The effects of feeding untreated and formic acid-treated colostrum ad libitum on intake and immunoglobulin levels in dairy calves

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1 SHORT COMMUNICATION The effects of feeding untreated and formic acid-treated colostrum ad libitum on intake and immunoglobulin levels in dairy calves L. K. M. Collings 1, K. L. Proudfoot 1, and D. M. Veira 2 1 Animal Welfare Program, Faculty of Land and Food Systems, University of British Columbia, 2357 Main Mall, Vancouver, British Columbia, Canada V6T 1Z4; and 2 Agriculture and Agri-Food Canada, PO Box 1000, Agassiz, British Columbia, Canada V0M 1A0 ( doug.veira@agr.gc.ca). Received 18 May 2010, accepted 20 September Collings, L. K. M., Proudfoot, K. L. and Veira, D. M The effects of feeding untreated and formic acid-treated colostrum ad libitum on intake and immunoglobulin levels in dairy calves. Can. J. Anim. Sci. 91: Newborn dairy bull calves were fed 4 L of colostrum by esophageal feeder or offered untreated or formic acid-treated colostrum ad libitum for 24 h; effects on intake, serum immunoglobulin G (IgG) levels and behavior were measured. The untreated ad libitum group consumed more colostrum (6 vs. 4 L) but had comparable serum IgG levels to the control (27.8 vs mg ml 1 ). Adding formic acid to colostrum successfully reduced total bacteria and coliform counts; however, it reduced 24 h ad libitum consumption. Formic acid is not a practical preservation agent for colostrum offered ad libitum to newborn calves. Key words: Colostrum, dairy calves, formic acid, immunoglobulin, bacteria Collings, L. K. M., Proudfoot, K. L. et Veira, D. M Incidence du colostrum traite ou pas a` l acide formique et servi à satie té sur la prise alimentaire et sur la concentration d immunoglobulines chez les veauxlaitiers. Can. J. Anim. Sci. 91: De jeunes veaux laitiers ont rec u 4 L de colostrum par une sonde œsophagienne ou du colostrum traité ou pas a` l acide formique à satie te pendant 24 h. Les auteurs ont mesure les effets du traitement sur la prise alimentaire, la concentration d IgG dans le sang et le comportement. Le groupe nourri a` satie té de colostrum non traite a absorbé plus de colostrum (6 c. 4 L), mais la concentration d IgG dans le sang était comparable a` celle releve e chez les sujets témoins (27,8 c. 28,1 mg ml 1 ). L addition d acide formique au colostrum réduit la nume ration totale de bacte ries et de coliformes, mais elle diminue la consommation de colostrum servi à satie té pendant 24 h. L acide formique n est pas un agent de conservation pratique pour le colostrum servi a` satie té aux veaux nouveau-ne s. Mots clés: Colostrum, veaux laitiers, acide formique, immunoglobuline, bacte ries There is no placental transfer of immunoglobulins in the bovine; dairy calves are born agammaglobulinemic. It is therefore vital that calves receive an adequate amount of good-quality colostrum within the first 24 h of life (Stott et al. 1979). Calves should consume at least 100 g of immunoglobulin G (IgG) in the first feeding of colostrum. Given the variation in colostrum quality, it is therefore suggested that in the first feeding of colostrum, calves receive at least 1012% of their body weight (Godden 2008). However, 8 to 11% of pre-weaned dairy calves die in the United States, indicating that improvements need to be made to both colostrum and calf management practices (Godden 2008). Although the natural method for a calf to consume colostrum is to allow it to suckle its dam, there are problems with this approach. First, it is not possible to assess the quantity and the immunoglobulin (Ig) content of the colostrum consumed. As a result calves allowed to suckle are at greater risk of a failure of passive transfer, i.e., serum IgG ofb10 mg ml 1 at 24 to 48 h after birth (Besser et al. 1991). Also, allowing dairy calves to suckle exposes them to a greater number of pathogens and thus increases their risk of illness and death (Wells et al. 1996). Dairy calves are typically offered colostrum as a fixed amount by nipple bottle or esophageal feeder soon after birth. In the wild, and in beef systems, calves would stay with their dam and suckle colostrum ad libitum, but because dairy calves are removed at birth they never have the opportunity to consume colostrum in ad libitum quantities or at the time that they choose. A simple system for offering milk ad libitum from a nipple to young calves has been successfully used by Abbreviations: AEA, apparent efficiency of IgG absorption; FA, formic acid; Ig, immunoglobulins; IgG, immunoglobulin G Can. J. Anim. Sci. (2011) 91: 5559 doi: /cjas

