The status of differentmacrominerals was appraised in soils, forages and blood plasma of two buffalo groups (lactating, non-lactating). Soilmg+ and K+ remained below their respective critical values, while Na+ and Ca+ exceeded their critical values in soil. Forage had high Ca andmg whereas, K and N concentrations stayed within prescribed limit. Blood plasma hadmean values higher than critical concentrations for Na, Ca, andmg. The observations recorded in current study implied thatmetals in toxic concentrations have been built up in animals through consuming forage contaminated withmetals both in tissues and at exposed plant parts.

Keywords: ruminant, pasture,mineral, nutrition,milk

One of themajor contributory sectors of agriculture in Pakistan is livestock sector. Contribution of livestock in GDP of the country is 12%.¹ Several products likemeat,milk,manure and leather hide are obtained from livestock.² Plants are the primary source of food for ruminants. Apart from fulfilling food needs, plants also provide adequate amounts ofminerals to the animals. Plants acquire theseminerals from soil. A complex interactive relationship occurs between soil, plants and animals. Hence,mineral status in these interdependent entitiesmust be ascertained for provision of balanced diet.³ Biomonitoring is necessary for regulating environmental stress triggered by upheavals inmineral concentrations.⁴

Macrominerals like calcium (Ca),magnesium (Mg), sodium (Na) and Potassium (K) are required in animals and plants for variousmetabolic processes.^5,6 Excess and dearth of theseminerals cause various physiological and cellular defects. High calcium levels cause slow bone growth and heart failure;⁷ elevated potassium level results inmembrane damage, tissue necrosis and hyperleukemia.^8,9magnesium shortage implicates in hypomagnesia.¹⁰ Forage digestibility is reduced deficiency of sodium.¹¹ An approach to devise a possible relationship ofmetal flow among soil, forages and blood plasma is the question of this study. Ascertainingmineral level in forages and their consumers was taken under consideration.

Study area

Livestock station at Shorkot city, district Jhang was taken as study area. Temperature of the area varies according to season withmean winter and summer temperatures dwindling between 12-35°C.

Soil and forage sampling

Various forages including some dominant plants like Sorghum bicolor, Zeamays, Pennisetum glaucum were grown in pasture at the time of sampling. Different sampling intervals like 5m, 10m, 15m and 20m were kept for collection of soil and forage. Forages were taken from 3-6cm at the feeding level of grazing animals from predefined sampling intervals. A total of 36 samples of soil and forages each were collected.

Blood sampling

Twenty buffaloes were grouped in two categories i.e. lactating (n=10) and non-lactating (n=10). Non-lactating buffaloes were below 4-5years and weighed between 70-100kg while lactating buffaloes were above 4-5years and weighed between 80-102kg. 15ml blood sample from jugular vein was collected in heparin containing syringes and plasma was separated via centrifugation.¹²

Sample preparation and analysis

After air drying, the forages and soil were oven dried for 3days to evademoisture. After grinding, the samples were passed through fine sieve. 1gm of each sample was taken for digestion with the help of H₂O₂ and H₂SO₄ using ratios defined by Vidovic et al.¹³ Themixture was heated for 3-4hours till transparency was achieved. The samples were diluted up to 50ml after filtration. H₂SO₄ (2ml) was added in 2ml of blood plasma and themixture was kept overnight; digestion was carried out till the dissolution of organicmatter. Final volume of 50ml wasmade using distilled water.

Analysis ofmetals

Atomic absorption spectrophotometer (Model #AA-6300, Shimadzu, Japan) was used for estimation of sodium, potassium,magnesium and calcium.

Statistical analysis

Metalmean values and statistical significant differences betweenmetal concentrations of all samples were determined using SPSS (version 21).

Soil potassium

ANOVA depicted significant (p≤0.05) K mean concentration differences with respect to sampling intervals for soil (Table 1). The concentration ranged between 83.40-106.20 mg/kg (Figure 1). Samples obtained from 15m distance from road side had highest K mean levels.

