Use of Every Ten-Day Criteria for Metabolic Profile Test after Calving and Dry Off in Dairy Herds

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1 FULL PAPER Clinical Pathology Use of Every Ten-Day Criteria for Metabolic Profile Test after Calving and Dry Off in Dairy Herds Katsuya KIDA 1) 1) Large Animal Clinic and Research Center, Motonopporo 612, Ebetsu, Hokkaido , Japan (Received 5 October 2001/Accepted 31 July 2002) ABSTRACT. The traditional metabolic profile test cannot be applied to peripartum dairy cows, because these cows are in a state of physiological abnormality making it difficult to interpret their blood components. This study aimed at establishing and evaluating the practicability of interpreting a metabolic profile test every 10 days (Ten-day criteria) during the dry and lactation periods in herds with high and no incidence of peripartum diseases. Data from 29,043 cows in 1,130 commercial dairy herds were used to establish standard values every 10 days, mean ± 1.0 standard deviation for the metabolic profile test. The practicability of these criteria was evaluated in herds with peripartum diseases. In the ten-day criteria, the body condition score, albumin, blood urea nitrogen, glucose, total chole sterol, nonesterified fatty acids, γ-glutamyl transpeptidase and aspartate aminotransferase, fluctuated during the dry and early lactation periods and there were very big changes in packed cell volume, blood urea nitrogen, total cholesterol and magnesium just after calving. The tenday criteria were able to detect overconditioned cows, low levels of albumin, total cholesterol and magnesium, and high nonesterified fatty acids in herds with a high incidence of peripartum diseases. In conclusion, the ten-day criteria can be successfully applied to peripartum cows, and is recommended because it is able to detect metabolic abnormalities not only in the herd, but also in individual cows. KEY WORDS: criterion, dairy cow, metabolic profile test. J. Vet. Med. Sci. 64(11): , 2002 The metabolic profile test (MPT) was first established by Payne et al. [19] as a tool for assessing metabolic status and helping in the diagnosis of metabolic disorders in dairy herds. Subsequently, many researchers have applied MPT to improve feeding management [1, 2, 4 7, 10, 12, 13, 18, 25], detect subclinical health problems and prevent production diseases [1, 6, 12, 16, 18]. MPT, in conjunction with animal and facility evaluation, body condition scoring and ration evaluation, is an extremely useful tool for nutritional evaluation in dairy herds [22,23]. MPT has also been applied in many commercial dairy herds in Japan [16,17]. To carry out the MPT, blood samples are collected from cows, divided into groups of several cows at two or three different stages of lactation and the dry period, according to milk production and feeding management [9, 23]. The levels of blood components reflect not only nutritional status, but also the health and physiological condition of each cow such as disease, age, lactation stage and milk production [4, 9, 11 13], so that cows selected for sampling should be representative of the herd [20, 23]. Normal MPT reference ranges are defined as mean values and different ranges of standard deviation (SD). Values from blood analysis are compared with the population average or ranges of standard values [6, 8, 9, 13, 16, 19]. Common dairy herds in Hokkaido consist of about 50 to 60 milking cows of various ages and health status. In such herds, it is often difficult to select an equal number of cows representative of each stage. Under these conditions, diagnosis by mean values at each stage could lead to wrong interpretation. This is because it is difficult to clearly determine whether changes in blood components are due to management problems in the herd, or specific problems in individual cows. This suggests that it is better to assess not only mean values at each stage, but also individual cow results for accurate diagnosis of MPT. It is generally recommended that peripartum cows should not be included in the MPT, because these cows are in a state of physiological abnormality making it difficult to interpret their blood components [19, 23], but the peripartum period is critical for the occurrence of metabolic disease [3, 14, 25]. Therefore, including these cows in the MPT would lead to early diagnosis and prevention of peripartum metabolic disorders. The aim of this study was to establish ten-day criteria for diagnosis of MPT in groups and individual peripartum cows. The practical application of these criteria was also assessed in commercial dairy herds, including the peripartum period. MATERIALS AND METHODS Dairy herds: In a total of 29,043 Holstein-Friesian