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SERUM ENZYME ACTIVITIES AND ELECTROLYTE CONCENTRATIONS OF BREEDING DONKEYS (Equus asinus): EFFECT OF SEX, AGE, LACTATION AND/OR PREGNANCY.

Byline: A. S. Qureshi and M. Ali

Abstract

Influence of sex, age in males and lactation and/or pregnancy in females on serum activities of four enzymes and concentrations of six electrolytes in 62 apparently healthy breeding donkeys comprising of 33 males and 29 females were studied. Statistical analysis revealed that out of 10 parameters: a) Sex affected activities of four enzymes namely, ALT, AST, LDH and y-GT significantly: Females showed significantly higher values of these enzymes; b) Age in males altered only two enzymes namely LDH and y-GT while electrolytes remained unaltered. Young foals aged less than 1 year showed highest values among these parameters whereas 3-7 year and above 10 year old donkeys showed the lowest values of LDH and y-GT, respectively; c) Lactation and/or pregnancy in females affected activities of 4 enzymes ALT, ALP, LDH and y-GT and concentrations of two electrolytes i.e., K and Cl.

Non-pregnant dry animals showed significantly (Pless than 0.05) lower activities of four serum enzymes i.e., ALT, ALP, LDH and y-GT than the rest of all physiological states of females under study except AST which remained unaltered. It is concluded that physiological states, performed by corresponding hormones, leading to mild or transitional stresses play the all important regulatory role which has substantial function in the maintenance of stable homeostasis.

Key words: Donkey; serum; enzymes; electrolytes; sex; age; lactation; pregnancy.

INTRODUCTION

Donkey constitutes an invaluable asset for farmers and industry in rural and urban communities in Pakistan. These animals are used for transport of wood, water, hay, tools needed for field work, pull carts are also ridden by their owners. Donkey may carry loads exceeding their body weight for long distances. It is the one of those beasts of burden which is tough enough to survive with such a workload in the harsh environments. The donkeys are immensely tolerant creatures, living intimately with humans and always working, quietly enduring their lot (Powell, 2004). The new possible uses of donkeys could be production of milk for children who are intolerant to cow's milk (Carroccio et al., 2000; Borioni et al., 2012) or in animal-assisted therapy and activity (Muraro et al., 2002; Qureshi and Enbergs, 2012).

Currently, Pakistan ranks third in the world with a population of about 4.9 million heads following China (11 million) and Ethiopia (5.2 million) which has been increased by 2.08 percent per annum (Anonymous, 2013). Despite this large number and multifarious importance of donkey in Pakistan economy, very little is known about the physiology and metabolism of this species as no baseline data is available about the serum biochemical norms of this specie in the natural ecology of Pakistan.

The present study was conducted to establish a baseline data of the serum biochemical values in local environment which may be useful in monitoring the health status of this animal as by improving the life of the donkeys we can also improve the lives of their impecunious owners.

MATERIALS AND METHODS

Donkeys: Sixty two healthy clinically healthy domestic donkeys (Equus asinus) of either sex were put into investigation for biochemical studies among sex, age and different physiological states. Experimental gut was comprised of 33 males and 29 females. Male animals were subjected to age related study while the female animals were investigated for different physiological states. Different groups of animals based on age in males and on physiological states in females are presented in Table 1.

The experimental donkeys belonged to different government breeding farms located in Punjab province of Pakistan. The management and nutritional conditions were intensive and comparable to a large extent. All donkeys were physically examined for rectal temperature, pulse rate and respiration rate prior to the collection of blood.

Laboratory Techniques: Jugular venipuncture method (Kolmer et al., 1959) was adopted for collection of blood using a 22 gauge needle. All samples were collected between 9am -11am before start of feeding. About 10 ml of blood was taken directly into a clean dry test tube without anticoagulant and it was left un-disturbed for clotting of blood for separation of serum. Centrifugation of the tube for 10 minutes at about 3000 rpm facilitated the separation of clear straw colored serum which was stored at -30 to -40C until analyzed.

Screening Panel: Serum activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), gamma-glutamyltransferase (y-GT), chloride (Cl) calcium (Ca), inorganic phosphorous (Pi) were determined by using commercial kits of DiaSys Diagnostic Systems GmbH, Holzheim (Germany) while sodium (Na) and potassium (K) by Centronic-GmbH, Germany. The absorbance of all these parameters were read with Spectronic-21 (Baush and Lomb, USA).