Figure 1 Variation in K mean concentration in soil at different sampling intervals.

Forage potassium

Significant (p≤0.05) variation in K forage concentration was exhibited at different sites (Table 1). Highest K level was found at the 20m distance while the range stayed between 0.088-0.102% (Figure 2).

Figure 2 Variation in K mean concentration in forage at different sampling intervals.

Soil Sodium

No significant difference in Na soil concentration was observed at different distances (Table 1). The range of Na level was 58.15-64.46 mg/kg (Figure 3). Farthest distance of 20m had highest Na level.

Figure 3 Variation in Na mean concentration in soil at different sampling intervals.

Forage Sodium

Non-significant difference in Na forage concentration was observed at different distances (Table 1). A range of 401.98-428.24 % was observed (Figure 4). Highest Na level was found at the 20m distance.

Figure 4 Variation in Na mean concentration in forage at different sampling intervals

Soil Calcium

ANOVA depicted significant (p≤0.05) Ca mean concentration differences with respect to sampling intervals for soil (Table 1). The concentration ranged between 63.35-82.22 mg/kg (Figure 5). Samples obtained from 15m roadside distance had highest Ca mean levels.

Figure 5 Variation in Ca mean concentration in soil at different sampling intervals

Forage Calcium

Variation in Ca forage level was not significant (Table 1). Farthest distance of 20m had highest Ca level with range between 2163.47-2416.87% at all sampling distances (Figure 6).

Figure 6 Variation in Ca mean concentration in forage at different sampling intervals

Soil Magnesium

ANOVA yielded significant (p≤0.05) Mg mean concentration differences with respect to sampling intervals for soil (Table 1). A range of 9.92-11.41 mg/kg was obtained while maximum level was found at 20m site (Figure 7).

Figure 7 Variation in Mg mean concentration in soil at different sampling intervals

Forages Magnesium

An insignificant difference in Mg forage level was observed at different distances (Table 1). Sampling point of 20m had highest level of Mg in plants (Figure 8).

Figure 8 Variation in Mg mean concentration in forage at different sampling intervals

Blood plasma potassium

Relatively similar concentrations of K were present in plasma of two buffalo categories (Table 2). Obtained range for samples was 17.24-17.27 mmol/L (Figure 9) and the highest level was ascertained in non-lactating buffaloes lower than the critical limit (2.20mmol/L) established by Grace.¹⁴

Figure 9 Variation in K mean concentration in blood plasma.

Blood plasma Sodium

Almost similar Na concentration was present in whole buffalo lot under study (Table 2). Non-lactating buffaloes, however, had somewhat higher Na mean level and the overall range was 1220.33-1311.65 mmol/L (Figure 10).

Figure 10 Variation in Na mean concentration in blood plasma.

Blood plasma Calcium

Relatively similar concentrations of Ca were present in plasma of two buffalo categories (Table 2). Obtained range for samples was83.70-90.92 mmol/L (Figure 11) and the highest level was ascertained in non-lactating buffaloes.

Figure 11 Variation in Ca mean concentration in blood plasma.

Blood plasma Magnesium

ANOVA yielded significant (p≤0.05) Mg mean concentration differences with respect to buffalo cohorts (Table 2). Lactating buffaloes had higher Mg mean level and the overall range was 6.81-8.09 mmol/L (Figure 12).

