CC BY
116
Journal of Stress Physiology & Biochemistry, Vol. 9 No. 1 2013, pp. 28-34 ISSN 1997-0838 Original Text Copyright © 2013 by Joshi, Kataria, Kataria, Pandey, Asopa, Sankhala, Pachaury and Khan
ORIGINAL ARTICLE
Stress related variations in serum vitamin E and C levels of Murrah buffaloes
Joshi A.1, N. Kataria*1, A.K. Kataria2, N. Pandey1, S. Asopa3, L.N. Sankhala4, R. Pachaury5 and S. Khan5
1 Department of Veterinary Physiology, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner - 334 001, Rajasthan, India
2 Apex Centre for Animal Disease Investigation, Monitoring and Surveillance, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner - 334 001, Rajasthan, India
3 Department of Veterinary Pathology, Apollo College of Veterinary Medicine, Jaipur, Rajasthan, India
4 Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner - 334 001, Rajasthan, India
5 Department of Animal Husbandry, Rajasthan, India
Phone 0091 - 151- 2546399 *E-Mail: nalinikataha@rediffmail. com
Received September 10, 2012
The study was designed to determine stress related variations in endogenous vitamin E and C levels of Murrah buffaloes. For this purpose, four hundred and fifty healthy adult female Murrah buffaloes between 4 and 12 years of age were sampled to harvest the sera during adverse ambiences viz. moderate, extreme hot and cold ambiences. Animals were broadly divided into non-pregnant milch, pregnant milch, pregnant dry, primipara and multipara. The mean values (^mol L"1) of serum vitamin E and C were 4.31±0.03 and 23.34±0.33, respectively during moderate ambience. The mean values of both the vitamin E and C depressed significantly (p<0.05) during hot and cold ambiences as compared to moderate ambience. It was observed that decline in each value during hot ambience was greater than that of respective cold ambience. A significant (p<0.05) variation was observed in the mean values of vitamin E and C in each ambience in the animals of all physiological states. The mean values of both the vitamins of non pregnant milch animals were highest (p<0.05) whereas they were lowest (p<0.05) in pregnant dry animals. It could be concluded that extreme ambiences produced oxidative stress in the buffaloes of all physiological states. The depressed levels of endogenous vitamin E and C in the serum showed their depletion in the body probably to combat free radical scavengers. Vitamin E and C should be supplemented to protect the animals from oxidative stress.
Key words: Ambience, vitamin E and C, buffalo, Murrah, serum
ORIGINAL ARTICLE
Stress related variations in serum vitamin E and C levels of Murrah buffaloes
Joshi A.1, N. Kataria*1, A.K. Kataria2, N. Pandey1, S. Asopa3, L.N. Sankhala4, R. Pachaury5 and S. Khan5
1 Department of Veterinary Physiology, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner - 334 001, Rajasthan, India
2 Apex Centre for Animal Disease Investigation, Monitoring and Surveillance, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner - 334 001, Rajasthan, India
3 Department of Veterinary Pathology, Apollo College of Veterinary Medicine, Jaipur, Rajasthan, India
4 Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner - 334 001, Rajasthan, India
5 Department of Animal Husbandry, Rajasthan, India
Phone 0091 - 151- 2546399 *E-Mail: nalinikataria@rediffmail. com
Received September 10, 2012
The study was designed to determine stress related variations in endogenous vitamin E and C levels of Murrah buffaloes. For this purpose, four hundred and fifty healthy adult female Murrah buffaloes between 4 and 12 years of age were sampled to harvest the sera during adverse ambiences viz. moderate, extreme hot and cold ambiences. Animals were broadly divided into non-pregnant milch, pregnant milch, pregnant dry, primipara and multipara. The mean values (^mol L-1) of serum vitamin E and C were 4.31±0.03 and 23.34±0.33, respectively during moderate ambience. The mean values of both the vitamin E and C depressed significantly (p<0.05) during hot and cold ambiences as compared to moderate ambience. It was observed that decline in each value during hot ambience was greater than that of respective cold ambience. A significant (p<0.05) variation was observed in the mean values of vitamin E and C in each ambience in the animals of all physiological states. The mean values of both the vitamins of non pregnant milch animals were highest (p<0.05) whereas they were lowest (p<0.05) in pregnant dry animals. It could be concluded that extreme ambiences produced oxidative stress in the buffaloes of all physiological states. The depressed levels of endogenous vitamin E and C in the serum showed their depletion in the body probably to combat free radical scavengers. Vitamin E and C should be supplemented to protect the animals from oxidative stress.
