Name: a versatile model of hard tick infestation on laboratory rabbits
Uploaded: 2018-10-06T15:00:00
Description: Watch this Scientific Journal Video about A Versatile Model of Hard Tick Infestation on Laboratory Rabbits at JoVE.com

We acknowledge the technical assistance of Evelyne Le Naour, French National Institute of Agricultural Research (INRA), Alain Bernier (INRA) and Oc&#233;ane Le Bidel (ANSES). Consuelo Almaz&#225;n was supported by a fellowship from the Laboratory of Excellence, Integrative Biology of Emerging Infectious Diseases (LabEx IBEID), Pasteur Institute. The rabbits and sheep were purchased by ANSES. Part of this work was funded by National Institutes of Health grant RO1AI090062 to Y. Park. Dr. Jeffrey L. Blair is acknowledged for reviewing the manuscript.

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 &#38; 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...

<ol>
	<li>Sonenshine, D. E., Roe, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ticks,+People+and+Animals.">Ticks, People and Animals.</a> Biology of Ticks, Vol I. , (2014).</li><li>Kr&#246;ber, T., Guerin, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+feeding+assays+for+hard+ticks.">In vitro feeding assays for hard ticks.</a> Trends in Parasitology. 23 (9), 445-449 (2007).</li><li>Bonnet, S., Jouglin, M., Malandrin, L., Becker, C., Agoulon, A., L'hostis, M., Chauvin, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transstadial+and+transovarial+persistence+of+Babesia+divergens+DNA+in+Ixodes+ricinus.ticks+fed+on+infected+blood+in+a+new+skin-feeding+technique.">Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus.ticks fed on infected blood in a new skin-feeding technique.</a> Parasitology. 134 (2), 197-207 (2007).</li><li>Bonnet, S., Liu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+artificial+infection+of+hard+ticks:+A+tool+for+the+analysis+of+tick-borne+pathogen+transmission.">Laboratory artificial infection of hard ticks: A tool for the analysis of tick-borne pathogen transmission.</a> Acarologia. 52 (4), 453-464 (2012).</li><li>Khols, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tick+rearing+methods+with+special+reference+to+the+Rocky+Mountain+Wood+Tick,+Dermacentor+andersoni+Stiles.">Tick rearing methods with special reference to the Rocky Mountain Wood Tick, Dermacentor andersoni Stiles.</a> Culture methods for invertebrate animals. , (1937).</li><li>Faccini, J. L. H., Chacon, S. C., Labruna, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rabbits+(Oryctolagus+cuniculus)+as+experimental+hosts+for+Amblyomma+dubitatum+Neumann+(Acari:+Ixodidae)+[Coelhos+(Oryctolagus+cuniculus)+como+hospedeiros+experimentais+de+Amblyomma+dubitatum+Neumann+(Acari:+Ixodidae).">Rabbits (Oryctolagus cuniculus) as experimental hosts for Amblyomma dubitatum Neumann (Acari: Ixodidae) [Coelhos (Oryctolagus cuniculus) como hospedeiros experimentais de Amblyomma dubitatum Neumann (Acari: Ixodidae).</a> Arquivo Brasileiro de Medicina Veterin&#225;ria e Zootecnia. 58 (6), 1236-1239 (2006).</li><li>Chacon, S. C., Freitas, L. H. T., Barbieri, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+weight+and+number+of+engorged+Amblyomma+cooperi.+Nuttal+(sic)+and+Warburton,+1908+(Acari:+Ixodidae)+larvae+and+nymphs+and+eggs+from+experimental+infestations+on+domestic+rabbits.">Relationship between weight and number of engorged Amblyomma cooperi. Nuttal (sic) and Warburton, 1908 (Acari: Ixodidae) larvae and nymphs and eggs from experimental infestations on domestic rabbits.</a> Brazilian Journal of Veterinary Parasitology. 13, 6-12 (2004).</li><li>Sonenshine, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+ticks+in+the+laboratory.">Maintenance of ticks in the laboratory.</a> Maintenance of Human, Animal, and Plant Pathogen Vectors. , (1999).</li><li>Levin, M. L., Schumacher, L. B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Manual+for+maintenance+of+multi-host+ixodid+ticks+in+the+laboratory.">Manual for maintenance of multi-host ixodid ticks in the laboratory.</a> Experimental and Applied Acarology. 70 (3), 343-367 (2016).</li><li>Bouchard, K. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+and+experimental+infestation+of+ticks+in+the+laboratory+setting.">Maintenance and experimental infestation of ticks in the laboratory setting.</a> Biology of Disease Vectors. , (2005).</li><li>Jones, L. D., Davies, C. R., Steele, G. M., Nutall, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rearing+and+maintenance+of+ixodid+and+argasid+ticks+in+the+laboratory.">The rearing and maintenance of ixodid and argasid ticks in the laboratory.</a> Animal Technology. 39, 99-106 (1988).</li><li>Slov&#225;k, M., Labuda, M., Marley, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mass+laboratory+rearing+of+Dermacentor+reticulatus+ticks+(Acarina,+Ixodidae).">Mass laboratory rearing of Dermacentor reticulatus ticks (Acarina, Ixodidae).</a> Biologia, Bratislava. 57 (2), 261-266 (2002).</li><li>Rechav, Y., Dauth, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+resistance+in+rabbits+to+immature+stages+of+the+Ixodid+tick+Rhipicephalus+appendiculatus.">Development of resistance in rabbits to immature stages of the Ixodid tick Rhipicephalus appendiculatus.</a> Medical and Veterinary Entomology. 1, 177-183 (1987).</li><li>Zacarias do Amaral, M. A., Azevedo Prata, M. C., Daemon, E., Furlong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+parameters+of+cattle+ticks+fed+on+rabbits.">Biological parameters of cattle ticks fed on rabbits.</a> Brazilian Journal of Veterinary Parasitology. 21 (1), 22-27 (2012).</li></ol>

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...

