Abstract

ISOLATION, PURIFICATION OF SALIVA TOXINS FROM THE INDIAN CATTLE TICK RHIPICEPHALUS (BOOPHILUS) MICROPLUS AND ITS EFFECTS ON VARIOUS HEMATOLOGICAL PARAMETERS IN ALBINO MICE

Nidhi Yadav and Ravi Kant Upadhyay*

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Abstract

In present research study tick saliva toxins were isolated and purified on Sepharose CL 6B 200 gel column at a constant flow rate of 5 mL/minute regular fractions. Eluted fractions were pooled lyophilized and its LD50 was determined in albino mice. The LD50 of the venom protein was Rhipicephalus microplus found 39.6 ± 0.047 mg/kilogram body weight of albino mice. For visualizing hematological effects physiological or sub-lethal dose was administered in laboratory reared albino mice In vivo administration of 40% and 80% of LD50 significantly decrease erythrocyte counts up to 59.86% and 60.6% in comparison to control. Contrary to this WBCs count was found to be increased up to 128% and 128.4% of the control after 10 hours of treatment of the same dose. Hemoglobin level was increased up to 103.64%, 106.44%, 110%, 105.7% and 104.57%, in blood in comparison to control at 2, 4, 6, 8, and 10 hour of treatment of 40% of 24-h LD50 respectively. The maximum increase in the packed cell volume (PCV) was obtained 2.0 times higher than the control at 10 hour of treatment Plasma hemoglobin level was increased 128.57% of the control after 10 hours of treatment of 80% of 24-h LD50 (Table 1 & 2). The level of mean corpuscular hemoglobin (MCH) was increased to 127.5% and 111.50% of the control at 8 hours of treatment with 40% and 80% of 24-h LD50 of Rhipicephalus microplus. All these effects on red blood cells, hemoglobin, MCH, total white blood cells, packed cell volume and plasma hemoglobin were time dependent and dose dependent. From comparative gel filtration elution pattern of known proteins tick saliva toxins obtained were low molecular weight toxic peptides ranging from 12-60 kDa. These show strong cytotoxic action on RBCs, leucocytes, platelets and vascular endothelium.

Keywords: Tick saliva toxins, hemoglobin, RBCs, WBCs, PCV, MCV, MCH and plasma hemoglobin.