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Evaluation of macrominerals concentrations in blood of lactating and dry Desi cows/Evaluacion de concentraciones de macrominerales en sangre de vacas Desi lactantes y secas.


Livestock needs the balanced and suitable levels of all the required nutrients for their animal health and production in any physiological stage (1). Minerals represent essential nutrients having a pronounced function in the life of organisms as their imbalances exert undesirable effects, especially in ruminants. Various microelements and some macro elements have been known to be very essential for livestock growth (2,3). Microelements function as cofactors are involved in many structural molecules in living organisms (2,4).

The elemental composition of body tissue fluids commonly reflects the dietary level of ruminants to a certain extent, depending on the animal tissue fluid and the mineral element in question. Chemical composition of animal tissue fluids can therefore be employed to assess the range of mineral elemental adequacies in ruminants. The blood plasma is an excellent indicator of minerals in animals, and also can be easily obtained from the animal without slaughtering procedures (2). Although some information is attainable on the normal mineral profile of Sahiwal cattle (5), however very little information is available on the mineral status of Desi cows. Therefore, the aim of this investigation is to evaluate the mineral status of lactating and dry cows comparatively. It is expected that the results will be useful for formulating the mineral mixtures to overcome the deficiency and toxicity of different minerals in this livestock species.


Study site. Pakistan is mainly an agricultural country with an arid and semi-arid continental subtropical climate. It is located between 23 and 36o N latitude and 60 and 75[degrees] E longitude.

The present investigation was conducted on Desi breed cows reared in the district of Sargodha, at a rural livestock farm during the winter season of 2013. The district of Sargodha is located between 30[degrees] 05' N longitude and 72[degrees] 67' E latitude and its altitude is 180 m above sea level. Different improved pastures as feeding sites for animals are possessed by the livestock farm. The pastures were fertilized and irrigated with tubewell and canal water. The main forage species available to animals during the winter were Medicago sativa and Trifolium alexandrinum as major forages, and Cichorium intybus, Brassica compestris, and Avena sativa as minor forages. The pasture was the main source of feeding for dairy cattle during the research period.

Animals, sample collection and analysis. Twenty lactating and twenty dry Desi cows, with an average body weight of 315 [+ or -] 37 kg (mean [+ or -] standard deviation) and having 2-5 years old of age, were chosen and considered as experimental groups. About 20 ml of blood from each animal was obtained in a clean sterilized glass test tube having heparin, as an anticoagulant, through jugular vein three times from October to December by one month interval at 11:00 am.

Blood was centrifuged (3.000 rpm x 15-30 min) and separated plasma was stored at -20[degrees]C until analysis. Wet digestion using perchloric acid and nitric acid (6). The plasma minerals including calcium (Ca), magnesium (Mg), copper (Cu), zinc (Zn) and iron (Fe) were determined using atomic absorption spectrophotometer (Perkin Elmer Corp.) according to the method of Akhtar et al (3).

Statistical analysis. Data were subjected to statistical analysis using the SPSS software and one-way analysis of variance (ANOVA) worked out. Statistical significance between the mean values was tested at p<0.05 level (7,8).


Mean plasma concentrations for Ca, Mg, Zn, Cu and Fe in lactating and dry cows are presented in table 1. Plasma Ca concentration was significantly higher in dry compared to lactating cows (7.6 versus 6.8 mg/dl; p<0.001). Plasma Mg level was higher, but not significantly, in lactating compared to dry cows (1.60 versus 1.20 mg/ dl; p>0.05). Significant higher blood plasma Zn concentration was found in the lactating cows compared to the dry cows (0.78 versus 0.60 mg/l; p<0.01). The Cu concentration of plasma was significantly lower in lactating compared to dry cows (0.56 versus 0.76 mg/l; p<0.001). The Fe concentration in blood plasma was significantly higher in lactating cows compared to dry cows (3.8 versus 2.6 mg/l; p<0.01).


Significant differences in plasma Ca concentrations in cows in different physiological stages, similarly to our results, have been previously reported by other Authors (5,9). In the present investigation, the low plasma Ca concentrations in lactating animals might be due to the physiological status of the animals; since blood Ca concentration is under the control of hormones, such as calcitonin and parathormone, acting on the kidneys, intestines and bones, and in dry cows, the blood levels of some vitamins which are under the control of these hormones increasing with the beginning of lactation (10). When ruminants start the lactation period, a reduction in plasma Ca concentration takes place due to high Ca requirements for milk production, and then Ca level starts to increase due to decreasing milk yield (11,12).

