4.1. Growth Performance
According to the results of our study, the PLS treatment had the lowest weaning BW and ADG compared with the other treatments (PSS, PWCSS and PWCLS). This finding could be partly explained by the lower starter feed intake for the PLS treatment than the PSS and PWCSS treatments, as the starter feed intake has been positively correlated with ADG in pre-weaning calves [
6,
22,
26]. However, although the PLS and PWCLS treatments had nearly identical feed intake, weaning BW and ADG remained lower in calves receiving the PLS treatment compared with the PWCLS treatment. This was likely attributable to reduced nutrient digestibility in the PLS treatment relative to the PWCLS treatment.
In dairy calf feeding, supplementation of forage to the starter feed has been a common practice. However, data are limited about how different particle sizes of forage affect calf performance in free-choice provision or the TMR feeding regimen [
8,
35,
36]. In the current study, weaning BW, ADG and starter feed intake were affected by the particle size of straw, regardless of physical forms of corn in starter diets. Omidi-Mirzaei et al. [
6] investigated the interactive effects of the forage source and forage particle size as a free-choice provision on the growth performance of calves fed texturized starters. In contrast to our results, Omidi-Mirzaei et al. [
6] reported that the particle size of forage did not affect weaning BW, ADG and total feed intake (forage intake and starter intake) during the pre-weaning period, which could be due to the differences in the forage provision method and the particle sizes of utilized forage between ours and that study [
6]. Also, Bagheri et al. [
8] observed that weaning BW, ADG, starter intake and forage intake were not influenced by the straw particle size as a free-choice provision to calves during the pre-weaning period. Similar to the previous studies mentioned above [
6,
8], feed efficiency was not affected by the straw particle size in our study, regardless of physical forms of corn in starter diets. Montoro et al. [
37] investigated the effects of 10% coarsely chopped grass hay or 10% finely ground grass hay in TMR containing 90% crumb starter concentrate on the performance and apparent digestibility of young calves from 2 to 56 days of age. In that study [
37], there was a tendency for improved feed efficiency in the calves fed chopped hay compared with those fed ground hay. Montoro et al. [
37] reported that the total tract apparent digestibility of dry matter, crude protein, NDF and ADF during the week after weaning were greater in calves offered the diet including chopped hay than in those fed the diet with ground hay. This result could explain the tendency for an improved feed efficiency in calves fed TMR with chopped hay. According to Montoro et al. [
37], the particle size of forage might play an important role in nutrient digestibility. Our result regarding feed efficiency was contrary to that of Montoro et al. [
37] when we evaluated the effects of the particle size of straw in the starter diets, regardless of corn physical form. These conflicting results were probably due to the differences in the type and particle sizes of utilized forages and forms (crumb starter vs. pelleted or textured starter) of utilized starter concentrates between ours and that study [
37].
It has been demonstrated that the physical form of the starter feeds—pelleted, texturized or ground [
5,
13,
38]—and processing methods of cereal grains used in starter diets [
13,
22,
24] can affect the starter intake and/or growth performance in young calves. In our study, weaning BW and ADG were affected by the physical form of corn despite there being no effect of ground versus whole corn on the starter feed intake, regardless of the forage particle size in TMR. This result could explain the effect of the physical form of corn on feed efficiency. Gholizadeh et al. [
26] investigated the interactive effects of forage provision (0 or 10% of chopped straw) and the physical form of corn (ground or whole) in the starter diets (mashed starter compared to a texturized starter containing whole grain) on growth performance of young calves. Contrary to our study, Gholizadeh et al. [
26] reported that weaning BW, ADG and feed efficiency during the pre-weaning period were not influenced by physical form of corn in the starter diets, regardless of forage provision. In addition, Gholizadeh et al. [
26] found that the starter feed intake was higher for the calves offered the diet with ground corn than those offered the diet with whole corn during the pre-weaning period, when the calves were fed TMR containing 10% of chopped wheat straw. These conflicting results between ours and that study [
26] were probably due to the differences in the particle sizes of straw used in starter diets because different particle sizes of forage could affect the growth performance and/or starter feed intake of young calves [
13,
18,
19]. The particle sizes of forage in our TMRs with both short and long particle sizes of straw were longer than those in the study conducted by Gholizadeh et al. [
26]. In contrast to our results, Kamyab-Fard et al. [
25] indicated that ADG and feed efficiency were similar for the calves fed the diets with or without whole corn when comparing forage-free starter diets containing 60% ground corn versus 40.2% ground corn and 19.8% whole corn. This discrepancy between ours and that study [
25] was probably due to the differences in the forage provision and/or form and composition of starter diets, such as amount of corn and the particle size distribution.