2 56 CANADIAN JOURNAL OF ANIMAL SCIENCE Jasper and Weary (2002). We used this system to offer colostrum to calves during the first day of life. Bacteria grow rapidly in untreated colostrum left at room temperature (Stewart et al. 2005). There are indications that bacteria in colostrum may depress Ig absorption (James et al. 1981) and thus colostrum offered ad libitum at ambient temperatures could result in reduced blood Ig levels because of bacterial numbers. Formic acid was added to colostrum to prevent bacterial growth and was compared with untreated colostrum offered ad libitum. The first objective of this trial was to determine the effect of offering colostrum ad libitum on the colostrum intake and Ig absorption of calves during the first day of life. A second objective was to test the effectiveness of formic acid as a means to reduce bacteria in colostrum while maintaining adequate intake and Ig absorption. All animals were managed according to the guidelines set by the Canadian Council on Animal Care (1993). Forty Holstein bull calves were initially screened for use in the experiment of which 29 were assigned to treatments. Eleven calves failed the first step of the experiment, i.e., to drink 1 L of colostrum in 25min, so they were removed from the experiment. Calves were removed from their dams within an hour of birth and moved to an individual calf pen. Calves were randomly assigned to one of three treatment groups. Blocks of three calves, one from each treatment, were offered colostrum from the same batch. To accomplish this, 10 batches of 25L of pooled colostrum from the first milking were accumulated, thoroughly mixed, divided into treatment groups and frozen (188C) for subsequent feeding. The batches of colostrum were prepared for the three treatments as follows: 1. Control, 4 L of untreated colostrum. 2. Untreated colostrum, 10 L. 3. Formic acid (FA) treated colostrum, 10 L of colostrum was acidified using FA (Fisher Scientific A119P-4, 29.3% Laboratory Grade FA), which had been diluted 1:2 with tap water. The FA was added very slowly to the colostrum while it was being mixed until the ph was between 4.3 and 4.4; this prevented precipitation of proteins. The control group of calves (n10) were fed 4 L of warm colostrum (358C) 2 h after birth via an esophageal feeder. Subsequently, the control calves were offered 3 L of milk at the next regular feeding time, which occurred approximately 8 to 16 h later depending on the time of the initial colostrum feeding. In the two other treatments, calves were offered colostrum ad libitum. In one group, calves were offered untreated colostrum ad libitum (n 10); in the other group, calves were offered colostrum treated with FA ad libitum (n9). For both treatments, one and a half hours after birth 1 L of warmed colostrum was weighed into a bucket attached to the outside of the stall. A Peach Teat nipple (FIL Agritech, New York) was secured on the front of each stall and was connected to the bucket with rubber tubing (0.50 m from the nipple to the bottom of bucket). The calf was encouraged to suckle the nipple by helping the calf stand at the nipple and placing its mouth on the nipple until the 1 L was consumed. Calves that did not consume 1 L in 25min were rejected from the experiment (n 11). If the calf finished the first liter, an additional 8 L of colostrum were thawed and warmed. Samples for determining Ig levels and bacterial counts were stored frozen (188C), and the ph was recorded in the stirred colostrum. The 8 L of colostrum were then weighed and the bucket placed outside the stall. The calf was shown the nipple, encouraged to drink for up to 6 min, and then left on its own. Four and a half hours after the first 1 L feeding, the colostrum was weighed, stirred and replaced. For a final time, the calf was shown the nipple and encouraged to drink for up to 6 min. Twenty-four hours after the first 1 L feeding, the bucket was removed from the stall and weighed. A sample of the remaining colostrum was frozen (188C) for IgG and bacterial analysis, and a sample was used to measure ph. A min-max thermometer placed in the calf stall indicated that the ambient temperature ranged from 5to 318C during the trial. All calves were weighed and blood sampled 24 h after the first offer of colostrum, serum was separated and frozen at 188C for subsequent analysis of IgG levels. Serum IgG content was determined using Bethyl Bovine IgG ELISA kits (#E10-118, Bethyl Laboratories, Montgomery, TX) following the kit instructions. The IgG content of the colostrum samples were determined by radial