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Evaluation of potential biomarkers to determine adequate colostrum provision in male dairy-beef calves upon arrival at the rearing facility beyond 14 days of age

Open AccessPublished:November 21, 2022DOI:https://doi.org/10.3168/jds.2022-22233

      ABSTRACT

      Colostrum consumption is crucial for passive immunization and development of the newborn calf. However, the incidence on failed transfer of passive immunity in male calves destined to dairy-beef production remains high to date. In addition, the lack of an automated procedure to validate the immunization status upon arrival at rearing facilities in calves beyond 14 d of age impedes the identification of failed transfer of passive immunity, and therefore, of those calves at high risk of suffering diseases. For this study, 82 newborn male Holstein calves (43.3 ± 0.86 kg of body weight; mean ± standard error) from a commercial dairy farm were used to investigate potential serum biomarkers of colostrum provision. The potential biomarkers selected were IgG, IgG1, cholesterol, alkaline phosphatase, gamma-glutamyl transferase (GGT), and total protein (TP). Treatments were as follows: high-colostrum (HC; n = 49), in which calves received 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth, for a total of 10 L of colostrum; and low-colostrum (LC; n = 33), in which calves received only 2 L of colostrum within the first 2 h after birth. After colostrum consumption, calves were allocated to individual hutches and fed 2 L of milk replacer twice daily at a concentration of 125 g/L as fed. Starter feed and water were offered ad libitum. At approximately 14 d of age (14.2 ± 0.81 d of age; mean ± standard error) calves were transported 2.5 h to a research unit at IRTA (Torre Marimon, Spain) simulating the arrival to a rearing facility. Blood samples were collected before feeding at birth, 48 h after birth, and at arrival to the rearing facility. Results on the serum concentrations of the potential biomarkers at arrival to the rearing facility showed that IgG, IgG1, GGT, and TP were greater for the HC calves compared with the LC calves. Serum concentrations of cholesterol and alkaline phosphatase did not show differences between treatment groups. Additionally, body weight losses from birth until arrival to the rearing facility were greater for the LC treatment compared with the HC. Because of their low cost, quickness, and ease of measurement, GGT and TP were good indicators of colostrum intake in calves arriving at rearing facilities beyond 14 d of age.

      Key words

      INTRODUCTION

      Male Holstein calves are considered by-products, or even waste products, from the dairy industry, and their postnatal care is not always a priority for producers (
      • Devant M.
      • Marti S.
      Strategies for feeding unweaned dairy beef cattle to improve their health.
      ). It has been estimated that, within a range of 12 to 43%, male calves entering the dairy-beef industry suffer failed transfer of passive immunity (FTPI;
      • Pardon B.
      • Alliët J.
      • Boone R.
      • Roelandt S.
      • Valgaeren B.
      • Deprez P.
      Prediction of respiratory disease and diarrhea in veal calves based on immunoglobulin levels and the serostatus for respiratory pathogens measured at arrival.
      ;
      • Wilson D.J.
      • Stojkov J.
      • Renaud D.L.
      • Fraser D.
      Risk factors for poor health outcomes for male dairy calves undergoing transportation in western Canada.
      ). Even though the importance of colostrum provision in newborn calves is globally understood, FTPI continues to be of great concern in both the veal and the dairy-beef production systems. Accordingly, previous studies have demonstrated that female calves receive higher volumes of less contaminated (average bacterial counts) colostrum compared with male calves (
      • Fecteau G.
      • Baillargeon P.
      • Higgins R.
      • Pare J.
      • Fortin M.
      Bacterial contamination of colostrum fed to newborn calves in Québec dairy herds.
      ) which, in some cases, do not receive colostrum at all (
      • Renaud D.L.
      • Duffield T.F.
      • LeBlanc S.J.
      • Haley D.B.
      • Kelton D.F.
      Management practices for male calves on Canadian dairy farms.
      ). Colostrum consumption is crucial for the correct development of the immune system by the ingestion of immunoglobulins, particularly IgG (
      • Godden S.
      Colostrum management for dairy calves.
      ) because newborn calves lack immunocompetence at birth (
      • Hulbert L.E.
      • Moisá S.J.
      Stress, immunity, and the management of calves.
      ). Additionally, nutrient and bioactive compounds found in the colostrum have also been shown to modulate the development and function of the gastrointestinal tract of newborn calves (
      • Blum J.W.
      Nutritional physiology of neonatal calves.
      ). The lack of information on the immunological background and health status of these male calves when they arrive at the rearing facilities reveals the large disconnection between the dairy and dairy-beef production industries. Also, the public is increasingly concerned about the health and welfare of male calves being marketed to the meat industry. Although the European Union has addressed some of these concerns by establishing minimal standards for the protection of calves reared for fattening, including housing, care, and nutrition (
      • European Union
      Council directive: Laying down minimum standards for protection of calves.
      ), other standards, such as fitness of young calves for transport, are still poorly described (Regulation 1/2005).
      Assessing FTPI in young calves at arrival to rearing facilities, when calves are normally older than 14 d of age, would provide important evidence on colostrum provision and, consequently, this information could be used to improve vaccination protocols, management, and nutrition considering calves' individual vulnerability, which in turn would affect antibiotics use. Several techniques are available for measuring FTPI; however, no one has developed a standardized method. Measurements of serum IgG concentration via radial immunodiffusion is considered the reference test (
      • Beam A.L.
      • Lombard J.E.
      • Kopral C.A.
      • Garber L.P.
      • Winter A.L.
      • Hicks J.A.
      • Schlater J.L.
      Prevalence of failure of passive transfer of immunity in newborn heifer calves and associated management practices on US dairy operations.
      ). The IgG concentration, especially IgG1 (the primary immunoglobulin in bovine colostrum) is a good indicator of FTPI but controversy exists regarding which value ranges should be applied in calves that are a few days old as well as the age up to which those ranges should apply (
      • Chigerwe M.
      • Hagey J.V.
      • Aly S.S.
      Determination of neonatal serum immunoglobulin G concentrations associated with mortality during the first 4 months of life in dairy heifer calves.
      ;
      • Pardon B.
      • Alliët J.
      • Boone R.
      • Roelandt S.
      • Valgaeren B.
      • Deprez P.
      Prediction of respiratory disease and diarrhea in veal calves based on immunoglobulin levels and the serostatus for respiratory pathogens measured at arrival.
      ;
      • Weaver D.M.
      • Tyler J.W.
      • Scott M.A.
      • Wallace L.M.
      • Marion R.S.
      • Holle J.M.
      Passive transfer of colostral immunoglobulin G in neonatal llamas and alpacas.
      ). However, samples for radial immunodiffusion must be sent to a referral laboratory, which delays diagnosis and increases costs. Using a refractometer is considered to be more practical because of the immediate responses and the lower cost. This technique indirectly measures total protein (TP) concentration, which correlates well with the concentration of IgG (
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      ;
      • Wilm J.
      • Costa J.H.C.
      • Neave H.W.
      • Weary D.M.
      • von Keyserlingk M.A.G.
      Technical note: Serum total protein and immunoglobulin G concentrations in neonatal dairy calves over the first 10 days of age.
      ). Concentration of maternal IgG has a half-life of approximately 10 d after colostrum provision (
      • Hassig M.
      • Stadler T.
      • Lutz H.
      Transition from maternal to endogenous antibodies in newborn calves.
      ), and previous research has shown than TP concentration measured up to a week (
      • Villarroel A.
      • Miller T.B.
      • Johnson E.D.
      • Noyes K.R.
      • Ward J.K.
      Factors Affecting Serum Total Protein and Immunoglobulin G Concentration in Replacement Dairy Calves.
      ) or 9 d (
      • Wilm J.
      • Costa J.H.C.
      • Neave H.W.
      • Weary D.M.
      • von Keyserlingk M.A.G.
      Technical note: Serum total protein and immunoglobulin G concentrations in neonatal dairy calves over the first 10 days of age.
      ) after colostrum consumption can be a good indicator of FPTI. However, the search for a biomarker able to detect colostrum intake in older calves remains necessary. Other serum biomarkers related to colostrum intake in preweaned calves are the plasma enzyme gamma-glutamyl transferase (GGT;
      • Weaver D.M.
      • Tyler J.W.
      • Scott M.A.
      • Wallace L.M.
      • Marion R.S.
      • Holle J.M.
      Passive transfer of colostral immunoglobulin G in neonatal llamas and alpacas.
      ), alkaline phosphatase (ALP;
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ), and cholesterol (CHOL;
      • Renaud D.L.
      • Duffield T.F.
      • LeBlanc S.J.
      • Ferguson S.
      • Haley D.B.
      • Kelton D.F.
      Risk factors associated with mortality at a milk fed veal calf facility: A prospective cohort study.
      ). Therefore, the objective of this study was to find a biomarker indicative of colostrum provision upon arrival at the rearing facility when calves are older than 14 d of age. This biomarker should potentially be economical and rapidly measurable to be practical at the farm level. We evaluated the use of IgG, IgG1, CHOL, ALP, GGT, and TP as potential biomarkers of colostrum provision.

