key: cord-0823552-68uimhq6
authors: Ort, S. B.; Aragona, K. M.; Chapman, C. E.; Shangraw, E.; Brito, A. F.; Schauff, D. J.; Erickson, P. S.
title: The impact of direct‐fed microbials and enzymes on the health and performance of dairy cows with emphasis on colostrum quality and serum immunoglobulin concentrations in calves
date: 2017-10-13
journal: J Anim Physiol Anim Nutr (Berl)
DOI: 10.1111/jpn.12806
sha: 110db606b7998896273c57e28d2d6601d4d015d2
doc_id: 823552
cord_uid: 68uimhq6

Thirty‐six cows were blocked by calving date and randomly assigned to one of three treatments. Cows were on treatments 3 weeks prepartum through 8 weeks post‐partum. Treatments were as follows: (i) no direct‐fed microbial (DFM) or cellulase and amylase enzymes (C), (ii) 45.4 g/day of DFM (D) or (iii) 45.4 g/day of DFM and 18.2 g/day of enzyme (DE). Total mixed ration fed and refused were measured daily to determine dry matter intake (DMI). Blood samples were taken three times weekly and analysed for β‐hydroxybutyrate, glucose and non‐esterified fatty acids. Body weight (BW) was measured weekly. Colostrum was weighed and analysed for IgA and IgG concentration. Calves were fed 4 L of colostrum within 2 hr of birth. Calf blood samples were taken at 0 and 24 hr for analysis of IgA and IgG concentrations and apparent efficiency of absorption. Milk yield was measured daily and samples collected weekly. Initial BW was different among treatments with D being lesser than C or DE treatments. Body weight, weight gain, efficiency of gain, DMI and blood parameters were unaffected. Treatment did not affect colostrum yield. Ash percentage of colostrum tended to increase with D and DE, while IgA and total solids yield decreased with D. Colostrum fat yield was decreased in D and DE. Treatments did not impact BW, serum IgA and IgG concentrations or apparent efficiency of absorption of calves. Post‐partum BW, DMI, blood parameters, milk production and composition were unaffected by treatment. However, cows on D gained more BW and tended to have greater efficiency of gain compared to those on DE, but were similar to C. Somatic cell scores were greatest for D. Results indicate that DFM and enzyme supplementation did not improve health and performance of dairy cattle during the pre‐ and post‐partum periods under conditions of this study.

and enzymes can impact health and performance of dairy cattle. However, no studies have evaluated the effect of prepartum DFM and enzyme supplementation of cows on colostrum quality and passive transfer of immunoglobulin to calves. Calves are born agammaglobulinaemic because of the synepitheliochorial placenta, which prevents transfer of immunoglobulin in utero (Akers, 2002) .

As a result, cattle have evolved to transfer immunoglobulin via colostrum.

There are contrary results with supplementation of DFM to animals. For example, Al-Saiady (2010) reported that bull calves fed probiotics had greater serum IgG concentrations than those on the control. Studies with poultry have also indicated that DFM can increase IgG and IgM concentrations and/or cells responsible for IgA, IgG and IgM production (Lee, Lillehoj, & Siragusa, 2010) .

However, Marakoudakis et al. (2010) found no impact of DFM on plasma IgA, IgG and IgM concentrations in dairy goats, but immunoglobulin in the mammary secretions was not evaluated. Queszada-Mendoza, Heinrichs, and Jones (2011) found no effect of probiotics on plasma, faecal or salivary IgG concentrations in calves. These differing outcomes suggest that there is potential for DFM to increase IgG content in dairy cattle colostrum, which could impact the health of the calf through more IgG available for absorption, as well as impact the health and production of the cow when fed post-partum.

The objectives of this study were to evaluate the impact of feeding DFM and enzymes (amylase and cellulase) to dairy cows on: (i) the quantity and composition of the colostrum with focus on IgG and IgA;

(ii) calf BW and IgG uptake; (iii) DMI both pre-and post-partum; (iv) blood glucose, BHB and NEFA concentrations; and (v) the quantity and quality of the milk produced during the first 8 weeks of lactation. Our hypothesis was that feeding DFM alone or in combination with enzymes during the transition period would increase colostral IgG and IgA, DMI pre-and post-partum, milk production, and serum concentrations and apparent efficiency of absorption of IgG and IgA in the calf.