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 feeding systems (avoiding the use of live animals) have been developed2,3,4 and they should be used whenever possible. However, these systems have not been able to completely replace tick feeding on live animals, and they are not suitable substitutes for many physiologic conditions required for scientific studies. Therefore, in some cases, the use of experimental animal hosts is crucial to guarantee the relevance of experimental results.
Laboratory New Zealand rabbits have shown to be the most suitable and accessible hosts for several ixodid tick species5,6,7,8,9. Two common strategies of tick feeding on rabbits have been frequently used: a) feeding on rabbit ears covered with cotton cloth or socks6,7, and b) feeding in cotton bags9, nylon bottles10 or neoprene chambers11&#160;glued to the rabbit&#39;s back. The feeding on rabbit&#39;s ears is not an elegant system, because ticks (especially early stages, larvae or nymphs) may crawl and attach deep in the ear canal, which is uncomfortable for the animal and makes the monitoring of tick feeding and/or the recovery of engorged ticks difficult. This system is also limited to only two tick groups on ears fully covered by socks protected by Elizabethan collars,representing a significant discomfort for the animal. Other systems9,10,11 are definitely more advanced and well suited for hard tick colony maintenance. However, they are limited in the number of experimental groups placed on the rabbit as well as in the modifiable sizes/shapes of the feeding chambers. In addition, these protocols often require hobbling the rear rabbit legs to avoid scratching and the use of Elizabethan collars to prevent grooming.
Herein we propose a simple, non-laborious and very effective method to feed multiple groups of hard ticks in closed chambers glued to the rabbit back covered by a jacket, eliminating the need for Elizabethan collars or hobbling during the experiment. Specifically, our system uses elastic capsules made from an ethylene-vinyl acetate (EVA) foam sheet covered by mosquito mesh and glued to the shaved rabbit back with fast solidifying (3 min) non-irritating latex glue. This technique allows the attachment of multiple capsules of desired size or shape, and few weeks after the experiment the rabbits are fully recovered. The system is suitable mainly for the nymphal and adult hard tick stages, but with a little modification it can be used for larvae feeding as well. The EVA foam-based methods for hard tick feeding may be adapted to other types of vertebrate hosts, for example sheep (which is shown as one of the alternatives in this paper).

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">New Zealand Rabbits (2.5-3.5 kg)</td>
			<td style="width:197px;">Charles River&#160;</td>
			<td style="width:399px;">Strain Code 571</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Rambouillet sheep</td>
			<td style="width:197px;">Local provider-tick free farm</td>
			<td>Female 6 months old</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">EVA foam 5 mm thick&#160;</td>
			<td style="width:197px;">Cosplay Shop</td>
			<td style="width:399px;">EVA-PE451kg (950mm x 450mm)</td>
			<td>10 mm PE45kg foam from the Cosplay Shop may be used for the large adult tick species</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Foam Sheet 9&#34; X 12&#34; 6 mm-White</td>
			<td style="width:197px;">Amazon</td>
			<td style="width:399px;">FOAMSHT6-20</td>
			<td>6 mm-EVA foam ca be ordered via Amazon as an alternative to the foam from Cosplay Shop</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Full length rabbit jackets&#160;</td>
			<td style="width:197px;">Harvard Apparatus, Inc.&#160;</td>
			<td style="width:399px;">620077- medium, 6270078 - large&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Non-toxic latex glue&#160;</td>
			<td style="width:197px;">Tear mender&#160;</td>
			<td style="width:399px;">Fabric &#38; Leather Adhesive</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Tubular cotton orthopedic stockinette</td>
			<td style="width:197px;">BSN Medical</td>
			<td style="width:399px;">9076 (12-15 cm wide)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Mosquito mesh&#160;</td>
			<td style="width:197px;">Loisirs Creatifs</td>
			<td style="width:399px;">Very fine filter nylon mesh fabric</td>
			<td>Any mosquito mesh, or curtain material with the mesh size less than 50 microns is suitable.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Leukoplast&#160;</td>
			<td style="width:197px;">BSN medical S.A.S</td>
			<td style="width:399px;">LF 72361-02</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:295px;">Adhesive hook-and-loop tape</td>
			<td style="width:197px;">AIEX store</td>
			<td style="width:399px;">AIEX 39.37 Feet/12m Hook and Loop Self Adhesive Tape Roll, 20 mm width, white colour</td>
			<td>Fullfiled by Amazon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fast drying glue &#160;</td>
			<td>Fixtout&#160;</td>
			<td>Superglue</td>
			<td></td>
		</tr>
	</tbody>
</table>

a versatile model of hard tick infestation on laboratory rabbits

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.

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&#39;s back, covered by a jacket (Figure&#160;1 and&#160;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 mo...

Watch this Scientific Journal Video about A Versatile Model of Hard Tick Infestation on Laboratory Rabbits at JoVE.com

A Versatile Model of Hard Tick Infestation on Laboratory Rabbits

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.

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 ...

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