immunodiffusion as described by Chelack et al. (1993). The colostrum samples were analyzed for both total plate count (Duncan et al. 2004) and coliform numbers (Davidson et al. 2004). The behavior of the calves offered colostrum ad libitum was video recorded for 24 h using one camera (Panasonic WV-BP334) and a time lapse video cassette recorder (Panasonic AG-6740). A red light (100 W) was placed beside the camera to provide adequate light for night-time recording. This video was used to record standing time, time at the nipple and visits to the nipple. The calf was considered to be at the nipple when its mouth was within one nose length of the nipple for at least 9 s without pulling away for more than 4 s. Animal was considered the experimental unit (n29). Two 22 contingency tables were used to determine if calving difficulty (hard pull or easy pull/unassisted calving as defined by the farm staff) or treatment affected whether a calf was rejected from the experiment or not. The following differences were tested for the 4.5h and 24 h after the first 1 L feeding of colostrum separately using SAS (version 9.1; SAS institute, Inc. 2003). Control calves were fed a fixed intake of 4 L per day, thus, differences in colostrum intake between control vs. untreated ad libitum were tested using a one-sample t-test (PROC TTEST). Differences in colostrum intake

3 COLLINGS ET AL. * UNTREATED AND FORMIC-ACID-TREATED COLOSTRUM FED AD LIBITUM 57 and colostrum IgG between untreated ad libitum and FA ad libitum groups were tested using PROC GLM. Differences in body weight, IgG intake, serum IgG (only 24 h) and apparent efficiency of absorption (AEA; only 24 h) between the treatments were determined using PROC GLM; contrasts were created for the following comparisons: control vs. untreated ad libitum and untreated ad libitum vs. FA ad libitum. Differences in the log 10 transformed total plate counts between ad libitum-fed calves with and without FA during the 0 h (directly after thaw) and 24 h period were tested using PROC GLM. Differences between the standing time, time at the nipple and frequency of visits to the nipple during the 4.5and 24 h after first feeding between untreated and FA ad libitum groups were tested using PROC GLM. Calves rejected from the experiment (n11; from ad libitum groups only) because they refused to drink 1 L in the allotted time had a 10 times greater odds of having had a difficult calving than an easy/unassisted calving (Fisher s exact P0.05), and three times greater odds of being in the FA-treated group than the non-fa group (i.e., the colostrum they rejected was treated with FA; Fisher s exact P 0.16). The effect of treatment on colostrum intake is shown in Table 1. In the first 4.5h, the control calves consumed more than those offered untreated colostrum ad libitum (P0.004). However, over 24 h this effect was reversed (P0.001). For the first 4.5h there was no difference in colostrum intake between the two ad libitum treatments (P0.96). However, by 24 h, the consumption of the untreated colostrum was greater than the FA-treated colostrum (P0.03). The control group and the calves offered the untreated colostrum had similar serum IgG levels (P0.95). In contrast, the calves consuming the FA-treated colostrum had lower serum IgG levels than those offered the untreated colostrum (P0.04). The AEA of IgG of the control calves was higher than calves offered the untreated colostrum (Table 1; P0.0005). The ad libitum-fed groups had similar AEA (P0.59). Bacterial counts in samples of the colostrum offered to both ad libitum treatments are also shown in Table 1. The total plate count was higher (PB0.0001) in the untreated compared with the FA-treated colostrum at both 0 and 24 h. Over the 24-h feeding period, coliforms increased in the untreated colostrum and in the FAtreated colostrum no coliforms were detected. Results for standing time, visits to the nipple and time at the nipple are shown in Table 2. There were no Table 1. Comparison of methods of offering colostrum to calves on colostrum and IgG intake, serum IgG level, apparent efficiency of absorption (AEA) of IgG and bacterial numbers Contrasts, P Control z (1) SE Ad lib y (2) SE FA ad lib x (3) SE 1 vs. 2 2 vs. 3 n Calf weight (kg) Colostrum IgG (g L 1 ) 4.5h h Colostrum intake (L) 4.5h h IgG intake (g) 4.5h h Serum IgG (mg L 1 ) AEA (%) Colostrum ph 0h B h B Plate count (log 10 ) 0h B h B Coliform (log 10 ) 0h ND w 24 h ND z Calves given colostrum by esophageal feeder. y Calves offered untreated colostrum ad libitum. x Calves offered formic acid treated colostrum ad libitum. w Not detected.