      MATERIALS AND METHODS

      Animals, Treatments, and Feeding

      All calves used in this study were managed following the principles and guidelines of the Animal Care Committee of Institut de Recerca i Tecnologia Agroalimentàries (Barcelona, Spain; RD 53/2013; project no. 11211). A total of 82 male Holstein calves (43.3 ± 0.86 kg of BW; mean ± SE) born at a commercial dairy farm (Granja Selergan, S.A., Lleida, Spain) in October 2020 [average ambient temperature of 9.3°C (range = 3.1–16.5°C) and 87% humidity (range = 81–100%)] were used in this study. Treatments were as follows: the high-colostrum (HC; n = 49; 42.4 ± 0.94 kg of BW) group received 4 L of colostrum within the first 2 h after birth, and 2 L of colostrum in the next 3 feedings within the first 24 h after birth; in total those calves received 10 L of colostrum (22.9 ± 0.30% of birth BW). The low-colostrum (LC; n = 33; 44.1 ± 0.77 kg of BW) group received only 2 L of colostrum within the first 2 h after birth (4.8 ± 0.36% of birth BW). All feedings were administered via esophageal tube. Only high-quality colostrum was used for all calves (average of 24.5% Brix). Colostrum samples of each pool given to the animals (a total of 43 pools) were collected. For the analysis of the composition, an equal quantity from different pools used during the study was mixed to create a single sample that was later analyzed for CP (56.28%), fat (29.48%), and lactose (10.07%). This colostrum pool was also analyzed for GGT (36,910 IU/L), lactoferrin (1,828.84 ng/mL), IgG (145.44 mg/mL), and IgG1 (33.11 mg/mL) concentrations. After colostrum consumption, calves were allocated to individual hutches and fed 2 L of milk replacer twice daily at a concentration of 125 g/L as fed (21.86% CP, 16.59% fat, 45.50% lactose; Schils). Calves had ad libitum access to starter feed (15.2% CP, 15.0% NDF, 5.0% ADF, 29.2% starch, 4.9% ether extract on a DM basis, with the main ingredients being 13.9% soybean meal, 38.9% corn, 9% wheat, 17% barley, 3% sunflower expeller, 1.9% palm oil, and 1.3% calcium carbonate), and water. At approximately 14 d of age (14.2 ± 0.81 d of age; mean ± SE), calves were transported 2.5 h from their origin dairy farm to an experimental research unit at IRTA (Torre Marimon, Spain) simulating the arrival at a rearing facility [average ambient temperature of 11°C (range = 4.4–17°C) and 87.5% humidity (range = 82–90%)]. Calves with respiratory or digestive problems or that were repeatedly treated with antibiotics at the dairy farm were not selected to be transported to the experimental research unit.

      Measurements and Sample Collection

      Body weight was recorded at birth and at arrival to the experimental research unit (simulating the arrival at a rearing facility) at approximately 14 d after birth. Blood samples were collected at birth before feeding and 48 h after birth to confirm if the colostrum administration in each treatment was correctly done and on arrival at the rearing facility before feeding to identify potential biomarkers of colostrum consumption. Samples were obtained from the jugular vein, using evacuated serum tubes (BD Vacutainer Plus Plastic Serum Tubes), and then centrifuged at 1,500 × g at 4°C for 15 min. A serum sample was used to measure TP by using a clinical refractometer (KERN refractometer, ORA-AL). The remaining serum was aliquoted and stored at −20°C until further analysis. As previously mentioned, colostrum samples from different pools used during the study were collected and mixed to obtain a single sample.

      Chemical Analysis

      Blood samples were processed to obtain serum for analysis of potential biomarkers of colostrum provision (IgG, IgG1, CHOL, ALP, GGT, and TP). Serum IgG and IgG1 concentrations were determined by an ELISA (species-specific Bovine IgG and Bovine IgG1, Bethyl Laboratories Inc.). Serum concentration of GGT, CHOL, ALP, and TP were analyzed by using a Beckman Coulter AU480 analyzer.
      The intra- and interassay coefficients of variations of IgG, IgG1, GGT, CHOL, ALP, and TP were 3.2 and 3.3%, 3.37 and 9.26%, 0.8 and 1%, 0.4 and 1.1%, 0.9 and 1.5%, and 0.38 and 0.74%, respectively. The starter feed was analyzed for DM (24 h at 103°C), ash (4 h at 550°C), CP by the Kjeldahl method (method 981.10;
      • AOAC International
      Official Methods of Analysis.
      ), and ADF and NDF (method 973.18;
      • AOAC International
      Official Methods of Analysis.
      ). Samples from the milk replacer used in this study were analyzed for DM (24 h at 103°C), ash (4 h at 550°C), CP by the Kjeldahl method (method 981.10;
      • AOAC International
      Official Methods of Analysis.
      ), and sugars (HPLC–Refractive Index; method 984.22;
      • AOAC International
      Official Methods of Analysis.
      ). Colostrum samples were analyzed for fat by the gravimetric method (ISO 1211/IDF 1: 2010;
      • ISO
      ISO 1211: Milk – Determination of Fat Content – Gavimetric Method.
      ), CP by the block-digestion method (ISO 8968–3/IDF 20–3:2004;
      • ISO
      ISO 8968-3: Milk – Determination of Nitrogen Content – Part 3: Block-digestion Method.
      ), and ashes (BOE-A-1997-1661;

      BOE. 1997. Orden de 31 de enero de 1997 por la que se setablecen los Metodos Oficinales de Analisis de ACeites y Grases, Cereales y Derivados, Productos Lacteos y productos derivados de la Uva. Agencia Estatal. Gobierno de Espana.

      ). Lactose was calculated removing the fat, CP, and ashes from the TS content.

      Statistical Analysis

      Based on the outcome of
      • Cuttance E.L.
      • Mason W.A.
      • Denholm K.S.
      • Laven R.A.
      Comparison of diagnostic tests for determining the prevalence of failure of passive transfer in New Zealand dairy calves.
      investigating diagnostic tests for FTPI in dairy calves, estimating prevalence of FTPI with a 95% confidence level at the herd level using different diagnostic tests required sample sizes varying from 42 to 61. Therefore, considering all the diagnostic tests with their sensitivity and specificity used in the current study it was estimated that between 30 and 54 calves/treatment would be required to detect differences. Calf was the experimental unit. The study design was a randomized unbalanced complete block design with a covariate adjustment. Animals were distributed using a stratified randomization where 1 of 3 animals balanced by parity and calving time was assigned to the LC treatment and 2 were assigned to the HC treatment. The model included the random effect of pen and the fixed effect of treatment (amount of colostrum), and initial BW and age as a covariate. The blocks were parity (primiparous and multiparous) and calving time (nighttime born or daytime born). Serum concentration of IgG, IgG1, CHOL, ALP, GGT, TP, and BW loss were analyzed using the MIXED procedure of SAS (version 9.4, SAS Institute Inc.). A normality test was conducted, and the nonnormal data were log-transformed to achieve normal distribution. Pearson correlation analyses were performed between serum IgG, IgG1, CHOL, ALP, GGT, and TP at 48 h after birth and at arrival. Additionally, the Pearson correlation analyses were also performed between serum IgG and GGT, between TP measured by refractometry and TP measured at the laboratory, and between TP measured by refractometry and serum IgG, all of them measured at arrival. Furthermore, IgG at 48 h was also correlated with TP measured at the laboratory, TP measured by refractometry, and GGT at arrival. All Pearson correlations were analyzed using JMP (version 16.0.0 SAS Institute Inc.). Lin's concordance correlation coefficient (CCC) was used to assess agreement between TP measured by refractometry and TP measured at the laboratory. The Lin's CCC was calculated using a SPSS package (IBM SPSS Statistics version 28.0) and an adapted syntax file from
      • Garcia-Granero M.
      Lin's Concordance Correlation Coefficient [Computer Programm: Syntax code SPSS].
      . Differences were declared significant at P ≤ 0.05, and trends were discussed at 0.05 ≤ P ≤ 0.10 for all models.