Figure 12 Variation in Mg mean concentration in blood plasma

The estimated concentration stayed within the critical limit (80mg/kg) described by Warncke & Robertson.¹⁵ Current study estimations were also lower than the other studies carried out in Florida¹⁶ and Pakistan.¹⁷ Leaching could be the possible reason of low K concentrations.¹⁸ The K levels did not exceed the critical limit of 0.8%.³ The current study results concurred with findings of Prabowo et al.¹⁹ in Indonesia, Ogebe et al.²⁰ in Nigeria, Tiffany et al.^21,22 in North Florida and²³ in Pakistan. However, these levels were lower than 8 g/kg, the recommended limit for grazing animals.⁶

Rhue & Kidde²⁴ estimated the critical level of 62mg/kg and the Na concentration in current study exceeded that. Similar values at other animal ranches were found in various other studies.^17,18,25,26 Optimal level of 0.06%²⁷ was not crossed in any sample. Several other studies in different parts of the world had reported similar results.^21,22,28 Na supplementation was not needed as the ruminants had adequate supply of Na from the forage. The results of Anon²⁹ and Khan et al.³⁰ were below the levels ascertained in current study.

The critical limit (72mg/kg) was surpassed.²³ Contrasting results were obtained by Ruan et al.³¹ for calcium in forages. The samples had higher levels than critical limit of 0.3%.³ Khan et al.^24,32 and Pastrana et al.³³ reportedmuch lower Ca levels than the current study. The level stayed within the critical range of 30mg/kg.²³ Ciećko et al.³⁴ and Tiffany et al.²² reported levels higher than the present investigation. The results suggested that soil was deficient ofmg and needed fertilization ormanuring.mineral lossmay occur through excretion³⁵ and it could be the probable cause of K level being devised asmarginally deficient. Present study levels were, however, higher than the findings of Grunwaldt et al.³⁶

Plasma had gathered Na higher than critical limit of 120mmol/L.³⁷ The values were above the findings of Rahman²⁸ but similar to Khan et al.³⁸ The values were above the critical limit of 2mmol/L.³⁹ Bloodmineral concentrations had been affected by themineral status in soil and forage.⁴⁰ The currently calculated values exceeded critical limit of 2.8mmol/L.¹⁴ Pasha et al.⁴¹ had reported similar findings but Gizachew⁴² reportedmg levels lower than the current study. Forages have the ability of fulfillment of essential nutrient requirements of the livestock.^38,43‒46

None.

Author declares that there is no conflict of interest.