cows from 1,130 commercial dairy herds in various districts of Hokkaido, the MPT was conducted from 1987 to Feeding and management were different in each herd. Generally all cows were housed indoors and grazed on pasture in summer. Feeding systems included continuous feeding of a totally mixed ration, or separate feeding of forage and concentrates. The MPT was conducted through four seasons. Cow selection: Cows included in the study were of various ages and had no apparent clinical problems. The cows were divided into 5 stages as shown in Table 1, and were subdivided into groups for 10-day intervals at each stage. Although the total number of cows tested varied in each herd, because of differences in the distribution of days in milk (DIM), about 30 cows per herd were selected from the

2 1004 K. KIDA Table 1. Classification of cows into dry period and lactation stages for metabolic profile test Stage Duration Dry period Dry off to calving Early lactation Calving to 49 DIM a) Peak lactation 50 to 109 DIM Mid lactation 110 to 209 DIM Late lactation 210 DIM to dry off a) Days in milk. 5 stages. The average number of calvings was 2.9 (range 1 to 13), and average milk production was 28.8 kg/cow/day (range 2.4 to 66.5 kg/cow/day). Body condition scoring: All MPT cows were body condition scored (BCS) on a scale of 1 to 5 (1= too thin, 5= too fat) [24] by the author. Blood collection and analytical methods: Blood was analyzed on the farm in a vehicle equipped with automatic biochemical analyzers. Blood samples were collected from the jugular vein in plain vacuum tubes between 09:00 a.m. and 11:00 a.m., 2 3 hr after morning feeding. Heparinized micro-capillary tubes were immediately filled for determination of the packed cell volume (PCV). Just after blood collection, to avoid a drop in glucose, serum was separated by centrifuging (750 g) for 5 min. As fibrin appears in the serum layer, a small quantity of plastic granules for serum separation (Blut-Z, TOHSHIN CHEMICAL Co., Ltd.) was then added into the sample tubes and further centrifuged (1,600 g) for 5 min. Within 1 2 hr of collection, serum components were analyzed with an automatic biochemical analyzer (VP-Super, DINABBOTT Co., Ltd. or TBA-40FR, TOSHIBA MEDICAL SYSTEMS Co., Ltd.) and an automated electrophoresis system (CTE-150 or CTE-700, JOKOH Co., Ltd.). MPT components: MPT components were chosen according to Payne et al. [19] and Ohgi et al. [16], as shown in Table 2. Indicators used for protein status were PCV, albumin (Alb), γ-globulin (Glb), and blood urea nitrogen (BUN), for energy status, glucose (Glc), total cholesterol (Cho) and nonesterified fatty acids (NEFA), for liver function, asparate aminotransferase (AST) and γ-glutamyl transpeptidase (GGT), and for mineral metabolism, calcium (Ca), inorganic phosphorus (ip) and magnesium (Mg). BCS was also estimated to assess energy balance at the time of blood sampling. Ten-day criteria: To define the ten-day criteria, population distributions of all the components of the MPT were evaluated by using the mean and SD. As distributions for Glb, NEFA and AST were skewed to the left, statistical analysis was done after logarithmic transformation. As for values which deviated from the range, the means ± 3 SD for all the cows were rejected. To establish the criteria, this process was repeated until no deviated value could be detected. For every 10 days, i.e. ten-day criteria (TDC) from dry off (60 days before calving) to calving and from calving to 300 DIM, the range of mean ± 1.0 SD for all MPT components was determined. For the control, stage criteria (STC) - mean ± 1.3 SD which is the practical and commonly used criterion [1, 16], was also determined. Variations in each component of TDC and STC during the dry and lactation periods were compared. Comparison of incidence of deviations in herds with high and no incidence of peripartum diseases: Two groups of herds were selected from the herds described above, as shown in Table 3. The diseased group (Group D) recorded a high incidence of prepartum diseases in the last 3 months before the MPT, and healthy group (Group H) no incidence of diseases. With the TDC, an individual cow s value was diagnosed as normal if its value was in the range of the upper (mean+1.0 SD) and lower limits (mean-1.0sd) or as abnormal if its value deviated from these ranges. In TDC and STC, the differences in incidence of deviations, during the dry and early lactation periods, in all components of Table 2. Analytical methods used for metabolic profile components Components Analysis method Protein status Packed cell volume (PCV) Micro capillary method Albumin (Alb) Electrophoretic analysis γ-globulin (Glb) Electrophoretic analysis Blood urea nitrogen (BUN) Urease-GLDH method Energy status Body condition score (BCS) 1(too thin) to 5(obese), 0.25 interval Glucose (Glc) HK-G6PDH method Cholesterol (Cho) Cholesterol oxidase method Nonesterified fatty acid (NEFA) Enzymatic colorimetric method Liver function Aspartate aminotransferase (AST) MDH UV method γ-glutamyl transpeptidase (GGT) Glu-3-CA-4-NA substrate method Minerals Calcium (Ca) O-CPC method Inorganic phosphorus (ip) Enzymatic UV method Magnesium (Mg) Enzymatic UV method

