Immunoglobulin transfer in the viable, newborn calf

Parturitions of 26 dairy-cows were observed and the viability of the newborn calves assessed. Immediately after birth as well as after the first and forth colostrum intake a blood probe was taken from the neonates and investigated for the following parameters: blood gas, blood picture, total-IG, concentrations of IGG1, IGM and IGE, total proteins and albumin. Before each feeding a colostrum sample was assayed for the same parameters with the exception of blood gas, blood picture and albumin. A transferindex (KLOBASA et al., 1994) was calculated to investigate the absorption of TP, IG, IGG1, IGM and IGE. In some of the blood samples there was no IGE detectable, even after colostrum intake. High individual differences were striking and blood concentrations were not related to the milk contents or the time until first feeding. The total proteins were strongly correlated to the serum concentrations of IGG1, but only in samples taken after first colostrum feeding. Vital calves showed the following tendency: colostrum intake of less than 1 l within the first 15 hours after birth and/or colostrum intake with TP-concentrations of 80 I.U./l was partly compensated by increased absorption of TP. 24 hours after birth serum-immunoglobulinconcentrations were 6-12 mg/ml; weight gain was normal and no calf became ill. In 60% of cases the transferindex for IGM was higher than for IGG1 or IGE.     

Effects of replacement of native fat in colostrum and milk with coconut oil on fat-soluble vitamins in serum and immune function in calves

Fat-soluble vitamins and their metabolites modulate immune function in a variety of animal species. The objective of the present study was to determine the role of fat-soluble vitamins in colostrum and milk in the development of specific aspects of immune function in the calf during the 1st wk postpartum. During this period, control calves (n = 6) were fed normal colostrum and milk, and calves in the treatment group (n = 6) were fed skimmed colostrum and skimmed milk supplemented with coconut oil. Treated calves did not experience the progressive increase in concentrations of retinol, beta-carotene, alpha-tocopherol, 1,25-dihydroxyvitamin D, or retinoic acids in serum that was observed in control calves. Acquisition of passive immunity, which is indicated by concentrations of immunoglobulin G1 in serum, was unaffected by treatment. Composition and functional capacities of populations of blood mononuclear leukocytes that were collected from birth to 7 d postpartum were also unaffected by treatment. Major changes in the function and composition of mononuclear leukocyte populations from all calves occurred during the experimental period and were unrelated to the concentrations of fat-soluble vitamins in serum. Populations of blood mononuclear leukocytes from calves were functionally hyporesponsive and compositionally different from populations of blood mononuclear leukocytes from adult nongravid cows. These differences likely reflected the immaturity of the immune system of the neonatal calf and may contribute to the increased susceptibility of the calf to infectious disease.     

Serum immunoglobulin G concentration in goat kids fed colostrum or a colostrum substitute

To determine the suitability of a new colostrum substitute derived from goat serum and to determine the amount of colostral IgG needed to achieve serum IgG concentration > 800 mg/dl, twin kids from 14 does were fed colostrum or a colostrum substitute. The volume of colostrum or colostrum substitute fed was calculated so that half the kids in each group received IgG at a low dosage (1.5 g/kg of body weight) and the other half received IgG at a high dosage (3 g/kg). Kids were bottle fed the colostrum or colostrum substitute and then fed pooled goat's milk until 18 hours old, at which time they were allowed to nurse their dams. Does were milked manually every 2 hours after parturition until specific gravity of mammary secretions was < 1.02, the specific gravity of goat's milk. Serum IgG concentration of each kid was determined by means of single radial immunodiffusion at birth and 12, 18, and 24 hours and 7, 21, and 42 days after birth. Kids were weighed at each blood collection and monitored for illness daily. None of the kids had measurable serum IgG concentrations at birth. Mean serum IgG concentration was significantly higher in kids fed colostrum than in kids fed colostrum substitute at all times, except days 7 and 42 (P < 0.05). By 24 hours after birth, serum IgG concentration was > 800 mg/dl in all kids fed colostrum, in 4 of 7 kids fed the substitute at the higher dosage, and in 2 of 7 kids fed the substitute at the lower dosage.(ABSTRACT TRUNCATED AT 250 WORDS)     

Examination of the total protein concentration and gamma-glutamyltransferase (gamma-GT) in calves during the first 14 days of life and their significance for predictions of health status in the first two postnatal weeks]