In both dry and lactating groups, the plasma Mg concentrations were lower than the suggested value of 2 mg/dl for ruminants as described by Xin et al (13). Low plasma Mg concentration would be expected due to low Mg concentration in the forage and the mineral supplement consumed. The findings of the present investigation are also consistent with those reported by Khan et al (14) who reported similar low Mg concentration in the blood plasma of cattle and buffalo indicating the deficiency of this element. The most plausible reasons for low plasma Mg concentration might be the low absorption capacity of Mg in the gastrointestinal tract of animals (15) and the low Mg concentration in forage and mineral supplement.

Plasma Zn concentrations of cows in both physiological stages were below the normal range of 0.80 mg/L suggested by McDowell (16). However, no animal in both groups exhibited clinical signs of Zn deficiency such as rough skin with lesions. Other elements are involved in Zn metabolism in non ruminants, although in ruminants no such interactions with other elements have been yet reported (16,17). It has also been reported that when Zn is bound to cell wall constituents, its bioavailability might reduce in livestock (18). Therefore, low Zn availability in animals in the present investigation might be due to such factors, hindering its availability. However, results of the current study are in agreement with the findings of other trials (19-21).

The Cu concentration in plasma of lactating cows was lower than the critical concentration of 0.65 mg/L suggested by McDowell (16). However, in dry animals, the higher plasma Cu concentration compared to the critical value suggests that these animals do suffer from Cu deficiency. Similar differences in plasma Cu concentration have already been reported between different physiological groups of ruminants by researchers in Pakistan (5,21), Colombia (22) and Nicaragua (23). These differences may be due to difference in geographical areas because of the blood mineral concentrations in animals have been reported to be differ from area to area (24). Contrary to our findings, Asif et al (5) reported significant lower Cu in plasma of cattle; however, the reason is still unclear.

The Fe plasma concentrations were sufficiently higher than the critical value of 1.10 mg/L suggested by McDowell (16). It has been reported that Fe deficiency is not a relevant problem in ruminant species and it only occurs when there is some loss of blood (15). Plasma Fe levels similar to the values obtained in our investigation have previously been reported for various classes of animals (22,25). Availability of Fe in blood plasma is dependent on many factors; for instance, in some cases, high concentrations of phosphorus and phytate in the diet may reduce its absorption due to formation of insoluble complexes in the intestinal tracts of animals (17). In fact, Fe deficiency is not commonly widespread for grazing ruminants due to high concentrations of Fe in pasture as well as contamination of forages by soil particles and both these factors contribute much to the Fe requirements of animals (12).

The Fe deficiency becomes a problem when livestock are kept on low Fe content forages, and supplementation of this element may reduce weight losses in lactating animals and cause very rapid weight compensation in suckling animals (12). The Fe deficiency has been reported in some regions of the world at a time when ruminants grazed forage plants grow on sandy soils and have heavy insects or parasite infestation, then Fe supplementation has been useful for the suffering animals. The blood levels, as well as the condition of the ruminants have been improved under supplementation regime (12,26). It has also been investigated that Fe supplementation shows no response in older animals consuming ordinary diets. However, Fe deficiency exists and in some cases, its supplementation exhibit marked improvement in the reproduction potential of the ruminants (10). In our investigation, the animals showed no Fe deficiency but it is possible that its supplementation would encourage reproduction capacity of the livestock grazing therein as Fe levels in blood plasma was lower than the suggested level.

The plasma concentrations of elements were not only different in animals with different physiological stages (i.elactating vs. dry cows), but were also different compared to the animals in other areas and zones, indicating the area effect. It is possible that forage plants grown in pastures grazed by animals may have lower mineral element levels, at least for some minerals, due to leaching soil minerals by water and this deficiency is shown in the plasma minerals profile of ruminants reared in these areas. It has already been reported the mineral profiles of plants vary with the soil chemistry and also by the climatic factors such as temperature, precipitation and drainage of the particular region (2,13).

In conclusion the results of this trial indicated that plasma Ca, Mg, Zn, Cu and Fe concentrations are influenced by physiological stage in dairy cows. In fact, the plasma Ca and Cu concentrations were higher whereas plasma Mg, Zn and Fe levels were lower in dry cows compared to lactating cows. Therefore, our results can be considered useful for formulating the mineral mixtures to overcome the deficiency and toxicity of different minerals in cows.


El ganado necesita niveles balanceados y adecuados de todos los nutrientes que se requieren para su salud zoosanitaria y para la produccion en cualquier etapa fisiologica (1). Los minerales representan nutrientes esenciales que tienen una funcion pronunciada en la vida de los organismos, ya que su desequilibrio ejerce efectos no deseados, especificamente en el caso de los rumiantes. Se han identificado varios microelementos y algunos macroelementos que son esenciales para el crecimiento del ganado (2,3). Los microelementos funcionan como cofactores involucrados en varias moleculas estructurales de los organismos vivos (2,4).