Key words: Ambience, vitamin E and C, buffalo, Murrah, serum
Numerous rapidly evolving methodologies for evaluating oxidative stress are available to researchers and clinicians, each with their own distinct advantages and disadvantages. Differences
in models and methodologies make it difficult to provide meaningful comparisons, even for studies that seem quite similar superficially. Clarity of understanding of the pathophysiology of oxidative
stress in ruminants will allow the design of specific antioxidant therapies. Heavy milk yielders like buffaloes have higher metabolic rate to derive energy involving oxidation reactions to meet the demand of production. These processes result in the formation of free radicals. To neutralize these free radicals body employs the system of endogenous antioxidants like vitamin E and vitamin
C. This is a part of physiological reactions where both remains in a state of balance. When rate of formation of free radicals overrides the rate of synthesis of endogenous vitamin E and C, problem arises in the form of oxidative stress.
Research in oxidative stress has been implicated in numerous disease processes in animals,
however, compared to human medicine, only a limited number of pathological conditions have been investigated in regard to the effects of oxidative stress in milch animals (Kataria et al., 2010a and Kataria et al., 2012). Studies in milch animals have been sporadic and mainly with mastitis, pneumonia, and retained placenta. More recently, studies have been focused on metabolic diseases that affect dairy animals during the peripartum period (Celi and Pietro, 2011).
Future research should focus on the establishment of a reference panel of biomarker of oxidative stress to be used in ruminant medicine. To help accelerate practical applications,
researchers have proposed the development of an oxidative stress index as an approach in veterinary medicine. Oxidative stress can result from diminished antioxidant protection as well as
increased free radical production. Therefore, investigating antioxidant depletion as a biomarker of oxidative stress may involve assessment of decreases in antioxidant concentrations or increases in their metabolites. However, such
changes may reflect a clinically significant or pathogenic event with the indication that the antioxidant defense system is functioning. Timely detection of endogenous antioxidant levels may be helpful in health management of stressed animal (Chirase et al., 2001). This can help in formulating the supplementation of vitamin E and C to heavy producers and protect the animals from various health disorders.
The Murrah is the most important premier milking breed of buffalo in India. They contribute greatly in enhancing economical status of farmers. The inevitability of exposure of these animals to extreme temperatures of arid and semiarid tracts results in development of oxidative stress and this makes these animals an easy target of a series of health disorders. Despite of immense quality characteristics of these animals very little scientific savoir faire is there about normal and clinical variations in the values of endogenous vitamin E and C. Therefore the present investigation was planned to establish physiological reference values of the endogenous vitamin E and C to be used in the field of veterinary clinical physiology and to find out the possibility of oxidative stress during extreme ambiences in Murrah buffaoes
MATERIALS AND METHODS
The investigation included four hundred and fifty healthy adult female Murrah buffaloes between 4 and 12 years of age to determine antioxidant status during moderate, extreme hot and cold ambient temperature periods. All the animals belonged to private dairy farms of Rajasthan state, India which were managed in similar conditions of feeding and watering. Animals were broadly divided into group A included nonpregnant milch (30); pregnant milch (30) and pregnant dry (30) animals and group B which
included primipara (30) and multipara (30) animals. The mean maximum ambient temperatures during moderate and hot periods were 30.33± 0.20 and 45.5 ± 0.08 °C, respectively, whereas mean minimum temperature during cold ambience was 4.88 ± 0.20 oC. In each ambience 150 blood samples were collected to harvest sera. Sampling was carried out in morning hours during moderate, hot and cold ambiences. Blood was collected directly into the clean, dry test tubes without any anticoagulant to harvest sera.
Vitamin E was determined by the basic spectrophotometric method (Nair and Magar,1955) with modifications ( Kataria et al., 2010b). It was a highly sensitive method based upon the colour reaction between phosphomolybdic acid and vitamin E. Serum vitamin C determination was carried out by a titration of serum ascorbate by 2,6-dichlorophenolindophenol dye (Varley,1988). Calculation was carried out as follows to convert the value in SI units:
100
Serum ascorbic acid (^mol L-1) =---X 2 X 0.008 X56.78
ml titration
200^l of the dye solution is equivalent to 0.008 mg ascorbic acid. 56.78 is the conversion factor for umol L-1 from mg/dl.
To test the significance, the changes in the means were measured by using multiple mean comparison procedures (Duncan, 1955 and Steel and Torrie, 1980). In each case, the moderate mean value served as the control.
RESULTS AND DISCUSSION
In the present study the mean values of serum vitamin E and vitamin C (Table 1) were significantly (p<0.05) lower during hot and cold ambiences as compared to respective moderate mean value. It was observed that decline in each value during hot
ambience was greater than that of respective cold ambience. Heat stress in animals reduces antioxidant activity (Calamari et al.,1999) through increased production of free (Ishikawa and Kanai, 1998) or due to insufficient endogenous synthesis (McDowell, 2000) resulting into a decrease in the antioxidant defense culminating in oxidative damage of biomolecules (Beckman and Ames, 1998).
