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Effect of mineral supplementation on serum calcium and phosphorus levels in pre and post-partum Marathwadi buffaloes.


Minerals are critical nutrients bearing significant effect in animal nutrition because both their excess and deficiency produce detrimental effects on health and productivity (Khan et. al., 2012). The relationship between reproduction and minerals has been recognized long back. Essential minerals are required for reproduction because of their cellular roles in metabolism, maintenance and growth. However, these nutrients may also have specific role and requirement in reproductive process. Recent studies suggests that borderline nutrient deficiencies may be manifested as impaired fertility before other clinical symptoms are apparent (Hadiya et al., 2010). The calcium effect on reproduction appears to be primarily and indirect one because it affects incidence of parturient paresis. Common sequel to parturient paresis includes dystocia, prolapse of uterus and retained placenta which tend to have negative effect on fertility. Due to phosphorus deficiency include a delayed onset of puberty and postpartum oestrus and milder deficiency have been reported to be associated with repeat breeding (Gaikwad et al., 2007). In reproductive health turn is not possible without proper mineral supplementation in their diet as most of roughages, green, concentrates and even most commercial feeds offered to Indian livestock are deficient in one or more of mineral element (Upadhyay et al. 2006). Previous research demonstrated that dietary minerals supplementation in ration of livestock is inevitable in amelioration of reproductive problems and quantity of mineral substances should be taken into consideration prior to parturition (Saghar, 2003; Akar and Yildiz, 2009; Tiwari et al., 2012; Pisal, 2012).

In India, mineral supplementation in dairy animals is not common practice. It is suggested that, trace element deficiencies are more common in combination than single element deficiency (Kankofer, 2000). It seems that low calcium, phosphorus, zinc, copper, iron, magnesium and selenium values before and after parturition predispose to retention of fetal membranes (Gupta et al., 2004; Akar and Yildiz, 2009; Han and Kim, 2005; Roche, 2006; Tillard et al., 2008), uterine prolapse (Risco, 2004; Bhatti et al., 2006; Mulligan et al., 2006; Pandey et al., 2007) and dystocia (Dufty et al., 1977; Pryor, 1976).

The present investigation was carried out to study the effect of mineral supplementation on serum calcium and phosphorus levels, before seven days of parturition, at the time of parturition and after seven days of parturition in Marathwadi buffaloes.

Materials and Methods

The fourteen advanced pregnant Marathwadi buffaloes, irrespective of their age and parity were selected from local buffalo farms in and around Parbhani. The present study was conducted in the year 2010-11. Animals were maintained under free-range system and were let loose for grazing in the field. The animal had free access to drinking water. Selection of buffaloes was based on records available from owner. Gynaeco-clinical examinations of all pregnant buffaloes were carried out for confirmation of advance pregnancy (after 10 months). A total of 14 animals were selected randomly. The selected buffaloes were calved within fifteen days. The selected buffaloes have good pedigree records. The 14 buffaloes were divided into two groups, those animals which shown serum Calcium and Phosphorus level below normal range i.e. (9.7-12.4 mg/dl and 5.6-6.5mg/dl) were taken in Group I (n=7) i.e. Treatment group and those animals which shown serum Calcium and Phosphorus level within normal range were taken in Group II (n=7) and were kept as untreated control group. Group I were given orally mineral feed supplement (Minfa (a)) daily 50 gm before and after 7 days of parturition. Blood sample from experimental animals were collected in the morning hours. Blood samples were collected aseptically from jugular venepuncture of all experimental buffaloes before and after 7 days of parturition. The all venous blood samples collected in a clean and sterilized test tube for serum separation. Serum samples were centrifuged at 300 RPM for 10 minutes. After separating from the clot and stored at -20oC until used for biochemical estimations. The serum calcium and phosphorus was estimated by-cpc End point method using Auto analyzer slim (SEAC), using standard biochemical kits (b). The values of serum Calcium and Phosphorus were expressed in mg/dl.

As the post-partum period is crucial where major metabolic flux occurs, there is sudden but significant depletion of nutrients during postpartum period, which affect the fertility. Hence, the levels of calcium and phosphorus were estimated in all experimental buffaloes.

Statistical analysis

The data analysed for mean and standard error (S.E.) and the significance was tested by using 't' test (Snedecor and Cochran, 1994).

