reduced DMI, ADG, and digestibility more than tallow, especially after weaning. These negative effects may well be due to altered rumen fermentation. Mirzaei et al. (2016) found that a texturized starter had greater total rumen VFA production than a mash form of starter. The similar VFA production among the treatments in the present study was in contrast to the finding of Gimeno et al. (2015) who reported that pelleted starter resulted in lower acetate: propionate ratio than ground starter. This may be attributed to the greater degree of gelatinization in pelleted starters. While the concentration of butyrate and acetate: propionate ratio was the greatest in this group, though DMI was the lowest in PSB calves, greater acetate production would result in lesser energy utilization because of hydrogen loss during fermentation (Ungerfeld, 2020). Fine grinding increases surface area of grain, increasing rate of fermentation and 8

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potential attachment sites for microbes or enzymes (Ribeiro et al., 2018), Thus, acetate:propionate ratio was less for PSB calves, which was also found by Bateman et al. (2009). Butyrate can stimulate the development of rumen papillae (Gorka et al., 2009), and the PSB calves had the greatest ruminal butyrate concentration, but the lesser DMI limited the growth of calves in this treatment. 4.3. Effect of the physical form of starter on blood serum metabolites Lower insulin concentration in blood serum of PSB than PSA and TS at d 49 and d 63 could be because PSA-fed calves had greater DMI than the PSB-fed calves, and this could indicate that more starch had been digested in the small intestine, mainly due to dif­ ferences in physical form (O’Dea et al., 1980). Greater rumen propionate concentration and greater blood glucose concentration often results in greater blood insulin concentration (Oh et al., 2015; Trefz et al., 2017). However, rumen propionate concentration and blood glucose during weaning (d 49) and after weaning (d 63) were not different in this trial. This may be because rumen propionate and blood glucose in this experiment were within the normal range of insulin. Also, rumen samples and blood samples were taken before morning feeding. Lower DM intake before morning feeding will not have much of an impact on rumen propionate and blood glucose concentrations. Serum IgG protects calves from infectious disease. Calves increase serum IgG levels after 3 wk of age when they produce their own IgG (Stilwell and Carvalho, 2011). High levels of blood total immunoglobulin have been found in high protein fed calves (Sharma et al., 2020). In this study, starters had no effects on blood IgG concentration because of similar chemical composition. Growth hormone axis has been found to be a marker of nutritional status and growth performance (P´erez-S´ anchez and Le Bail, 1999). ´nchez et al., Ration intake and protein intake affected circulating growth hormone concentration in gilthead sea bream (P´ erez-Sa 1995). In our study, TS fed calves had greater starter intake than PSB fed calves after weaning, but had similar blood growth hormone concentration. 4.4. Effect of the physical form of starter on digestibility of nutrients The two analysis methods (acid insoluble ash and indigestible NDF) produced similar digestibility data differences in this study. Digestibility of DM, CP, and ADF were greater for PSB calves than TS calves. This may be due to corn being very hard, reducing TS digestibility. It was reported that DM digestibility was greater for texturized than pelleted starters (Porter et al., 2007). It is also possible that calves in TS group could have disproportionately eaten the cracked corn, which would have reduced overall digestibility as these calves were observed having visible particles of corn in their feces. The PSA calves had greater ADF digestibility than PSB and TS fed calves, which coincided with numerically greater propionate and lower acetate:propionate ratio. This may be due to corn being very hard, reducing TS digestibility. 5. Conclusion Our results confirmed that physical form of starters can influence digestibility of starter, starter intake and growth. Texturized starter fed calves had greater starter intake after weaning but lower digestibility due to hard, flinty corn which was in cracked form in the texturized starter versus ground in pelleted starters. Also, texturized starter aided calf rumen development by prompting rumi­ nation behaviour 2 d earlier than the pelleted starter fed calves. Soybean and distillers dried grains ingredients in pelleted starter decreased DMI and ADG because of their fat content and level. Ethics statement All animal care and procedures were approved by the Animal Welfare and Ethical Committee of Institute of Animal Science, Chinese Academy of Agricultural Sciences (No. IAS20180115). CRediT authorship contribution statement C. Du: Conceptualization, Methodology, Formal analysis, Investigation, Writing - original draft. L. Ma: Writing - review & editing. Y.G. Zhen: Investigation, Software. A.F. Kertz: Writing - review & editing. W.J. Zhang: Writing - review & editing. D.P. Bu: Project administration, Resources, Supervision, Validation. Declaration of Competing Interest The authors declare no conflicts of interest. Acknowledgements This research was supported by National Key Research and Development Program of China (2017YFD0500502, 2018YFD0501600) The Agriculture Science and Technology Innovation Program (ASTIP-IAS07-1, CAAS-XTCX2016011-01), Beijing Dairy Industry Innovation Team (BAIC06-2020). Calf starters were produced and donated for the trial by Apichat Wantanasak of Chia Tai Group, Beijing, China. Software and data repository resources. Data are not deposited in an official repository. 9

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