Ansar Mahmood,Dr.Asma Hassan,Iram Maqsood,Tang Li-jie,Syed Mohsin Bukhari,Khalil-Ur-Rehman,and Shahla Andleeb

1 Department of Wildlife and Fisheries,Government College University,Faisalabad,Pakistan

2 College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China

Introduction

Infertility,non-infectious abortions,anemia and bone abnormalities are some of the clinical signs suggested mineral deficiencies in the livestock (McDowell,1985;Bicknell,1995).The quality and quantity of mineral nutrients in the forages mainly depend upon the factors like texture of the soil,fertilization of the soils and irrigation water.Mineral availability,particularly trace elements,varies largely from soil to plants and animals.Imbalances in minerals profiles in soil and forages in arid and arid-semi regions are main constraints in achieving the maximum production and reproduction potential in livestock (Khan et al.,2008,2009;McDowell and Arthington,2005).Mineral imbalances in soil and forage are responsible for impaired performance among the ruminants.During the harsh and dry seasons,important components of forage,such as protein,energy,and fiber are often not available to grazing livestock,which in turn reduce the intake of different minerals in animals in these pastureland conditions (McDowell et al.,1983;2003). Feed contains milk,bones are high in both P and Ca2+.Their absorption is dependent on their solubility at the portion of contact with the absorption membranes (Maynard et al.,1979).Hupocalcaemia,milk fever,and hypomagnesaemia,grass staggers,are the common problems of sheep and cattle than the goats (Mowlem,1988).Sheep feed on roughage diets usually excretes little P in their urine (Scot and McLean,1981).Whole blood contains 35-45 mg • 100 mL-1,most of it is in the cells.Plasma P levels are more easily changed by diet than Ca2+levels depending upon the age and the species.Calcium and P requirements are highly dependent on the levels of productivity and physiological state of animals (Anonumous,1984b;1989a).Phosphorus is the 2nd most important plant nutrient.Most of the soils and plants of live stock grazing areas of the world especially in the tropical areas are low in P (McDowell et al.,1983).It involves in most of the important physiological processes like respiration,photosynthesis and energy storage.Milk is the natural product involved having high quantity of P and Ca2+for well grazing animals (McDowell,1992).In herbivores major pathway for excretion are the feces,while urine in the carnivores.Deficiencies of P cause the slow growth,weakening of bones,tetany and low milk production (McDowell,1985).For the determination of mineral imbalances,it is important to locate the areas where these imbalances are present.Conclusive results will help the livestock growers in formulations of mineral nutrients for the better performance,development and financial benefits of animals.It will also help the small livestock holders for more productivity and economic benefits by improving mineral nutrition.