Hot ambience associated decrease in serum vitamin E showed its depletion in an attempt to reduce the production of reactive oxygen species and to combat increased oxidative threats (Walwadkar et al., 2006). Insufficient endogenous synthesis of vitamin E under stressful conditions could be another cause for depletion (Kataria et al., 2010c). Decreased levels of vitamin C confirm the presence of oxidative stress because repletion is reported after its supplementation (Weiss, 2001). Kataria et al., (2010c) also recommended the use of vitamin E and C in the disease conditions causing oxidative stress.
In group A, serum mean value of each vitamin in non pregnant milch, pregnant milch and pregnant dry animals differed significantly (p<0.05) from each other in all the ambiences and the mean value of each vitamin of non pregnant milch animals was highest whereas it was lowest in pregnant dry animals. Probably in these animals greater stress resulted in depletion of levels of vitamin vitamin E and C. In group B, the mean value of each serum vitamin was significantly (p<0.05) lower in multipara animals than primipara in each ambience. Pregnancy associated oxidative stress in reference to vitamin E has been discussed in water buffaloes (Dimri et al., 2010). Age related variation in antioxidant level could be hypothesised on the basis of relationship of free radicals with age (Sastre
et al., 2000). Vitamin E is also important in the management of stressed animals (Chirase et al., 2001), therefore its use is suggested to combat oxidative stress (Bourdel-Marchasson et al., 2001). Probably in these animals greater stress resulted in depletion of levels of vitamin C. Higher vitamin C level in animals reflected towards its higher synthesis to combat free radicals (Long, 1963). Bhadula et al. (1970) studied the effect of
physiological states on plasma vitamin C level in buffaloes. Nazifi et al. (2009) discussed about the influence of age on free radical generation and consequently, the enzyme antioxidant defense. Based on the results, the assumption can be made that the endogenous antioxidant defense systems were altered to adapt and prevent oxidative stress effects (Kataria et al., 2010d).
Table 1. Mean ± SEM values of serum vitamin E and vitamin C in Murrah buffalo
Effects Serum vitamins
Vitamin E, ^mol L-1 Vitamin C, ^mol L-1
Ambience
Moderate/ Control (150) 4.31±0.03 a 23.34±0.33a
Group A
Non-pregnant milch (30) 4.85±0.02 b 26.92±0.22 b
Pregnant milch (30) 4.30±0.02 b 23.00±0.30b
Pregnant dry(30) 3.80±0.03 b 20.10±0.40 b
Group B
Primipara (30) 4.88±0.02c 25.01±0.30c
Multipara (30) 3.74±0.01c 21.67±0.32c
Hot (150) 2.43±0.02a 16.00±0.21a
Group A
Non-pregnant milch(30) 2.90±0.02 b 18.00±0.30 b
Pregnant milch (30) 2.40±0.01b 16.20±0.40b
Pregnant dry(30) 2.00±0.02 b 13.80±0.20 b
Group B
Primipara (30) 3.45±0.02c 17.50±0.23 c
Multipara (30) 1.41±0.01c 14.50±0.25c
Cold (150) 3.57±0.02a 20.00±0.30a
Group A
Non-pregnant milch (30) 3.67±0.02 b 23.50±0.30 b
Pregnant milch (30) 3.58±0.02b 20.00±0.40b
Pregnant dry(30) 3.46±0.01 b 16.50±0.30 b
Group B
Primipara (30) 3.69±0.01c 22.30±0.30c
Multipara (30) 3.45±0.02c 17.70±0.40c
Figures in the parenthesis indicate number of animals. column differ significantly (p<0.05) from each other.
The findings clearly reflected the presence of oxidative stress during extreme ambiences in buffaloes of all the physiological states.The depressed levels of endogenous vitamin E and C in the serum showed their depletion in the body probably to combat free radical scavengers. It can be recommended that the vitamin E and should be
Means superscribed by same superscripts within a
supplemented during adverse conditions in the animals of all the physiological states to protect them from oxidative stress.
REFRENCES
Beckman, K.B., Ames, B.N. (1998).The free radical theory of ageing matures. Physiol. Rev. 78,
547-581.
