effect of infection of the filarial parasite brugia malayi - Pondicherry ...

To 1.5 ml of carbonate buffer, 1.5 ml of disodium phenyl phosphate, 0.1 ml of
magnesium chlorideand 0.1 ml of 1:20 diluted tissue homogenate was added and incubated
for 15 minutes. The reaction was stopped by adding 1.0 ml of 10 % TCA. The contents were
then centrifuged and the residue was discarded. To the supernatant, 1.0 ml of sodium
carbonate solution and 0.5 ml of Folin's reagent was added and the absorbance was read at
640 nm after 10 minutes. The activity was expressed in terms of pmoles of phenol
liberatedimg protein4 5 minutes.
4.2.1.7 Haemoglobin
Austin (1932).
Reagents
Haemoglobin was estimated by the cyanmethaemoglobin method of Drabkin and
1. Fenicyanide-cyanide reagent: This was prepared by dissolving 200 mg potassium
fenicyanide, 50 mg potassium cyanide and 140 mg potassium dihydrogen phosphate in a
litre of water.
2. Cyanmethaemoglobin standard: Purchased from Span Diagnostics Pvt. Ltd., Surat, India.
This was kept in the dark at 4'C. It had an equivalent hemoglobin concentration of 60 mg%.
Twenty p1 of blood was added to 4.0 ml of the fenicyanide-cyanide reagent. This
was allowed to stand for 15 minutes and read against a reagent blank at 540 nm. The
standards were diluted in fenicyanide-cyanide solution to obtain a range of concentrations
and read in the same manner.
Blood hemoglobin values were expressed as &dl.
4.2.1.8 Total RBC counts
Lewis, 1975).
Total RBC counts was determined according to the standard procedure (Dacie and