La composicion elemental de los fluidos del tejido corporal normalmente refleja en cierta medida los niveles dieteticos de los rumiantes, dependiendo del fluido de tejido animal y del elemento mineral en cuestion. Por lo tanto, la composicion quimica de los fluidos de tejido animal puede ser empleada para evaluar el que tan adecuado es el nivel de minerales elementales en los rumiantes. El plasma sanguineo es un excelente indicador de los minerales presentes en animales y a la vez puede obtenerse facilmente sin necesidad de procedimientos de matanza (2). Aunque es posible acceder a cierta informacion del perfil mineral del ganado Sahiwal (5), hay muy poca informacion disponible acerca del estado de los minerales en las vacas Desi. Por lo tanto, el objetivo de esta investigacion es evaluar comparativamente el estado de los minerales en vacas secas y lactantes. Se espera que los resultados sean utiles para formular las mezclas minerales que permitan superar la deficiencia y la toxicidad de distintos minerales en esta especie de ganado.


Lugar del estudio. Pakistan es un pais principalmente agricola con clima continental subtropical arido y semiarido. Esta ubicado entre las latitudes 23 y 36[degrees] norte y las longitudes 60 y 75[degrees] este.

La presente investigacion se realizo en vacas de raza Desi criadas en el distrito de Sargodha, en una finca ganadera rural durante el invierno de 2013. El distrito de Sargodha esta ubicado entre las latitudes 30[degrees] 05'norte y 72[degrees] 67' este, con una altitud de 180 m sobre el nivel del mar. La finca ganadera posee distintos pastizales mejorados que sirven como lugar de alimentacion para los animales. Los pastizales fueron fertilizados y regados con agua de pozos entubados y de canales. Las principales especies de forraje disponibles para los animales durante el invierno fueron Medicago sativa y Trifolium alexandrinum en mayor medida, y en menor medida Cichorium intybus, Brassica compestris y Avena sativa. Los pastizales fueron la principal fuente de alimentacion para el ganado lechero durante el periodo de investigacion.

Animales, recoleccion de muestras y analisis. Se eligieron veinte vacas Desi lactantes y veinte secas, con un peso corporal promedio de 315 [+ or -] 37 kg (media [+ or -] desviacion estandar) y una edad de 2-5 anos, que fueron consideradas como grupos experimentales. Se obtuvieron 20 ml de sangre de cada animal en un tubo de ensayo de vidrio limpio y esterilizado, con heparina como anticoagulante, extraidos de la yugular tres veces de octubre a diciembre a intervalos de un mes a las 11:00 am.

La sangre se centrifugo (3.000 rpm x 15-30 min) y el plasma separado se almaceno a una temperatura de -20[degrees]C hasta ser analizado. Luego se hizo una digestion humeda utilizando acido perclorico y acido nitrico (6). Se determinaron los materiales del plasma, incluyendo calcio (Ca), magnesio (Mg), cobre (Cu), zinc (Zn) y hierro (Fe), utilizando un espectrometro de absorcion atomica (Perkin Elmer Corp.) segun el metodo de Akhatar et al (3).

Analisis estadistico. Los datos fueron sometidos a un analisis estadistico utilizando software SPSS y se realizo un analisis de varianza en un solo sentido (ANOVA). La prueba de significacion estadistica entre los valores de medios se hizo a un nivel de p<0.05 (7,8).


La concentracion media de Ca, Mg, Zn, Cu y Fe en plasma de vacas lactantes y secas se relaciona en la tabla 1. La concentracion de Ca en plasma fue significativamente mayor en vacas secas en comparacion con vacas lactantes (7.6 frente a 6.8 mg/dl; p<0.001). La concentracion del nivel de Mg en plasma fue mayor, pero no de manera significativa, en vacas lactantes en comparacion con vacas secas (1.60 frente a 1.20 mg/dl; p>0.05). Se encontro una concentracion en plasma de Zn significativamente mayor en las vacas lactantes en comparacion con las vacas secas (0.78 frente a 0.60 mg/l; p<0.01). La concentracion de Cu en plasma fue significativamente menor en vacas lactantes en comparacion con vacas secas (0.56 frente a 0.76 mg/l; p<0.001). La concentracion de Fe en plasma fue significativamente mayor en vacas lactantes en comparacion con vacas secas (3.8 frente a 2.6 mg/l; p<0.01).


