Global Advanced Research Journal of Medicine and Medical Sciences (GARJMMS) ISSN: 2315-5159
June 2016 Issue Vol. 5(6), pp. 188-193
Copyright © 2016 Global Advanced Research Journals

 

Full Length Research Paper

Evaluation of the hemolymph extraction technique in Rhipicephalus sanguineus sensu lato for detecting Anaplasma platys

Glenda C Velásquez-Serra

Instituto Nacional de Investigación en Salud Pública. Proyecto Prometeo. Quito. Ecuador.

E-mail: glenticks@gmail.com, gvelasquez@inspi.gob.ec

Accepted 06 June, 2016

Abstract

Diagnosis of Anaplasma platys, can be performed in the laboratory by using hemolymph test vector organs or tissues using serological colorants and molecular methods. The purpose of the research was to evaluate the hemolymph extraction technique in the brown tick of the dog (Rhipicephalus sanguineus sl) for the detection of the rickettsia, identify the canine anatomical region of most collection of specimens, the site for greater fluidity of hemolymph, rickettsial infection prevalence in the tick and compare the reference method (Test hemolymph) with indirect immunofluorescence test (IIF). The vector was placed in a petriplate in ventral position; an incision in the coxal suture (intersegmental membrane), between the front and rear coxotrocanteral joints, was performed to visualize the spontaneous release of hemolymph; this was collected in a calibrated capillary glass (10µL). Hemolymph samples were performed with the following procedures: they were stained with rapid staining Hemacolor®. Antigens were prepared to practice test (IIF) and dilutions were performed at the hemolymph in PBS phosphate buffer, by placing them in contact with plates prepared with canine antigens of A. platys.157 ticks in 21 infected dogs were counted. Majority of specimens were collected in the head / neck (36.9%) region. The largest volume of hemolymph was obtained from the intersegmental membrane of the coxa. Inclusion bodies were shown in platelets. The prevalence of infection with A. platys in R. sanguineus, using hemolymph test was 27.39%. IIF technique revealed the presence of antigens of A. platys in the hemolymph of R. sanguineus and macerated. By comparing the Hemolymph Test with IIF, it showed a specificity of 100%. The sensitivity for the detection of A. platys was 37.50%. It was shown for the first time in the country, by extracting hemolymph of R. sanguineus the presence of A. platys using both methods.

Keywords: Estimation, test, hemolymph, Rhipicephalus sanguineus, Anaplasma platys

 

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