This experiment was reviewed and approved by the University of New Hampshire Institutional Animal Care and Use Committee (Protocol #141107).

Thirty-six multiparous Holstein cows were used in this study.

They were grouped into 12 blocks based on expected calving date.

Parity number ranged from 2 to 7. There were 12, 11, seven, three and two cows in their second, third, fourth, fifth and sixth lactation respectively. Within each block, cows were randomly assigned to one of three treatments: (i) 0 g/day DFM and enzymes (C); (ii) 45.4 g/day of DFM product (D); and (iii) 45.4 g/day of D plus 18.2 g/ day of enzyme product (DE) . The DFM (D) contained Enterococcus faecium and Saccharomyces cerevisiae at 1.323 billion cfu/g. The enzyme (E) was a combination of both DFM and enzymes, which contained 0.88 billion cfu/g of E. faecium and S. cerevisiae. The enzyme portion of E contained both amylase and cellulase. The ingredients of D were calcium carbonate, rice hulls, active dry yeast, mineral oil, fructooligosaccharides, dried E. faecium fermentation product, sodium silico aluminate, and natural and artificial flavours. The ingredients of E contained rice hull extract, sodium silico aluminate, dried Trichoderma longibrachiatum fermentation extract, dried Bacillus subtilis fermentation extract, dried Aspergillus niger fermentation extract, torula dried yeast, dried E. faecium fermentation product, Yucca schidigera extract, riboflavin, calcium gluconate, nicotinic acid, biotin, pyridoxine hydrochloride, thiamine mononitrate, vitamin B 12 , citric acid, natural and artificial flavours, and mineral oil. The experimental treatments were being evaluated for combination and for eventual commercial application.

Cows began the study 3 weeks prior to the expected calving date and continued through 8 weeks of lactation. All feed ingredients and nutrient composition are reported in Table 1 . Feed ingredients and nutrient composition of the prepartum, transition totally mixed ration (TMR) and TMR are found in Tables 2, 3, 4 and 5 respectively.

Orts analyses are found in Tables 6, 7 and 8. Nutrient composition of orts indicated that there were similar values and little sorting by treatment. All cows were fed the prepartum TMR prior to calving and then switched to the transition TMR immediately post-partum, which helped them to transition to the post-partum TMR at 2 weeks after calving. All cows were kept in tie stalls with mattresses and bedded with kiln dried sawdust from day −21 to 56. Each cow was fed in separate wooden feed tubs (90 × 90 × 90 cm). Three days prior to the expected calving date, cows were moved to individual maternity pens (4.3 × 3.7 m) until parturition and placenta delivery. At all times, the cows had access to automated water bowls (Delaval, Tumba, Sweden).

Daily DMI was recorded for each cow throughout the study. Samples of TMR and orts were taken each day and then frozen at −20°C until further processing. All samples were then dried at 55°C in forced air and converted to somatic cell score (SCS, DairyOne Cooperative, Inc.).

Body weight was also recorded every Friday from day −21 to 56 for each cow using a platform scale (Cardinal Scale Manufacturing Co., Hooksett, NH, USA). Calving ease was documented at calving with the scores of 1, 2 or 3 for unassisted calving, some assistance easy calving or assisted difficult calving respectively. There were 37 calves born alive on this study and 32 of those calves were born with a calving score of 1 (10 C, 11 D and 11 DE). Three other calves were born with a calving score of 2 (2 C and 1 D), and two calves were born with a score of 3 (1 C and 1 D).

All calves were removed from the cow prior to nursing immediately after calving or as early as possible. After taking the 0-hr blood samples, 4 L of maternal colostrum was fed to the calves via bottle or oesophageal tubing. A total of 13 calves did not receive 4 L of maternal colostrum. Six of these 13 calves did not receive the amount of colostrum intended because of difficulties in bottle feeding and stomach tubing 4 L of colostrum. Another three of these 13 calves did not receive 4 L of colostrum due to the cow producing less than the amount intended. Finally, four calves did not receive any maternal colostrum because of dam being leukosis positive or produced <3 L of colostrum. As a result, these four calves did not have any blood samples taken at 24 hr of age as they received good quality colostrum from other cows not on the study or colostrum replacer.