In our study, weaning wither height and the change in wither height were lower for the calves fed PLS than those fed PSS, PWCSS or PWCLS, and were similar for PSS, PWCSS and PWCLS. Our results related to wither height were congruent with the findings of ADG and weaning BW. Rincker et al. [
39] found that wither height at weaning was positively correlated with weaning BW and ADG during the pre-weaning period in Holstein female calves. In the diets including short straw, weaning heart girth and heart girth changes were greater for the PWCSS treatment (with whole corn) than the PSS treatment. Similarly, in the diets including long straw, weaning heart girth and heart girth changes were higher in the PWCLS treatment (with whole corn) than the PLS treatment. Similar results were observed for weaning BW. Silva et al. [
40] indicated that body weight could be reliably estimated using heart girth measurements in pre-weaned Holstein–Friesian calves aged 1 to 90 days, which showed the relationship between body weight and heart girth.
4.2. Ruminal pH and Volatile Fatty Acids
In the current study, the mean ruminal pH at 68 days of age ranged from 5.76 to 6.06 in neonatal calves and did not differ among the treatments. Supplementation of forage to the starter feeds prevents decreases in ruminal pH and ruminal acidosis in young calves [
5,
8,
10,
16]. Similar to studies in mature ruminants, changes in the particle size of forage have been mainly investigated as one of the most known physical characteristics of forage to affect the ruminal pH of young calves. Nemati et al. [
14] observed that ruminal pH was affected by the particle size of alfalfa hay mixed with finely ground concentrates, regardless of the level of forage (12.5 or 25%) in TMR, and ruminal pH was higher for the calves offered a medium particle size of forage than those offered a fine particle size of forage. In contrast with the results of Nemati et al. [
14], in our study, ruminal pH was not influenced by the particle size of straw, which was probably due to the differences in the level of utilized forages and the form of starter concentrates between ours and that study [
14]. Similar to the result of our study, Omidi-Mirzaei et al. [
6] reported that the particle size of forage as a free-choice provision did not affect ruminal pH values in dairy calves. In addition, Bagheri et al. [
8] also found that ruminal pH at weaning was not influenced by different particle sizes of wheat straw provided as a free-choice provision to dairy calves. Under the conditions of our experimental design, differences in the particle size distribution of wheat straw between treatments had no effects on ruminal pH at weaning, which might be related to the greater chewing activity in calves compared to cows, and consequently a decrease in the particle size of forage entering the rumen. Also, the calves fed the starter diets including different particle sizes of straw might have sorted for similar particle sizes of straw, thereby showing similar ruminal pH values [
8].
Whole corn grain can lead to increased particle sizes in the starter diet and probably result in better rumen health by preventing decreases in the ruminal pH of young calves. Therefore, whole corn grain inclusion in calf starter diets can be considered as a potential alternative of forage feeding to modulate ruminal pH and improve rumen health [
25,
28]. Kamyab-Fard et al. [
25] observed that ruminal pH was influenced by different levels of whole corn in starter feed and increasing the amount of whole corn replaced with ground corn led to increased ruminal pH in young calves fed forage-free starter feeds. In our study, the presence of a forage source in experimental starter diets may have caused the lack of effect of whole corn on ruminal pH. Similar to the result of our study, Gholizadeh et al. [
26] reported that ruminal pH values were not affected by different physical forms of corn (ground or whole) in the starter diets mixed with 10% chopped wheat straw in Holstein calves. The high level of milk feeding could lead to lower mean solid feed intake during the pre-weaning period [
26,
41], which may have caused the lack of possible effect of the straw particle size and physical form of corn in starter diets on ruminal pH in our study, where calves were fed 7.5 L/day milk from 15 to 42 days of age. For instance, the particle size of forage did not influence ruminal pH in the calves fed 6 L/day milk from 3 to 44 days of age [
6]. In addition, Gholizadeh et al. [
26] found that ruminal pH was not affected by wheat straw inclusion or corn grain physical form in the calves fed 8 L/day milk from 11 to 45 days of age and 6 L/day milk from 46 to 55 days of age.