The objective of the present paper was to examine the development of the total protein concentration in blood serum and the gamma-GT activity in 49 calves in relation to the health status during the first 14 days of life. For this purpose, blood samples were taken immediately after birth of the animals, after 6, 12, 18 and 24 hours and from then onwards once a day with the calves being subject to a thorough clinical examination each time. All calves received pooled colostrum. At the end of the experiment, the calves were subdivided into three groups on the basis of their health status, i.e. control calves, calves with a slight or moderately severe disease and calves suffering from a severe disease. In all three groups the total protein concentration increased significantly after the colostrum intake and decreased slowly from the 2nd day of life onwards. The gamma-GT activity as well increased significantly after the colostrum intake and decreased rapidly after 12th hour of life and progressively in the further course of the experiment. Both parameters varied considerably from one calf to the next and no statistically significant differences in these parameters between the three groups could be found. It can thus be deduced that the total protein concentration in blood serum and the gamma-GT activity do not allow any predictions about the development of the health status during the first 14 days of life to be made.     

Effects of a colostrum replacement product derived from serum on immunoglobulin G absorption by calves

Calves are born hypogammaglobulinemic and rely on immunoglobulin (Ig) from colostrum to obtain passive immunity. Previous research has indicated that colostrum supplements derived from milk are less effective than is maternal colostrum in providing adequate IgG to neonatal calves. Our objective was to determine the absorption of IgG by newborn calves fed a USDA food-grade colostrum supplement derived from bovine serum or fed pooled maternal colostrum. Holstein calves (n = 20; 10 bulls) were removed from the dam within 1 h of birth and were housed in individual stalls for the 24-h study. Calves were fed 2 L of colostrum or colostrum replacer at 1.5 and 13.5 h (+/- 0.1 h). Calves were blocked by colostrum pool, and replacer was fed to provide equal intakes of IgG within blocks. Jugular blood was collected at 1 and 24 h (+/- 0.1 h) for analysis of IgG by radial immunodiffusion. At 24 h, calves were injected with 1.5 ml of Evans blue dye to estimate plasma volume. Mean plasma IgG at 24 h of age was 7.3 +/- 0.4 g/L and was affected by an interaction of block and treatment. Apparent efficiency of IgG absorption of 24 h was reduced when 750 g of the colostrum replacement product were fed but was increased when 266 g of colostrum replacement product were fed. Mean plasma volume was unaffected by treatment and was 3.5 +/- 0.2 L or 9.1% of BW. These data indicate that efficiency of IgG absorption from the colostrum replacement product may be affected by amount of material fed. Proteins other than IgG in the colostrum replacement product might have reduced the efficiency of IgG absorption.     

Effects of housing and colostrum feeding on the prevalence of selected infectious organisms in feces of Jersey calves

Neonatal Jersey calves (n = 96) were used to evaluate effects of housing (individual hutches or wooden pens in a barn) and colostrum feeding (calves were separated from the dam and fed 2 L of colostrum in nipple-bottles or allowed to nurse the dam for 3 d) on the prevalence of selected organisms in feces. Prevalence of Cryptosporidium and Eimeria were reduced, and prevalence of rotavirus tended to be reduced, when calves were housed in hutches. Prevalence of coronavirus was unaffected by treatment. Weekly prevalence of Giardia was increased when calves were left to nurse the dam for 3 d. Mean prevalence of Cryptosporidia (wk 1 to 4), Eimeria (wk 4 to 5), Giardia, rotavirus, and coronavirus (wk 1 to 5) were 34.7, 20.6, 27.1, 15.8, and 4.9%, respectively. Escherichia coli (K99 positive) were observed in 3 of 174 samples cultured. Methods of housing and colostrum feeding affected acquisition of enteropathogens in this study.     