      RESULTS AND DISCUSSION

      Failed Transfer of Passive Immunity

      The total amount of IgG consumed by the HC calves was 1,450 g in 10 L of colostrum administered in 4 feedings, and the total amount of IgG consumed by the LC calves was 290 g in 2 L of colostrum administered in 1 feeding. All potential biomarkers except for CHOL and ALP showed greater (P < 0.01; Table 1) concentrations for the HC calves compared with the LC by 48 h after birth, confirming that the amounts of colostrum given were different between treatments. However, the LC treatment did not achieve what is considered to be an FTPI condition. Failed transfer of passive immunity has been described as <1.0 g/dL serum IgG concentration in calves that are between 1 and 7 d old (
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      ). In a more recent study, 4 categories have been proposed to assess FTPI as follows: excellent, good, fair, and poor with serum IgG levels of ≥25.0, 18.0–24.9, 10.0–17.9, and <10 g/L, respectively (
      • Lombard J.
      • Urie N.
      • Garry F.
      • Godden S.
      • Quigley J.
      • Earleywine T.
      • McGuirk S.
      • Moore D.
      • Branan M.
      • Chamorro M.
      • Smith G.
      • Shivley C.
      • Catherman D.
      • Haines D.
      • Heinrichs A.J.
      • James R.
      • Maas J.
      • Sterner K.
      Consensus recommendations on calf- and herd-level passive immunity in dairy calves in the United States.
      ). In the present study, results showed that IgG serum concentrations 48 h after birth were 72.6 ± 3.61 mg/mL and 29.9 ± 4.35 mg/mL (mean ± SE) for the HC and LC groups, respectively. Based on
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      our calves did not suffer FTPI and based on
      • Lombard J.
      • Urie N.
      • Garry F.
      • Godden S.
      • Quigley J.
      • Earleywine T.
      • McGuirk S.
      • Moore D.
      • Branan M.
      • Chamorro M.
      • Smith G.
      • Shivley C.
      • Catherman D.
      • Haines D.
      • Heinrichs A.J.
      • James R.
      • Maas J.
      • Sterner K.
      Consensus recommendations on calf- and herd-level passive immunity in dairy calves in the United States.
      both treatments fell into the “excellent” category. Even though we reached a high immunoglobulin level in both treatments, 2 L of colostrum may not be sufficient to provide the additional benefits of nutrients and bioactive substances contained in the colostrum that modulate for, for example, the development of the gastrointestinal tract (
      • Hammon H.M.
      • Liermann W.
      • Frieten D.
      • Koch C.
      Review: Importance of colostrum supply and milk feeding intensity on gastrointestinal and systemic development in calves.
      ). To achieve FTPI in the LC calves it might be necessary to increase the time between birth and colostrum administration or to provide a low-quality colostrum. The LC treatment was selected based on commercial practices with the limitation of not achieving an FTPI condition. However, the amount of colostrum fed to the HC calves compared with commercial practices for research purposes allowed us to see differences between treatments.
      Table 1Serum IgG, IgG1, cholesterol (CHOL), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), and total protein (TP) concentrations at birth and 48 h after birth in male Holstein calves
      Calves in the high-colostrum group (HC) were fed 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth. Calves in the low-colostrum group (LC) were fed only 2 L of colostrum within the first 2 h after birth.
      ItemTreatmentSEMP-value
      Trt = effect of the amount of colostrum provision on serum concentration of potential biomarkers.
      HCLCTrt
      Potential biomarker (at birth)
       IgG, mg/mL0.230.800.2410.15
       IgG1, mg/mL0.090.290.0830.21
       CHOL, mg/dL18.719.81.460.58
       ALP, ng/mL5.135.180.0920.71
       GGT, ng/mL11.0012.10.790.49
       TP, g/dL4.224.20.0640.89
      Potential biomarker (48 h after birth)
       IgG, mg/mL72.6
      Values with different superscript within a row differ at P ≤ 0.05.
      29.9
      Values with different superscript within a row differ at P ≤ 0.05.
      3.98<0.01
       IgG1, mg/mL27.0
      Values with different superscript within a row differ at P ≤ 0.05.
      13.2
      Values with different superscript within a row differ at P ≤ 0.05.
      1.71<0.01
       CHOL, mg/dL29.427.51.250.30
       ALP, ng/mL5.535.440.0820.43
       GGT, ng/mL1,946
      Values with different superscript within a row differ at P ≤ 0.05.
      562
      Values with different superscript within a row differ at P ≤ 0.05.
      148.5<0.01
       TP, g/dL7.14
      Values with different superscript within a row differ at P ≤ 0.05.
      5.63
      Values with different superscript within a row differ at P ≤ 0.05.
      0.143<0.01
      a,b Values with different superscript within a row differ at P ≤ 0.05.
      1 Calves in the high-colostrum group (HC) were fed 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth. Calves in the low-colostrum group (LC) were fed only 2 L of colostrum within the first 2 h after birth.
      2 Trt = effect of the amount of colostrum provision on serum concentration of potential biomarkers.
      The generally accepted cut-off points for measuring FTPI using TP is between 5 and 5.5 g/dL in calves from 1 to 8 d old (
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      ). In this study, TP concentrations by 48 h after birth were 7.1 ± 0.09 g/dL and 5.6 ± 0.11 g/dL (mean ± SE) for the HC and LC group, respectively (Table 1), suggesting that TP is probably the most sensitive of all biomarkers to define FTPI because the LC calves had a TP concentration close to the limit described by
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      .
      As mentioned, although the treatments applied were different enough to see differences on the concentration of IgG, IgG1, GGT and TP, it seems that providing 2 L of good-quality colostrum within 2 h after birth did not cause FTPI. However, adequate colostrum provision (amount, time, quality) is a concept that goes beyond FTPI. Colostrum contains nutrients (carbohydrates, proteins, lipids), minerals, vitamins, and growth promoters among other bioactive compounds that exert important morphological and functional changes in calves (
      • Blum J.W.
      Nutritional physiology of neonatal calves.
      ). This means that colostrum provision can exert positive effects on calves' lifetimes beyond the immune protection at an early age resulting from immunoglobulins. These positive effects are relevant for the development of the gastrointestinal tract in the newborn calf (
      • Blum J.W.
      • Hammon H.
      Colostrum effects on the gastrointestinal tract, and on nutritional, endocrine and metabolic parameters in neonatal calves.
      ); it has been demonstrated that high amounts of consumed colostrum increased the intestinal epithelial cells' growth, promoting intestinal absorption (
      • Blättler U.
      • Hammon H.M.
      • Morel C.
      • Philipona C.
      • Rauprich A.
      • Romé V.
      • le Huërou-Luron I.
      • Guilloteau P.
      • Blum J.W.
      Feeding colostrum, its composition and feeding duration variably modify proliferation and morphology of the intestine and digestive enzyme activities of neonatal calves.
      ). Ingestion of colostrum has also been shown to increase crypt cell proliferation, decrease apoptosis, and stimulate villus growth in repeatedly colostrum-fed calves (
      • Blum J.W.
      Nutritional physiology of neonatal calves.
      ). Additionally, the interactions of the many different bioactive substances (vs. their individual actions) contained in colostrum have been described to promote intestinal growth and cell proliferation (
      • Hammon H.M.
      • Liermann W.
      • Frieten D.
      • Koch C.
      Review: Importance of colostrum supply and milk feeding intensity on gastrointestinal and systemic development in calves.
      ). In the present study, even if biomarkers were not indicative of FTPI, they might be indicative of calves' colostrum consumption at their farm of origin. In this scenario, being able to detect colostrum provision even when the quality remains unknown would still provide valuable information that might allow producers to make decisions over management, vaccination, and nutritional protocols for the newly arrived calves.

