SciELO - Scientific Electronic Library Online

 
vol.71 issue1Evaluation of tear production, intraocular pressure, retinography and ocular biometry in bovine Nelore and Gir breedsBovine alphaherpesvirus 1 and 5 in semen from bulls presenting genital lesions under field conditions in Brazil author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

Indicators

Related links

Share


Arquivo Brasileiro de Medicina Veterinária e Zootecnia

Print version ISSN 0102-0935On-line version ISSN 1678-4162

Arq. Bras. Med. Vet. Zootec. vol.71 no.1 Belo Horizonte Jan./Feb. 2019

http://dx.doi.org/10.1590/1678-4162-10176 

Veterinary Medicine

Serum biochemical profile of neonatal buffalo calves

Perfil bioquímico sérico de bezerros bubalinos no período neonatal

D.C. Souza1 

D.G. Silva1 

T.G. Rocha1 

B.M. Monteiro2 

G.T. Pereira1 

L.C. Fiori1 

R.B. Viana3 

J.J. Fagliari1 

1Universidade Estadual Paulista ˗ Jaboticabal, SP

2Universidade de São Paulo ˗ São Paulo, SP

3Universidade Federal Rural da Amazônia ˗ Belém, PA

ABSTRACT

Serum blood samples from 50 Murrah buffalo calves were examined in this study. The animals were allocated into three groups according to the number of parturitions of their mothers: G1 (n= 15) calves from primiparous buffaloes, G2 (n= 19) calves from buffaloes with two to four parturitions, and G3 (n= 16) calves from buffaloes with five or more parturitions. Blood samples were taken at birth, before colostrum ingestion, at 24h, 48h, and 72h after birth, and at 7, 14, 21, and 30 days after birth for determination of levels of gammaglutamyl transferase (GGT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), creatine kinase, total protein, albumin, globulins (including immunoglobulin G), iron, total calcium, ionized calcium, phosphorus, sodium, and potassium. The age of the calves was found to influence all of the biochemical parameters, with the exception of ionized calcium and potassium in the calves in groups G1 and G3. The calving order was found to influence AST, GGT, total protein, albumin, and globulins, including IgG. The high serum ALP activity in the first two days after birth indicates that measurement of the levels of this enzyme may be used as an indirect method of assessing passive immunity transfer.

Keywords: Bubalus bubalis; newborn; number of parturitions; murrah; passive immunity transfer (PIT)

RESUMO

Amostras de sangue de 50 bezerros de búfalo Murrah foram examinados nesse estudo. Os animais foram distribuídos em três grupos de acordo com a paridade de suas genitoras: G1 (n=15) bezerros de búfalas primíparas, G2 (n=19) bezerros de búfalas com 2 a 4 gestações, e G3 (n=16) bezerros de búfalas com cinco ou mais gestações. Amostras de sangue foram colhidas ao nascimento, antes da ingestão de colostro e 24h, 48h, e 72h após o nascimento e 7, 14, 21 e 30 dias após nascimento para determinar níveis de gammaglutamil transferase (GGT), fosfatase alcalina (ALP), aspartato aminotrasferase (AST), creatina quinase, proteínas totais, albumina, globulina (inclusive imunoglobulina G), ferro, cálcio total, cálcio ionizado, fósforo, sódio e potássio. A idade dos bezerros influenciou todos os parâmetros bioquímicos, exceto cálcio ionizado e potássio nos bezerros dos grupos G1 e G3. A ordem de nascimento influenciou AST, GGT, proteínas totais, albumina e globulinas, inclusive IgG. Intensa atividade ALP no soro nos primeiros dois dias após nascimento indica que medidas dos níveis dessa enzima podem ser utilizados como método indireto de avaliar transferência passiva de imunidade.

Palavras chave: Bubalus bubalis; neonato; número de gestações; murrah; transferência passiva de imunidade

INTRODUCTION

The buffalo (Bubalus bubalis) is an animal of great economic importance; the global buffalo population has been estimated to exceed 194 million animals (Food…, 2017). One of the critical points in buffalo production is the neonatal period, wherein high mortality rates occur, reaching up to 17% in Murrah animals (Shivahre et al., 2014). This is relevant because a 20% neonatal mortality rate results in a 38% reduction in farm profit (Radostits et al., 2007). In all species, the neonatal period represents a critical moment in which organs must adapt to extrauterine life. This is a difficult transition from intrauterine protection to the many challenges presented by the environment (Piccione et al., 2009).

Knowledge of normal values of serum biochemical parameters is important when evaluating injuries to organs and tissues caused by various diseases and in the determination of animal welfare. It also allows for monitoring the metabolic condition, functional abnormalities of the organs, and the adaptation of the organism to nutritional and physiological changes. Factors such as species, breed, age, rearing systems, feeding, and number of parturitions influence serum biochemical values; thus, the identification of these factors and their interactions is crucial for the correct interpretation of the blood parameters (Klinkon and Jezek, 2012).