Nutrient and immunity transfer from cow to calf pre- and postcalving

Nutritional and management strategies for dairy cattle are designed to prepare the cow for lactation and to minimize the incidence of metabolic diseases around calving. However, strategies initiated during the dry period should also consider the potential effects on the calf prior to and after calving. Fetal requirements for energy and protein are significant, particularly during the last trimester of gestation. Energy requirements increase to 1.3 to 1.5 times maintenance in late pregnancy; therefore, the formulation of rations for dry cows must contain sufficient energy to support fetal growth plus maintenance. Protein requirements during pregnancy increase, particularly during the last 2 mo. Colostrum is a source of immune components and nutrients to the neonate and contains more protein, immunoglobulins (Ig), nonprotein nitrogen, fat, ash, vitamins, and minerals than does milk. Because some vitamins do not cross the placental barrier, colostrum is the primary source of these nutrients for the calf after birth. Colostrum from cows that are not supplemented with vitamin E during the dry period may provide inadequate vitamin E to calves after birth. The Ig concentration in colostrum is not markedly affected by prepartum protein nutrition; diets containing high crude protein (CP) generally increase the nonprotein fraction of colostrum, but low CP diets do not affect the CP or Ig concentration of colostrum. However, data from beef calves suggest that absorption of IgG may be impaired when low protein diets are fed during the dry period. Diets for dry cows may be balanced to reduce the cation to anion ratio, which may reduce the incidence of parturient paresis. Recent research also suggests that these diets might increase the incidence of calves born in respiratory acidosis, which may impair the acquisition of passive immunity.     

Delaying colostrum intake by one day impairs plasma lipid, essential fatty acid, carotene, retinol and alpha-tocopherol status in neonatal calves

To study whether a delayed start of colostrum feeding in calves affects plasma lipids, fatty acids and fat-soluble vitamins, one group was fed colostrum (milkings 1-4) on d 1 and 2, then mature milk up to d 7, whereas two other groups were fed glucose or water on d 1, colostrum (milkings 1-4) on d 2 and 3 and then mature milk up to d 7. In calves fed colostrum on d 1, starting 5-7 h after birth, plasma concentrations of triglycerides, phospholipids, total cholesterol and of essential and nonessential fatty acids in triglyceride, phospholipid and cholesterol ester fractions as well as of carotene, retinol and alpha-tocopherol up to d 7 were significantly higher than in calves in which colostrum feeding started after >24 h of life. On the other hand, plasma concentrations of vitamin B-6, vitamin B-12 and folic acid were not influenced. Results indicated reduced efficiency of absorption of colostral fatty acids and of fat-soluble vitamins, but not of (selected) water-soluble vitamins, if colostrum is not fed on d 1 of life. In conclusion, colostrum intake within the first 24 h of life is required for an adequate plasma lipid, essential fatty acid, carotene, retinol and alpha-tocopherol status in the first week of life of calves.     

Influence of environment on passive immunity in calves

Passive immunity in neonatal calves is influenced by environment. Placing newly born Holstein calves (108 head) in three different housing environments (shade, cooled shade, hutch) during hot weather produced differences in body temperature, serum cortocosteroids, immunoglobulin IgG1 concentrations, and mortality. Experimental design permitted examination of effects due to treatments, time, differences in colostrum, and climatic environment in an analysis of variance. Calves exposed to the hotter, less desirable environment responded by having a higher mortality, higher serum corticosteroid concentration, and lower serum immunoglobulin IgG1 at 2 and 10 days after birth. All of these were correlated. Calves that died had serum immunoglobulin IgG1 falling below the mean for all experimental calves.     

Delaying colostrum intake by one day has important effects on metabolic traits and on gastrointestinal and metabolic hormones in neonatal calves

Effects on metabolic and endocrine traits of feeding colostrum on d 1 and 2, then mature milk up to d 7, or glucose or water on d 1, colostrum on d 2 and 3 and then mature milk up to d 7 were studied in calves. Calves fed colostrum within the first 24 h after birth had significantly higher rectal temperatures, heart rates and respiratory frequencies than calves provided only water or glucose. Significantly elevated plasma nonesterified fatty acid and bilirubin concentrations on d 1 and 2 of life in calves fed only water on d 1 compared with calves of the other groups mirrored reduced energy intake. Fecal consistency was significantly higher during wk 1 of life, and gastrin and glucose-dependent insulinotropic polypeptide increased only on d 1 and/or 2 of life in calves already fed colostrum on d 1, expressing improved functioning of the gastrointestinal tract. Significantly higher plasma globulin levels up to d 7 in calves fed colostrum on d 1 than in those starting colostrum intake only on d 2 demonstrated significantly enhanced efficiency of gamma-globulin absorption. Furthermore, significantly higher circulating glucose, albumin, insulin, insulin-like growth factor-I concentrations and significantly lower urea levels in calves fed colostrum on d 1 compared with those fed colostrum starting on d 2 of life indicated stimulation of anabolic processes. In conclusion, colostrum intake by calves within the first 24 h of life is needed not only for an adequate immune status, but also to produce the additional important and favorable effects on metabolic and endocrine traits and on vitality.     