4 58 CANADIAN JOURNAL OF ANIMAL SCIENCE differences (P0.05) in any of the variables between the two ad libitum treatments during either the 4.5or 24 h period. The two most important factors influencing successful passive transfer in the calf are the length of time from birth to first colostrum consumption and the quantity of Ig consumed. Immunoglobulins from colostrum are most effectively absorbed by the calf very early in life (Stott et al. 1979). The control calves fed 4 L by esophageal feeder met the criteria for quick delivery of large amounts of immunoglobulins resulting in excellent blood levels of IgG and provide a good contrast to experimental ad libitum colostrum treatments. The calves offered untreated colostrum ad libitum consumed 6 L during the first 24 h of life. However, adding FA significantly reduced voluntary consumption of colostrum to 4.3 L. Calves visited the nipple a similar number of times and spent the same time at the nipple on both ad libitum treatments, which suggests that acidtreated colostrum was less palatable, resulting in a reduced amount of colostrum being consumed. This is further supported by the large number of calves on the FA treatment compared with the untreated colostrum (8 vs. 3) that were rejected from the experiment for failing to voluntarily drink 1 L in 25min. Feeding acid-treated milk or milk replacer to older calves has not been associated with reduced intake when offered in less than ad libitum quantities (Gu ler et al. 2006). However, when milk was offered ad libitum, both FA and propionic acid depressed milk consumption relative to untreated milk (Bush and Nicholson 1987). There do not appear to be other reports of acidified colostrum being fed to calves immediately after birth. It is possible that the low ph of colostrum offered in this experiment had an adverse effect on palatability for such young calves. Polzin et al. (1977) fed acetic, propionic and formic-acid-treated colostrum in limited quantities to 4-d-old calves and reported that refusals occurred during the first 2 d, but were not subsequently a problem. All calves in this experiment consumed at least 200 g of IgG, except for one calf offered FA colostrum (99.4 g) and one control calf (198.4 g). All calves on the control and untreated colostrum had successful passive transfer of IgG; however, two of the FA colostrum calves had failed passive transfer (IgGB10 mg ml 1 ), which is the result of the amount and time when colostrum was consumed. There are problems with the ad libitum colostrum feeding system as executed in the present trial. Although several calves consumed a large amount of colostrum, there was a lot of variation between individual calves in both ad libitum treatments. The amount of colostrum consumed during the 24 h period ranged from 4.24 to 8.38 L in calves offered the untreated colostrum ad libitum, and 1.27 to 7.09 L for calves on the FA colostrum. Eleven calves failed to drink the first 1 L of colostrum, and were not entered into the study. Lidfors (1996) observed that many calves failed to drink for at least 3 h after birth and had to be given manual help to suckle. Perhaps the criterion used to reject calves used in this experiment was too rigorous and more calves might have been recruited if a 3 or 4 h interval after birth had been used. There was a concern that leaving colostrum unrefrigerated for 24 h would result in excessive growth of bacteria, which could inhibit Ig absorption (James et al. 1981), and thus the FA colostrum treatment was included in the experiment. The initial total plate count in the untreated colostrum increased from at 0 h to after 24 h. The current guidelines suggest that the total plate count in colostrum fed to calves should be less than CFU ml 1 and the coliform count less than CFU ml 1 (McGuirk and Collins 2004). In spite of the much higher levels observed in the untreated group in this experiment, there was more than adequate absorption of IgG to ensure successful passive transfer; whether future health of the calves would be compromised by consuming large numbers of potentially pathogenic bacteria could not be determined in this experiment. Formic acid reduced the total plate count to an average of CFU ml 1 and the coliform to undetectable levels in the colostrum sampled at 24 h. Offering untreated colostrum ad libitum to calves during the first day of life, as done in this trial, had no advantage in ensuring adequate Ig absorption. Table 2. The time spent standing, number of visits to the nipple and time spent at the nipple for calves offered untreated or FA treated colostrum ad libitum for 24 h Ad libitum SE FA ad libitum SE P Standing time min 4.5h min 24 h Time at the nipple min 4.5h min 24 h Visits to the nipple no. 4.5h no. 24 h

