Anti-tick monoclonal antibody applied by artificial capillary feeding in Rhipicephalus (Boophilus) microplus females

Abstract

The tick Rhipicephalus microplus is an ectoparasite harmful to livestock, a vector of disease agents that affects meat and milk production. However, resistance to acaricides reflects the need for alternative tick control methods, among which vaccines have gained increasing relevance. In this scenario, monoclonal antibodies can be used to identify and characterize antigens that can be used as vaccine immunogens. Capillary tube artificial feeding of partially engorged R. microplus females with monoclonal antibodies against proteins from the gut of tick were used to test the effects of immunoglobulins in the physiology of the parasite. The results of artificial feeding showed that female ticks over 25mg and under 60 mg in weight performed better in the artificial feeding process, with a 94-168% weight increase after 24h of feeding. Results showed that artificial feeding of ticks proved to be a viable technique to study the effects of antibodies or drugs in the physiology of the parasite. One monoclonal antibody (BrBm2) induced decreased oviposition. Moreover, the antigen recognized by BrBm2 was identified as a 27-kDa protein and immunolabeled on digestive vesicles membranes of digestive cells of partially and fully engorged females.

Artificial feeding of Rhipicephalus microplus female ticks with anti calreticulin serum do not influence tick and Babesia bigemina acquisition

“…The present study was designed in agreement to previous similar ones Gonsioroski et al, 2012) that used similar methodology. Herein, groups of adult female ticks (partially engorged R. microplus) were fed with serum enriched with anti-CRT antibodies supplemented-bovine blood characterizing CRT potential as antigen for the control of cattle tick infestations and infection with B. bigemina.…”

“…Having this in mind, 15-20 L of anti-CRT serum was presented to each tick during capillary feeding assays in an attempt to surpass the low antibodies titers. Semi-engorged females were used in accordance with Gonsioroski et al (2012) and Sonenshine (1991) that demonstrate that females within a weight range accept artificial feeding better. Table 2 summarizes the results acquired relatively to parameters associated with artificial feeding.…”

“…Microhematocrit capillary tubes (75 mm × ∅1.5 mm) were filled with the blood meal and placed over the ticks' mouthparts. Tubes were replaced every 2-3 h, as described previously (Gonsioroski et al, 2012). After feeding, ticks were detached from the double-sided tape and weighed again to determine tick weight increase during feeding.…”

“…With the exception of the Bm86 antigen, directed against R. microplus, already included in a commercial vaccine (Canales et al, 2009;Rodriguez et al, 1994;Willadsen, 2006;Willadsen et al, 1989), identification of efficient antigens is still a major limitation in the development of new vaccines (Parizi et al, 2012) stressing the need for new experimental approaches or the combination of different approaches to develop effective vaccines (de la Fuente and . A low cost alternative for testing new compounds directed to tick inoculation of medicines or antibodies against a specific protein, while not completely abolishing the need of cattle use, would significantly reduce it (Gonsioroski et al, 2012;LewTabor et al, 2014). One approach to increase the possibilities of identifying tick protective antigens is to combine RNAi functional studies with in vitro tick feeding.…”

“…Tick artificial feeding is a broadly used technique for different purposes Antunes et al, 2014;Broadwater et al, 2002;Gonsioroski et al, 2012;Inokuma and Kemp, 1998;Matsuo et al, 2004), besides mimicking the natural process of feeding, allows blood meal manipulation once the polyclonal or monoclonal antibodies can be supplemented in the meal Antunes et al, 2014;Gonsioroski et al, 2012;Lew-Tabor et al, 2014), with their effects being therefore assessed.…”