The available data on buffaloes are scarce, and this leads to the use of parameters obtained for cattle, which may lead to misinterpretations of the data, particularly those obtained in the first week of life that are affected by changes related to birth and by colostrum intake (Pérez-Santos et al., 2015). Therefore, it is necessary to determine the serum biochemical values of healthy buffaloes to evaluate whether any alterations are due to physiological changes or pathological processes. The aim of this study was to determine the serum biochemical profile of neonatal Murrah buffalo calves and to observe the influence of the age of the calves and the number of parturitions of their dams on the values of the biochemical parameters.

MATERIAL AND METHODS

Fifty healthy Murrah buffalo calves, of both sexes, born from eutocic births, from clinically healthy buffalo cows, reared on property located in the State of São Paulo, were included in this study. The animals were raised in a semi-extensive system, with annual milk production of 300,000kg, an average of 79 lactating animals, and an average individual production of 2,765kg of milk in 300 days of lactation. After birth and natural colostrum intake, the umbilical cords of the calves were treated with 2% iodine solution. The calves remained with their mothers during the first five days of age. The calves then entered a collective pen, and were fed milk from a mammary quarter, not milked, twice a day during the milking of their dams, until weaning at 90 days old. In addition to the dairy diet, calves had access to Brachiaria spp. pastures and were fed a diet composed of soybeans and maize.

The calves used in the experiment were examined and considered to be clinically healthy when they did not present any alterations upon physical examination (Dirksen et al., 1993). The calves were allocated to three experimental groups according to the number of parturitions of their mothers: G1 (n= 15) calves (10 females and 5 males) from primiparous buffaloes, G2 (n= 19) calves (7 females and 12 males) from buffaloes with two to four parturitions, and G3 (n= 16) calves (6 females and 10 males) from buffaloes with five or more parturitions.

The venous blood samples were collected at the following moments: at birth, before colostrum intake (M0), at 24h (M1), 48h (M2) and 72h (M3) after birth and at 7 (M4), at 14 (M5), 21 (M6) and 30 (M7) days after birth. Blood samples (10mL) were collected by jugular venipuncture after local antisepsis. A vacuum collection system in siliconized tubes without anticoagulant (Vacutainer, Bencton Dickinson, Franklin Lakes, USA) was used. Blood samples were centrifuged at 2,000g for 10min, and 2.0mL aliquots of blood serum were separated and stored in plastic tubes, labeled and maintained at -20°C until the time of the laboratory tests.

Serum activities of gammaglutamyl transferase (GGT) (modified Szasz method), aspartate aminotransferase (AST) (UV-IFCC kinetics method), creatine kinase (CK) (UV method), and alkaline phosphatase (ALP) (modified Bowers and McComb method) were determined, as well as serum concentrations of total protein (biuret method), albumin (bromocresol green method), total calcium (CPC method), phosphorus (modified Daly and Ertinghausen method), and iron (modified Goodwin method), using a set of commercial reagents (Labtest Diagnóstica, Lagoa Santa, Minas Gerais, Brazil). The samples were analyzed in a semiautomatic spectrophotometer (Labquest, Labtest Diagnóstica, Lagoa Santa, Minas Gerais, Brazil), using light of appropriate wavelength for each test.

Globulins were calculated from the arithmetic difference between the total protein and albumin concentrations. Immunoglobulin G (IgG) concentrations were determined by protein fractionation using polyacrylamide gel electrophoresis containing sodium dodecyl sulfate (SDS-PAGE), according to the technique proposed by Laemmli (1970). After fractionation, the gel was stained for 10min in 0.25% Coomassie blue solution and then bleached in 7% acetic acid solution to remove excess dye until the protein fractions were clear. IgG concentrations were determined by computerized densitometer (Shimadzu CS-9301PC, Tokyo, Japan). For reference, a marker solution (Sigma, St Louis, MO, USA) with different molecular weights was used in addition to the purified bovine IgG protein.

The serum concentrations of ionized calcium, sodium, and potassium were determined in an ion analyzer by the ion-selective electrode method (9180 Electrolyte Analyzer, Roche Diagnostics, Mannheim, Germany).

The results were evaluated by analysis of variance (ANOVA), using the Tukey test to compare the means, after verification of the homogeneity of the samples (Zar, 1999). Significance was verified at 5% probability. Statistical analyses were performed using the SAS (Statistical Analysis System) statistical program. This study was evaluated by the Ethical Committee on the Use of Animals (CEUA) of FCAV / UNESP / Jaboticabal campus and approved under protocol number 010028/2014.

RESULTS AND DISCUSSION

The serum activities of the enzymes in G1, G2, and G3 calves, from birth to 30 days old, are presented in Table 1 and Figure 1.