Bhadula, S.K., Sharma, H.R., Desai, R.N. (1970). Observations on vitamin C levels in the blood plasma of sahiwal and murrah heifers during oestrus. Zeitschrift fur Tierphysiologie Tierernahrung und Futtermittelkunde. 27, 234-236
Bourdel-Marchasson, I., Christine, M., Beauvieux,
D., Peuchant, E., Richard, H.S., Decamps, A., Reignier B., Paul Emeriau, J., Rainfray, M. (2001). Antioxidant defences and oxidative stress markers in erythrocytes and plasma from normally nourished elderly alzheimer patients. Age Ageing. 30, 235-241.
Calamari, L., Mianti, M.G., Amendola, F., Lombardi, G. (1999). On some aspects of the oxidative status and on vitamin E and C in blood of dairy cows during summer. Proceedings of 13th Associazione Scientifica Produzioni Animali Congress. Piacenza,Italy, 449-451.
Celi, P., Pietro, N. (2011). Biomarkers of oxidative stress in ruminant medicine. Immuopaharma. Immunotoxicol. 33, 233-240.
Chirase, N.K., Greene, L.W., Purdy, C.W., Loan, R.W., Briggs, R.E. McDowell, L.R. (2001). Effect of environmental stressors on ADG, serum retinal and alpha-tocopherol concentrations, and incidence of bovine respiratory disease of feeder steers. J. Anim. Sci. 79, 188.
Dimri, U., Ranjan, R., Sharma, M.C., Varshney, V.P. (2010). Effect of vitamin E and selenium supplementation on oxidative stress indices and cortisol level in blood in water buffaloes during pregnancy and early postpartum period. Trop. Anim. Hlth. Prod. 42, 405-410.
Duncan, D.B. (1955). Multiple range and multiple F tests. Biomet. 11, 1-42
Ishikawa, N., Kanai, Y. (1998). Nitro blue tetrazolium
reduction of neutrophils in heat stressed goats is not influenced by selenium and vitamin E injections. J. Vet. Med. Sci. 60, 12431249.
Kataria, N., Kataria, A.K., Maan, R. (2010a). Correlation of serum IgE with stress in Indian dromedaries affected with skin wounds. J. Stress Physiol. Biochem. 6, 17-24.
Kataria, N., Kataria, A.K., Maan, R. (2010b). Evaluation of oxidative stress due to hot environmental condition in healthy Marwari goats from arid tract in India. Philipp. J. Vet. Anim. Sci. 36 (2), 175-184.
Kataria, N., Kataria, A.K., Maan, R. Gahlot, A.K. (2010c). Evaluation of oxidative stress in brucella infected cows. J. Stress Physiol. Biochem. 6 (2), 19-31.
Kataria, N., Kataria, A.K., Pandey, N., Gupta, P. (2010d). Serum biomarkers of physiological defense against reactive oxygen species during environmental stress in Indian dromedaries. HVM Bioflux. 2, 55-60.
Kataria, N., Kataria, A.K., Joshi, A., Pandey, N. Khan, S. (2012). Serum antioxidant status to assess oxidative stress in brucella Infected buffaloes. J. Stress Physiol. Biochem. 8 (2), 5-9.
Long, C.H., Ullrey, D.E., Miller, E.R., Vincent, B.H. Zutaut, C.L. (1963). Sheep haematology from birth to maturity III serum calcium, phosphorus, magnesium, sodium and potassium. J. Anim. Sci. 24, 145-150.
McDowell , L.R. (2000). Vitamins in animals and human nutrition. Iowa state university press, Ames, I.A.
Nair, P.P., Magar, N.G. (1955). Detemination of vitamin E in blood. J.Biol.chem. 220(1), 157159.
Nazifi, S., Saeb, M., Ghafari, N., Razeghian, I.,
Razavi, S.M., Vosoughi, F., Dehghani, F., Orangi, H. (2009). Reference values of oxidative stress parameters in adult native Iranian goats. Bulg. Vet. Med. 12, 119-124.
Sastre, J., Pallardo, F.V., Garcia-de-la-Asuncion, J., Vina, J. (2000). Mitochondria, oxidative stress and aging. Free Radic. Res. 32, 189.
Steel, R.G.D., Torrie, J.H. (1980). In: Principles and procedures of statistics. A biometrical approach. 2nd ed. McGraw -Hill Kogakusha, Ltd. Tokyo. pp. 187-190.
Varley, H. (1988). In: Practical Clinical Biochemistry. 4th edn. CBS publishers, New Delhi. pp. 349393.
Walwadkar, S.D., Suryakar, A.N., Katkam, R.V., Kumbar, K.M., Ankush R.D. (2006). Oxidative stress and calcium-phosphorus levels in rheumatoid arthritis. Indian J. Clin. Biochem. 21, 134-137.
Weiss, W.P. (2001). Effect of dietary vitamin C on concentrations of ascorbic acid in plasma and milk. J. Dairy Sci. 84, 2302-2307.