In the present study, ruminal acetate concentrations did not differ among the treatments. Molaei et al. [
42] reported that ruminal acetate concentrations were not affected by the physical form (10% medium chopped vs. 10% pelleted) of alfalfa hay mixed with starter concentrate in Holstein calves. In addition, Omidi-Mirzaei et al. [
6] observed that the particle size of forage did not affect ruminal acetate concentrations in dairy calves. Gholizadeh et al. [
26] reported that the ruminal acetate concentrations of dairy calves were not influenced by different physical forms of corn (ground or whole) in the starter diets containing 10% chopped wheat straw. Additionally, Gholizadeh et al. [
26] found that ruminal acetate concentrations were higher for calves fed diets with straw compared to those fed diets without straw when considering forage inclusion (0 or 10% of chopped straw) into the starter concentrate. Forage provision during the pre-weaning period can promote cellulolytic microbial growth and results in increased acetate production in the rumen of calves [
43,
44]. In our study, similar ruminal acetate concentrations for all treatments could be explained by the same level of wheat straw used in the experimental diets.
In the present study, ruminal propionate, butyrate and total VFA concentrations at weaning were affected by the interaction between the straw particle size and corn physical form. In the diets including short straw, ruminal propionate, butyrate and total VFA concentrations were higher for the treatment with whole corn (PWCSS) than the treatment without whole corn (PSS). On the other hand, in the diets including long straw, the ruminal propionate concentration was greater in the calves fed the diet without whole corn (PLS) compared to the calves fed the diet with whole corn (PWCLS), and ruminal butyrate and total VFA concentrations were similar for the PLS and PWCLS treatments. These results suggest that the effect of the physical form of corn (ground or whole) on ruminal propionate, butyrate and total VFA concentrations could vary depending on the particle size of straw in starter diets. This could be due to the differences in the particle size distribution of starter diets, resulting in feed sorting. In contrast to the results of our study, Gholizadeh et al. [
26] reported that ruminal propionate, butyrate and total VFA concentrations of Holstein calves were not influenced by the different physical forms of corn (ground or whole) in the starter diets containing 10% chopped straw with the same particle size. These conflicting results between ours and that study [
26] were probably due to the effect of the forage particle size, as mentioned above. In the study by Kamyab-Fard et al. [
25], when comparing forage-free starter diets containing 60% ground corn versus 40.2% ground corn and 19.8% whole corn, ruminal butyrate and total VFA concentrations were similar for the calves fed the diets with or without whole corn, while there was a tendency for a lower ruminal propionate concentration in the calves offered the diet including whole corn compared to those fed the diet without whole corn. The results of ruminal propionate, butyrate and total VFA concentrations in that study [
25] were similar to our results for the diets including long straw. However, in the diets including short straw, our results for ruminal propionate, butyrate and total VFA concentrations were different from the findings of the study by Kamyab-Fard et al. [
25], and the reasons for the different results could not be explained. The different results for whole corn inclusion in calf starter diets on ruminal VFA concentrations might be related to the forage provision (forage-free or offering forage) and/or the absorption rate of VFAs produced in the rumen of young calves, since the absorption of VFA can be a factor affecting its concentration in the rumen fluid [
25,
45]. In fact, the numerically lower ruminal pH (5.80) for the PWCSS treatment may have negatively influenced the VFA absorption compared with the PSS treatment (6.06). Therefore, higher ruminal propionate, butyrate and total VFA concentrations may have been found in the PWCSS treatment than the PSS treatment in the spot sampling of rumen fluid 4 h after feeding, which was probably due to their slow absorption by the rumen wall [
25,
46]. Similarly, Kamyab-Fard et al. [
25] indicated that the concentrations of ruminal total VFA were increased when ruminal pH values decreased in Holstein calves. In addition, Kitkas et al. [
47] reported that the ruminal propionate concentration was elevated when ruminal pH values decreased in dairy cows.