For all data, differences among treatments were determined using the LSMEANS option for all procedures in SAS 9.4 (2013 

Colostrum concentrations of IgG (g/L) and IgA (g/L), as well as concentrations and yields of total protein, fat, lactose and ash, were analysed where Y ijk = the dependent variable, μ = the overall mean; B i = the random effect of block (I = 1,…12), T j = the effect of treatment (j = C, D, DE), P k = the covariate measure of expected parity (k = 2,…7) and E ijk = the residual error.

Initial urine pH data were analysed using the MIXED procedure in SAS 9.4 according to the following model:

where Y ij = the dependent variable, μ = the overall mean, B i = the ran- 

Calf data for serum IgG, IgA, AEA and BW were analysed using the MIXED procedure of SAS 9.4 according to the following model:

where Y ijk = the dependent variable, μ = the overall mean, B i = the random effect of block (I = 1,…12), T j = the effect of dam treatment (j = C, D, DE), P k = the covariate measure of parity (k = 2,…7) and E ijk = the residual error.

Body weight, BW gain, efficiency of gain, DMI, serum BHBA, serum NEFA and serum glucose were analysed using the REPEATED procedure of SAS 9.4 according to the following model:

where Y ijkl = the dependent variable, μ = the overall mean, B i = the random effect of block (I = 1,…12), T j = the effect of treatment (j = C, D, DE), W k = the effect of week (k = −3, −2, and −1); TW jk = the treatment by week interaction, P l = the covariate measure of parity (k = 1,…6) and E ijkl = the residual error.

Post-partum DMI, milk yield, energy-corrected milk (ECM) yield, milk efficiency, total protein yield (kg), total protein content (%), fat yield (kg), fat content (%), lactose yield (kg), lactose content (%), ash yield (kg) and ash content (%), SCS, serum NEFA, serum BHBA and serum glucose were analysed using the REPEATED procedure of SAS 9.4 according to the following model:

where Y ijkl = the dependent variable, μ = the overall mean, B i = the ran- and E ijkl = the residual error. 

Prepartum cow data are presented in Table 9 . Initial BW was different among treatments where cows supplemented with D weighed less at the start of the study compared to C or DE cows (p = .003). However, there were no differences in average BW or BW gain during the prepartum period. No effect of treatments was found on efficiency of gain during the prepartum period. There was no effect of treatments on DMI of cows 21 days prior to parturition. There was no effect of treatments on prepar- 

Calf results are presented in Table 10 . No treatment effects were observed for calf BW. There was also no impact of treatments on the T A B L E 9 Body weight (BW), dry matter intake (DMI), urine pH, blood metabolites, and colostrum yield and colostrum composition for cows fed control, direct-fed microbial (DFM) (D) or DFM + enzymes (DE) treatments during the 21 days prior to parturition 

Post-partum cow results are presented in Table 11 . No effect of treatments was observed for overall BW. Body weight change was different among treatments with cows on the D treatment showing lesser BW losses than those on the DE treatment (p = .03). Both treatments were similar to cows on C. Dry matter intake during the 8-week post-partum period was not different among treatments. Efficiency of gain tended to be greater (p = .09) for cows on the D treatment in comparison with those on the DE treatment. Treatments did not alter serum glucose, BHBA and NEFA concentrations. Milk and ECM yields were not affected by the D or DE treatments, likely due to similar DMI across treatments. Milk efficiency of cows on the D and DE treatments was similar to those of C cows. There also was no effect of treatments on milk composition, with the exception of SCS. The SCS of cows on the D and DE treatments were not different from cows on the C treatment. However, D was different from the DE treatment (p = .04).

Dry matter intake was similar among treatments during the prepartum period. This coincides with previous studies which found that DFM containing S. cerevisiae and E. faecium fed at either 2 g/day or 90 g/ day during the 21-ays prepartum period had no impact on DMI (Nocek et al., 2003; Oetzel et al., 2007) . Prepartum DMI data in this study do not agree with that of Nocek and Kautz (2006) who found that the supplementation of DFM containing both S. cerevisiae and E. faecium at a rate of 2 g/day increased DMI during the prepartum period. Also, Dann, Drackley, McCoy, Hutjens, and Garrett (2000) found that supplementation of S. cerevisiae culture at 60 g/day increased DMI of prepartum Jersey cows.