1. Chandio AA, Yuansheng J, Magsi H. Agricultural sub-sectors performance: An analysis of sector-wise share in agriculture GDP of Pakistan. International Journal of Economics and Finance. 2016;8(2):156‒162.
2. Economic Survey of Pakistan. Pakistan; 2008-2009:67‒68.
3. Mcdowell LR, Conrad JH. Trace mineral nutrition in Latin America. World Animal Review. 1977;24:24‒33.
4. Caggiano R, Sabia S, Emilio MD, et al. Metal levels in fodder milk dairy products and tissues sampled in ovine farms of Southern Italy. Environ Res. 2005;99(1):48‒57.
5. Masters DG, Purser DB, Yu SX, et al. Mineral nutrition of grazing sheep in northern China. I. Macro-minerals in pasture feed supplements and sheep. Asian-Australian Journal of Animal Sciences. 1993;6(1):99‒105.
6. Underwood EJ, Suttle NF. The Mineral Nutrition of Livestock. Common wealth Agricultural Bureaux, London; 1981. p. 223‒225.
7. Soetan KO, Olaiva CO, Oyewole OE. The importance of mineral elements for humans domestic animals and plants: a review. African Journal of Food Science. 2010;4(5):200‒222.
8. Singh SP, Kumar N, Sharma KB, et al. Serum and CSF mineral profile of Himalayan yak (Bos grunniens) in their natural habitat. Asian-Australian Journal of Animal Science. 1999;12(2):189‒191.
9. Latimer KS, Mahaffey EA, Prasse KW. Duncan and Prasse’s Veterinary Laboratory Medicine: Clinical Pathology. 4th ed. Ames, Iowa: Iowa State Press; 2003. p. 450.
10. Ram L, Schonewille JT, Martens H, et al. Magnesium absorption by wethers fed potassium bicarbonate in combination with different dietary magnesium concentrations. J Dairy Sci. 1998;81(9):2485‒2492.
11. Grusak MA, Dellapenna D. Improving the nutrient composition of plants to enhance human nutrition and health. Annu Rev Plant Physiol Plant Mol Biol 1999;50:133‒161.
12. Fick KR, Mcdowell LR, Miles PH, et al. Method of Mineral Analysis for Plant and Animal Tissues. 2nd ed. USA: Animal Science Department University of Florida Gainesville; 1979.
13. Vidovic M, Sadibasic A, Cupic S. et al. Perceived family cohesion adaptability and communication in eating disorders. Environmental Research. 2005;97(1):26‒31.
14. Grace ND. The mineral requirement of grazing ruminants. The Oslo Centre for Critical Architectural Studies Publications. 9.9. New Zealand Society of animal production. 1983:56‒66 p.
15. Warncke DD, Robertson LS. Understanding the MSU soil test report: results and recommendations. Extension bulletin 937 MSU AG Facts. Cooperative Extension Service Michigan State University, USA; 1976.
16. Mooso GD. Warm-season grass production and nutritive uptake with liquid and solid N-P-K fertilizers. USA: M.S. Thesis Brigham Young University, 1982.
17. Khan ZI, Ashraf M, Javed I, et al. Transfer of sodium from soil and forage to sheep and goats grazing in a semiarid region of Pakistan. Influence of the seasons. Trace Elements and Electrolytes. 2007;24:49‒54.
18. Espinoza JE, Mcdowell LR, Wilkinson NS, et al. Monthly variation of forage and soil minerals in Central Florida. Communications in Soil Science and Plant Analysis. 1991;22(11‒12):1137‒1149.
19. Prabowo A, Mcdowell LR, Wilkinson NS, et al. Mineral Status of Grazzinmg cattle in South Sulawesi Indonesia; I. Macrominerals. Am. Journal of Animal Science. 1990;4(2):111‒120.
20. Ogebe PO, Ayoade JA, Mcdowell LR, et al. Mineral concentrations of forages and soils in Benue State Nigeria. I. Macro minerals and forage in vitro organic matter digestibility and crude protein concentrations. Communication in Soil Science and Plant Analysis. 1995;26(13‒14):1989‒2007.
21. Tiffany ME, Mcdowell LR, Connor GAO, et al. Effects of pasture applied bio solids on forage and soil concentrations over a grazing season in north Florida. I. Macrominerals. Crude Protein and In Vitro Digestibility. Communication in Soil Science and Plant Analysis. 2000;31(1‒2):201‒213.
22. Tiffany ME, Mcdowell LR, Martin FG, et al. Effects of residual and reapplied bio solids on forage and soil concentrations over a grazing season in north Florida. II. Microminerals. Communication in Soil Science and Plant Analysis. 2001;32(13‒14):2211‒2226.
23. Khan ZI, Ahmad K, Ashraf M, et al. Mineral status of forage and its relationship with that of plasma of farm animals in southern Punjab, Pakistan. Pakistan Journal of Botany. 2009;41(1):67‒72.