Table 1 Mean ± standard deviation of aspartate aminotransferase (AST), creatine kinase (CK), gammaglutamyl transferase (GGT), and alkaline phosphatase (ALP), serum activities of neonatal buffalo calves born from primiparous buffaloes (G1), multiparous with two to four parturitions (G2), and multiparous buffaloes with more than five parturitions (G3) at birth, before colostrum intake (M0), at 24h (M1), 48h (M2), and 72h (M3) after birth, and at 7 (M4), 14 (M5), 21 (M6) and 30 (M7) days after birth 

Group Moments
M0 (0h) M1 (24h) M2 (48h) M3 (72h) M4 (7 days) M5 (14 days) M6 (21 days) M7 (30 days)
Aspartate aminotransferase (U/L)
G1 41.0±3.94Ad 108±16.3Aa 103±11.84Aab 92.5±16.4Abc 90.1±11.7Abc 87.0±14.1Ac 88.7±28.5Ac 82.8±10.7Ac
G2 41.9±15.2Ae 105±18.3Aa 97.3±16.4Aab 87.3±14.9Abc 75.3±7.85Bcd 73.9±40.1Bd 64.5±11.0Bd 67.5±13.5Bd
G3 44.9±17.3Ad 108±16.7Aa 101±17.2Aab 91.7±12.7Abc 79.9±9.66ABc 74.3±17.0ABc 76.0±28.0ABc 75.0±11.6ABc
Creatine kinase (U/L)
G1 107±51.0Ad 285±106Aa 179± 45.0Aabc 124±28.4Acd 170±79.8Abc 213±55.6Aab 251±92.9Aab 215±92.8Aab
G2 129±66.9Abc 231±95Aa 158±102Abc 147±113Ac 128±42.6Ac 253±143Aab 207±48.0Aa 261±128Aa
G3 115±45.8Ae 324±116Aa 170±52.6Acde 123±29.4Ae 150±51.9Ade 228±108Abcd 293±162Aabc 247±45.2Aab
Gammaglutamyltransferase (U/L)
G1 15.3±4.84Af 1,253±797Ba 464±297Bb 319±210Ab 139±87.4Bc 58.1±32.4ABd 32.6±17.0ABe 18.9±7.58Aaf
G2 22.9±7.65Afg 1,714±814Aa 714±376Ab 403±173Ac 200±106Ad 78.9±51.8Ae 43.2±34.6Af 20.5±9.21Ag
G3 16.4±2.89Af 1,074±655Ba 341±181Bb 205±110Bc 97.1±52.3Bd 44.0±20.6Be 24.4±8.48Bf 15.3±3.95Afg
Alkaline phosphatase (U/L)
G1 205±102Ade 1,579±901Aa 600±283Ab 338±143Ac 191±40.8Ae 130±24.6Adef 119±28.7Adf 101±32.8Af
G2 124±38.5Ade 1,250±510Aa 530±301Ab 322±174Ac 217±136Ad 143±59.5Ae 117±41.5Ae 107±46.8Ae
G3 158±45.8Abd 1,297±780Aa 465±233Aa 275±81.2Ab 194±57.9Abc 137±44.3Acde 112±32.6Ade 97.9±37.5Ae

Mean values followed by the same upper case letters in the same column and lower case letters on the same line do not differ significantly according to Tukey’s test (P> 0.05).

Figure 1 Serum activities of aspartate aminotransferase (AST), creatine kinase (CK), gammaglutamyl transferase (GGT), and alkaline phosphatase (ALP) of buffalo calves born from primiparous buffaloes (G1), multiparous buffaloes with two to four parturitions (G2), and multiparous buffaloes with more than five parturitions (G3) at birth, before colostrum intake (M0), at 24h (M1), 48h (M2), and 72h (M3) after birth, and at 7 (M4), 14 (M5), 21 (M6) and 30 (M7) days after birth. 

The maximum value of AST and CK was observed 24h (M1) after birth, with significant difference between the three groups (Table 1). The abrupt elevation of the values of these two enzymes between birth (M0) and 24h (M1) after birth can be attributed to injuries to the skeletal muscle during birth and by the increase in calf muscular activity, since the animal moves to a stationary position and begins to move immediately after birth (Boyd, 1989; Kaneko et al., 2008). The serum activity of AST was influenced by the number of parturitions from 7 (M4) to 30 (M7) days after birth, when calves in group G1 exhibited higher serum activity of the enzyme. The same did not occur with the serum activity of creatine kinase (Table 1), which originates primarily from skeletal and cardiac muscles and is used as marker of muscle injury; its activity may be increased after exercise, long recumbence, trauma, or damage to the musculature (Kaneko et al., 2008). Serum activities of CK decreased until 72h (M3) after birth and increased from 7 (M4) days after birth, maintaining this value until 30 (M7) days after birth (Figure 1). This increase is expected, since the growth of the calf is accelerated in the first month of life and the animal begins to exhibit greater social interaction with the other calves of the herd, such as running, jumping, and nodding (Klinkon and Jezek, 2012).

