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Assessment of therapeutic effect of buffered phosphorus in post parturient haemoglobinuria (PPH) in buffaloes.

Introduction

Post-parturient haemoglobinuria (PPH) is emerging as a potent threat to buffaloes in India and other countries of world (Gahlawat et al., 2007). It is a sporadic disease in high producing dairy cattle characterised by intravascular haemolysis, haemoglobinuria and anemia (Ok et al., 2009). Deficiency of inorganic phosphorus and oxidative stress have been recorded as consistent finding in the disease and it does respond to sodium acid phosphate and ascorbic therapy (Gahlawat et al., 2007 and Bhikane et al., 2011). Therefore, it was considered appropriate to investigate the status of oxidative stress within the erythrocytes of haemoglobinuric buffaloes during therapy with buffered inorganic phosphorus with inosine and sodium pyruvate by measuring erythrocytic MDA, GSH and erythrocytic fragility.

Materials and Methods

The present investigation was carried out on 6 clinical cases of PPH in buffaloes showing signs of coffee coloured urine and straining during defecation. For therapeutic studies, buffaloes suffering from clinical PPH were administered buffered inorganic phosphorus along with insoine and sodium pyruvate injection (Novizaca)@ 100 ml, 50 ml and 25 ml respectively for 3-4days.

Blood samples were collected in a vial with anticoagulant before and after treatment to assess therapeutic effect of drugs by estimating erythrocytic MDA (Ohkawa et al., 1979), GSH (Beutler, 1971) and erythrocytic osmotic fragility (Schalm et al., 1975).

Earlier these animals were given other treatment such as Vitamin B complex, Vitamin E plus Se and Berenil etc. by the practioners for 2-4 days.

Blood samples from six clinically healthy buffaloes parturited within 45 days were also collected once to keep them as healthy control.

Results and Discussion

For therapeutic studies, buffaloes suffering from post parturient heamoglobinuria (PPH) brought for treatment were selected for study.

For assesment of therapeutic efficacy, the injectable preparation consisting of combination of buffered phosphorus along with inosine and sodium pyruvate (Novizaca) was given to animals. The drug combination given to these animals indicated better response as in all the six animals there was stoppage of passing coffee coloured urine. Post therapy, cure rate was 100 percent i.e. all the animals recovered fully after treatment. Bhikane et al. (2011) reported that treatment with buffered phosphorus preparation showed early recovery (2.5 [+ or -] 0.22 days) with minimum concentration and no untoward reaction. Goel et al. (1988) reported that Inosine is exclusively used as oxygen releaser in buffaloes (with low levels of haemoglobin) recovered from post parturient haemoglobinuria, as it reduced the respiratory distress and concluded that buffaloes with very low levels of haemoglobin (3.63 [+ or -] 0.06 g%) could be saved with the use of inosine.

Following parameters were assessed to measure the efficacy of preparation consisting of combination of buffered phosphorus along with inosine and sodium pyruvate:

a) Erythrocytic malondialdehyde

In these animals, MDA level (Table 2) decreased significantly (P<0.05) post treatment. It was probably due to administration of buffered phosphorus along with inosine and sodium pyruvate. Buffered phosphorus restores plasma inorganic phosphorus levels within normal limits, inosine helped in reducing respiratory distress and sodium pyruvate provided energy supplementation (Goel et al., 1988; Gahlawat et al., 2007 and Bhikane et al., 2011). Mata and Bhardwaj (1985) proposed a hypothesis that in PPH of buffaloes, phosphorus deficiency decreases glucose utilization rate and ATP production by erythrocytes leading to decrease in synthesis as well as reduction of level of reduced glutathione which predisposes the erythrocytes to adverse effect of oxidants. The resultant oxidative stress probably lead to lipid peroxidation of red cell membrane with eventual intravascular haemolysis.

b) Erythrocytic reduced glutathione

Post-treatment mean GSH activity (Table 2) increased significantly (P<0.05) in these animals, it might be due to the fact that phosphorus might be acting as antioxidant (Gahlawat et al., 2007) and further reduction in oxidative stress by reduction of respiratory distress by inosine and energy supplementation by sodium pyruvate.

Glucose-6-phosphate dehydrogenase enzyme maintains a continuous supply of reduced nicotinamide adenine dinucleotide phosphate (NADPH) which in turn is essential for regeneration of reduced glutathione from oxidized glutathione (Gahlawat et al., 2007).

c) Erythrocytic osmotic fragility

Mean erythrocytic osmotic fragility (Table 1 and Fig. 1) decreased significantly (P<0.05) post treatement it was probably due to supplementation of phosphorus by buffered phosphorus as low plasma inorganic phosphorus leads to slowing down of Embden Meyerhoff pathway which subsequently leads to building of oxidative stress on erythrocytes leading to precipitation of haemoglobin and Heinz body formation. Thus subnormal concentration of ATP predisposes RBCs to altered structures and function, a loss of normal deformability, increase in fragility and haemolysis resulting in haemoglobinemia and haemoglobinuria (Bhikane et al., 2011).

References

Beutler, E. (1971). Red cell metabolism. In: E. Beutler ed. A Manual of Biochemical Methods. New York, G.S. pp. 146.

Bhikane, A.U., Ghoke, S.S., Masare, P.S., Salunke, V.M. and Syed, A.M. (2011). A new approach in Clinical management of Phosphorus Deficiency Haemoblobinuria (PDH) in Buffaloes. Intas Polivet. 12: 56-58.