Materials and Methods

Studies were carried out by selecting lactating,nonlactating and male,three animals of each,and healthy goats reared at private goat farm in the vicinity of Faisalabad,Pakistan.The city is situated in the rolling plains of northeast Punjab between longitude 73°74 East,latitude 30°31.5 North,with an elevation of 184 m above the sea level.The soil of Faisalabad comprise alluvial deposits mixed having variable calcareous characteristics,making it very fertile.Due to its high evapo-transpiration,it features an arid climate.The mean maximum and minimum temperature in summer are (39±2.14)℃ and (27±1.07)℃,respectively.In winter,it peaks at around 21℃ and 6℃,respectively.The average yearly rainfall lies only at about 200 mm and is highly seasonal with approximately half of the yearly rainfall in two months,July and August.Studies were carried out by selecting lactating,non-lactating and male,three animals of each,and healthy goats.Four samples were collected after the interval of each fortnight from all the groups of animals.The animals were allowed for free grazing for 4-5 h daily.All the experimental animals remained the same throughout the study period.Soil samples upto 15-20 cm depth were collected with soil samples from 10 different sites of the study area.Soil samples were air dried,ground,thoroughly mixed and sieved through mesh size ＜2 mm for chemical analysis.Forage plants were gathered from the same site from where the soil samples were collected.Washed with 1% HCl followed by four washings with distill water in order to remove the dust and unwanted particles.Samples were oven dried (THELCO,model 6,Germany) at (70±2)℃ and ground to powder stored in brown paper bags (5 cm×8 cm).Blood samples were collected from jugular vein by the disposable syringes.A pinch of anticoagulant (EDTA) was added in it in order to avoid the clotting of blood sample.Samples were centrifuged (HERMLE,Z-233 M-2,abnet,Germany) at 3 000 r • min-1for 20 min to separate the blood plasma.Samples were stored in freezer at –20℃ for further analyses.Morning and evening samples of milk were collected from lactating goats mix to make them uniform sample.One mL of K2Cr2O7(30%) added in them as preservative.Samples were stored at –20℃in freezer.Fortnightly urine samples were collected in a plastic bowl (5 cm×8 cm).Shifted them in nalgene bottles (50 mL) rinsed with 1 N-HCl to remove the dust resides before collection.Samples were stored in freezer at –20℃ (SANYO,Biochemical Freezer,Model MDF-U333,Japan).Water samples from tube well and canal waters were collected from where animals drank and used to irrigate the animal grazing areas.Blood samples were centrifuged (HERMLE,Z-233 M-2,Abnet,Germany) for 20 min at 3 500 r • min-1in order to separate the blood plasma after filtering stored at –20℃.Samples were digested with perchloric acid and nitric acids (2 : 1).After digestion require volumes were made with double distilled water.Milk was digested with mixture of perchloric acid (HClO4) and nitric acid (HNO3) in the ratio of 2 : 1.Samples were filtered through Whatman filter paper # 42 for half an hour final volume made 25 mL with distilled water.Total phosphorus forage plants,fecal matter,blood and milk were determined by the method developed by Jackson (1662).AB-DTPA (Ammonium Bi-carbonate Diethylene Triamine Penta Acetic acid) extractable phosphorus was analyzed by adopting the method developed by Olsen and Sommer (1982).Phosphorus concentrations were determined by spectrophotometer (U-2800 Model No.122-0003,Hitachi,Japan.Total dissolved salts (TDS),salinity (SAL),electrical conductivity (E.C.) and pH of water and soil (1 : 1 ratio of soil and water) were performed by E.C.meter (SENSO Direct Con 200,Germany) and pH meter (HANNA Instruments,pH 210,Romania).The statistical analysis of data was performed by following Steel and Torri (1980).Differences among means were assessed using Duncan's new multiple ranges test (Duncan,1955).

Results

Feces

Fecal phosphorus ranged from (33±0.66) mg • kg-1to(55±0.74) mg • kg-1,whereas in winter it ranged from 13 mg • kg-1to 100 mg • kg-1.Their means were observed as (46±0.12) mg • kg-1and (59±0.56) mg • kg-1for summer and winter,respectively.In male animals,inconsistent patterns were recorded for both seasons.In summer,it ranged from (13±0.55) mg • kg-1to (90±0.54) mg • kg-1with seasonal mean (64±0.45) mg • kg-1and 23 mg • kg-1to 108 mg • kg-1with mean (78±0.78) mg • kg-1(Table 1).Phosphorus levels in all types of animals differed significantly (p＜0.01).Similarly fortnights,and seasons had significantly effects (p＜0.01) on its concentrations in fecal matter (Table 2).

Table1 Concentrations of phosphorus (mean±SD) in feces,urine,milk and blood plasma of lactating,non-lactating and male goats in forage plants,soil,canal and tube well water

Urine

In lactating animals,urine levels ranged from (7.0±0.32) mg • L-1to (11.5±0.47) mg • L-1with mean (9±0.89) mg • L-1and (8.0±0.55) mg • L-1to (10.0±0.25) mg • L-1with mean (9.0±0.12) mg • L-1for summer and winter,respectively.For non-lactating and male animals their means were (8.0±0.23) mg • L-1,and(9.5±0.60) mg • L-1and 7(.75±0.66) mg • L-1and (9.87±0.99) mg • L-1for summer and winter (Table 1).Among three types of animals,its concentrations did not vary significantly (p＞0.01),but fortnights and seasons had significant variations (p＜0.01) (Table 2).For lactating animals,their means were (71.0±0.88)mg • L-1and (65.0±0.44) mg • L-1,for non-lactating animals (67±0.21) mg • L-1and (62±1.025) mg • L-1and for male (71±0.22) mg • L-1and (33±0.36) mg • L-1for summer and winter,respectively.Levels of phosphorus did not vary among all three types of animals and seasons had no significant effects (p＞0.01) on its levels while fortnights had significant effects on its concentrations (p＜0.01) (Table 2).