The serum activities of GGT and ALP were influenced by age, throughout the experimental period, in all groups (Figure 1). There was an influence of the number of parturitions throughout the study, aside from the moment before the colostrum intake (M0) and at 30 (M7) days after birth (Table 1). This result differs from that reported by Feitosa et al. (2010), who did not find a difference in GGT values in the first 48h of life of calves of multiparous and primiparous cows, but is similar to that reported by Rocha et al. (2012), who observed a difference in the mean values of GGT and ALP between bovine calves born from primiparous and multiparous cows in the first 24h after birth. The buffalo calves in this study presented higher serum activity of GGT and ALP at 24 h (M1) after birth, which significantly differed from the serum activity at other moments; this result is in agreement with the results of previous studies (Knowles et al., 2000; Rocha et al., 2012; Pérez-Santos et al., 2015). The significant increase in the values of these enzymes between M0 and M1 is due to the absorption of GGT and ALP present in colostrum, since both are produced in large quantities by the mammary gland cells of ruminants (Kaneko et al., 2008). The serum GGT activity in calves after colostrum feeding is 60 to 160 times greater than the activity observed in cows; a significant correlation was previously described between the activity of this enzyme and the serum concentration of IgG (Radostits et al., 2007; Feitosa et al., 2010). The measurement of serum ALP activity is not recommended as an indirect form of evaluation of passive immunity transfer (PIT) in cattle due to the low correlation with serum IgG levels (Rocha et al., 2012). However, the elevation of the enzyme values at 24h (M1) and 48h (M2) after birth in buffalo calves is markedly higher than those reported in cattle (Rocha et al., 2012; Pérez-Santos et al., 2015). Thus, the measurement of ALP in this species should be studied as an alternative to the evaluation of PIT, provided that the origin of the enzyme has been identified, since there is an increase in activity of bone isoenzyme ALP in animals with high osteoblastic activity, as in neonates (Kaneko et al., 2008). The mean values of GGT and ALP enzymes decreased from 24h (M1) until 30 (M7) days after birth, at which point the enzymatic activity resembled that at the moment prior to colostrum consumption (Table 1). This reduction is a result of the degradation of the enzymes in the calf intestine over time and the process of calcification of bone epiphyses, with lower serum activity of bone isoenzyme ALP (Kaneko et al., 2008).

The serum concentrations of proteins in G1, G2, and G3 calves, from birth to 30 days old, are presented in Table 2 and Figure 2.

Table 2 Mean ± standard deviation of the total protein, albumin, globulins, and immunoglobulin G (IgG) concentrations of buffalo calves born from primiparous buffaloes (G1), multiparous with two to four parturitions (G2), and multiparous buffaloes with more than five parturitions (G3) at birth, before colostrum intake (M0), at 24h (M1), 48h (M2) and 72h (M3) after birth, and at 7 (M4), 14 (M5), 21 (M6), and 30 (M7) days after birth 

Group Moments
M0 (0h) M1 (24h) M2 (48h) M3 (72h) M4 (7 days) M5 (14 days) M6 (21 days) M7 (30 days)
Total Protein (g/dL)
G1 4.68±0.17Ae 9.21±2.00Aa 8.94±1.49Aa 8.83±1.67Aab 8.34±1.30Aab 8.06±0.93Abc 7.55±0.93Acd 7.21±0.79Ad
G2 4.70±0.13Ae 8.36±1.03ABab 8.43±1.01ABa 8.37±1.03ABab 8.10±1.04Aab 7.68±0.90Abc 7.31±0.79Acd 6.77±0.94Ad
G3 4.88±0.32Ad 7.78±1.56Bab 7.97±1.65Ba 7.87±1.72Bab 7.60±1.35Aab 7.41±1.03Aabc 7.08±0.79Abc 6.77±0.59Ac
Albumin (g/dL)
G1 2.49±0.19Aab 1.95±0.24Bd 2.06±0.28Ad 2.09±0,21Bcd 2.28±0.24Bbc 2.44±0.28Aab 2.46±0.20Bab 2.51±0.26Aa
G2 2.52±0.21Aab 2.14±0.20Ad 2.22±0.22Acd 2.34±0.25Abc 2.51±0.23Aab 2.55±0.17Aa 2.65±0.24ABa 2.58±0.37Aa
G3 2.44±0.26Abc 2.06±0.26ABd 2.17±0.26Acd 2.29±0.24Ac 2.46±0.17ABb 2.57±0.22Aab 2.66±0.24Aa 2.55±0.18Aab
Globulins (g/dL)
G1 2.19±0.08Af 7.26±2.19Aa 6.88±1.71Aa 6.73±1.82Aab 6.09±1.43Abc 5.62±1.12Acd 5.09±1.05Ade 4.70±0.82Ae
G2 2.26±0.18Ae 6.22±1.12Ba 6.16±1.07ABa 6.04±1.05ABa 5.60±1.05ABab 5.13±0.98ABbc 4.66±0.71Acd 4.19±0.83Ad
G3 2.42±0.42Ae 5.72±1.65Ba 5.80±1.70Ba 5.58±1.79Bab 5.15±1.31Babc 4.84±1.06Bbcd 4.42±0.85Acd 4.22±0.59Ad
Immunoglobulin G (mg/dL)
G1 296±205Ae 4,093±1,558Aa 3,682±1,207Aa 3,642±1,289Aa 2,913±972Ab 2,441±732Abc 1,957±647Acd 1,575±521Ad
G2 341±184Af 3,532±858ABa 3,399±770ABa 3,107±793ABab 2,730±699Abc 2,168±577Acd 1,703±456Ade 1,387±478Aef
G3 274±203Ae 3,125±1,484Ba 3,009±1,441Bab 2,822±1,375Bab 2,424±1,076Abc 1,964±859Acd 1,556±660Ade 1,356±389Ade