Previous studies have not indicated that prepartum supplementation of DFM and enzymes impacts urine pH. This might be due to the fact that both of these additives do not have enough of an anion capacity to bring about a change in the metabolic pH. Our results for glucose and NEFA concentrations are supported by previous studies which found no impact of DFM supplementation on prepartum glucose and NEFA (Nocek & Kautz, 2006; Oetzel et al., 2007) . However, DFM supplementation has been reported to decrease prepartum BHBA which is in contrast to what was reported in this study (Nocek & Kautz, 2006; Oetzel et al., 2007) . Defrain, Hippen, Kalscheur, and Tricarico (2005) also reported increased BHB concentrations for cows supplemented with enzymes which is in contrast to what was reported in this study.

No studies to date have reported the impact of DFM and enzymes on colostrum yield, quality and composition. Colostrum yield was similar among treatments. The lack of impact of the D and DE treatments in this study on colostrum yield is perhaps due to the fact that there was no effect on prepartum DMI. There was a decrease in IgA yield, fat yield and total solids yield of colostrum in cattle fed the D treatment in comparison with those fed either the C or DE treatments. Also, the ash concentration of colostrum increased in cattle fed the D and DE in comparison with those cows fed C. However, it is not fully understood why these results were observed and further research is needed. It can be hypothesized that decreased yields of IgA, fat and total solids may be linked to increased post-partum SCC in cows fed the D treatment.

Specifically, milk components such as IgA, fat and other solids may have been more degraded due to a greater enzymatic activity of the somatic cells or the presence of bacteria within the mammary gland.

There were no treatment effects on calves in this experiment. A previous study showed that calves born from cows supplemented with a 105 mg of Se-yeast/day had greater serum IgG and absorption of IgG than those from cows on the control diet (Hall et al., 2014) . Al-Saiady (2010) observed greater serum IgG concentration and greater 5-week BW in calves supplemented with a probiotic. However, there were no impacts on serum IgG concentrations in calves provided with a prebiotic supplement (Queszada-Mendoza et al., 2011) . In both these studies, calves were over 24 hr old and AEA was not evaluated. Efficiency of gain tended (p = .09) to be greater for cows on the D treatment in comparison with cows on the DE treatment. This confirms that there were more nutrients being partitioned to the body condition of these cows and that cows on the D treatment lost less BW during the post-partum period. Under the conditions of this experiment, feeding enzymes in combination with DFM were not beneficial. The underlying mechanism responsible for this is not known and further research will need to be performed.

Dry matter intake was similar among treatments. This lack of effect of treatments on DMI is in contrast to previous studies with DFM

supplementation. An array of studies with various types and levels of DFM supplementation have been shown to increase DMI during early lactation (Dann et al., 2000; Moallem, Lehrer, Livshitz, Zachut, & Yakoby, 2009; Nocek & Kautz, 2006; Nocek et al., 2003; Oetzel et al., 2007) . However, Schingoethe et al. (2004) found that supplementation of S. cerevisiae culture did not affect DMI of lactating dairy cows.

As for studies with enzyme supplementation, several researchers have shown that there was no impact of enzymes supplemented at various rates on DMI of lactating cows (Reddish & Kung, 2007; Rode et al., 1999; Yang et al., 2000) . This suggests that treatment did not have an impact on the digestibility of the TMR.

Blood metabolite concentrations were similar among treatments. In contrast, Nocek et al. (2003) reported that post-partum concentrations of glucose increased and NEFA decreased with DFM supplementation. Defrain et al. (2005) also reported that enzyme supplementation increased post-partum glucose concentrations.

However, no effect of DFM and enzyme supplementation on postpartum BHB and NEFA concentrations was reported by Defrain et al. (2005) and Oetzel et al. (2007) . Conversely, Nocek and Kautz (2006) reported that DFM increased post-partum BHBA concentrations in transition cows, which is undesirable due to possible development of ketosis. Means in same row with superscripts a, b significantly differ (p < .05).

Efficiency of gain = body weight gain (kg)/dry matter intake (kg); Means in same row with superscripts x, y differ (p < .10).