Otros Autores (5,9) ya han reportado diferencias significativas en las concentraciones de Ca en plasma en las distintas etapas fisiologicas de las vacas. En la presente investigacion, las bajas concentraciones de Ca en animales lactantes puede deberse al estado fisiologico de los animales; dado que la concentracion de Ca en sangre esta bajo el control de las hormonas, tales como la calcitonina y la paratirina, que actuan sobre los rinones, intestinos y huesos, y en vacas secas, los niveles en sangre de algunas vitaminas que estan controladas aumentan con el inicio de la lactancia (10). Cuando los rumiantes inician el periodo de lactancia, ocurre una reduccion en la concentracion de Ca en plasma debida a los altas exigencias de Ca para producir leche; posteriormente, el nivel de Ca empieza a aumentar conforme disminuye la produccion de leche (11,12).

Tanto en los grupos secos como en los lactantes, las concentraciones de Mg en plasma fueron menores al valor de 2 mg/dl sugerido para rumiantes segun Xin et al (13). Se podria esperar una baja concentracion de Mg en plasma debido a la baja concentracion de Mg en el forraje y en el suplemento mineral consumidos. Los hallazgos de la presente investigacion tambien son consistentes con los hallazgos reportados por Khan et al (14), quien reporto una baja concentracion de Mg en el plasma sanguineo de vacunos y bufalos similar, indicando la deficiencia de este elemento. Las razones mas plausibles para explicar la baja concentracion de Mg en plasma pueden ser la baja capacidad de absorcion de Mg en el tracto gastrointestinal de los animales (15) y la baja concentracion de Mg en el forraje y en el suplemento mineral.

Las concentraciones de Zn en las vacas de ambos estados fisiologicos estuvieron por debajo del rango normal de 0.80 mg/L sugerido por McDowell (16). Sin embargo, en ambos grupos ningun animal presento signos clinicos de deficiencia de Zn tales como una textura de piel aspera con lesiones. En no rumiantes, hay otros elementos involucrados en la metabolizacion de Zn, aunque en rumiantes no se han reportado aun interacciones tales con otros elementos (16,17). Tambien se ha reportado que cuando el Zn esta ligado a los componentes de la pared celular, su biodisponibilidad se puede ver reducida en el ganado (18). Por lo tanto, la baja disponibilidad de Zn en los animales que se observo en la presente investigacion puede ser debida a dichos factores, los cuales dificultan su disponibilidad.

Las concentraciones de Cu en vacas lactantes estuvieron por debajo de la concentracion critica de 0.65 mg/L sugerida por McDowell (16). Sin embargo, en el caso de vacas secas, el mayor nivel en la concentracion de Cu comparado con el valor critico sugiere que estos animales sufren de deficiencia de Cu. Investigadores en Paquistan (5,21), Colombia (22) y Nicaragua ya han report diferencias similares en la concentracion de Cu en plasma entre distintos grupos fisiologicos de rumiantes. Dichas diferencias pueden deberse a las diferencias de las zonas geograficas, ya se ha reportado que las concentraciones de minerales en sangre de los animales difieren de una zona a la otra (24). A diferencia de nuestros hallazgos, Asif et al (5) reporto un nivel de Cu en plasma significativamente menor en el ganado; sin embrago, la razon aun no es clara.

Las concentraciones de Fe en plasma fueron suficientemente mayores al valor critico de 1.10 mg/L sugerido por McDowell (16). Se ha reportado que la deficiencia de Fe no es un problema pertinente en las especies rumiantes y que ocurre unicamente cuando hay una perdida de sangre (15). Para varias clases de animales, niveles de Fe en plasma similares a los valores obtenidos en nuestra investigacion ya han sido reportados (22,25). La disponibilidad de Fe en plasma depende de multiples factores; por ejemplo, en algunos casos las altas concentraciones de fosforo y fitato en la dieta pueden reducir su absorcion debido a la formacion de complejos insolubles en el tracto intestinal de los animales (17). De hecho, la deficiencia de Fe comunmente no se ve extendida en rumiantes de pastoreo debido a las altas concentraciones de Fe en los pastos, como tambien por la contaminacion de particulas de tierra en los forrajes. Ambos factores contribuyen en gran medida a satisfacer los requerimientos de Fe en animales (12).