Statistical Analysis: Grand means, group means and their standard errors and ranges were calculated for each parameter separately. In addition, effect of sex was compared by the student't' test, the four age groups among males and four lactation and/or pregnancy states in females were tested by one way analysis of variance. Significantly different group means were compared by Duncan's multiple range Test (Steel et al., 1997). All computations were done with the help of Microsoft excel program.

RESULTS

Grand means, group means and standard error means (SEM) with ranges of different groups of four enzymes and six electrolytes under investigation are depicted in Table 2. Statistical analysis revealed that out of 10 parameters studied: Sex affected serum activities of four enzymes namely: ALT, AST, LDH and y-GT. Females showed significantly (Pless than 0.05) higher values of ALT, AST, LDH and y-GT whereas concentrations of electrolytes remained unaltered in different sexes.

Age made significant effect on serum activities of ALT, LDH and y-GT. Male foals (aged less than 1 year) showed significantly (Pless than 0.05) higher values as compared with older age groups under investigation i.e., 7-10 years and above 10 years.

Lactation and/or Pregnancy in females significantly (Pless than 0.05) affected six parameters: i) Non-pregnant dry animals showed significantly (Pless than 0.05) lower activities of four serum enzymes i.e., ALT, ALP, LDH and y-GT than the rest of all physiological states of females under study except AST which remained unaltered. Statistically higher serum activities of these enzymes were however, recorded among females having different physiological states like, ALT (15.90+1.85 IU/L) in pregnant dry females, ALP (424.07+ 18.29 IU/L) in immature females, LDH (640.96+ 45.51 IU/L) in pregnant dry, and y-GT (64.45+23.25 IU/L) in pregnant lactating females.

Amongst electrolytes, serum Cl was found significantly (Pless than 0.05) higher in heifers (111.88+1.67mmol/L) than the immature (97.17+1.67mmol/L) and non pregnant dry (103.66+1.60 mmol/) females while serum K was found significantly higher in pregnant dry (5.20+ 0.21 mmol/L) than the pregnant lactating (4.10+ 0.40 mmol/L), non pregnant dry (4.14+ 0.23 mmol/L) and non pregnant lactating (3.90+ 0.11 mmol/L) and female.

Table 1. Distribution of animals on the basis of sex, age in males and pregnancy and/or lactation in this study

Males:###Number of Animals

Less than 1 year 4###4

3-6 years###10

7-10 years###12

Above 10 years###7

Females:

Immature females###4

Heifers###7

Pregnant Dry###4

Pregnant Lactating###4

Non Pregnant Dry###8

Non Pregnant Lactating###5

Table 2. Grand means + SE, and ranges of serum activities of enzymes and of electrolytes and mineral levels in domestic donkeys (Equus asinus).

Groups###n###AST###ALT###AP###LDH###GGT###Na###K###Cl###Ca###P

###U/L###mmol/L

a. Grand Means

Mean###62###244.397.4###9.700.58###348.137.72###430.8919.18###39.371.70###143.850.93###4.240.07###107.231.62###11.970.25###3.670.14

SEM

Range###62###110.20-354.30###3.50-22.40###203.70-514.30###205.50-720.10###16.60-87.70###121-161###3.10-5.90###40.20-148.30###6.90-19.70###0.50-6.70

. Effect of sex

Males###32###225.1310.67a###8.440.48a###352.799.31###331.7517.44a###33.761.37a###142.241.34###4.290.09###108.212.84###11.780.44###3.800.19

Females###29###266.308.67b###11.131.06b###342.8312.78###543.7221.54b###45.762.87b###143.961.20###4.190.11###106.121.26###12.180.18###3.500.20

c. Effect of age in males

Less than###4###227.8740.61###7.300.51###379.1514.39###460.8087.23a###43.021.46b###146.751.84###4.500.54###100.072.46###12.320.27###4.100.23

1 year

3-7 years###10###229.2320.23###7.350.83###357.9525.14###280.3120.00b###35.072.77ab###144.103.30###4.360.12###106.132.70###11.790.91###3.920.45