3 TEN-DAY CRITERIA FOR METABOLIC PROFILE TEST 1005 Table 3. Profiles of the groups with high and no incidence of peripartum diseases Group D a) Group H b) Number of herds Number of cows in dry period and early lactation Number of peripartum diseases treated in the last 3 month before MPT Milk production as the adjusted corrected milk c) (kg/cow/day) a) Diseased group - occurrence of peripartum diseases such as retained placenta, milk fever, ketosis, fatty liver and abomasal displacement etc. b) Healthy group - no incidence of prepartum diseases which required veterinary attention. c) 3.5% fat corrected milk produced by 150 DIM cows in a herd, with 1/3 of the herd consisting of primiparous cows [15]. MPT in groups D and H were evaluated by chi-square analysis. The components in which significant differences were observed between groups D and H in TDC, distribution of the number of deviations every 10 days during the peripartum period was further evaluated. Feeding and management of the cows during the peripartum period were also evaluated. RESULTS For several components of the MPT, upper limits (mean SD) or lower limits (mean SD) in every ten-day period from dry off through lactation, compared to upper limits (mean SD) or lower limits (mean SD) of each stage, are shown in Fig. 1. Protein status: In TDC, Glb went down before calving but Alb, BUN and PCV remained stable during the dry period. Alb, Glb and BUN went up in the early lactation stage. In particular, the TDC BUN lower limit during the first 10 days after calving was lower (7.9 mg/100 ml) than in STC (8.5 mg/100 ml). The TDC PCV upper limit, during the first 10 days after calving was higher (35.5%) than the STC (34.0%) and then went down during early lactation. Between the peak and mid lactation, TDC PCV fluctuated between the lower (27.3%) and upper (35.2%) limits of STC. Except for PCV, all the other protein indicators were stable after peak lactation. Energy status: During the dry period in TDC, BCS and NEFA went up but Cho gradually went down. Just after calving, there were drastic changes in BCS and NEFA. BCS went down and NEFA went up. TDC BCS and NEFA went down until 70 to 100 DIM, BCS then gradually went up and NEFA remained stable until dry off. In the TDC, Cho upper limits were higher (171 mg/100 ml) than in the STC (148 mg/100 ml) just after dry off, but were lower (115 mg/100 ml) than in the STC just before calving. After calving the level of TDC Cho went up to 80 DIM and remained high until late lactation when it started to go down. During the first 10 days after calving, the lower limits of TDC Cho were lower (58 mg/100 ml) than STC Cho (83 mg/100 ml) whereas the TDC Cho upper limits were higher (226 mg/ 100 ml) than the STC Cho (212 mg/100 ml) at 40 to 49 DIM. Liver function: In TDC, AST was stably fluctuating between 44 to 73 IU/l during the dry period. It then went up rapidly during the first 10 days after calving and went down thereafter until 50 DIM. After 50 DIM, it remained stable fluctuating between 49 and 82 IU/l. TDC GGT went down during the dry period, and gradually went up after calving reaching a light peak (28 IU/l) during mid lactation. Mineral status: In TDC, except during the first 10 days after calving when values were low, there was little variation in Ca and ip during all the stages. TDC Mg was stable during the dry period fluctuating between 2.1 and 2.7 mg/ 100 ml. During the first 10 days after calving, TDC Mg was at the lowest value and was also lower than STC Mg. It then went up until 30 DIM remaining stable and fluctuating between 2.3 and 2.8 mg/100 ml thereafter. Incidence of deviations: For the results of the MPT, the method of diagnosis by TDC or STC is shown in Fig. 2. The results of the MPT were classified into any one of four diagnoses, such as normal by both criteria, abnormal by both criteria, normal by TDC but not by STC or normal by STC but not by TDC. In TDC, there were significant differences between groups D and H in the lower limit (mean SD) incidence of deviations of Alb, Cho, NEFA, AST and Mg (Table 4). In group D, the incidence of Alb (p<0.01), Cho (p<0.01), AST (p<0.05) and Mg (p<0.05) deviations was higher and NEFA (p<0.01) lower than in group H. On the other hand, there