      Potential Biomarkers of Colostrum Provision at Arrival

      Differences in IgG and IgG1 concentrations were found between HC and LC calves. Serum concentrations of IgG and IgG1 were greater (P < 0.01; Figure 1A and B, respectively) on arrival at the rearing facility for the HC treatment compared with the LC treatment. Three main classes of immunoglobulins are found in serum, milk, and colostrum in bovines: IgG, IgM, and IgA. Additionally, IgG is subdivided in 2 subclasses: IgG1 and IgG2. Of these 2, IgG1 represents more than 75% of the immunoglobulins in colostrum (
      • Korhonen H.
      • Marnila P.
      • Gill H.S.
      Milk immunoglobulins and complement factors.
      ). Both immunoglobulins can be used as indicators of colostrum consumption; however, IgG1 could be a better estimate based on its predominance in colostrum. Several methods have been developed to determine IgG concentration in the blood of newborn calves as an approach to detect FTPI or colostrum consumption. However, even though results are promising in differentiating calves with different colostrum provision using IgG or IgG1, direct measurement of immunoglobulins by radial immunodiffusion, turbidimetric immunoassay, or ELISA is expensive because these tests require laboratory interpretation and trained personnel. Therefore, the lack of less costly and automated techniques makes it difficult to control colostrum provision in herds based on these measurements.
      Figure thumbnail gr1
      Figure 1Serum concentrations (mean ± SE) of IgG (A), and IgG1 (B) in male Holstein in 2 treatment groups. Calves in the high-colostrum group (HC) were fed 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth. Calves in the low-colostrum group (LC) were fed only 2 L of colostrum within the first 2 h after birth. Different letters within a time point denote differences among treatments (P < 0.05); order of the letters denotes the treatment with the highest value. Trt = treatment.
      The use of CHOL as a biomarker of colostrum consumption has been previously explored (
      • Marcato F.
      • van den Brand H.
      • Kemp B.
      • van Reenen K.
      Evaluating potential biomarkers of health and performance in veal calves.
      ). Cholesterol can be found in higher concentrations in the colostrum compared with mature milk (
      • Shope R.E.
      • Gowen J.W.
      Cholesterol and cholesterol ester content of bovine colostrum.
      ). Consequently, neonatal calves' serum CHOL concentration after birth might be indicative of the amount of colostrum consumed (
      • Renaud D.L.
      • Duffield T.F.
      • LeBlanc S.J.
      • Ferguson S.
      • Haley D.B.
      • Kelton D.F.
      Risk factors associated with mortality at a milk fed veal calf facility: A prospective cohort study.
      ). Results from this study showed no differences (P = 0.39; Figure 2 A) for serum CHOL concentration between HC or LC calves. Cholesterol was first described as a marker of calves' mortality risk during the first 21 d after arrival at rearing facilities by
      • Renaud D.L.
      • Duffield T.F.
      • LeBlanc S.J.
      • Ferguson S.
      • Haley D.B.
      • Kelton D.F.
      Risk factors associated with mortality at a milk fed veal calf facility: A prospective cohort study.
      . Cholesterol plays an important role in intestinal signaling, mediating intestinal lipid absorption, and regulating plasma lipid concentration (
      • Thurnhofer H.
      • Hauser H.
      Uptake of cholesterol by small intestinal brush border membrane is protein-mediated.
      ), which in neonatal calves represent the major energy source to satisfy the demands of tissue growth (
      • Ontsouka E.C.
      • Albrecht C.
      • Bruckmaier R.M.
      Invited review: Growth promoting effects of colostrum in calves based on interaction with intestinal cell surface receptors and receptor-like transporters.
      ). Additionally, CHOL regulates lactase activity, an enzyme facilitating lactose absorption in the intestine and whose low concentration has been associated with the risk of morbidity and diarrhea in the newborn calf (
      • Kien C.L.
      • McClead R.E.
      • Cordero Jr., L.
      In vivo lactose digestion in preterm infants.
      ). In the present study we were expecting to see differences in CHOL concentration between treatments based on the amounts of colostrum consumed. However, it seems that even when feeding different amounts of colostrum, CHOL concentration did not vary significantly. Because we were unable to generate an actual FTPI condition, it remains unclear how this could have affected CHOL concentration.
      Figure thumbnail gr2
      Figure 2Serum concentrations (mean ± SE) of cholesterol (CHOL, A), and alkaline phosphatase (ALP, B) in male Holstein calves in 2 treatment groups. Calves in the high-colostrum group (HC) were fed 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth. Calves in the low-colostrum group (LC) were fed only 2 L of colostrum within the first 2 h after birth. Cov BW d-4 = BW on d 4 as covariate; Trt = treatment.
      Increased serum ALP concentration has been previously correlated with increases in serum GGT in calves (
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ;
      • Zanker I.A.
      • Hammon H.M.
      • Blum J.W.
      Activities of γ-Glutamyltransferase, alkaline phosphatase and aspartate-aminotransferase in colostrum, milk and blood plasma of calves fed first colostrum at 0–2, 6–7, 12–13 and 24–25h after birth.
      ). Additionally,
      • Britti D.
      • Massimini G.
      • Peli A.
      • Luciani A.
      • Boari A.
      Evaluation of serum enzyme activities as predictors of passive transfer status in lambs.
      found an increase in ALP concentration after colostrum consumption and a positive correlation between ALP and IgG in newborn lambs. In disagreement, the present study showed no differences between treatments for serum ALP concentration (P = 0.86; Figure 2B).
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      described a large initial drop followed by a gradual decline in ALP concentration after colostrum intake in calves. This pattern seems to act similarly in lambs. However, because the activity of ALP in ewe's colostrum is lower than in lambs' serum, ALP appears to be derived from the brush border of the intestine and its concentration might be associated with feeding but not necessarily ingestion of colostrum (
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ).
      • Pauli J.V.
      Colostral transfer of gamma glutamyl transferase in lambs.
      proposed that a similar phenomenon exists in calves. However, based on the results from the present study, serum ALP cannot be considered a good biomarker of colostrum consumption in calves older than 14 d of age. Inconsistencies in the literature to date make it difficult to validate ALP serum concentration as an indicator of colostrum consumption.
      Measurements of serum GGT concentration have been previously studied as a potential biomarker for FTPI (
      • Perino L.J.
      • Sutherland R.L.
      • Woollen N.E.
      Serum gamma glutamyltransferase activity and protein concentration at birth and after suckling in calves with adequate and inadequate passive transfer of immunoglobulin G.
      ;
      • Parish S.M.
      • Tyler J.W.
      • Besser T.E.
      • Gay C.C.
      • Krytenberg D.
      Prediction of serum IgG1 concentration in Holstein calves using serum gamma glutamyltransferase activity.
      ;
      • Buczinski S.
      • Dubuc J.
      • Bourgeois V.
      • Baillargeon P.
      • Côté N.
      • Fecteau G.
      Validation of serum gamma-glutamyl transferase activity and body weight information for identifying dairy calves that are too young to be transported to auction markets in Canada.
      ). In the present study, GGT serum concentration was greater (P < 0.01; Figure 3A) in the HC group compared with the LC group upon arrival at the rearing facility. Gamma-glutamyl transferase is an enzyme involved in amino acid transport and produced in the mammary gland ducts during colostrogenesis (
      • Baumrucker C.R.
      γ-Glutamyl transpeptidase of bovine milk membranes: Distribution and characterization.
      ;
      • Blum J.W.
      • Hammon H.
      Colostrum effects on the gastrointestinal tract, and on nutritional, endocrine and metabolic parameters in neonatal calves.
      ). For this reason, its concentration in bovine colostrum is higher than in milk (
      • Cuttance E.L.
      • Regnerus C.
      • Laven R.A.
      A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand.
      ). After colostrum consumption, GGT is absorbed in the small intestine of the calf via the same nonselective passage that is used by IgG (
      • Parish S.M.
      • Tyler J.W.
      • Besser T.E.
      • Gay C.C.
      • Krytenberg D.
      Prediction of serum IgG1 concentration in Holstein calves using serum gamma glutamyltransferase activity.
      ). After absorption, GGT can be found in calves' serum at concentrations 60–160 times greater than in adult cattle, declining to adult activities by 5 wk of age (
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ;
      • Yu K.
      • Canalias F.
      • Solà-Oriol D.
      • Arroyo L.
      • Pato R.
      • Saco Y.
      • Terré M.
      • Bassols A.
      Age-related serum biochemical reference intervals established for unweaned calves and piglets in the post-weaning period.
      ). Some studies have shown positive correlations between serum GGT and serum immunoglobulin concentrations (
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ;
      • Perino L.J.
      • Sutherland R.L.
      • Woollen N.E.
      Serum gamma glutamyltransferase activity and protein concentration at birth and after suckling in calves with adequate and inadequate passive transfer of immunoglobulin G.
      ;
      • Parish S.M.
      • Tyler J.W.
      • Besser T.E.
      • Gay C.C.
      • Krytenberg D.
      Prediction of serum IgG1 concentration in Holstein calves using serum gamma glutamyltransferase activity.
      ;
      • Weaver D.M.
      • Tyler J.W.
      • Scott M.A.
      • Wallace L.M.
      • Marion R.S.
      • Holle J.M.
      Passive transfer of colostral immunoglobulin G in neonatal llamas and alpacas.
      ). Results from the present study showed a positive correlation (P < 0.01) between serum IgG and GGT concentrations at arrival, although with a low coefficient of determination (R2 = 0.53; CI 95% = 0.60–0.82; P < 0.001). However, a high concentration of GGT in a calf is not necessarily indicative of having been fed a good-quality colostrum because there is no biological relation between GGT and colostrum IgG concentration (
      • Weaver D.M.