7-10###12###213.5316.14###8.850.99###353.0610.46###325.0626.12ab###31.551.66a###143.831.46###4.250.16###108.727.22###11.890.35###3.740.31

years

Above###7###237.6224.96###9.960.65###329.8816.18###342.9014.87ab###30.393.10a###141.424.08###4.170.18###114.944.08###11.271.65###3.580.39

10 years

d. Effect of lactation and/or pregnancy in females

Immature###4###244.4030.34###7.500.59ab###424.0718.29a###521.8039.29ab###51.403.92a###144.751.75###4.380.18ab###97.171.67a###12.370.32###3.750.54

Heifers###7###258.7015.91###14.722.58a###356.2732.14ab###637.1727.68a###48.433.42a###145.002.75###3.940.21a###111.882.74b###12.890.36###3.670.66

Preg. Dry###4###294.2321.53###15.901.85a###316.1757.16ab###640.9645.51a###52.837.05a###141.672.18###5.200.21b###108.702.19ab###11.460.47###3.600.32

Preg.###4###318.309.50###12.150.35ab###322.6022.80ab###628.4518.35a###64.4523.25a###142.501.50###4.100.40a###107.951.65ab###11.100.60###3.750.15

Lactating

Non preg###8###258.7521.10###5.480.93b###304.7018.49b###417.7934.78b###29.323.05b###144.253.42###4.140.23a###103.661.60a###12.210.32###3.180.31

Dry

Non preg###5###269.006.63###14.800.85a###343.7812.64ab###539.7025.23ab###52.126.17a###143.402.25###3.900.11a###106.862.69ab###11.880.45###3.460.40

Lactating

DISCUSSION

Irrespective of sex, age and physiological status, grand means + SE with ranges (maximum-minimum) of all 62 donkeys (Equus asinus) comprising of 33 males and 29 females recorded in the present study are in general comparable to the previous authors who reported from different parts of the world (Zinkle et al., 1990; French and Patrick 1995, Jordana, et al., 1998; Mori, et al. 2003, Caldin et al., 2005; Aluja et al. 2006; Lemma and Moges, 2009; Simenew et al. 2011). Sow et al., 2012 however, recorded relatively lower values of AST, Ca, iP whereas higher value of ALT. In addition, Simenew et al. 2011 reported an enormously low mean value of calcium. It is assumed that minor differences observed in some parameters may be due to difference in methodologies, laboratory equipments, ecological conditions and management practices of donkeys.

Females showed significantly (Pless than 0.05) higher serum activities of ALT, AST, LDH and y-GT. Serum electrolytes remained unaltered in different sexes. These findings were in line with Caldin et al., (2005) and Lopez et al. (2006). Jordana et al. (1998) recorded no effect of sex on these parameters in Catalonian donkey breeds. Possible explanations are not available in literature, also when other species have been considered.

Young male foals aged less than 1 year showed highest activities of two enzymes LDH and y-GT which gradually decreased with progression in age. This effect might be due to less stable biological membranes which allow more leakage of cellular enzymes into the blood. Four age groups in males under investigation followed a decreasing trend with progressive age in ALP which was however not statistically secured. A similar trend was determined by various workers in donkeys of various origins (Dinev and Khubenov, 1986; Zinkle et al 1990; Jordana et al, 1998 and Caldin et al, 2005). It might be due to decreased bone metabolism in adults and/or due to growth hormone which increases renal phosphate re-absorption (Kaneko, 2008).

To authors' knowledge none of the researcher has so far reported the effect of lactation and/or pregnancy on clinical chemistry of donkeys. Gul et al. (2007) compared merely lactating and non lactating donkeys. The most striking observation in the present data was the lowest enzymatic activities of ALT, AP, LDH and y-GT in non pregnant females when dry, which might be due to the relatively depressed metabolic activity among non pregnant and dry animals. Serum concentrations of two electrolytes i.e., K and Cl were found highest among pregnant dry and heifers, respectively.

It is assumed that physiological states, performed by corresponding hormones, leading to mild or transitional stresses play the all important regulatory role which has substantial function in the maintenance of stable homeostasis.

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Publication:Journal of Animal and Plant Sciences
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Date:Feb 29, 2016
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