Mean values followed by the same upper case letters in the same column and lower case letters on the same line do not differ significantly according to Tukey’s test (P> 0.05).

Figure 2 Serum concentrations of total protein, albumin, globulins, and immunoglobulin G (IgG) of buffalo calves born from primiparous buffaloes (G1), multiparous with two to four parturitions (G2), and multiparous buffaloes with more than five parturitions (G3) at birth, before colostrum intake (M0), at 24h (M1), 48h (M2), and 72h (M3) after birth, and at 7 (M4), 14 (M5), 21 (M6) and 30 (M7) days after birth. 

Total protein and globulin mean serum concentrations varied throughout the study period (Table 2); both parameters exhibited an abrupt increase between the time before the colostrum intake (M0) and 24h (M1) after birth. The determination of the serum total protein concentration of calves is useful as an indirect indicator of PIT, since the increase in values after the first day is primarily due to the absorption of globulins, particularly IgG, demonstrating a significant correlation with the values of total protein and globulins (Feitosa et al., 2010; Rocha et al., 2012). The serum concentrations of total protein and globulins decreased from 24h (M1) to 30 (M7) days after birth (Figure 2); this result was in agreement with previous reports (Fagliari et al., 1998; Rocha et al., 2012; Pérez-Santos et al., 2015). Such reduction is due to the degradation of the immunoglobulins acquired by colostrum ingestion. The number of parturitions influenced total protein concentrations at 24h (M1), 48h (M2) and 72h (M3) after birth, with the highest total protein levels occurring in G1 calves (Table 2). The number of parturitions also influenced the results of globulins, since the G1 calves presented higher serum globulin levels from 24h (M1) after birth until 14 (M5) days after birth. This differs from the results in bovine calves, where calves born from multiparous cows exhibited greater levels of globulins during the first week of life (Rocha et al., 2012).

Serum albumin concentration was influenced by the order of calving at 24h (M1) and 72h (M3) after birth, and at 7 (M4) and 21 (M6) days after birth, and the mean concentration was found to be higher in G2 and G3 calves than in G1 calves (Table 2). The buffalo calves presented variation of serum albumin concentration throughout the neonatal period, with a significant reduction in albumin concentration in the first hours after calving, most likely due to rapid plasma expansion after colostrum intake and by the absorption of immunoglobulins present in the colostrum. The serum albumin values obtained were similar to those reported in bovine calves from 24h after birth (Knowles et al., 2000; Piccione et al., 2009), when there was a gradual increase in albumin concentrations until 21 days after birth, with a subsequent decrease at the end of the first month (Figure 2). Variations in serum albumin concentrations observed over time are most likely related to the maturation of hepatic tissue and to the intake of solid food (Knowles et al., 2000; Birgele and Ilgaza, 2003; Kaneko et al., 2008).

Serum IgG concentrations peaked at 24h (M1) after birth (Table 2), with values considered optimal for PIT (Feitosa et al., 2010). In contrast to previous reports in cattle (Feitosa et al., 2010; Rocha et al., 2012), the highest serum concentrations of IgG were observed in G1 calves (Figure 2). G3 calves exhibited the lowest serum concentrations of IgG, most likely due to the lower immune capacity of the older animals, which results in a lower amount of immunoglobulin in the colostrum.

The serum concentrations of total calcium, iron, phosphorus, sodium, potassium, and ionized calcium in G1, G2, and G3 calves, from birth to 30 days old, are presented in Table 3 and Figure 3.

