A Versatile Model of Hard Tick Infestation on Laboratory Rabbits

Summary

We have developed a simple and versatile system to feed hard ticks on laboratory rabbits. Our non-laborious protocol uses easily accessible materials and can be adjusted depending on the requirements of the various experimental settings. The method allows comfortable monitoring and/or sampling of ticks during the entire feeding period.

Abstract

The use of live animals in tick research is crucial for a variety of experimental purposes including the maintenance of hard tick colonies in the laboratory. In ticks, all developmental stages (except egg) are hematophagous, and acquiring a blood-meal when attached to their vertebrate hosts is essential for the successful completion of their life cycle. Here we demonstrate a simple method that uses easily openable capsules for feeding of hard ticks on rabbits. The advantages of the proposed method include its simplicity, short duration and most importantly versatile adjustment to the needs of specific experimental requirements. The method makes possible the use of multiple chambers (of various sizes) on the same animal, which permits feeding of multiple stages or different experimental groups while reducing the overall animal requirement. The non-irritating and easily accessible materials used minimizes discomfort to the animals, which can be easily recovered from an experiment and offered for adoption or reused if the ethical protocol allows it.

Introduction

The hard ticks (Ixodidae) are well known as slow feeding arthropods and can be attached on a host for several days, or weeks, depending on the species and developmental stage¹. These obligatory hematophagous arthropods are vectors of a wide variety of infectious agents, such as bacteria, protozoa, and viruses, and thus present a significant risk to humans and animal health¹. When studying tick biology or evaluating new control methods, the establishment of an effective tick feeding system is crucial in order to effectively design the experiments and accomplish the goal(s) of the study. Recently, several artificial tick f....

Protocol

NOTE: In this study, rabbits were maintained in standard cages with food and water offered ad libitum at the French Agency for Food, Environmental and Occupational Health & Safety (ANSES) accredited animal facilities in Maisons-Alfort, France. Animals were monitored twice daily by two experienced technicians for any abnormal skin reactions, health problems or complications. The experimental room was secured by framing the interior of the doorway with double-sided tape to avoid accidental escape of ticks. The method works best if two people work as a team, but it is possible to complete single handedly by one experienced person. Although most rabbits are d....

Results

Here we propose for the first time a detailed step-by step method of hard tick feeding in EVA foam capsules applied to a shaved rabbit's back, covered by a jacket (Figure 1 and Figure 2). This protocol is suitable for various types of experiments when different tick groups on the same host are required and can be also used for mass rearing of hard ticks. The tick feeding success in the laboratory mostly relies on.......

Discussion

The most important step in this entire protocol is to glue the capsule firmly to the shaved skin. For this reason, constant pressure for at least 3 minutes is critical, especially at the corners. When inoculating the ticks into the capsule, it is important to apply them deep into the opposite corner from the open one to avoid tick escape during the sealing. When planning the experiments, make sure that all the capsules are covered by the jacket to avoid damage by chewing or scratching. Make sure that the neck region of t.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
New Zealand Rabbits (2.5-3.5 kg)	Charles River	Strain Code 571
Rambouillet sheep	Local provider-tick free farm	Female 6 months old
EVA foam 5 mm thick	Cosplay Shop	EVA-PE451kg (950mm x 450mm)	10 mm PE45 kg foam from the Cosplay Shop may be used for the large adult tick species
Foam Sheet 9" X 12" 6 mm-White	Amazon	FOAMSHT6-20	6 mm-EVA foam ca be ordered via Amazon as an alternative to the foam from Cosplay Shop
Full length rabbit jackets	Harvard Apparatus, Inc.	620077- medium, 6270078 - large
Non-toxic latex glue	Tear mender	Fabric & Leather Adhesive
Tubular cotton orthopedic stockinette	BSN Medical	9076 (12-15 cm wide)
Mosquito mesh	Loisirs Creatifs	Very fine filter nylon mesh fabric	Any mosquito mesh, or curtain material with the mesh size less than 50 μm is suitable.
Leukoplast	BSN medical S.A.S	LF 72361-02
Adhesive hook-and-loop tape	AIEX store	AIEX 39.37 Feet/12m Hook and Loop Self Adhesive Tape Roll, 20 mm width, white colour	Fullfiled by Amazon
Fast drying glue	Fixtout	Superglue