5 COLLINGS ET AL. * UNTREATED AND FORMIC-ACID-TREATED COLOSTRUM FED AD LIBITUM 59 Although it provides a more natural way of feeding, the delay in colostrum intake in the ad libitum treatments prevented any potential improvements in serum IgG concentrations. Adding FA to colostrum was effective in suppressing bacterial growth in colostrum, but it had a negative effect on intake and, therefore, cannot be recommended. The authors thank Lori Vickers for her help collecting data in the barn and Audrey Nadalin for her support in the barn and with the laboratory procedures. Besser, T. E., Gay, C. C. and Pritchett, L Comparison of three methods of feeding colostrum to dairy calves. J. Am. Vet. Med. Assoc. 198: Bush, R. S. and Nicholson, J. W. G Effect of two acids and formalin in calves milk on feed consumption and performance. Can. J. Anim. Sci. 67: Canadian Council on Animal Care Guide to the care and use of experimental animals. Vol. 1. E. D. Olfert, B. M. Cross, and A. A. McWilliams, eds. 2nd ed. CCAC, Ottawa, ON. Chelack, B. J., Morley, P. S. and Haines, D. M Evaluation of methods for dehydration of bovine colostrum for total replacement of normal colostrum in calves. Can. Vet. J. 34: Davidson, P. M., Roth, L. A. and Gambrel-Lenarz, S. A Coliform and other indicator bacteria. Pages in H. M. Wehr and J. F. Frank, eds. Standard methods for the examination of dairy products. American Public Health Association, Washington, DC. Duncan, S. E., Yaun, B. R. and Sumner, S. S Microbiological count methods. Pages in H. M. Wehr and J. F. Frank, eds. Standard methods for the examination of dairy products. American Public Health Association, Washington, DC. Godden, S Colostrum management for dairy calves. Vet. Clin. North Am.: Food Anim. Pract. 24: Gu ler, O., Yanar, M., Bayram, B. and Metin, J Performance and health of dairy calves fed limited amounts of acidified milk replacer. S. Afr. J. Anim. Sci. 36: James, R. E., Polan, C. E. and Cummins, K. A Influence of administered indigenous microorganisms on uptake of [iodine-125] gamma-globulin in vivo by intestinal segments of neonatal calves. J. Dairy Sci. 64: Jasper, J. and Weary, D. M Effects of ad libitum milk intake on dairy calves. J. Dairy Sci. 85: Lidfors, L. M Behavioural effects of separating the dairy calf immediately or after 4 days post-partum. Appl. Anim. Behav. Sci. 49: McGuirk, S. M. and Collins, M Managing the production, storage, and delivery of colostrum. Vet. Clin. North Am.: Food Anim. Pract. 20: Polzin, H. W., Otterby, D. E. and Johnson, D. G Responses of calves fed fermented or acidified colostrum. J. Dairy Sci. 60: SAS Institute, Inc SAS/STAT user s guide. V SAS Institute, Inc., Cary, NC. Stewart, S., Godden, S., Bey, R., Rapnicki, P., Fetrow, J., Farnsworth, R., Scanlon, M., Arnold, Y., Clow, L., Mueller, K. and Ferrouillet, C Preventing bacterial contamination and proliferation during the harvest, storage, and feeding of fresh bovine colostrum. J. Dairy Sci. 88: Stott, G. H., Marx, D. B., Menefee, B. E. and Nightengale, G. T Colostral immunoglobulin transfer in calves I. Period of absorption. J. Dairy Sci. 62: Wells, S. J., Dargatz, D. A. and Ott, S. L Factors associated with mortality to 21 days of life in dairy heifers in the United States. Prev. Vet. Med. 29: 919.

6 This article has been cited by: 1. Cynthia G. Todd, Ken E. Leslie, Suzanne T. Millman, Jan M. Sargeant, Heather Migdal, Kathleen Shore, Neil G. Anderson, Trevor J. DeVries Milk Replacer Acidification for Free-Access Feeding: Effects on the Performance and Health of Veal Calves. Open Journal of Animal Sciences 06:03, [Crossref]