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Serum Concentrations of Iron, Copper, Zinc, Manganese and Cobalt in Anoestrus Cattle and Buffaloes under Farm Condition

Vishal Mudgal Vishnu Kumar Gupta Shalini Srivastava Ajaz Ahmed Ganai
Vol 2(1), 150-159

The present study was planned to determine serum micro-mineral status (iron, copper, zinc, manganese, cobalt) in 50 anoestrus Murrah buffalo, Sahiwal and Crossbred cows and heifers. Serum micro-mineral contents were determined by atomic absorption spectrophotometer. Mean serum concentrations of iron, copper, zinc, manganese and cobalt taken from 14 Murrah buffalo cows and heifers were 2.82 ± 0.65, 0.48 ± 0.20, 2.65 ± 1.46, 0.24 ± 0.06 and 0.49 ± 0.24ppm, respectively. These mean values were 2.47 ± 1.35, 0.52 ± 0.16, 3.69 ± 2.57, 0.18 ± 0.09 and 0.56 ± 0.22ppm, respectively in 20 Sahiwal cow and heifers, while, 3.16 ± 1.05, 0.45 ± 0.13, 2.76 ± 1.51, 0.21 ± 0.07 and 0.51 ± 0.22ppm, respectively in 16 crossbred cow and heifers. Compared with normal range of these trace elements in healthy animals the level of copper was on a border way to show the signs of clinical deficiency symptoms, while the levels of other elements was in quite safe range to encounter any problem. It was concluded that deficiency of copper, could be responsible for anoestrus condition in these animals. Thus, to overcome the deficiency, strategic dietary supplementation of copper with better bioavailability could be a suitable approach.

Keywords : Buffalo Sahiwal Crossbred Cows Heifers Anoestrus Serum Trace elements


Normal productive and reproductive behavior in domestic animals is closely associated with nutritional status of the animals. In this regard, minerals in particular play very crucial role to exploit the actual genetic potential of the animals. Animals can’t perform to their genetic potential if their mineral needs are not meet even if they receive 100% of their protein and energy needs. Mineral deficiency exists widely in livestock and the severity of the deficiency depends upon the type of feed, physiological status of the animals and the agro-climatic conditions of the region. Deficiency of a single or multiple minerals or their imbalances may cause various reproductive failures, such as infertility, poor conception, anoestrous etc. (Hidiroglou, 1979). Several trace elements acts as cofactor and helps in hormone synthesis and hence influences biochemical functions associated with reproduction (Valee and Wacker, 1976). Because of their role in the endocrine system and in tissue integrity, mineral have a beneficial role to play in resumption of follicular growth and fertility in dairy cows and buffaloes.

Mineral that affects reproduction in ruminants are generally found within the trace elements group (Cu, Co, Zn, Fe and Mn). The role of copper and zinc are known to have a significant correlation with reproductive hormones (Progesterone and esteridol) (Prasad et al., 1989) and hence with normal reproduction, as they are specific activators of enzyme systems that assist in maintaining the activity of hyperphysical hormones in blood (Georgievskii, 1982). Zinc (Zn) deficiency may reduce GnRH secretion that eventually leads to the arrest of ovulation (Kaswan and Bedwal 1995). Lack of manganese (Mn) may inhibit the synthesis of cholesterol and its precursors that in turn may limit the synthesis of sex hormones and possibly other steroids (Doisey 1973) and thus have a significant role to play in maintaining the optimum fertility to the animals (Underwood, 1981). Copper (Cu) has its role in maintaining optimum fertility, as it is involved in FSH, LH and estrogen activity (Desai et al 1982). A low level of serum iron (Fe) has been found to influence normal gonadal activity possibly by causing improper hormonal output and deterioration of general physiological function (Sharma et al 1986).

Dietary deficiency of micronutrients is well combated by commercially available feeds which usually are deficient in trace elements and hence optimum supplementation of all the micro minerals results as stimulus for ovarian rebound and initiation of ovarian activity (Markandaya et al., 2002) and the most devastating economic loss in dairy industry is caused due to suboptimal production and infertility and that’s why supplementation of deficient mineral/s had dramatically increased the fertility status of the grazing cattle (Mc Donald, 1986).