4.3. Blood BHB and Fecal Score
In our study, the blood BHB concentration at 68 days of age was not influenced by the particle size of straw, while the straw particle size affected the starter feed intake. This was in contrast to previous studies [
48,
49,
50] that reported blood BHB concentration positively correlated with solid feed intake in calves. There was no effect of the physical form of corn in starter diets on both the blood BHB concentration and starter feed intake, regardless of the forage particle size. Similar to our study, Bagheri et al. [
8] reported that the particle size of wheat straw offered as a free-choice provision to calves did not affect the blood BHB concentration during the pre-weaning or post-weaning periods. In contrast to our result, Nemati et al. [
14] found that blood BHB concentrations at 70 days of age were affected by the particle size of forage in the diets offered as a TMR and were greater in calves fed medium compared to fine particle sizes of forage, regardless of the level of forage. These different results regarding the BHB concentration might be due to the fact that calves were weaned earlier, thus the greater solid feed intake in the study conducted by Nemati et al. [
14] compared with our study. Similar to our study, Gholizadeh et al. [
26] found that the physical form of corn in starter diet (ground vs. whole) did not affect the blood BHB concentration during the pre-weaning or post-weaning periods when the calves were fed the TMR containing 10% of chopped wheat straw. In addition, Kamyab-Fard et al. [
25] also reported that the serum BHB concentration was similar for the calves fed the diets with or without whole corn when comparing forage-free starter diets containing 60% ground corn versus 40.2% ground corn and 19.8% whole corn.
Blood BHB is a measure of rumen epithelial metabolic activity and indicates the conversion of ruminal butyrate to β-hydroxy butyrate. Butyrate is absorbed by the ruminal epithelium and oxidized into BHB [
20,
49]. Deelen et al. [
49] reported that blood BHB results were positively associated with ruminal butyrate in dairy calves around weaning. According to the results of our study, there was a consistent relationship between blood BHB and ruminal butyrate concentrations for all treatments except for PWCSS. Although the ruminal butyrate concentration at 68 days of age was higher in the PWCSS treatment than in the other treatments (PSS, PLS and PWCLS), the blood BHB concentration at 68 days of age did not differ among the treatments, which might suggest that increased ruminal butyrate concentration for the PWCSS treatment had a limited effect on rumen epithelium maturation when considering the results of the blood BHB concentration as an indicator of rumen epithelium development. Similar results for feed efficiency between the PWCSS treatment and the other treatments might confirm this suggestion, since rumen papillae development is important for nutrient and VFA absorption [
43,
51].
In the current study, a higher fecal score indicated the formation of softer feces. Mean fecal scores were higher in the calves fed straw with a long particle size than those fed straw with a short particle size. Calves fed the starter diets with whole corn (both PWCSS and PWCLS) had a higher mean fecal score. The highest mean fecal score was observed in the PWCLS treatment. In the present study, the number of calves treated for diarrhea was one for the PSS, PWCSS and PLS treatments and two for the PWCLS treatment. The increase in the mean fecal score for the PWCLS treatment had no clinical importance when considering the number of calves treated for diarrhea. In contrast to our results, Mirzaei et al. [
19] observed that the fecal score was not influenced by the particle size of alfalfa hay in dairy calves during the pre-weaning period, when alfalfa hay was mixed with finely ground concentrates and offered as a TMR. This conflicting result between ours and that study [
19] might be due to the differences in the type and particle sizes of used forage and/or the differences in the level of forage inclusion and the form of concentrates in starter diets. In our study, the mean fecal score was affected by the physical form of corn in starter diets. Contrary to the findings of our study, Gholizadeh et al. [
26] found that the fecal score was not affected by the corn grain physical form (ground or whole) in the starter diets of Holstein calves fed TMR containing 10% of chopped wheat straw during the pre-weaning period. Kamyab-Fard et al. [
25] also reported that the fecal score was not associated with different levels of whole corn in forage-free starter feeds. This discrepancy between the result of our study and that of others [
25,
26] was probably due to the differences in the form and composition of starter diets, such as amount of corn and the particle size distribution.
This study has some limitations due to the relatively short experimental period and the use of specific straw particle sizes and a specific corn type in the experimental diets. Further research with longer durations and/or different straw particle sizes and corn varieties is needed to evaluate and confirm these findings.