Table 3 Mean ± standard deviation of the total calcium, iron, phosphorus, sodium, potassium, and ionized calcium, of buffalo calves born from primiparous buffaloes (G1), multiparous with two to four parturitions (G2), and multiparous buffaloes with more than five parturitions (G3) at birth, before colostrum intake (M0), at 24h (M1), 48 h (M2), and 72h (M3) after birth, and at 7 (M4), 14 (M5), 21 (M6) and 30 (M7) days after birth 

Group Moments
M0 (0h) M1 (24h) M2 (48h) M3 (72h) M4 (7 days) M5 (14 days) M6 (21 days) M7 (30 days)
Total calcium (mg/dL)
G1 9.44±0.72Aab 9.83±0.44Aab 10.1±0.90Aab 10.7±0.85Aa 10.5±0.85Aa 10.1±0.80Aab 10.2±0.96Aab 9.50±0.65Ab
G2 9.69±1.47Aabc 9.80±1.53Ac 10.6±1.07Aab 10.8±1.37Aa 10.9±1.28Aa 10.3±1.24Aabc 9.96±1.47Abc 9.70±1.64Ac
G3 9.98±0.82Aabc 9.48±1.06Abc 9.16±1.02Aab 10.4±0.88Aab 10.4±0.86Aa 9.76±0.96Aabc 9.87±1.16Aabc 9.19±0.85Ac
Iron (µg/dL)
G1 81.5±24.3Aab 98.9±32.3Ab 119±41.5Aab 112±49.9Ab 102±51.1Ab 147±94.0Aab 156±70.2Aab 214±89.2Aa
G2 89.3±41.6Aabcd 74.6±39.0Ab 106±45Abcd 101±76Ab 137±133Abd 131±92.0Abcd 172±104Aac 194±75.9Aa
G3 81.3±20.7Ac 88.0±47.2Ab 135±81.9Aabc 133±65.4Aabc 160±141Aabc 146±70.5Aabc 213±97.0Aa 178±85.8Aac
Phosphorus (mg/dL)
G1 5.34±0.32Ab 7.70±1.88Aa 8.40±1.60Aa 8.94±1.44Aa 8.99±1.29Aa 8.70±0.77Aa 8.33±1.05Aa 8.13±0.60Aa
G2 5.31±0.64Ac 6.47±1.13Ac 7.77±1.84Ab 8.98±1.59Aa 9.44±0.98Aa 8.65±0.74Aab 8.53±1.02Aab 8.44±1.04Aab
G3 5.46±0.83Ad 6.70±1.83Acd 7.36±1.96Abc 9.10±1.96Aa 9.30±1.30Aa 8.61±1.04Aab 8.38±0.75Aab 8.09±1.16Aab
Sodium (mMol/L)
G1 136±3.78Aab 138±2.37Aa 135±1.41Aab 135±2.44Ab 136±5.53Ab 136±1.68Aab 134±2.16Ab 133±3.37Ab
G2 133±6.00Aab 141±12.8Aa 138±4.00Aa 138±8.25Aab 136±2.97Aa 133±2.59Abc 133±3.27Abc 132±4.96Ac
G3 137±2.19Aabc 137±1.24Aa 136±1.74Aab 135±2.25Aabc 136±3.17Aa 134±3.21Aabc 134±3.01Abc 132±4.98Ac
Potassium (mMol/L)
G1 4.70±0.28Aa 4.59±0.40Aa 4.73±0.33Aa 4.81±0.39Aa 5.03±0.38Aa 4.83±0.53Aa 4.76±0.62Aa 4.79±0.33Aa
G2 4.38±0.31Aab 4.69±0.64Ab 4.81±0.50Aab 4.99±0.55Aab 5.17±0.40Aa 4.92±0.47Aab 4.99±0.46Aab 4.96±0.37Aab
G3 4.67±0.43Aa 4.72±0.35Aa 4.73±0.25Aa 4.89±0.36Aa 5.15±0.33Aa 5.00±0.49Aa 4.98±0.54Aa 4.89±0.46Aa
Ionized calcium (mMol/L)
G1 0.90±0.10Aa 1.02±0.09Aa 0.97±0.07Aa 0.90±0.11Aa 0.95±0.09Aa 0.93±0.07Aa 0.94±0.09Aa 0.94±0.08Aa
G2 1.01±0.09Aa 1.03±0.13Aa 0.98±0.13Aa 0.94±0.12Aa 0.93±0.09Aa 0.94±0.11Aa 0.93±0.10Aa 0.94±0.07Aa
G3 0.97±0.08Aa 1.00±0.14Aa 0.99±0.12Aa 0.90±0.10Aa 0.94±0.09Aa 0.97±0.07Aa 0.93±0.09Aa 0.97±0.07Aa

Mean values followed by the same upper case letters in the same column and lower case letters on the same line do not differ significantly according to Tukey’s test (P> 0.05).

