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

4. MTT : 5 mg in 1 ml water
5. Phenazine metho suiphate (PMS) : 2.5 rng in 0.5 ml water
6. Agar (approx., 2%) :20ml .
Electrophoresis conditions
1. Bridge buffer: 0.2 M phosphate buffer, pH 7.0
Gel buffer : 0.01 M phosphate buffer. pH 7.0
2. I2 Vlcm for 5 b, with cooling.
One percent agarose gel was prepared in 0.01 M phosphate buffer and fvted on a
horizontal electrophoretic system. About 10 p1 of each sample were added into individual
well along with the control sample. Whatman filter paper was used as the wick between the
bridge buffer and electrophoresis was carried out at constant current (I2 V) for 5 hrs. The
isoenzymes were visualised by activity staining as follows. The unfixed gel was rinsed with
cold his buffer (0.1 M, pH 8.3) and incubated for 10 minutes at 37C in dark in a staining
solution containing 200 p1 calcium lactate, 10 mg NAD, 5 mglml m, 2.5 md0.5 ml PMS
in 2% agar made upto 25 ml. The isoenzymes LDH 1-4 migrated anodally, while LDH-5
migrated cathodally. The LDH-X isoenzyme migrates between LDH-3 and LDH 4.
43.2. RESULTS
The effect of B. ma lay^ on testosterone, y -GTP and LDH-X of testes was studied and
the data are presented in Figs. 114 8~115. The testosterone levels were observed to increase
initially (upto 60 days) and decreased significantly w0.05) thereafter. And althrough the
study period. the activity of this hormone was always lower than the control.
y-GTP level increased significantly WO.05) in the infected group during the
parasite development than in control throughout the study period.
The LDH-X mzyme was visualized on agarose gel by activity staining (Platel). The
LDH-X bands in the infected animals were clearly visible on 30th day and faintly visible on
60th day but absenthot visible on 90th and 120th day, whereas. in control the enzyme bands
were clearly visible throughout the study period.
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