Gahlawat, I., Singh, K. and Kumar, R. (2007). Investigations on oxidative stress in post-parturient haemoglobinuria in Buffaloes receiving Sodium Acid Phosphate Therapy. Ital. J. Anim. Sci. 6: 974-977.

Goel, P., Malik, K.S., Dwarkanath, P.K. and Chugh, S.K. (1988). Role of oxygen releasers in post parturient haemoglobinuria of buffaloes. Indian Vet. J. 65: 823-26.

Ohkawa, H., Ohishi, N. and Yagi, K. (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 95: 351-58.

Ok, M., Guzelbektes, H., Sen, I., Coskun.A. and Ozturk, A.S. (2009). Post-parturient haemoglobinuria in three dairy cows. A case report. Bull. Vet. Inst. Pulawy 53: 421-23.

Mata, M. M. and Bhardwaj, R.M. (1985). Possible alterations in erythrocytes metabolism and integrity in post parturient haemoglobinuria. Indian J. Vet. Med. 5: 67-72.

Schalm, O.W., Jain, N.C. and Carrol, E.J. (1975). Materials and methods for the study of blood. In: Veterinary Hematology. Lea and Febiger, Philadelphia, pp. 15-81.

Sushma Yadav (1), V. K. Jain (2), Rakesh Kumar (2) and Sridhar (2)

Department of Veterinary Medicine College of Veterinary Sciences Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS) Hisar--125004 (Haryana)

(1.) Post Graduate Scholar and Corresponding author. E-mail: sysunilrao@gmail.com

(2.) Professor

(a)--Brand of Intas Animal Health, Ahmedabad

Table 1: Effect of Novizac (a) combination
on mean osmotic fragility during treatment
of post-parturient haemoglobinuria (n=6)

Salt Solution       Mean osmotic fragility (%)
concentration (%)

                        Healthy control

0.85                 0.16 (b) [+ or -] 0.02
0.80                0.24 (b) [+ or -]  0.02
0.75                 0.58 (b) [+ or -] 0.08
0.70                 2.33 (b) [+ or -] 0.09
0.65                 5.18 (b) [+ or -] 0.06
0.60                15.24 (c) [+ or -] 0.08
0.55                40.18 (c) [+ or -] 0.04
0.50                60.22 (c) [+ or -] 0.05
0.45                90.09 (c) [+ or -] 0.03
0.40                94.42 (c)  [+ or -] 0.11
0.35                97.13 (b) [+ or -] 0.07
0.30                99.36 (c) [+ or -] 0.06
0.25                  99.61 [+ or -] 0.07
0.20                99.64 (a) [+ or -] 0.09
0.10                  100.00 [+ or -] 0.00
0.00                  100.00 [+ or -] 0.00

Salt Solution       Mean osmotic fragility (%)
concentration (%)

                    Buffered phosphorus combination

                         Pre-treatment            Post-treatment

0.85                1.19 (a) [+ or -] 0.01    0.14 (b) [+ or -] 0.01
0.80                3.33 (a) [+ or -] 0.06    0.32 (b) [+ or -] 0.02
0.75                5.32 (a) [+ or -] 0.07    0.75 (b) [+ or -] 0.01
0.70                13.84 (a) [+ or -] 0.32   3.31 (c) [+ or -] 0.17
0.65                30.35 (a) [+ or -] 0.12   7.14 (c) [+ or -] 0.18
0.60                50.41 (a) [+ or -] 0.02   19.35 (b) [+ or -] 0.09
0.55                85.51 (a) [+ or -] 0.08   45.52 (b) [+ or -] 0.07
0.50                93.39 (a) [+ or -] 0.06   65.78 (b) [+ or -] 0.06
0.45                95.76 (a) [+ or -] 0.06   93.42 (b) [+ or -] 0.06
0.40                97.58 (a) [+ or -] 0.03   95.84 (b) [+ or -] 0.04
0.35                98.44 (a) [+ or -] 0.09   97.09 (b) [+ or -] 0.02
0.30                99.79 (a) [+ or -] 0.03   99.52 (b) [+ or -] 0.03
0.25                  99.79 [+ or -] 0.03       99.69 [+ or -] 0.05
0.20                99.20 (b) [+ or -] 0.05   99.66 (a) [+ or -] 0.13
0.10                 100.00 [+ or -] 0.00      100.00 [+ or -] 0.00
0.00                 100.00 [+ or -] 0.00      100.00 [+ or -] 0.00

Means values bearing different superscripts in a row differ
significantly (p<0.05)

Table 2:  Effect of Novizac on erythrocytic
MDA & GSH during treatment of post-parturient
haemoglobinuria in buffaloes (n=6)

Sr.   Groups                       MDA
No.                           n mol/ml RBC

1     Healthy Control   259.24 (c) [+ or -] 2.83
2     Pre-treatment     528.69 (a) [+ or -] 0.76
PPH   Post-treatment    290.44 (b) [+ or -] 2.05

Sr.   Groups                      GSH
No.                               mg%

1     Healthy Control   62.20 (a) [+ or -] 1.89
2     Pre-treatment     32.40 (c) [+ or -] 0.92
PPH   Post-treatment    53.53 (b) [+ or -] 0.71

Means values bearing different superscripts
in a column differ significantly (p<0.05)
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Article Details
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Title Annotation:Research Article
Author:Yadav, Sushma; Jain, V. K.; Kumar, Rakesh; Sridhar
Publication:Intas Polivet
Article Type:Report
Geographic Code:9INDI
Date:Jul 1, 2014
Words:1514
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