      • Tyler J.W.
      • Scott M.A.
      • Wallace L.M.
      • Marion R.S.
      • Holle J.M.
      Passive transfer of colostral immunoglobulin G in neonatal llamas and alpacas.
      ;
      • de Souza R.S.
      • dos Santos L.B.C.
      • Melo I.O.
      • Cerqueira D.M.
      • Dumas J.V.
      • Leme F.O.P.
      • Moreira T.F.
      • Meneses R.M.
      • de Carvalho A.U.
      • Facury-Filho E.J.
      Current diagnostic methods for assessing transfer of passive immunity in calves and possible improvements: A literature review.
      ). In this scenario, measurements of GGT could be used only as an indicator of colostrum intake (
      • Blum J.W.
      • Hammon H.
      Colostrum effects on the gastrointestinal tract, and on nutritional, endocrine and metabolic parameters in neonatal calves.
      ). Further studies evaluating GGT upon arrival at rearing facilities and correlating the amount, timing, and quality of colostrum offered may provide more information on the potential of GGT as a biomarker of colostrum provision in calves older than 14 d of age because, as mentioned previously, measurements of IgG and IgG1 are expensive and tedious, whereas GGT determination is cheaper, quicker, and can be conveniently automated (
      • Hogan I.
      • Doherty M.
      • Fagan J.
      • Kennedy E.
      • Conneely M.
      • Brady P.
      • Ryan C.
      • Lorenz I.
      Comparison of rapid laboratory tests for failure of passive transfer in the bovine.
      ).
      Figure thumbnail gr3
      Figure 3Serum concentrations (mean ± SE) of gamma-glutamyl transferase (GGT, A), total protein (TP, B), and TP calculated with a refractometer (C) in male Holstein calves in 2 treatment groups. Calves in the high-colostrum group (HC) were fed 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth. Calves in the low-colostrum group (LC) were fed only 2 L of colostrum within the first 2 h after birth. Different letters within a time point denote differences among treatments (P < 0.05); order of the letters denotes the treatment with the highest value. Cov BW d-4 = BW on d 4 as covariate; Trt = treatment.
      At arrival to the rearing facility, the concentration of TP was greater (P < 0.01; Figure 3 B) for the HC group compared with the LC group when TP concentration was determined with a biochemical analyzer. As expected, serum concentration of TP exhibited a similar pattern compared with IgG1 (
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      ;
      • Dawes M.E.
      • Tyler J.W.
      • Hostetler D.
      • Lakritz J.
      • Tessman R.
      Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves.
      ;
      • Godden S.
      Colostrum management for dairy calves.
      ). Determining FTPI by measuring TP is based on the fact that the ingestion of immunoglobulins via colostrum intake (major contributor of serum proteins in newborn calves) increases TP concentration in serum (
      • Hogan I.
      • Doherty M.
      • Fagan J.
      • Kennedy E.
      • Conneely M.
      • Brady P.
      • Ryan C.
      • Lorenz I.
      Comparison of rapid laboratory tests for failure of passive transfer in the bovine.
      ). Concentration of TP was also measured by using a clinical refractometer. This technique is inexpensive, easy for farm personnel to perform and interpret, and offers immediate results. Likewise, results showed greater (P < 0.01; Figure 3 C) serum concentration of TP measured by refractometry for the HC calves compared with the LC calves at arrival to the rearing facility. However, when assessing colostrum provision by measuring TP some aspects need to be considered. The first consideration is the age of the animals. Previous studies have demonstrated that age can influence IgG serum concentration mainly based on a protein catabolism process (
      • Cuttance E.L.
      • Regnerus C.
      • Laven R.A.
      A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand.
      ;
      • de Souza R.S.
      • dos Santos L.B.C.
      • Melo I.O.
      • Cerqueira D.M.
      • Dumas J.V.
      • Leme F.O.P.
      • Moreira T.F.
      • Meneses R.M.
      • de Carvalho A.U.
      • Facury-Filho E.J.
      Current diagnostic methods for assessing transfer of passive immunity in calves and possible improvements: A literature review.
      ).
      • Cuttance E.L.
      • Regnerus C.
      • Laven R.A.
      A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand.
      suggested that calves should not be older than 1 wk of age,
      • de Souza R.S.
      • dos Santos L.B.C.
      • Melo I.O.
      • Cerqueira D.M.
      • Dumas J.V.
      • Leme F.O.P.
      • Moreira T.F.
      • Meneses R.M.
      • de Carvalho A.U.
      • Facury-Filho E.J.
      Current diagnostic methods for assessing transfer of passive immunity in calves and possible improvements: A literature review.
      proposes 24 to 48 h, if possible, and
      • Wilm J.
      • Costa J.H.C.
      • Neave H.W.
      • Weary D.M.
      • von Keyserlingk M.A.G.
      Technical note: Serum total protein and immunoglobulin G concentrations in neonatal dairy calves over the first 10 days of age.
      described that serum TP concentration up to 9 d of age can provide reliable estimates of FPTI in calves. Second, this test should be performed in healthy nondehydrated calves because a process of dehydration or protein-losing enteropathy will alter protein concentration in serum (
      • Hogan I.
      • Doherty M.
      • Fagan J.
      • Kennedy E.
      • Conneely M.
      • Brady P.
      • Ryan C.
      • Lorenz I.
      Comparison of rapid laboratory tests for failure of passive transfer in the bovine.
      ) leading to misinterpretations of TP concentration. This is an important consideration because transported calves arrive at the rearing facilities dehydrated (
      • Renaud D.L.
      • Duffield T.F.
      • LeBlanc S.J.
      • Ferguson S.
      • Haley D.B.
      • Kelton D.F.
      Risk factors associated with mortality at a milk fed veal calf facility: A prospective cohort study.
      ;
      • Wilson D.J.
      • Stojkov J.
      • Renaud D.L.
      • Fraser D.
      Risk factors for poor health outcomes for male dairy calves undergoing transportation in western Canada.
      ). Therefore, the time when TP should be measured is crucial for the reliability of the results. In this study, calves were transported only 2.5 h from their origin dairy farm to the rearing facility and did not exhibit signs of dehydration (e.g., sunken eyes or skin turgor). Thus, the differences in TP concentration found between HC and LC calves can be considered reliable. Measuring TP 24 h after arrival when calves have been rehydrated and fed after a long-distance transportation could be an optimal time to perform the test. However, further studies should investigate the interactions between transport duration, dehydration, and TP concentration in unweaned calves to better understand the veracity of TP concentration as a tool to estimate FTPI.
      Results of the Pearson correlation analysis between the different biomarkers at 48 h after birth and at arrival when calves are approximately 14 d of age showed that IgG, IgG1, ALP, and TP had a positive significant correlation but with a low coefficient of correlation or determination (Table 2). A higher coefficient of determination was observed for GGT (Table 2); however, an R2 of 49% is still low as a prediction of the concentration of GGT over time. The Pearson correlation analysis performed between serum TP concentration obtained by refractometry and TP measured at the laboratory at arrival showed a significant positive correlation (P < 0.01) between techniques although the coefficient of determination was lower than expected (R2 = 0.35; CI 95% = 0.43–0.59; P < 0.001). Similar results were obtained with Lin's CCC (CCC = 0.185; CI 95% = 0.113–0.255), demonstrating that the agreement of the TP analyzed by refractometry and by laboratory techniques was low.
      Table 2Correlation between serum IgG, IgG1, cholesterol (CHOL), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), and total protein (TP) concentrations 48 h after birth and at arrival to the rearing facility in male Holstein calves
      Calves were fed 4 L of colostrum within the first 2 h after birth, and 2 L of colostrum in the next 3 feedings within the first 24 h after birth (HC) and fed only 2 L of colostrum within the first 2 h after birth (LC)
      ItemR2CorrelationCI 95%P-value
      IgG, mg/mL0.290.530.36–0.68<0.001
      IgG1, mg/mL0.090.300.09–0.49<0.01
      CHOL, mg/dL<0.0010.01−0.21–0.230.91
      ALP, ng/mL0.120.340.13–0.52<0.01
      GGT, ng/mL0.490.700.57–0.80<0.001
      TP, g/dL0.220.460.28–0.62<0.001
      1 Calves were fed 4 L of colostrum within the first 2 h after birth, and 2 L of colostrum in the next 3 feedings within the first 24 h after birth (HC) and fed only 2 L of colostrum within the first 2 h after birth (LC)
      The reason why the correlation between those measurements was low is unknown and further comparisons should be done by sending the samples to different laboratories or using other lab techniques. When the correlation between the IgG at 48 h and TP measured at the laboratory at arrival (R2 = 0.08; CI 95% = 0.06–0.47; P = 0.01) was compared with the correlation between IgG at 48 h and TP concentration obtained by refractometry at arrival (R2 = 0.26; CI 95% = 0.32–0.66; P < 0.001), the association was improved. On the other hand, a Pearson correlation with an R2 = 0.61 (CI 95% = 0.67–0.85; P < 0.001) was observed between serum TP calculated by refractometry and serum IgG concentration at arrival. The high correlation between serum IgG and serum TP concentration measured by refractometry has been studied in newborn calves (
      • McBeath D.G.
      • Penhale W.J.
      • Logan E.F.
      An examination of the influence of husbandry on the plasma immunoglobulin level of the newborn calf, using a rapid refractometer test for assessing immunoglobulin content.
      ; R2 = 0.72). However, in our study TP values measured either at the laboratory or with the refractometer at arrival were not good enough to predict the level of IgG 48 h after birth and may be reliable only when compared with IgG at the same time point. However, most of the literature has only shown levels of IgG or TP until 7 d after birth (
      • Rauprich A.B.E.
      • Hammon H.M.
      • Blum J.W.
      Influence of feeding different amounts of first colostrum on metabolic, endocrine, and health status and on growth performance in neonatal calves.
      ;
      • Renaud D.
      • Pardon B.
      Preparing male dairy calves for the veal and dairy beef industry.
      ), and comparisons when calves are older could not be made.