were significant differences between groups D and H only in Alb (p<0.01) and Cho (p<0.01) in the lower limits of STC (mean SD). In TDC upper limits, there were significant differences in 4 components, i.e. a higher incidence of deviations of BCS (p<0.01) and NEFA (p<0.01) and a lower incidence of Alb (p<0.01) and Mg (p<0.01) in group D than in group H. In STC upper limits, significantly higher incidences of deviations in Glb (p<0.05), BCS (p<0.01), NEFA (p<0.05) and GGT (p<0.05) were observed in group D than in group H. The proportions of cows with deviated values in TDC or STC to the total number of cows during the dry and early lactation periods, are shown in Fig. 3. Except for BCS, all the other components in which significant differences were observed between groups D and H, the distribution pattern of the number of deviations by TDC was similar to that of the total number of cows tested. On the other hand, in the

4 1006 K. KIDA distribution of the number of deviations by STC, it was observed that there were more cows with deviated values in upper BCS, lower Cho, high NEFA and lower Mg from calving to 20 DIM, and fewer deviated from 40 to 50 DIM. The proportion of herds in each group that had some feeding and related problems is shown in Table 5. The proportion of herds in which dry cows had depression of the left paralumbar fossa, was higher in group D than in group H (p<0.01). Furthermore, no lead feeding was given to prepartum cows in more herds in group D than in group H (p<0.05). After calving, though feeding of concentrate reached over 15 kg/day in about half the herds in each group, the rate of daily increase in concentrate feeding was higher in group D than in group H (p<0.01). The proportion of the herds in which cows were allowed to graze for more than 6 hr a day (p<0.05), and in which cows were fed moldy hay, grass or corn silage (p<0.01), was higher in group D than in group H. DISCUSSION Fig. 1. Standard values for blood metabolites and body condition score by every 10-day and stage criteria during the dry and lactation periods. PCV: packed cell volume, BUN: blood urea nitrogen, BCS: body condition score, Glc: glucose, NEFA: nonesterified fatty acid, Cho: total cholesterol, Mg: magnesium. Long lines: mean ± 1.3 SD in each stage. Short lines: mean ± 1.0 SD every 10 days. Since Payne et al. [19] explained the principle of MPT, its criteria and interpretation have been widely studied. Standard values in each stage or group of milk production which are used to interpret group data, have also been constituted [9, 23]. STC values observed in this study were similar to those reported by Ohgi et al. [16]. This study found that by using TDC, all the MPT components varied according to days relative to calving during the dry and early lactation periods. Furthermore, there were also variations in BCS, PCV, BUN and Cho during mid and late lactation. Cows DIMs are not uniform at any particular stage, so that the estimations of average values might be affected when STC is used. This is further supported by the different distribution patterns of the number of deviations between TDC and STC during the peripartum period. The dramatic changes in almost all MPT components except PCV, GGT, Ca and ip, particularly during the early lactation stage, suggest that diagnosis by a single criterion such as STC would be difficult. To apply the MPT to peripartum cows, it would be better to set the criterion at shorter intervals such as every day, rather than every 10 days, but establishing practical everyday criteria requires data on a large number of cows, which was impossible in this study. On the other hand, there was little variation in ten-day criteria of Alb, Glob, NEFA and minerals during the late lactation stage, so that it may not be necessary to use TDC in late lactation for these components. There are big differences in milk production and feeding management between the beginning (up to 210 DIM) and the end of late lactation. Although the level of blood components may not be different, the reason for the result is different and the interpretation of the result should also be different. Therefore, in this study, the criteria were established at 10-day intervals for all components during all stages. To avoid misinterpretation, it is recommended that MPT