On the basis of above literature it may be hypothesized that the deficiency of some micro minerals in the blood may cause anoestrus in animals and hence the present study was, therefore, designed to determine levels of micro minerals (iron, copper, zinc, manganese and cobalt) in the serum of anoestrus Murrah buffalo, Sahiwal and crossbred cows and heifers.

Materials and Methods

The experiment was conducted at a government Livestock Farm at Jabalpur (MP). The animals were free from any anatomical abnormality of reproductive organs and were maintained under stall fed condition with green fodders, straw, concentrate and ad libitum drinking water.

The study was conducted on 50 adult anestrous cattle and buffaloes (cows and heifers), including 14 Murrah buffalo, 16 crossbred cows and 20 Sahiwal cows and heifers. About 20 ml of blood from each animal was collected in a clean sterilized glass test tube through jugular venipuncture in the morning (before watering and feeding), and kept in slanting position for 45 min, followed by centrifugation at 700×g for 15 min to separate out the serum. Serum samples were stored at -20oC till their analysis. Wet digestion of samples was done using double acid mixture (Nitric acid + Perchloric acid). The serum iron, copper, zinc, manganese and cobalt were determined using atomic absorption spectrophotometer. The mean values (± SE) for iron, copper, zinc, manganese and cobalt were calculated.

Results and Discussion

Delayed onset of puberty as well as anestrous remains a very common problem affecting animal’s economy. The mechanism of mineral reproduction interaction is not fully understood because of the complexity of the neuro hormonal dialogue. Some mineral acts directly on gonads while others act through hypophyseal pituitary gonadal axis. Several trace elements acts as cofactor or activate enzymes and helps in hormone synthesis and hence influences biochemical functions associated with reproduction. Because of their role in the endocrine system and in tissue integrity, mineral have a beneficial role to play in resumption of follicular growth and fertility in dairy cows and buffaloes. Hence, afford was made to find out the cause of the same to overcome it. The serum’s trace mineral status was assessed in Murrah buffalo, Sahiwal and Crossbred cows and heifers has been represented in the tables (1-3).

Table 1: Serum trace elements status (ppm) of Murrah buffalo cows and heifers

Animal Fe Cu Zn Mn Co
1 3.1 0.275 1.125 0.2 0.35
2 2.95 0.35 1.475 0.175 0.5
3 1.2 0.3 2.375 0.25 0.6
4 3.8 0.225 1.8 0.325 0.25
5 2.275 0.225 4 0.25 0.325
6 3.1 0.625 2.375 0.15 0.3
7 2.275 0.45 3.15 0.325 0.675
8 2.025 0.4 2.15 0.25 0.15
9 2.2 0.575 3.25 0.25 0.2
10 3.375 0.45 6.5 0.1 0.5
11 3.7 0.3 5.425 0.325 0.275
12 2.7 0.825 1.175 0.25 0.9
13 3.5 0.925 1.2 0.175 0.95
14 3.325 0.775 1.125 0.325 0.925
Average 2.82 ± 0.65 0.48 ± 0.20 2.65 ± 1.46 0.24 ± 0.06 0.49 ± 0.24


Table 2: Serum trace elements status (ppm) of Sahiwal cows and heifers

Animal No Fe Cu Zn Mn Co
1 2.675 0.3 2.05 0.2 0.575
2 8.025 0.25 1.575 0.25 0.45
3 2.35 0.4 1.575 0.15 0.325
4 4.325 0.75 1.075 0.25 0.625
5 2.4 0.225 3.975 0.125 0.825
6 1.55 0.35 8.475 0.075 0.4
7 1.875 0.375 3.725 0.05 0.525
8 2.025 0.3 11.95 0.1 0.425
9 1.45 0.525 6.225 0.1 0.8
10 1.8 0.675 6.25 0.075 0.325
11 1.025 0.4 4.75 0.075 0.6
12 0.85 0.6 6.025 0.15 0.55
13 1.975 0.65 5.75 0.3 0.15
14 1.325 0.55 2.2 0.325 0.75
15 2.075 0.7 1.675 0.125 1.025
16 1.9 0.775 1.825 0.175 1.175
17 1.9 0.625 1.1 0.475 0.5
18 2.925 0.55 1.35 0.125 0.225
19 3.8 0.65 1.3 0.3 0.4
20 3.125 0.8 0.875 0.2 0.6
Average 2.47 ± 1.35 0.52 ± 0.16 3.69 ± 2.57 0.18 ± 0.09 0.56 ± 0.22