24. Rhue RD, Kidder G. Analytical procedures used by the IFAS extension soil laboratory and the interpretation of results. USA: Soil Science 438-446 Department, University of Florida Gainesville, 1983.
25. Khan ZI Hussain A. Mcdowell LR, Ashraf MY. Seasonal variation of calcium in soil and forage minerals in plant animal system at a semiarid sheep ranch Punjab Pakistan. International Journal of Biotechnology. 2005;2:163‒174.
26. Khan ZI, Ahmad K, Ashraf M, et al. A comparative study on mineral status of blood plasma of small ruminants and pastures in Punjab Pakistan. Pakistan Journal of Botany. 2008;41:67‒72.
27. National Research Council. Mineral tolerance of domestic animals. Washington DC: National Academy Press; 1980.
28. Rehman A, Kinacid MM, Elzubejr PIEA. Mineral deficiencies in grazing cattle in Kardofan and Darfur regions in the eastern Sudan. Tropical Animal Health and Production. 1998;30:123‒135.
29. Anonymous. Nutrient requirements of domestic animals. National Research Council. Nutrient Requirements of Beef Cattle. 6th Rev ed. Washington DC: National Academic Science; 1985. 1p.
30. Khan ZI, Ashraf M, Hussain A. Evaluation of manganese status of grazing sheep in a semi-arid region of Pakistan. Trace Elements and Electrolytes. 2007;24:55‒60.
31. Ruan MH, Chen SH, Li WQ. Relationship between supply and absorption of potassium calcium and magnesium in flue-cured tobacco. Subtropical Agriculture Research 2006;2(2):97‒101.
32. Khan ZI, Hussain A, Ashraf M, et al. Mineral status of soils and forages in South Western Punjab-Pakistan: Micro-minerals. Asian-Australian Journal of Animal Science. 2006;19(8):1139‒1147.
33. Pastrana R, Mcdowell LR, Cornad JH, et al. Mineral status of sheep in the Paramo region of Colombia. II. Trace Mineral Small Ruminant Research. 1991;5(1‒2):23‒34.
34. Ciecko Z, Wyszkowski M, Krajewski W, et al. Effect of organic matter and liming on the reduction of cadmium uptake from soil by triticale and spring oilseed rape. Sci Total Environ 2001;281(1‒3):37‒45.
35. Mcmahon FG, Ryan JR, Akdamar K, et al. Effect of potassium chloride supplements on upper gastrointestinal mucosa. Clin Pharmacol Ther. 1984;35(6):852‒855.
36. Grunwaldt EG, Guevara JC, Estevez OR, et al. Instituto Argentino Investigations delas Zonas Aridas (IADIZA) CC507(5500). Tropical animal health and Production. 2005;37:527‒540.
37. Dighe AS, Rao A, Coakley AB. Analysis of laboratory critical value reporting at a large academic medical center. American Journal Clinical Pathology. 2006;125(5):758‒764.
38. Khan ZI. Effect of seasonal variation on the availability of macro-and micro nutrients to animals (sheep and goats) through forage from soil. Ph.D Thesis University of Agriculture Faisalabad Pakistan. 2003. 286p.
39. Radostits OM, Gay CC, Hinchliff KW. Veterinary Medicine a textbook of disease of cattle sheep pigs goats and horses. 9th ed. WB Saunders; 2000. p. 1599‒1603.
40. Mtimuni JP, Mfitilodze MW, Mcdowell LR. Interrelationships of minerals in soil-plant-animal system at Kuti Branch Malawi. Communication in Soil Science and Plant Analysis. 1990;21:415‒427.
41. Pasha TN, Khan ZM, Farooq U. Assessment of micro minerals status in soil water feed resources and its influence on blood plasma of sheep and goats in central Mix cropping zpone of Punjab Pakistan. Tropical and subtropical Agro ecosystems. 2009;11:249‒253.
42. Gizachew L, Hirpha A, Jalata E, et al. Mineral animal status of soils native pastures and cattle blood serum in the mid-altitude of Western Ethiopia. African Journal of Ranger and forage sciences. 2002;19(3):147‒155.
43. Fujihara T, Matsui T, Hyashi S, et al. Mineral Status of Grazing Philippine goats. II The Nutrition of selenium copper and zinc of goats in Luzon Island. Asian- Australian Journal of animal science. 1992;5(2):389‒395.
44. Grace ND, Lee J. Effect of increasing Fe intake and Cu content of tissues in grazing sheep. Proceedings of the New Zealand Society of Animal Production. 2005;50:265‒268.
45. Khan ZI, Hussain A, Ashraf M, et al. Evaluation of variation of soil and forage minerals in pasture in a semiarid region of Pakistan. Pakistan Journal of Botany. 2005;37(4):912‒931.
46. Meschy F. Recent progress in the assessment of mineral requirements of goats. Livestock Production Science. 2000;64(1):9‒14.