Rubella antibodies in women of childbearing age during an epidemic and the two years thereafter

Blood leukocyte patterns, neutrophil phagocytosis of killed Escherichia coli in vitro, and plasma corticosteroids were studied in colostrum-fed (CF) and colostrum-deprived (CD) calves during the 1st 144 hours after birth. There was a marked increase in neutrophil numbers between 6 and 12 hours in CF but not CD calves, apparently as a result of colostrum ingestion. Phagocytosis was inactive at birth but increased quickly thereafter. Phagocytosis was more efficient in CF than in CD calves. Plasma corticosteroid concentrations were high at birth and decreased quickly thereafter in both groups of calves.     

Systemic availability of bovine immunoglobulin G and chicken immunoglobulin Y after feeding colostrum and whole egg powder to newborn calves

In connection with a study on the prophylaxis of infectious diarrhea with specific egg yolk antibodies, the systemic availability of colostral bovine immunoglobulin G (bIgG) and chicken immunoglobulin Y (IgY) after feeding egg powder was investigated on 26 newborn calves from 23 different farms. Blood was sampled daily and at the same day time from these calves in the first 14 days of life. During the feeding of colostrum, the mean bIgG concentration was highest at day 1 post natum with a value of 9.3 mg/ml serum. Thereafter, the mean bIgG level was reduced continuously to a significant lower concentration of 4.9 mg/ml serum at day 12 post natum and remained nearly constant at 5.2 mg/ml till to the end of the observation period. Total protein concentrations in the serum did not change and plateaued at a mean value of 56.2 mg/ml (SD 11.2). The number of colostrum meals had no significant effect on the mean bIgG concentrations during that period. The individual variation of bIgG concentrations was very high on every day of the sampling period. The mean coefficient of variation was at 52.1 % (SD 5.7). After having described the individual bIgG concentration curves mathematically with a regression curve, two groups with significantly different bIgG elimination constants (k) could be obtained. Thus in one group (n = 10) with k-values of < -0.02 a mean half time of serum bIgG of 24.3 days (SD 4.6) was calculated. In the other group of calves (n = 16) with elimination constants of k > -0.02, a mean half time of 68.5 days (SD 36.7) could be calculated, possibly because these calves started earlier with their endogenous bIgG production. Additionally, to 18 of these calves 20 g egg powder with an IgY concentration of 15 mg/g was fed up to day 14. Calves had a maximal mean IgY concentration of 1.9 micrograms/ml serum if egg powder feeding started already during the first 12 hours of life. Starting at a later time resulted in a significant reduction of IgY levels. For example, the mean initial IgY concentration dropped to 0.035 micrograms/ml serum after having had the first egg powder application between 25 and 48 hours post natum. Using the individual IgY elimination constant derived from a regression analysis (r2 = 0.84) of the IgY concentration curve, a mean IgY half time of 5.0 days (SD 2.5) could be calculated. To prevent the absorption of heterologous antibodies and consecutively, also to prevent a possible systemic effect, egg powder for prophylactic purposes in newborn calves should be fed after the first 24, better 48 hour, post natum. Most important for the prophylactic effect of specific antibodies on infectious diarrhea is not their systemic but their high local intestinal availability.     

Collecting duct carcinomas of the kidney: a comparative loss of heterozygosity study with clear cell renal cell carcinoma

Lyophilizing was compared to freezing as a method of colostrum storage. Eight lots of colostrum from the first milking were divided into two equal parts; one was frozen, and the other was lyophilized. Twenty-two newborn calves were divided into two groups and fed either 2 L of frozen and thawed colostrum or 2 L of reconstituted lyophilized colostrum. The calves were bled at 12, 18, 24, and 72 h after feeding, and levels of the immunoglobulins IgG1, IgG2, IgM, and IgA were determined with a radial immunodiffusion assay, in colostrum and sera. The mean concentration of individual immunoglobulin isotypes in the sera of calves fed either frozen or lyophilized colostrum did not differ significantly. Calves fed from the same lots of colostrum had similar immunoglobulin concentrations in their sera, irrespective of the method of storage. All immunoglobulin isotypes were absorbed with equal efficiency from frozen and lyophilized colostrum as determined by calculation of the absorption coefficient.     