Table 3: Serum trace elements status (ppm) of Crossbred cows and heifers

Animal No Fe Cu Zn Mn Co
1 1.975 0.275 3.175 0.15 0.1
2 2.375 0.425 2.575 0.15 0.3
3 1.71 0.545 5.47 0.075 0.15
4 3.7 0.675 5.45 0.175 0.625
5 3.1 0.375 1.525 0.275 0.65
6 3.125 0.35 1.65 0.25 0.55
7 3.575 0.3 2 0.25 0.85
8 2.89 0.3 1.6 0.275 0.85
9 6.65 0.475 1.25 0.125 0.45
10 4.875 0.275 1.375 0.15 0.275
11 2.275 0.45 3.4 0.15 0.725
12 2.3 0.5 5.75 0.175 0.575
13 3.2 0.8 5.075 0.325 0.9
14 2.775 0.525 1.6 0.225 0.5
15 2.575 0.625 0.95 0.375 0.35
16 3.475 0.325 1.25 0.3 0.325
Average 3.16 ± 1.05 0.45 ± 0.13 2.76 ± 1.51 0.21 ± 0.07 0.51 ±0.22

When the values were compared to that of standard values of physiologically normal animals (Geogievskii, 1982, Mc Donald, 1986) it indicated that the serum levels of Zinc (Zn), Manganese (Mn), Iron (Fe) and cobalt (Co) were remained in the normal range of healthy adult ruminants, while, a wide variation among the values in different animals were observed, but overall the level found to be on higher side of the normal values reported for these trace minerals, which indicated the good status of the animals raised under the standard conditions in an organized manner.

The level of copper was found below the critical level of 0.65ppm (Miles and Mc Dowell, 1983) in all the cows and heifers irrespective of the breed and species, which remains a sign of hypocupraemia because in general the normal serum copper concentration, studied under different physiological conditions were found to be with in the range of 0.65 to 1ppm under farm conditions (Geogievskii, 1982). Deficiency of copper is generally complicated by anemia, debility, lack of appetite and subsequently results in failure of estrous and delayed onset of puberty (Underwood, 1981). It indicates that the reproductive problems may be associated with the reduced copper level.

Similar to the present findings anestrous was also been reported in the animals having lower blood copper level by Munro (1957). Ramana et al. (2000) reported reduced plasma level of copper (0.31ppm) and correlated it with the reproductive problems of dairy animals. Similarly, Phillippo et al. (1987) also found that the copper deficient heifers (due to high Mo levels) had a decreased pregnancy rate. Lower copper concentration in anoestrus cattle has also been reported in various studies (Deshpande et al., 1981; Dabas et al., 1987; Dutta et al., 2001). Estrogen hormone has been reported to increase copper level (Sato and Henkin, 1973) and the lower level of copper in anoestrus buffaloes in the present study may be due to lower estrogen level in anoestrus animals (Rajkumar et al., 2006).

While, Muehlenbein et al. (2001) observed positive effect of copper supplementation on the conception rate in cows, which were very much Cu deficient before and during the breeding season. Rmmana et al. (2000) also reported a correlation of reduced level of serum copper (0.31ppm) with the reproductive problems of dairy animals.

Lall et al. (2000) when supplemented mineral mixture in the diet of adult buffaloes and heifer, they also observed that the animals came into heat and conceived within a period of 2-4 weeks after starting the supplementation. Koley and Biswas (2004) observed the onset of estrous in 6 anestrous heifers within 14-21 days, 4 animals within 28-35 days and 4 animals within 35-42 days of mineral mixture supplementation. In their study 70% of the animals exhibited estrous during the experimental period. Identical findings were also observed by Tiwari et al. (1999) and Kumar et al. (2002).

The results may be concluded that copper is an essential trace element for normal reproduction performance of the animals and strategic dietary supplementation of copper through it’s highly bio-available source could be a suitable approach to counteract the problem.


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