Figure 3 Serum concentrations of total calcium, iron, phosphorus, sodium (Na), potassium (K), and ionized calcium (Cai) of buffalo calves born from primiparous buffaloes (G1), multiparous with two to four parturitions (G2), and multiparous buffaloes with more than five parturitions (G3) at birth; before colostrum intake (M0); at 24h (M1), 48h (M2), and 72h (M3) after birth; and at 7 (M4), 14 (M5), 21 (M6), and 30 (M7) days after birth. 

Serum concentration of total calcium was not influenced by the number of parturitions (Table 3), as previously reported in bovine calves (Rocha et al., 2012). The mineral concentrations varied over time (Figure 3). An increase in calcium concentration was observed from birth (M0) to 7 (M4) days old in all groups, which can be attributed to the absorption of calcium from the colostrum (Jezek et al., 2006). From 14 (M5) days after birth, the results changed slightly, with a reduction observed at 30 (M7) days after birth. Evaluation of serum total calcium concentration is important for the adjustment of this element in the diet and in detecting possible metabolic and kidney problems (Kaneko et al., 2008).

Serum iron levels increased throughout the study (Table 3), mainly after 7 (M4) days after birth (Figure 3). The buffalo calves exhibited an increase in serum iron levels during the first month of life, in contrast to observations in cattle, wherein a decrease in iron levels can be observed until 30 days after birth with lower mean values than those of buffalo calves (Knowles et al., 2000). This difference is due to the higher concentration of iron in buffalo milk than in cow milk (61 ppm vs. 37ppm) (Verruma and Salgado, 1994). The serum iron concentration was not influenced by the number of parturitions (Table 3). The evaluation of serum iron levels is of great importance because this mineral participates in several metabolic processes, such as hematopoiesis, hemoglobin synthesis, activation of the cellular immune response, and pathogen-host interactions (Kaneko et al., 2008). Iron levels are also relevant to type 1 insulin-like growth factor (IGF-1), which is directly related to animal weight gain and growth (Prodanovic et al., 2014). While iron supplementation is required in bovine calves (Atyabi et al., 2006), it is not recommended in buffaloes during the neonatal period due to the risk of intoxication.

The number of parturitions did not influence the serum concentration of inorganic phosphorus (Table 3). The serum concentration of the mineral increased from birth (M0) to 7 (M4) days after birth (Figure 3), most likely due to the increased renal reabsorption of phosphate by young animals due to the action of growth hormone (Rosol and Capen, 1997). From 7 (M4) days after birth, there was a decrease in phosphorus levels until 30 (M7) days after birth. The serum concentrations of inorganic phosphorus of studied animals were higher than those previously reported in buffalo calves (Fagliari et al., 1998) and lower than those of bovine calves (Rocha et al., 2012).

The number of parturitions did not influence the serum concentrations of the measured electrolytes (Table 3). The serum concentrations of sodium, potassium, and ionized calcium exhibited slight variation over the study period (Figure 3). The ionized calcium and potassium concentrations were not affected by age in the G1 and G3 groups. Electrolyte dosing is of great importance in assessment of animal health, since electrolyte imbalances lead to changes in the pH of body fluids, blood volume, heart rate, muscle contractions, and stability of cell membranes; it is also essential for the correction of acid-base imbalances and for cation-anion balance of diets (Kaneko et al., 2008).

CONCLUSIONS

The age of the calves influenced the biochemical parameters, with the exception of ionized calcium and potassium concentrations in calves of primiparous and multiparous buffaloes with more than five parturitions. The number of parturitions influenced the serum activities of AST and GGT and the serum concentrations of total protein, albumin, globulins, and IgG in neonatal buffalo calves. Such variations are important because they allow for discrimination of physiological and pathological processes. The high serum activity of ALP in the first two days after birth, after colostrum intake, indicates that measurement of this parameter may be used as an indirect method to determine PIT failure. The serum iron concentrations of neonatal buffalo calves were high, and therefore contraindicate the supplementation of iron in these animals.

ACKNOWLEDGEMENTS

The authors acknowledge the contributions of the São Paulo Research Foundation (FAPESP) in terms of the financial support and scholarship granted (Process: 2014/09246-5).