      • Rauprich A.B.E.
      • Hammon H.M.
      • Blum J.W.
      Influence of feeding different amounts of first colostrum on metabolic, endocrine, and health status and on growth performance in neonatal calves.
      showed how IgG concentration decreased rapidly 3 d after birth, while the decrease in TP was slower. Additionally, a recent study showed inconsistencies in the use of TP measured by refractometry as an estimate of IgG concentration in calves due to the existence of residual nonprotein solutes contained in colostrum, which accounted for approximately 36% of the variation in results (
      • Schalich K.M.
      • Reiff O.M.
      • Nguyen B.T.
      • Lamb C.L.
      • Mondoza C.R.
      • Selvaraj V.
      Temporal kinetics of bovine mammary IgG secretion into colostrum and transition milk.
      ). Recently, measuring TP by using a refractometer has been questioned and other methods have been proposed (
      • Lombard J.
      • Quigley J.
      • Haines D.
      • Garry F.
      • Earleywine T.
      • Urie N.
      • Chamorro M.
      • Godden S.
      • McGuirk S.
      • Smith G.
      • Shivley C.
      • Catherman D.
      • Heinrichs A.J.
      • James R.
      • Maas J.
      • Sterner K.
      • Sockett D.
      Letter to the editor: Comments on Schalich et al. 2021. Colostrum testing with Brix is a valuable on farm tool. 10.193/jas/skab083.
      ;
      • Schalich K.M.
      • Selvaraj V.
      Contradictions on colostrum IgG levels and Brix values are real and can be explained. Response to letter by Lombard et al. 2022.
      ). In the present study, Pearson correlations between TP by refractometer at arrival and IgG at 48 h (R2 = 0.26; CI 95% = 0.32–0.66; P < 0.001) had a higher coefficient of determination than the correlation between GGT at arrival and IgG at 48 h (R2 = 0.17; CI 95% = 0.21–0.58; P < 0.001). The sensitivity and specificity (Sp) analysis using IgG (
      • Lombard J.
      • Urie N.
      • Garry F.
      • Godden S.
      • Quigley J.
      • Earleywine T.
      • McGuirk S.
      • Moore D.
      • Branan M.
      • Chamorro M.
      • Smith G.
      • Shivley C.
      • Catherman D.
      • Haines D.
      • Heinrichs A.J.
      • James R.
      • Maas J.
      • Sterner K.
      Consensus recommendations on calf- and herd-level passive immunity in dairy calves in the United States.
      ) and TP (
      • Tyler J.W.
      • Hancock D.D.
      • Parish S.M.
      • Rea D.E.
      • Besser T.E.
      • Sanders S.G.
      • Wilson L.K.
      Evaluation of 3 assays for failure of passive transfer in calves.
      ) at 48 h after birth showed a sensitivity of 0.9%, Sp of 0.31%, and an accuracy of 0.81%. The sensitivity and Sp analysis using IgG (
      • Lombard J.
      • Urie N.
      • Garry F.
      • Godden S.
      • Quigley J.
      • Earleywine T.
      • McGuirk S.
      • Moore D.
      • Branan M.
      • Chamorro M.
      • Smith G.
      • Shivley C.
      • Catherman D.
      • Haines D.
      • Heinrichs A.J.
      • James R.
      • Maas J.
      • Sterner K.
      Consensus recommendations on calf- and herd-level passive immunity in dairy calves in the United States.
      ) and GGT (
      • Parish S.M.
      • Tyler J.W.
      • Besser T.E.
      • Gay C.C.
      • Krytenberg D.
      Prediction of serum IgG1 concentration in Holstein calves using serum gamma glutamyltransferase activity.
      ) at 48 h after birth showed a sensitivity of 0.91%, Sp of 1%, and an accuracy of 0.91%. The greater Pearson correlation between GGT at 48 h and GGT at arrival (Table 2) and the high accuracy and Sp between IgG at 48 h and GGT at 48 h could make the GGT concentration as good as TP as a biomarker of colostrum consumption and with the advantage that it might not be affected by the dehydration status of the animals.
      As mentioned previously, colostrum has potential beneficial effects on gut development beyond providing immunity through immunoglobulins. These additional effects that colostrum has on calves' gastrointestinal tracts and physiology might be responsible for the differences observed for BW losses in the present study. Body weight loss was calculated as the difference between BW at birth and BW at arrival to the rearing facility. Results showed greater (P < 0.01; Figure 4) BW losses for the LC calves compared with the HC calves. At dairy farms, when male calves are fed a low plane of nutrition, their dietary requirements are not fully covered, which causes BW losses (
      • Winder C.B.
      • Kelton D.F.
      • Duffield T.F.
      Mortality risk factors for calves entering a multi-location white veal farm in Ontario, Canada.
      ). Based on the results on BW losses, it could be assumed that administrating lower amounts of colostrum at birth directly affects calves' performance at arrival. Low BW at arrival to veal farms has been previously associated with an increase of mortality risk (
      • Fallon R.J.
      • Harte F.J.
      • Harrington D.
      The Effect of calf purchase weight, serum IG level and feeding systems on the incidence of diarrhoea, respiratory disease, and mortality.
      ;
      • Moore D.A.
      • Sischo W.M.
      • Festa D.M.
      • Reynolds J.P.
      • Atwill E.R.
      • Holmberg C.A.
      Influence of arrival weight, season, and calf supplier on survival in Holstein beef calves on a calf ranch in California, USA.
      ;
      • Winder C.B.
      • Kelton D.F.
      • Duffield T.F.
      Mortality risk factors for calves entering a multi-location white veal farm in Ontario, Canada.
      ). Additionally, it has been demonstrated that malnourished calves have increased cortisol levels, which can affect their immune response (
      • Drackley J.
      Does early growth affect subsequent health and performance of heifers?.
      ) and their capacity to deal with pathogens, especially those provoking diarrhea (
      • Fallon R.J.
      • Harte F.J.
      • Harrington D.
      The Effect of calf purchase weight, serum IG level and feeding systems on the incidence of diarrhoea, respiratory disease, and mortality.
      ;
      • Paré J.
      • Thurmond M.C.
      • Gardner I.A.
      • Picanso J.P.
      Effect of birthweight, total protein, serum IgG and packed cell volume on risk of neonatal diarrhea in calves on two California dairies.
      ), a disease of great incidence in young calves. All together, these conditions aggravate calves' health and performance at a time in their lives when they are overexposed to stressful situations like transportation, commingling, feed restriction, and undeveloped immunity, among others (
      • Roland L.
      • Drillich M.
      • Klein-Jöbstl D.
      • Iwersen M.
      Invited review: Influence of climatic conditions on the development, performance, and health of calves.
      ). For this reason, dairy farmers should be encouraged to ensure an adequate plane of nutrition for male calves from birth until arrival at the rearing facilities and to collect BW records to avoid transportation of unfit calves at risk of suffering from disease in order to decrease morbidity and mortality rates upon arrival at the rearing facilities.
      Figure thumbnail gr4
      Figure 4Body weight losses in male Holstein calves in 2 treatment groups. Calves in the high-colostrum group (HC) were fed 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth. Calves in the low-colostrum group (LC) were fed only 2 L of colostrum within the first 2 h after birth. Differences were calculated between BW at birth and BW at arrival to the rearing facility. Different letters within a time point denote differences among treatments (P < 0.05); order of the letters denotes the treatment with the highest value. Trt = treatment.
      In summary, the use of biomarkers that could detect colostrum consumption will offer important information that can be used by veal and dairy-beef producers to classify, isolate, and treat calves depending on their individual risk. Previous studies have evaluated the concentration of these biomarkers in newborn calves to assess colostrum consumption or FTPI (
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ;
      • Blum J.W.
      • Hammon H.
      Colostrum effects on the gastrointestinal tract, and on nutritional, endocrine and metabolic parameters in neonatal calves.
      ;
      • Marcato F.
      • van den Brand H.
      • Kemp B.
      • van Reenen K.
      Evaluating potential biomarkers of health and performance in veal calves.
      ). However, their application in older calves arriving at rearing facilities remains unclear. Among the potential biomarkers of colostrum consumption considered in this study, GGT and TP measured by refractometer might be good indicators of colostrum intake in calves arriving at rearing facilities because they are faster, less expensive, easier to automate, and more consistent techniques compared with measuring immunoglobulins. The age at arrival to a rearing facility for dairy-beef calves varies widely but is normally beyond 14 d of age. One of the benefits of using serum GGT concentration as an alternative to TP as a marker of colostrum consumption is that, based on the literature, GGT concentration decreases to adult activities by 5 wk of age (
      • Thompson J.C.
      • Pauli J.
      Colostral transfer of gamma glutamyl transpeptidase in calves.
      ). This could allow the detection of differences in concentration in calves arriving at rearing facilities until approximately 1 mo of age. Additionally, it is still unknown until what age TP could be detected in serum to have a good correlation with IgG. Another advantage is that GGT concentration in serum is not affected by the hydration status of the calves upon arrival at the rearing facility. And, as previously mentioned, dehydration after long-distance transportation periods is quite common in dairy-beef calves. On the other hand, TP provides more information on the quality of the colostrum and can be easily measured by using a refractometer, a test that can be performed at the farm. Additionally, even though BW is not routinely recorded at the dairy farms, measurements of BW loss could also be a good estimator of risk during the first weeks after arrival. Even though these biomarkers might seem promising in assessing colostrum consumption in dairy-beef calves, further investigation should be conducted on stablishing valid ranges in serum considering if calves were HC or LC at the farm of origin. Additionally, the most appropriate time to measure the concentration of these biomarkers in addition to the study of better correlations needs to be further defined in the search of a validated tool to assess colostrum consumption upon arrival at rearing facilities.