Results and Discussions Calcium

On perusal of Table 1 it reveals that, the average serum calcium levels before 7 days of parturition, at time of parturition and after 7 days of parturition were (8.85 [+ or -] 0.27, 10.14 [+ or -] 0.64), (10.92 [+ or -] 0.38, 8.06 [+ or -] 0.60) and (11.40 [+ or -] 1.35, 9.59 [+ or -] 0.51) from group-I and group-II, respectively. The statistical analysis revealed that serum calcium levels before 7 days of parturition and after 7 days of parturition were non-significant. However, at the time of parturition these levels were found significant (P<0.01) between the groups.

Rise in serum calcium level in post partum period has also been reported by Blum et al. (1972), Rowland et al. (1975), Shrikhande et al. (1999) and El-Rainbow and Shamav (2006) which is in line with the present results. In present study it was observed that, calcium level declined about the time of birth and remained depressed for next 7 days in control group. Post-parturient hypocalcaemia was related to sudden demand for calcium to synthesize colostrums and recorded significant (P<0.01) rise in serum calcium level after 7 days of parturition (Nale, 2003). The calcium level was 8.15 [+ or -] 0.61 mg per cent on day of parturition, which increased significantly by day 15 postpartum and again by day 60 postpartum and remained more or less static thereafter (Hadiya et al., 2010). Moddie and Robertson (1962) reported abrupt reduction in plasma calcium levels around parturition, which recovered by 14th to 30th day postpartum in dairy cows. However, lower calcium level in cows with retained placenta than in non-retaining cases (Mohanty et al., 1994).


On perusal of T able 2 it reveals that, average serum phosphorus levels before 7 days of parturition, at the time of parturition and after 7 days of parturition in buffaloes were (4.90 [+ or -] 0.27, 5.79 [+ or -] 0.08), (5.50 [+ or -] 0.17, 4.79 [+ or -] 0.25) and (5.89 [+ or -] 0.14, 5.70 [+ or -] 0.23) from group-I and group-II, respectively. The statistical analysis revealed that serum phosphorus levels were non-significant after 7 days of parturition. However, before 7 days of parturition and at time of parturition these levels were found significant (P<0.01) between the groups.

Similar results were obtained by Blum et al. (1972), Wilson et al. (1977), Shrikande et al. (1999) and Nale (2003). In accordance with present results, the similar result were obtained by El-Rainbow and Shamav (2006) who reported maternal phosphorus declined about the time of birth and remained depressed for the next 7-8 days in control group. The level of phosphorus increased significantly by day 30-45 postpartum (Hadiya et al., 2010). Ullah et al. (2010) suggested that the increased levels of minerals were due to the supplementation which have not reached to a significant level. Sane (1977) opined that infertility due to nutritional deficiencies particularly of phosphorus is usually characterized by failure of oestrus or cessation of estrous cycle. Asthana et al. (2007) reported that repeat breeding cows supplemented with mineral mixture revealed significant difference of serum inorganic phosphorous at different stages of observation i.e. day 0, 21 and 45. It also differed significantly with other treatment groups at day 45, but not on day 0 and day 21.

Kirchner et al. (1977) also reported decreased value of calcium and phosphorus after parturition and calcium values resumed towards normalcy after 7 days of parturition. Deshpande et al. (1998) reported significant increase in calcium and inorganic phosphorus levels from day of calving to 7th, 14th and 21st day postpartum. Singh and Vadnere (1987) recorded significantly higher plasma calcium and phosphorus levels at induced oestrus in anoestrus cows supplemented with minerals as compared to pre treatment and control values. The lower level of plasma calcium and phosphorus at parturition and subsequently at oestrus or early pregnancy was thought to be due to high oestrogen levels, as oestrogen may change appetite in cows, hence diminished calcium and phosphorus intake and absorption (Sahukar et al., 1984). Patel et al. (2006) did not find any variation in the overall means of plasma calcium, inorganic phosphorus and magnesium levels in fertile and infertile groups of cows. Serum calcium and phosphorus in cows with retention of placenta were lower as compared to cows with non-retention of placenta (Bari et al., 1996). Begum et al., 2010 was indicated that 120 per cent Ca and P supplementation increased milk production and reproductive performance of lactating buffaloes. Lactation tends to lower the level of blood calcium as a result of the transfer of blood calcium to the milk. It is due to the dietary calcium may reduce the calcium mobilization from bones for metabolism, resulting in lower calcium level in blood serum (Boda et al., 1980).