REFERENCES

ATYABI, N.; GHARAGOZLOO, F.; NASSIRI, S.M. The necessity of iron supplementation for normal development of commercially reared suckling calves. Comp. Clin. Pathol., v.15, p.165-168, 2006. [ Links ]

BIRGELE, E.; ILGAZA, A. Age and feed effect on the dynamics of animal blood biochemical values in postnatal ontogenesis in calves. Vet. Zoot., v.22, p.5-10, 2003. [ Links ]

BOYD, J.W. Serum enzyme changes in newborn calves fed colostrum. Vet. Clin. Pathol., v.18, p.47-51, 1989. [ Links ]

DIRKSEN, G.; GRÜNDER, H.D.; STÖBER, M. Exame clínico dos bovinos. Rio de Janeiro: Guanabara Koogan, 1993. 419p. [ Links ]

FAGLIARI, J.J.; SANTANA, A.E.; LUCAS, F.A. et al. Constituintes sanguíneos de bovinos recém-nascidos da raça Nelore (Bos indicus) e Holandesa (Bos taurus) e de bubalinos (Bubalis bubalus) raça Murrah. Arq. Bras. Med. Vet. Zootec., v.50, p.253-262, 1998. [ Links ]

FEITOSA, F.L.F.; CAMARGO, D.G.; YANAKA, R. et al. Índices de falha de transferência de imunidade passiva (FTIP) em bezerros holandeses e nelores, às 24 e 48 horas de vidas: valores de proteína total, de gamaglobulina, de imunoglobulina G e da atividade sérica de gamaglutamiltransferase, para o diagnóstico de FTIP. Pesqui. Vet. Bras., v.30, p.696-704, 2010. [ Links ]

FOOD and Agriculture Organization of the United Nations. FAOSTAT 2017. Available in: <http://faostat.fao.org/site/573/DesktopDefault.aspx?PageID=573#ancor>. Accessed in: 24 Jan. 2017. [ Links ]

JEZEK, J.; KLOPCIC, M.; KLINKON, M. Influence of age on biochemical parameters in calves. Bull. Vet. Inst. Pulawy, v.50, p.211-214, 2006. [ Links ]

KANEKO, J.J.; HARVEY, J.W.; BRUSS, M.L. Clinical biochemistry of domestic animals. 6.ed. San Diego: Academic Press, 2008. 916p. [ Links ]

KLINKON, M.; JEZEK, J. Values of blood variable in calves. A bird’s-eye view of veterinary medicine. intech. 2012. Available in: <http://www.intechopen.com/books/a-bird-s-eye-view-of-veterinary-medicine/values-of-blood-variables-in-calves>. Accessed in: 4 Nov. 2015. [ Links ]

KNOWLES, T.G.; EDWARDS, J.E.; BAZELEY, K.J. et al. Changes in the blood biochemical and haematological profile of neonatal calves with age. Vet. Rec., v.147, p.593-598, 2000. [ Links ]

LAEMMLI, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, v.227, p.680-685, 1970. [ Links ]

PÉREZ-SANTOS, M.; CASTILLO, C.; HERNÁNDEZ, J. et al. Biochemical variables from Holstein-Friesian calves older than one week are comparable to those obtained from adult animals of stable metabolic status on the same farm. Vet. Clin. Pathol., v.44, p.145-151, 2015. [ Links ]

PICCIONE, G.; CASELLA, S.; GIANNETTO, C. et al. Influences of age on profile of serum proteins in calf. Acta Vet. Beograd, v.59, p.413-422, 2009. [ Links ]

PRODANOVIC, R.; KIROVSKI, D.; VUJANAC, I. et al. Relationship between serum iron and insulin-like growth factor-I concentrations in 10-day-old calves. Acta Vet. Brno, v.83, p.133-137, 2014. [ Links ]

RADOSTITS, O.M.; GAY, C.C.; HINCHCLIFF, K.W. et al. Veterinary medicine: a textbook of the diseases of cattle, horses, sheep, pigs and goats. 10.ed. Londres: W.B. Saunders, 2007. 2065p. [ Links ]

ROCHA, T.G.; NOCITI, R.P.; SAMPAIO, A.M. et al. Passive immunity transfer and serum constituents of crossbred calves. Pesqui. Vet. Bras., v.32, p.515-522, 2012. [ Links ]

ROSOL, T.J.; CAPEN C.C. Calcium regulating hormones and diseases of abnormal mineral (calcium, phosphorus, magnesium) metabolism. In: KANEKO, J.J.; HARVEY, J.W.; BRUSS, M.L. et al. (Eds.). Clinical biochemistry of domestic animals. 5.ed. San Diego: Academic Press, 1997. p.619-702. [ Links ]

SHIVAHRE, P.R.; GUPTA, A.K.; PANMEI, A. et al. Mortality pattern of Murrah buffalo males in an organised herd. Vet. World, v.7, p.356-359, 2014. [ Links ]

VERRUMA, M.R.; SALGADO, J.M. Análise química do leite de búfala em comparação ao leite de vaca. Sci. Agric., v.51, p.131-137, 1994. [ Links ]

ZAR, J.H. Biostatistical analysis. 4.ed. New Jersey: Prentice Hall, 1999. 663p. [ Links ]

Received: July 06, 2017; Accepted: April 17, 2018

E-mail: campos.damazio@gmail.com

Creative Commons License This is an open-access article distributed under the terms of the Creative Commons Attribution License