      CONCLUSIONS

      Serum concentrations of GGT and TP appear to be the most reliable indicators of colostrum consumption in calves beyond 14 d of age. In the present study, even though calves were fed high-quality colostrum and did not suffer FTPI, HC calves showed greater concentrations of most of the biomarkers evaluated. These results obtained under experimental conditions might show even greater differences when evaluating colostrum consumption in the practice where transported male calves are vulnerable to stressors like commingling, mixing, contagious diseases, environmental conditions, and feed restriction, among many others that increase their risk. Further research should be undertaken to investigate if these additional transport stressors confirm the results observed in the present study.

      ACKNOWLEDGMENTS

      This work was funded by Minisiterio de Ciencia e Innovación grant (PID2019-104021RB-I00). We are also grateful to CERCA Programme (Generalitat de Catalunya). We are thankful to the personnel of Granja Selergan, S.A. (Lleida, Spain), and Maria Vidal, Marina Tortadès, Xavier Vergara, Irene Pinyol, and Anna Solé (IRTA, Caldes de Montbui, Spain) for their technical assistance. The authors have not stated any conflicts of interest.

      REFERENCES

        • AOAC International
        Official Methods of Analysis.
        15th ed. AOAC International, 1995
        • AOAC International
        Official Methods of Analysis.
        16th ed. AOAC International, 1996
        • AOAC International
        Official Methods of Analysis.
        17th ed. AOAC International, 2002
        • Baumrucker C.R.
        γ-Glutamyl transpeptidase of bovine milk membranes: Distribution and characterization.
        J. Dairy Sci. 1979; 62 (37261): 253-258
        • Beam A.L.
        • Lombard J.E.
        • Kopral C.A.
        • Garber L.P.
        • Winter A.L.
        • Hicks J.A.
        • Schlater J.L.
        Prevalence of failure of passive transfer of immunity in newborn heifer calves and associated management practices on US dairy operations.
        J. Dairy Sci. 2009; 92 (19620681): 3973-3980
        • Blättler U.
        • Hammon H.M.
        • Morel C.
        • Philipona C.
        • Rauprich A.
        • Romé V.
        • le Huërou-Luron I.
        • Guilloteau P.
        • Blum J.W.
        Feeding colostrum, its composition and feeding duration variably modify proliferation and morphology of the intestine and digestive enzyme activities of neonatal calves.
        J. Nutr. 2001; 131 (11285335): 1256-1263
        • Blum J.W.
        Nutritional physiology of neonatal calves.
        J. Anim. Physiol. Anim. Nutr. (Berl.). 2006; 90 (16422763): 1-11
        • Blum J.W.
        • Hammon H.
        Colostrum effects on the gastrointestinal tract, and on nutritional, endocrine and metabolic parameters in neonatal calves.
        Livest. Prod. Sci. 2000; 66: 151-159
      1. BOE. 1997. Orden de 31 de enero de 1997 por la que se setablecen los Metodos Oficinales de Analisis de ACeites y Grases, Cereales y Derivados, Productos Lacteos y productos derivados de la Uva. Agencia Estatal. Gobierno de Espana.