ECM yield = energy-corrected milk yield; ECM yield = (12.86 × fat kg) + (7.04 × protein kg) + (0.3246 × milk kg).

Milk efficiency = energy-corrected milk yield (kg)/dry matter intake (kg).

Means in same row with superscripts a, b significantly differ (p < .05).

T A B L E 1 1 Post-partum BW, dry matter intake (DMI), blood metabolites, and milk yield and composition for cows fed control, direct-fed microbial (DFM) (D) or DFM + enzymes (DE) treatments during the first 8 weeks of lactation In this study, there was no effect of treatment on milk yield. This is probably due to the fact that there was no effect of treatment on DMI. Previous studies with DFM and enzymes have shown similar results. Dann et al. (2000) and Oetzel et al. (2007) found no difference in milk yield of cows supplemented with or without DFM. In addition, AlZahal, Dionissopoulos, Laarman, Walker, and McBride (2014) found no impact of DFM on milk yield during the first 6 weeks of lactation. Between week 7 and 10, an increase in milk yield of cows supplemented with DFM was observed (AlZahal et al., 2014) . Other studies have found increases in milk yield with DFM supplementation during early lactation (Moallem et al., 2009; Nocek & Kautz, 2006; Nocek et al., 2003; Wohlt et al., 1991) . In addition, it has been shown that enzyme supplementation increased milk production . However, most studies with enzyme supplementation have indicated that it does not impact milk production Holtshausen, Chung, Gerardo-Cuervo, Oba, & Beauchemin, 2011; Reddish & Kung, 2007) .

Our results were corroborated by Schingoethe et al. (2004) who reported no impact on ECM yield with yeast supplementation. Boyd, West, and Bernard (2011) found an increase in ECM with supplementation of DFM containing Lactobacillus acidophilus and Propionibacterium freudenreichii, which was likely due to an increase in the yield of milk components such as fat and protein. However, these bacteria were not present in the DFM supplement used in the current study.

There was no impact of treatment on milk efficiency. This was expected because treatment did not impact ECM yields or DMI. Beauchemin et al. (1999) and Boyd et al. (2011) found no effect of DFM supplementation on milk efficiency as well. In contrast, Schingoethe et al. (2004) did find a 7% increase in milk efficiency with yeast supplementation. In addition, Holtshausen et al. (2011) found that cows in early lactation fed an enzyme product had greater milk production efficiency than those fed the control. However, in the study by Holtshausen et al. (2011) , the enzyme product contained mainly xylanase and endoglucanase activity, whereas the enzyme product used in present study primarily contained amylase and cellulase activity. In this study, there was no impact of treatment on milk components, with the exception of SCS. Our data concur with those of other researchers regarding milk components. Dann et al. (2000) found no impact of DFM on the concentrations of fat, protein, lactose, total solids and MUN, which agree with our results. Moallem et al. (2009) found no impact on milk fat and protein concentrations between cattle supplemented with DFM or without DFM. Nocek and Kautz (2006) found no impact of DFM supplementation on milk fat and protein yields, as well as milk protein concentration. A similar response has been observed for enzyme supplementation (Reddish & Kung, 2007 ). Oetzel et al. (2007 found that DFM supplementation only increased milk fat concentration for cows in their first lactation. Dann et al. (2000) did not find any impact of DFM supplementation on SCC. However, SCS was reduced in the milk of cows when the enzymes were fed in combination with the DFM alone, but similar to control cows. These data suggest that adding cellulase and amylase to the diet of a cow fed DFM reduces SCS. However, Reddish and Kung (2007) reported no impact of enzyme supplementation containing cellulase and xylanase activity on SCC. As a result, it can be concluded that there is some other underlying factor contributing to the differences between treatments and further research would need to be performed to determine these discrepancies.

Data from this experiment show no benefits of supplementing DFM or enzymes or a combination of both to pre-or post-partum dairy cows. As a result, it can be concluded that DFM and enzyme supplementation was not beneficial for improving the health and performance of dairy cattle during the transition period and early lactation under the conditions of the present experiment.

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The authors thank Agri-King, Inc., Fulton, IL, USA, for support of this study. The authors also thank the farm staff of the Fairchild Dairy