Estimation of plasma volume in Holstein and Jersey calves

The concentration of immunoglobulin (Ig) G in the blood of neonatal calves shortly after birth is a widely used criterion to determine the degree of acquisition of passive immunity. Another method used to determine the biological mechanisms of IgG absorption is calculation of the apparent efficiency of IgG absorption. Estimation of the efficiency of IgG absorption requires the estimation of plasma volume in neonatal calves. Previous estimates of plasma volume in a few calves of varying breeds have been made; the estimates ranged from 7 to 14.5% of body weight (BW). Holstein (n = 97 from four farms) and Jersey (n = 49 from one farm) calves were fed fresh maternal colostrum or colostrum that had been previously frozen. Calves were fed 2 L of colostrum at 4.1 h (SE = 0.2; range = 0.3 to 11.0 h) and 12 h later. Plasma volume was measured by determining the concentration of Evans' blue dye in a jugular blood sample collected 10 min after injection of approximately 1.5 ml of 1.5% Evans' blue dye. Factors that affected plasma volume (milliliters) were BW, breed, and age at sampling; r2 of the regression was 0.60. Factors that affected plasma volume (percentage of BW) were BW, breed, and age at sampling; r2 of the regression was 0.08. Mean plasma volume for all calves was 3162 ml (SE = 79) and was 9.86% of birth BW (SE = 0.15%). Mean plasma volume was 2250 ml (9.71% of BW) and 3623 ml (9.94% of BW) for Jersey and Holstein calves, respectively. Body weight was the best predictor of plasma volume.     

Prolonged colostrum feeding enhances xylose absorption in neonatal calves

Colostrum contains numerous components that influence gastrointestinal development in neonates. To test the influence of differences in duration of colostrum feeding in newborn calves on gastrointestinal absorptive capacity, .5 g xylose/kg BW was administered on d 5 of life. Calves of group GrC6 were fed colostrum from the first six milkings on the first 3 d and then milk replacer. Calves of group GrC1 were fed first colostrum only and then milk replacer in the same amounts as calves of group GrC6. Calves of group GrM were fed only milk replacer: they received no colostrum. The rise of plasma xylose after xylose intake was greater (P < .05) in GrC6 and GrC1 than in GrM but not significantly greater in GrC6 than in GrC1. Basal and mean plasma glucose concentrations on d 5 of life were higher in GrC6 than in GrM, but there was no difference in the magnitude of its postprandial rise. The data indicate greater xylose absorptive capacity in calves after prolonged colostrum intake compared with calves fed only milk replacer.     

Long-term outcome after the mustard repair for simple transposition of the great arteries. 28-year follow-up

The effects of feeding different amounts of colostrum or only milk replacer and the effects of Long-R3-IGF-I (administered s.c. or orally; 50 microg/[kg BW x d] for 7 d), and of s.c. injected recombinant bovine GH (rbGH; 1 mg/[kg BW x d] for 7 d) on small intestinal mucosal morphology in newborn calves were studied by histomorphometry. Neonatal calves fed colostrum six times exhibited greater (P < .01) villus circumferences, areas, and heights in total small intestine and especially in the duodenum than calves fed only milk replacer. Furthermore, villus circumferences and areas in total small intestine were greater (P < .05) in calves fed colostrum once than in calves fed no colostrum. Villus size in total small intestine was smaller (P < .05) in rbGH-treated than in control calves; jejunum villus circumferences and heights were especially reduced (P < .05). Crypt depths in ileum were greater (P < .05) in rbGH-treated calves. In conclusion, prolonged colostrum supply significantly enhanced small intestinal villus size in neonatal calves. In contrast, Long-R3-IGF-I had no significant influence on small intestinal morphology, and rbGH in supraphysiological amounts even reduced small intestinal mucosal variables after 1 wk of treatment. The study demonstrated enhanced postnatal development of the gastrointestinal tract by prolonged colostrum feeding, but not by Long-R3-IGF-I or GH.     