5 TEN-DAY CRITERIA FOR METABOLIC PROFILE TEST 1007 should exclude peripartum cows which have physiologically wide variations in levels of blood components [9, 19, 22, 23], but this period is very important for disease prevention. For example, even though high NEFA levels about Fig. 2. Example of diagnosis by TDC and STC. TDC:, STC: , area a : normal by both criteria, area b : normal by STC and abnormal by TDC, area c ; normal by both criteria, area d : normal by TDC and abnormal by STC are shown. Y axis indicates mean ± 3 SD in all stages of criterion. calving time are common, they are associated with problems of off feed or calving stress [21]. A high increase in NEFA is also an obvious sign of ketosis and fatty liver [14]. There was also a gradual decrease in NEFA during the early lactation period. This is difficult to diagnose from the average value for several cows. In this study, similar variations in cows pre and post calving were observed in Alb, BUN, BCS, Glc, Cho, AST and Mg. Therefore, TDC reflects physiological changes in peripartum cows, and would more correctly interpret MPT results than STC. Adams et al. [1] reported that the distribution of values for a given parameter is important for the interpretation of MPT. This study found that interpretation of MPT by distribution of values in TDC was effective for detection not only in a herd but also of individual abnormalities. Adams et al. [1] suggested that using 1.0 SD from the mean in establishing normal ranges leads to a high incidence of deviate values in apparently normal herds, so that it is recommended to use 1.3 SD from the mean [1, 16]. By this criterion, 19% of cows tested are diagnosed as having deviate values. On the other hand, with 1.0 SD from the mean judges 32% of cows tested as having deviated values. There is a theoretical difference of 13% incidence of deviations between 1.3 and 1.0 SD criteria. This was confirmed in this study in which a difference of 10 to 15% incidence of deviations was observed between TDC and STC. One SD from the mean with TDC reflected physiological changes in cows better than STC during the dry and lactation periods. Furthermore considering the incidence of deviations, this study found that TDC detects abnormal in every ten-day during peripartum period, and TDC discriminates traits of herds with peripartum diseases better than STC. For example, Table 4. Incidence of deviations by every 10-day and stage criteria during the dry and early lactation periods in dairy herds with high and no incidence of peripartum diseases a) Deviations (%) in lower limits Deviations (%) in upper limits STC c) TDC b) TDC STC Components D d) H e) D H D H D H Packed cell volume Albumin ** ** ** γ-globulin * Blood urea nitrogen Body condition score ** ** Glucose Cholesterol ** ** Nonesterified fatty acids ** ** * Aspartate aminotransferase * γ-glutamyl transpeptidase * Calcium Inorganic phosphorus Magnesium * ** a) Rate (%) of cows out of the criteria. b) Ten-day criteria during dry and early lactation periods. c) Stage criteria in the dry and early lactation periods. d) Herds with high incidence of peripartum diseases. e) Herds with no incidence of diseases. * Significant difference (p< 0.05) between diseased and healthy herds for each criterion. ** Significant difference (p< 0.01) between diseased and healthy herds for each criterion.

6 1008 K. KIDA Fig. 3. Proportion of cow distribution every 10 days during the dry and early lactation periods. Lines indicate proportion of cows to total number of cows in the periods. Cows tested: + +, cows with deviated values from TDC: and STC: are shown. TDC detected a significantly higher incidence of hypomagnesemia which indicates dry matter intake (DMI) depression [21] during the dry and early lactation periods in herds with peripartum diseases, whereas STC showed no difference. This DMI depression was supported by results for many dry cows in diseased herds having depression of the left paralumbar fossa. Depression of the left paralumbar fossa indicates reduced rumen capacity and low DMI. TDC also detected high NEFA levels (energy deficiency) [9, 20, 23] better than STC (p<0.01 vs p<0.05). These high NEFA concentrations, particularly in group D, were because of inappropriate feeding such as no lead feeding, rapid increase in concentrate feeding and feeding moldy forage. This kind of management may lead to energy deficiency. Both TDC and STC correctly detected a high incidence of deviations in lower Alb and Cho. These indicate chronic protein deficiency or liver dysfunction in herds with peripartum diseases. It was considered that low Alb and Cho values were also a result of the inappropriate management described above. Blood parameters in the cow are affected by various factors such as age [2, 23], milk production [2, 11, 23], season [2, 8, 23], place of cow management [16] and in particular days relative to calving. These factors must be considered when establishing criteria for MPT. The criteria used in this study were set up on the basis of data from cows in many areas of Hokkaido with different management systems, season, age and milk production. Criteria based on the above factors, i.e. management systems, season, age and milk production, may be useful in improving accuracy, but there are wide differences in feeding management and housing systems between seasons in Hokkaido, making it very difficult to interpret MPT. These differences are sometimes observed between cows in the same herd. Depending on the difference in individual cow status, it is difficult to interpret MPT in different situations. It would therefore be better to