There was incidence of retention of placenta in one buffaloes, uterine prolapse in one buffaloes and dystocia in two buffaloes from the control group, where as no any incidence of uterine prolapse and dystocia in the treatment Group. The variation of present finding with those reported by various authors may be due to different feeding practices, post-partum period, age and breed of buffaloes.

From the present study it can be concluded that rise in serum levels of calcium and phosphorus at parturition and 1 week later of post parturient period after given pre-partum dietary mineral supplementation. Due to pre-partum dietary mineral supplementation there was no any incidence of calving disorders in the treatment group.


Authors wish to grateful acknowledge the facilities rendered by the Associate Dean of College of Veterinary and Animal Sciences, MAFSU, Parbhani (M.S.), India. We also thank the staff of Department of Animal Reproduction, College of Veterinary and Animal Sciences, Parbhani (M.S.), India for their care of the animals and their technical assistance.


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P.M. Mane (1), R.L. Dhoble (2), P. R. Suryawanshi (3), S.M. Gaikwad (4) and R.J. Chaudhari (4)

Department of Animal Reproduction

College of Veterinary and Animal Sciences

Maharashtra Animal Fishery Sciences University

Parbhani--431402 (Maharashtra)

(1.) Post Graduate Scholar and Corresponding Author Email:

(2.) Professor

(3.) Post Graduate Scholar

(4.) Assistant Professor

(a)--Brand of Intas Animal Health, Ahmedabad

(b)--Brand of Ambica Diagnostics, Parbhani
Table 1. Serum blood calcium levels (mg/dl)

Sr. No.     Before 7 days          At the time of
            parturition (mg/dl)    parturition (mg/dl)

            Group-I     Group-II   Group-I   Group-II

1           8.07        10.02      10.13     6.92
2           8.9         10.08      11.04     8.34
3           9.51        8.68       9.83      10.98
4           7.82        12.94      10.19     9.09
5           8.63        11.9       12.08     7.11
6           9.42        8.41       10.66     6.24
7           9.64        8.99       12.49     7.73
Mean        8.85        10.14      10.92     8.06
SE          0.27        0.64       0.38      0.60
't" value        1.46 (NS)              3.39 **

Sr. No.     After 7 days
            parturition (mg/dl)

            Group-I   Group-II

1           9.48      9.15
2           10.09     12.06
3           8.56      8.46
4           10.42     10.37
5           12.13     10.01
6           19.11     8.14
7           10.04     8.93
Mean        11.40     9.59
SE          1.35      0.51
't" value       1.13 (NS)

(NS) = Non-significant, ** P<0.01

Table 2. Serum blood phosphorus levels (mg/dl)

Sr.         Before 7 days        At the time of
No.         parturition (mg/dl)  parturition (mg/dl)

            Group-I   Group-II   Group-I   Group-II

1            5.49       5.7       5.59       4.82
2            3.38       5.9        6.1       4.87
3            4.78       5.58       4.9       3.89
4            5.09       6.01      5.78       5.22
5            5.34       5.71      5.31       4.91
6             4.9       5.59      4.99       4.01
7            5.31       6.08      5.83       5.85
Mean         4.90       5.79      5.50       4.79
SE           0.27       0.08      0.17       0.25
't" value         3.13 **              4.37 **

Sr.         After 7 days
No.         parturition (mg/dl)

            Group-I     Group-II

1             5.69        5.83
2             6.02        6.53
3             5.91        4.83
4             6.03        6.09
5             6.41        5.9
6             5.21        4.95
7               6         5.78
Mean          5.89        5.70
SE            0.14        0.23
't" value          1.01 (NS)

(NS) = Non-significant, ** P<0.01
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Title Annotation:Research Article
Author:Mane, P.M.; Dhoble, R.L.; Suryawanshi, P.R.; Gaikwad, S.M.; Chaudhari, R.J.
Publication:Intas Polivet
Article Type:Report
Geographic Code:9INDI
Date:Jan 1, 2014
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