Randomized controlled trial of effects of Escherichia coli antiserum on serum immunoglobulin G concentrations and morbidity and mortality rates in foals

OBJECTIVE: To determine whether administration of commercially available Escherichia coli antiserum to neonatal foals would affect serum IgG concentration or morbidity and mortality rates during the first 60 days of life. DESIGN: Randomized controlled trial. ANIMALS: 271 neonatal foals on 4 well-managed farms. PROCEDURE: Foals were randomly assigned to a treatment or control group. All foals were allowed to suckle colostrum normally. In addition, treatment-group foals were given E coli antiserum (10 micromilligrams) orally between 0 and 8 hours after birth. Serum samples were obtained between 18 and 36 hours after birth, and serum IgG concentration was determined. Foals were monitored for the first 60 days after birth, and causes of disease or death were recorded. RESULTS: Groups did not differ significantly in regard to breed, sex, month of birth, season of birth, age of dams, parity of dams, duration of gestation, or specific gravity of colostrum before suckling. In addition, groups did not differ significantly in regard to mean serum IgG concentration, prevalence of complete or partial failure of passive transfer of immunity, frequency or causes of disease, or frequency of death from infectious causes. CLINICAL IMPLICATIONS: In this group of foals on well-managed farms, administration of E coli antiserum did not alter serum IgG concentrations or morbidity and mortality rates during the first 60 days of life.     

Experimental oral transmission of Ehrlichia phagocytophila to calves

Three groups of four calves were used to determine whether Ehrlichia phagocytophila could be transmitted orally to calves via infected milk. Groups 1 and 2 consisted of four-week-old calves and group 3 of newborn calves. The calves in group 1 were fed for several days with milk from cows infected experimentally with E phagocytophila. The calves in groups 2 and 3 were fed 200 ml of whole blood containing E phagocytophila organisms; for group 2 the blood was added to milk before being fed, and for group 3 the blood was added to colostrum before being fed within three hours after birth. Blood samples for haematological, serological and cytological examination, and for polymerase chain reaction (PCR) were collected from all the calves, starting on the first or only day of administration and then every four days for four weeks. The calves of groups 1 and 2 showed no clinical, haematological or serological changes, and there was no direct or indirect evidence of the agent. In contrast, all the calves in group 3 had mild pyrexia and seroconverted on day 8, and in one of them E phagocytophila organisms were visible in leucocytes, and the PCR on the buffy coat was positive on day 8.     

Thyroid hormones in dairy calves after birth and in the first days of life. Amplitude of variation, maternal-fetal relationships and reaction patterns specific to the individual

Dairy calves and their dams from the dairy herd of the institute were used for this investigation. Calvings were watch carefully and jugular venous blood samples taken immediately after delivery from the cow and the calf and in the calf at 24 hours and at 48 hours of postnatal age as well were analyzed for T4, FT4, T3 and FT3 by luminescence enzyme immunoassay (LEIA). Higher thyroid values in calves than in their dams could be found increasing short after birth and reaching their highest level within 24 hours of postnatal life. The individual thyroid hormone values at birth and at 24 hours and at 48 hours of age are well correlated meaning the changes during the first days are specific for each calf. Strong correlation between the thyroid hormone values of the cows and those of the calves could be found. Mean values of thyroid hormones were significantly higher in female than in male calves all time but not different in the dams of these calves.     

Physiologic differences between twin and single born beef calves in the first two days of life

Behavioral observations and hematological, serum biochemical and blood gas measurements were made on 8 naturally occurring twin calves during the first 48 hours of life. These values were compared to similar measurements collected from 30 single born calves, born under the same calving conditions. All calves survived to at least 3 weeks of age without physically detectable disease. Although the gestational age of the twins and singles were not different, the twins had a lower mean birth weight. Calving difficulty score, time interval to standing and time interval to nursing were not different between the 2 groups. Twin calves had significantly lower rectal temperatures, arterial oxygen tensions and blood glucose concentrations than the single calves through the first 12 hours of life. Hct, Hgb concentration, and RBC were lower in twin calves throughout the 48 hour period. The N:L ratio was lower in the twins at birth. Mean serum IgG1 concentrations were lower in twins only at 24 hours whereas IgM concentrations were lower at both 24 and 48 hours in twins. Serum chemistry value differences between twin and single calves were most numerous at 24 hours of age when serum proteins, urea nitrogen, total bilirubin, sodium, chloride, and total calcium concentrations were higher in the twins and serum phosphorus concentration was lower in the twins.