        • Britti D.
        • Massimini G.
        • Peli A.
        • Luciani A.
        • Boari A.
        Evaluation of serum enzyme activities as predictors of passive transfer status in lambs.
        J. Am. Vet. Med. Assoc. 2005; 226 (15786999): 951-955
        • Buczinski S.
        • Dubuc J.
        • Bourgeois V.
        • Baillargeon P.
        • Côté N.
        • Fecteau G.
        Validation of serum gamma-glutamyl transferase activity and body weight information for identifying dairy calves that are too young to be transported to auction markets in Canada.
        J. Dairy Sci. 2020; 103 (31864751): 2567-2577
        • Chigerwe M.
        • Hagey J.V.
        • Aly S.S.
        Determination of neonatal serum immunoglobulin G concentrations associated with mortality during the first 4 months of life in dairy heifer calves.
        J. Dairy Res. 2015; 82 (26383079): 400-406
        • Cuttance E.L.
        • Mason W.A.
        • Denholm K.S.
        • Laven R.A.
        Comparison of diagnostic tests for determining the prevalence of failure of passive transfer in New Zealand dairy calves.
        N. Z. Vet. J. 2017; 65 (27580795): 6-13
        • Cuttance E.L.
        • Regnerus C.
        • Laven R.A.
        A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand.
        N. Z. Vet. J. 2019; 67 (31401943): 277-286
        • Dawes M.E.
        • Tyler J.W.
        • Hostetler D.
        • Lakritz J.
        • Tessman R.
        Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves.
        J. Am. Vet. Med. Assoc. 2002; 220 (11918273): 791-793
        • de Souza R.S.
        • dos Santos L.B.C.
        • Melo I.O.
        • Cerqueira D.M.
        • Dumas J.V.
        • Leme F.O.P.
        • Moreira T.F.
        • Meneses R.M.
        • de Carvalho A.U.
        • Facury-Filho E.J.
        Current diagnostic methods for assessing transfer of passive immunity in calves and possible improvements: A literature review.
        Animals (Basel). 2021; 11 (34679982)2963
        • Devant M.
        • Marti S.
        Strategies for feeding unweaned dairy beef cattle to improve their health.
        Animals (Basel). 2020; 10 (33080998)1908
        • Drackley J.
        Does early growth affect subsequent health and performance of heifers?.
        Adv. Dairy Technol. 2005; 17: 189-205
        • European Union
        Council directive: Laying down minimum standards for protection of calves.
        Off. J. Eur. Union. 2008; 10: 7-13
        • Fallon R.J.
        • Harte F.J.
        • Harrington D.
        The Effect of calf purchase weight, serum IG level and feeding systems on the incidence of diarrhoea, respiratory disease, and mortality.
        Bov. Pract. 1987; 22: 104-106
        • Fecteau G.
        • Baillargeon P.
        • Higgins R.
        • Pare J.
        • Fortin M.
        Bacterial contamination of colostrum fed to newborn calves in Québec dairy herds.
        Can. Vet. J. 2002; 43 (12125183): 523-527
        • Garcia-Granero M.
        Lin's Concordance Correlation Coefficient [Computer Programm: Syntax code SPSS].
        https://tinyurl.com/yc38wynf
        Date: 2009
        Date accessed: July 25, 2022
        • Godden S.
        Colostrum management for dairy calves.
        Vet. Clin. North Am. Food Anim. Pract. 2008; 24 (18299030): 19-39
        • Hammon H.M.
        • Liermann W.
        • Frieten D.
        • Koch C.
        Review: Importance of colostrum supply and milk feeding intensity on gastrointestinal and systemic development in calves.
        Animal. 2020; 14 (32024575): s133-s143
        • Hassig M.
        • Stadler T.
        • Lutz H.
        Transition from maternal to endogenous antibodies in newborn calves.
        Vet. Rec. 2007; 160 (17308022): 234-235
        • Hogan I.
        • Doherty M.
        • Fagan J.
        • Kennedy E.
        • Conneely M.
        • Brady P.
        • Ryan C.
        • Lorenz I.
        Comparison of rapid laboratory tests for failure of passive transfer in the bovine.
        Ir. Vet. J. 2015; 68 (26309724): 18
        • Hulbert L.E.
        • Moisá S.J.
        Stress, immunity, and the management of calves.
        J. Dairy Sci. 2016; 99 (26805993): 3199-3216
        • ISO
        ISO 8968-3: Milk – Determination of Nitrogen Content – Part 3: Block-digestion Method.
        International Organization for Standardization, 2004
        • ISO
        ISO 1211: Milk – Determination of Fat Content – Gavimetric Method.
        International Organization for Standardization, 2010
        • Kien C.L.
        • McClead R.E.
        • Cordero Jr., L.
        In vivo lactose digestion in preterm infants.
        Am. J. Clin. Nutr. 1996; 64 (8901788): 700-705
        • Korhonen H.
        • Marnila P.
        • Gill H.S.
        Milk immunoglobulins and complement factors.
        Br. J. Nutr. 2000; 84 (11242450): 75-80
        • Lombard J.
        • Quigley J.
        • Haines D.
        • Garry F.
        • Earleywine T.
        • Urie N.
        • Chamorro M.
        • Godden S.
        • McGuirk S.
        • Smith G.
        • Shivley C.
        • Catherman D.
        • Heinrichs A.J.
        • James R.
        • Maas J.
        • Sterner K.
        • Sockett D.
        Letter to the editor: Comments on Schalich et al. 2021. Colostrum testing with Brix is a valuable on farm tool. 10.193/jas/skab083.
        J. Anim. Sci. 2022; 100: 1-3
        • Lombard J.
        • Urie N.
        • Garry F.
        • Godden S.
        • Quigley J.
        • Earleywine T.
        • McGuirk S.
        • Moore D.
        • Branan M.
        • Chamorro M.
        • Smith G.
        • Shivley C.
        • Catherman D.
        • Haines D.
        • Heinrichs A.J.
        • James R.
        • Maas J.
        • Sterner K.
        Consensus recommendations on calf- and herd-level passive immunity in dairy calves in the United States.
        J. Dairy Sci. 2020; 103 (32448583): 7611-7624
        • Marcato F.
        • van den Brand H.
        • Kemp B.
        • van Reenen K.
        Evaluating potential biomarkers of health and performance in veal calves.
        Front. Vet. Sci. 2018; 5 (29977895): 133
        • McBeath D.G.
        • Penhale W.J.
        • Logan E.F.
        An examination of the influence of husbandry on the plasma immunoglobulin level of the newborn calf, using a rapid refractometer test for assessing immunoglobulin content.
        Vet. Rec. 1971; 88 (4994654): 266-270
        • Moore D.A.
        • Sischo W.M.
        • Festa D.M.
        • Reynolds J.P.
        • Atwill E.R.
        • Holmberg C.A.
        Influence of arrival weight, season, and calf supplier on survival in Holstein beef calves on a calf ranch in California, USA.
        Prev. Vet. Med. 2002; 53 (11821140): 103-115
        • Ontsouka E.C.
        • Albrecht C.
        • Bruckmaier R.M.
        Invited review: Growth promoting effects of colostrum in calves based on interaction with intestinal cell surface receptors and receptor-like transporters.
        J. Dairy Sci. 2016; 99 (26874414): 4111-4123
        • Pardon B.
        • Alliët J.
        • Boone R.
        • Roelandt S.
        • Valgaeren B.
        • Deprez P.
        Prediction of respiratory disease and diarrhea in veal calves based on immunoglobulin levels and the serostatus for respiratory pathogens measured at arrival.
        Prev. Vet. Med. 2015; 120 (25937168): 169-176
        • Paré J.
        • Thurmond M.C.
        • Gardner I.A.
        • Picanso J.P.
        Effect of birthweight, total protein, serum IgG and packed cell volume on risk of neonatal diarrhea in calves on two California dairies.
        Can. J. Vet. Res. 1993; 57 (8269362): 241-246
        • Parish S.M.
        • Tyler J.W.
        • Besser T.E.
        • Gay C.C.
        • Krytenberg D.
        Prediction of serum IgG1 concentration in Holstein calves using serum gamma glutamyltransferase activity.
        J. Vet. Intern. Med. 1997; 11 (9470159): 344-347
        • Pauli J.V.
        Colostral transfer of gamma glutamyl transferase in lambs.
        N. Z. Vet. J. 1983; 31 (16030994): 150-151
        • Perino L.J.
        • Sutherland R.L.
        • Woollen N.E.
        Serum gamma glutamyltransferase activity and protein concentration at birth and after suckling in calves with adequate and inadequate passive transfer of immunoglobulin G.
        Am. J. Vet. Res. 1993; 54 (8093994): 56-59
        • Rauprich A.B.E.
        • Hammon H.M.
        • Blum J.W.
        Influence of feeding different amounts of first colostrum on metabolic, endocrine, and health status and on growth performance in neonatal calves.
        J. Anim. Sci. 2000; 78 (10784179): 896-908
        • Renaud D.
        • Pardon B.
        Preparing male dairy calves for the veal and dairy beef industry.
        Vet. Clin. North Am. Food Anim. Pract. 2022; 38 (35219487): 77-92
        • Renaud D.L.
        • Duffield T.F.
        • LeBlanc S.J.
        • Ferguson S.
        • Haley D.B.
        • Kelton D.F.
        Risk factors associated with mortality at a milk fed veal calf facility: A prospective cohort study.
        J. Dairy Sci. 2018; 101 (29290439): 2659-2668
        • Renaud D.L.
        • Duffield T.F.
        • LeBlanc S.J.
        • Haley D.B.
        • Kelton D.F.
        Management practices for male calves on Canadian dairy farms.
        J. Dairy Sci. 2017; 100 (28551179): 6862-6871
        • Roland L.
        • Drillich M.
        • Klein-Jöbstl D.
        • Iwersen M.
        Invited review: Influence of climatic conditions on the development, performance, and health of calves.
        J. Dairy Sci. 2016; 99 (26874416): 2438-2452
        • Schalich K.M.
        • Reiff O.M.
        • Nguyen B.T.
        • Lamb C.L.
        • Mondoza C.R.
        • Selvaraj V.
        Temporal kinetics of bovine mammary IgG secretion into colostrum and transition milk.
        J. Anim. Sci. 2021; 99 (33715013)skab083
        • Schalich K.M.
        • Selvaraj V.
        Contradictions on colostrum IgG levels and Brix values are real and can be explained. Response to letter by Lombard et al. 2022.
        J. Anim. Sci. 2022; 100 (35483038)skac120
        • Shope R.E.
        • Gowen J.W.
        Cholesterol and cholesterol ester content of bovine colostrum.
        J. Exp. Med. 1928; 48 (19869468): 21-24
        • Thompson J.C.
        • Pauli J.
        Colostral transfer of gamma glutamyl transpeptidase in calves.
        N. Z. Vet. J. 1981; 29 (16030805): 223-226
        • Thurnhofer H.
        • Hauser H.
        Uptake of cholesterol by small intestinal brush border membrane is protein-mediated.
        Biochemistry. 1990; 29 (2328246): 2142-2148
        • Tyler J.W.
        • Hancock D.D.
        • Parish S.M.
        • Rea D.E.
        • Besser T.E.
        • Sanders S.G.
        • Wilson L.K.
        Evaluation of 3 assays for failure of passive transfer in calves.
        J. Vet. Intern. Med. 1996; 10 (8884716): 304-307
        • Villarroel A.
        • Miller T.B.
        • Johnson E.D.
        • Noyes K.R.
        • Ward J.K.
        Factors Affecting Serum Total Protein and Immunoglobulin G Concentration in Replacement Dairy Calves.
        J. Adv. Dairy Res. 2013; 1: 1-5
        • Weaver D.M.
        • Tyler J.W.
        • Scott M.A.
        • Wallace L.M.
        • Marion R.S.
        • Holle J.M.
        Passive transfer of colostral immunoglobulin G in neonatal llamas and alpacas.
        Am. J. Vet. Res. 2000; 61 (10895892): 738-741
        • Wilm J.
        • Costa J.H.C.
        • Neave H.W.
        • Weary D.M.
        • von Keyserlingk M.A.G.
        Technical note: Serum total protein and immunoglobulin G concentrations in neonatal dairy calves over the first 10 days of age.
        J. Dairy Sci. 2018; 101: 6430-6436
        • Wilson D.J.
        • Stojkov J.
        • Renaud D.L.
        • Fraser D.
        Risk factors for poor health outcomes for male dairy calves undergoing transportation in western Canada.
        Can. Vet. J. 2020; 61 (33299241): 1265-1272
        • Winder C.B.
        • Kelton D.F.
        • Duffield T.F.
        Mortality risk factors for calves entering a multi-location white veal farm in Ontario, Canada.
        J. Dairy Sci. 2016; 99 (27720158): 10174-10181
        • Yu K.
        • Canalias F.
        • Solà-Oriol D.
        • Arroyo L.
        • Pato R.
        • Saco Y.
        • Terré M.
        • Bassols A.
        Age-related serum biochemical reference intervals established for unweaned calves and piglets in the post-weaning period.
        Front. Vet. Sci. 2019; 6 (31069239): 123
        • Zanker I.A.
        • Hammon H.M.
        • Blum J.W.
        Activities of γ-Glutamyltransferase, alkaline phosphatase and aspartate-aminotransferase in colostrum, milk and blood plasma of calves fed first colostrum at 0–2, 6–7, 12–13 and 24–25h after birth.
        J. Vet. Med. A Physiol. Pathol. Clin. Med. 2001; 48 (11379391): 179-185