Blood

Non-significant (p＞0.1) variations were observed among the animals seasons and FNXS while significant (p＜0.1) differences among FN,AXS,AXFN and AXFNXS.It ranged from (33.01±0.36) mg • L-1to(71.0±0.88) mg • L-1for both seasons.Lactating (114±0.36 mg • L-1) and males (114±6.54 mg • L-1) depicted about similar levels at the 4th fortnight in summer.The highest (71±0.88 mg • L-1) concentrations were on the 2nd whereas the lowest (33±0.36 mg • L-1)on the 1st fortnight in lactating animals during summer.

Milk

Phosphorus concentrations in milk varied from (11±0.36) mg • L-1to (40±0.58) mg • L-1with mean (25±0.54) mg • L-1and from (12±1.25) mg • L-1to (40±0.63) mg • L-1with mean for summer and winter,respectively.The highest (40±0.58 mg • L-1) levels were found on the 1st and the 2nd fortnights (40±0.63 mg • L-1) for summer and winter (Table 1).Seasons and fortnights had significant variations (p＜0.01) in the phosphorus concentrations.

Table2 Analysis of variance (ANOVA) of data for P concentration in feces,urine,blood plasma and milk at different fortnights during summer and winter

Forages

In forages it varied from (11±0.36) mg • kg-1to (40±0.58) mg • kg-1with seasonal mean (755±1.98) mg • kg-1and (52±0.63) mg • kg-1to (40±0.63) mg • kg-1with seasonal mean (500±3.65) mg • kg-1for summer and winter,respectively (Table 1).Its concentrations varied significantly (p＜0.01) due to the effects of seasons,fortnights and their interactions (Table 3).Seasonal means for canal in summer were (0.97±8.78) mg • L-1and (0.65±0.21) mg • L-1;and (4.12±0.55) mg • L-1and (3.73±0.54) mg • L-1for winter (Table 1).

Soil

Significant (p＜0.1) variations were recorded during seasons,FXN,AND SXFS.Overall means were(785±4.98) mg • kg-1and (752±9.87) mg • kg-1for summer and winter,respectively.In-consistent patterns were recorded in its concentrations during both seasons.It's lowest (445±8.44 mg • kg-1) levels and the highest (1060±54.2 mg • kg-1) levels were on the 1st and the 4th fortnights in winter.

Water characteristics

SAL in all types of waters in both seasons ranged from (0.37±0.42) g • L-1to (0.60±0.114) g • L-1.TDS in waters ranged from (523±9.14) mg • L-1to (811±3.14) mg • L-1.E.C.and pH varied from (523±6.25) μS • cm-3to (1003.7±4.04) μS • cm-3and 8.13±1.01 to 8.39±1.04,respectively.

Seasons,fortnights and their interactions had significant differences (p＜0.01) on the phosphorus levels except sampling periods (p＞0.01) for canal water (Table 3).

Table3 Analysis of variance (ANOVA) of data for P concentration in soil,forage,tube well water and milk at different fortnights during summer and winter