La deficiencia de Fe se vuelve problematica cuando el ganado se mantiene con forrajes de baja concentracion de Fe, en donde el suplemento de este elemento puede reducir la perdida de peso en animales lactantes y puede causar una muy rapida compensacion en el aumento de peso de los animales amamantados (12). Se ha reportado una deficiencia de Fe en algunas regiones del mundo cuando los rumiantes se alimentan de forraje proveniente de plantas que crecen en suelos arenosos y que estan altamente infestadas con insectos o parasitos, en donde el suplemento de de Fe ha sido util para los animales afectados. Tanto los niveles en sangre como la condicion de los rumiantes han mejorado bajo el regimen de suplemento (12,26). Tambien se ha investigado que el suplemento de Fe no demuestra respuesta alguna en animales mayores que consumen dietas normales. Sin embargo, la deficiencia de Fe existe y en algunos casos la administracion de suplementos ha demostrado una mejoria notoria en el potencial de reproduccion de los rumiantes (10). En nuestra investigacion, los animales no presentaron una deficiencia de Fe, aunque es posible que el aporte de suplementos hubiera podido favorecer la capacidad de reproduccion del ganado que alli pastaba, ya que los niveles de Fe en el plasma sanguineo fueron menores al nivel sugerido.

Las concentraciones en plasma de los elementos fueron distintos no solo en animales que estaban en distintas etapas fisiologicas (es decir, vacas lactantes frente a vacas secas), sino que tambien fueron distintos entre animales de distintas areas y zonas, lo cual indica que existe un efecto de area. Es posible que las plantas de las cuales se compone el forraje que crecen en pastizales en donde pastan los animales tengan contenidos menores de los elementos minerales, al menos en el caso de algunos minerales, debido a la lixiviacion de los minerales en el suelo por accion del agua, deficiencia que se evidencia en el perfil de minerales en plasma de los rumiantes criados en estas zonas. Ya ha sido reportado que los perfiles de minerales de las plantas varian con la quimica del suelo, como tambien por factores climaticos particulares de esta region tales como temperatura, precipitacion y drenaje (2,13).

En conclusion, los resultados de este estudio indicaron que las concentraciones en plasma de Ca, Mg, Zn, Cu y Fe se ven influenciadas por la etapa fisiologica de las vacas. De hecho, las concentraciones en plasma de Ca y Cu fueron mayores, mientras que los niveles de Mg, Zn y Fe fueron menores en vacas secas comparadas con vacas lactantes. Por lo tanto, nuestros resultados pueden considerarse utiles para formular las mezclas de minerales para superar la deficiencia y la toxicidad de los distintos minerales en las vacas.


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Zafar Iqbal Khan, [1] Ph.D, Alireza Bayat, [2] Ph.D, Kafeel Ahmad, [1] Ph.D, Muhammad Sher, [3] Ph.D, Muhammad Khalid Mukhtar, [1] Ph.D, Zafar Hayat, [4] Ph.D, Vincenzo Tufarelli, [5] * Ph.D.

[1] University of Sargodha, Department of Biological Sciences, Pakistan. [2] Animal Production Research, MTT, FI 31600, Jokioinen, Finland. [3] University of Sargodha, Department of Chemistry, Pakistan. [4] University of Sargodha, Department of Animal Sciences, Pakistan. [5] University of Bari 'Aldo Moro', Department of Emergency and Organ Transplantation, Section of Veterinary Science and Animal Production, Bari, Italy. * Correspondence:

Received: October 2014; Accepted: March 2015.
Table 1. Mineral plasma concentrations of lactating and dry
Desi cows.

Mineral     value (1)     Lactating cows          Dry cows

Ca, mg/dl     8.00      6.8 [+ or -] 0.153   7.6 [+ or -] 0.09
Mg, mg/dl     2.00      1.6 [+ or -] 0.04    1.2 [+ or -] 0.03
Zn, mg/L      0.80      0.78 [+ or -] 0.08   0.60 [+ or -] 0.05
Cu, mg/L      0.65      0.56 [+ or -] 0.07   0.76 [+ or -] 0.04
Fe, mg/L      1.10      3.8 [+ or -] 0.03    2.6 [+ or -] 0.10

Mineral     Significance (2)

Ca, mg/dl          ***
Mg, mg/dl          NS
Zn, mg/L           **
Cu, mg/L           ***
Fe, mg/L           **

(1) According to McDowell and Arthington (13) and McDowell (16);

(2) **, p < 0.01;

***, p < 0.001;

NS, not significant ;

(3) mean [+ or -] standard error.
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Author:Khan, Zafar Iqbal; Bayat, Alireza; Ahmad, Kafeel; Sher, Muhammad; Mukhtar, Muhammad Khalid; Hayat, Z
Publication:Revista MVZ (Medicina Veterinaria y Zootecnia)
Date:May 1, 2015
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