7 TEN-DAY CRITERIA FOR METABOLIC PROFILE TEST 1009 Table 5. Proportion of herds that had some feeding and related management problems during the dry and early lactation periods in dairy herds with high and no incidence of peripartum diseases Management problems Group D a) Group H b) Prepartum Depression of left paralumbar fossa c) ** No lead feeding * Postpartum Rapid increase in daily concentrate feeding d) ** Much concentrate feeding to early lactation cows e) ns Grazing f) * Feeding of totally mixed ration ns Moldy forage ** Low quality grass hay and silage g) ns Total number of herds a) % of herd observed in herds with high incidence of peripartum diseases. b) % of herd observed in herds with no incidence of diseases. c) Sign of DMI depression observed in dry cow in >14 days prepartum periods. d) >0.5 kg/day increasing of concentrate feeding until 21 DIM. e) >15kg/day of concentrate fed to early lactation cow. f) >6 hours gazing on pasture. g) Harvested too late. * Significant difference (P<0.05) between healthy herds and herds with high incidence of diseases. ** Significant difference (P<0.01) between healthy herds and herds with high incidence of diseases. use TDC in interpreting MPT as it accurately reflects physiological changes in cows during all stages of the dry and lactation periods. Evaluation of factors affecting an individual cow would be helpful and practical in the interpretation of MPT. This study established and used TDC to interpret MPT in cows during the peripartum period. TDC is not only able to detect management problems in a herd but also abnormal individual values, and in this respect is superior to STC. ACKNOWLEDGEMENTS. The author gratefully acknowledges the staff of the Large Animal Clinic and Research Center, Federation of Hokkaido Agricultural Mutual Aid Associations for their help in analyzing blood, and the bovine practitioners in local clinics of the Agricultural Mutual Aid Association in Hokkaido for their assistance in field work. REFERENCES 1. Adams, R. S., Stout, W. L., Kradel, D. C., Guss, S. B. Jr., Moser, B. L. and Jung, G. A Use and limitations of profiles in assessing health or nutritional status of dairy herds. J. Dairy Sci. 61: Amano, H., Takesima, Y., Nitta, M., Mabuti, T., Tokuti, T. and Yagi, T Relationship of haematocrit values with age, lactation stage, nutrition levels of dairy cows and temperature. J. Jpn. Vet. Med. Assoc. 45: Bertics, A. J., Grummer, R. R., Cadorniga-Valino, C. and Stoddard, E Effect of prepartum dry matter intake on liver triglyceride concentration and early lactation. J. Dairy Sci. 75: Blowey, R. W., Wood, D. W. and Davis, J. R A nutritional monitoring system for dairy herds based on blood glucose, urea and albumin levels. Vet. Rec. 92: Blowey, R. W A practical application of metabolic profiles. Vet. Rec. 97: Cote, J. F. and Hoff, B Clinical effects of low dietary phosphorus concentrations in feed given to lactating dairy cows. The Bovine Practitioner 26: Dyk, P. B., Emery, R. S., Liesman, J. L., Bucholtz, H. F. and Vande Haar, M. J Prepartum nonesterified fatty acids in plasma are higher in cows developing periparturient health problems. J. Dairy Sci. 78 (Suppl. 1): Ghergariu, S., Rowlands, G. J., Pop, A. L., Danielescu, N. and Moldovan, N. A A comparative study of metabolic profiles obtained in dairy herds in Romania. Brit. Vet. J. 140: Herdt, T. H Variability characteristics and test selection in herd-level nutritional and metabolic profile testing. Vet. Clin. North Am. Food Anim. Pract. 16: Hiratsuka, H., Takahashi, K., Kurosawa, T., Sonoda, M., Ataku, K. and Narasaki, N Correlation between nutrient intake and blood components of dairy cows in early lactation stage. J. Jpn. Vet. Med. Assoc. 40: Jones, G. M., Wildman, E. E., Troutt, H. F. Jr. Lesch, T. N., Wagner, P. E., Boman, R. L. and Lanning, N. M Metabolic profiles in Virginia dairy herds of different milk yields. J. Dairy Sci. 65: Kronfeld, D. S., Donoghue, S., Copp, R. L., Sterns, F. M. and Engle, R. H Nutritional status of dairy cows indicated by analysis of blood. J. Dairy Sci. 65: Lee, A. J., Twardock, A. R., Bubar, R. H., Hall, J. E. and Davis, C. L Blood metabolic profiles, their use and rela-

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