Discussion

Levels of total phosphorus in soil found lower during the months of summer as compared to winter.Lower levels of forage P might be due to the low levels of P in soils during dry seasons (Pasha et al.,2009).Orden et al.(1999) demonstrated that variations in minerals could be related to fluctuations in environmental changes and P should be in the lowest levels due to dry conditions of summer season.The pH of the most Pakistani soil ranged from 7.5 to 8.5.In present study,P concentrations of soil were much higher than findings of Perveen et al.(2011) who found soil extractable P 1.58 mg • kg-1at Nahaqi and 34.26 mg • kg-1at Urmarbala,in surface soils of vegetable growing areas of Peshawar,Pakistan.Water properties like SAL,TDS,EC and pH were within the ranges of normal water quality (Rashid and Memon,2001).Tube well water had lower and canal water higher levels of P than the values recorded by Khan et al.(2010a) in surveying the water quality of Ravi and Chenab rivers,Pakistan.Higher concentrations of P in canal water might be due to the mixing of effluent of industry because it is flowing nearby the industrial area of Faisalabad.Phosphorus plays vital role in many physiological processes like photosynthesis,respiration,energy storage and cell division.Energy rich compounds which play important role are ADP and ATP.Many variations in P concentrations of forage plants were observed during both the months.

Higher P contents were evaluated during summer than winter.Present values of forages P were lower from the findings of Ahmad et al.(2008) who found 15-28 mg • g-1in leaves and 20-29 mg • g-1in pods collected from Soone valley,Pakistan.Prabowo et al.(2011) evaluated 0.17% and 0.18% P in forage plants in dry and wet seasons,respectively.The differences might be due to the environmental conditions,soil types and genetical differences in forage plants.Forages must be fed to animals after a sudden harvest otherwise reasonable amount may be lost due to environmental factors.Tuen and Dahan (1991) demonstrated in a study at Malaysia that fresh rice straw retained 1.3 g • kg-1P and exposed to weather,for 10 days,retained from 0.6 to 0.7 g • kg-1,he also elaborated that about 50% of P was lost when exposed to sunlight and rainfall.Stored straw had 0.7% P as compared to 0.25% in fresh rice straw.Phosphorus concentration in fecal matter for all groups of animals was lower in summer as compared to winter season.Furthermore,it was concluded that phosphorus excretion through feces was not in association with the above researchers when they introduced rice straw to goats at Malaysia for the assessment of nutritive value of rice straw.It was evaluated that non-lactating animals excreted more P through urine during summer than winter.Similarly they also evaluated 2.3 mg • L-1P excreted through urine in goats fed with rice straw at Malaysia.In all animal groups major portion forage plants was transferred to blood plasma in the months of summer than winter.Variations in blood plasma P might be due to the animal types,efficiency of absorptions and chemical forms of minerals.Present findings of blood plasma were in agreement with the results of McDowell (1992) who assessed 45-60 mg • kg-1P for adult animals and lower than 60-80 mg • kg-1for young animals.He also concluded that when blood plasma P falls to levels of 10-20 mg • kg-1in milking cows,animals were suffering from severe deficiency of P.

Interrelations of minerals among soil and forage and animals could gave the exact profile of blood plasma minerals in developing countries (Mtimuni et al.,1990).Phosphorus levels in plasma were higher in both seasons than the findings of Pasha et al.(2009) who observed 31 mg • L-1and 36 mg • L-1P in adult goats during summer and winter at mix cropping zone of Punjab,Pakistan.Present results of milk P during both seasons were less than values recorded by McDowell (1992) who demonstrated that P＜45 mg • kg-1in cattle and sheep indicated P deficiency.Similarly Borys et al.(2006) evaluated 1.67 g • kg-1of P in sheep's milk and Celik et al.(2001) obtained 16.18%,dry matter,6.6% fats,9.6 solids not fat,4.54% protein,0.76% ash and 960 mg • kg-1P in the domestic buffalo's milk breeded in Erzurum plain,Turkey.Mellado et al.(2006) investigated the relationship between secretion of milk and excretion of fecal matter and evaluated that fecal matter P was the good predictor of milk yield,lactation length and milk composition.He found feces P＞21% produced 21 L milk and ＜21% produced 18 L milk.

Conclusions

It was concluded from present study that fecal matter,forage,milk,tube well and canal water contained lower amounts of P which could not satisfy the phosphorus requirements for ruminants.Phosphorus concentrations in blood plasma were within the critical limits which,to some extent,satisfied its levels in animals.So phosphorus supplementations were required in the areas under experimentation to fulfill their requirements.

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