Name: a capsule-based model for immature hard tick stages infestation on laboratory mice
Uploaded: 2020-07-09T14:00:00
Description: Watch this Scientific Journal Video about A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice at JoVE.com

We acknowledge the technical assistance of Alain Bernier French National Institute of Agricultural Research (INRAE), and Oc&#233;ane Le Bidel (ANSES). The study was supported by the DIM One Health - R&#233;gion &#206;le-de-France (Acronym of the project: NeuroPaTick). The mice were purchased by ANSES. Dr. Jeffrey L. Blair is acknowledged for reviewing the earlier version of the manuscript.

Please note that this protocol can be only applied when all welfare and safety measures are met in the laboratory. This protocol received permission to use mice for tick feeding by the Ethics Committee for Animal ExperimentsComEth Anses/ENVA/UPEC, Permit Numbers E 94 046 08. For the endpoint, the animals were exposed to CO2 for 9 min in two phases of 4 and 5 min each one.
1. Preparation of the capsule
<ol>
	<li>Stick 2 mm thick EVA-foam and the adhesive double sticky foam together...

<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=.">.</a> Biology of Ticks. , (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., 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=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>Kohls, G. M., Galtsoff, P. S., Lutz, F. E., Welch, P. S., Needham, J. G. <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.">Tick rearing methods with special reference to the Rocky Mountain wood tick, Dermacentor andersoni.</a> Culture methods for invertebrate animals. , 246-256 (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).">Rabbits (Oryctolagus cuniculus) as experimental hosts for 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+rabbit.">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 rabbit.</a> Brazilian Journal of Veterinary Parasitology. 13, 6-12 (2004).</li><li>Sonenshine, D. E., Maramorsch, K., Mahmood, F. <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. , 57-82 (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>Almaz&#225;n, C., 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=Identification+of+protective+antigens+for+the+control+of+Ixodes+scapularis+infestations+using+cDNA+expression+library+immunization.">Identification of protective antigens for the control of Ixodes scapularis infestations using cDNA expression library immunization.</a> Vaccine. 21 (13-14), 1492-1501 (2003).</li><li>Banks, C. W., Oliver, J. H., Hopla, C. E., Dotson, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+life+cycle+of+Ixodes+woodi.+(Acari:Ixodidae).">Laboratory life cycle of Ixodes woodi. (Acari:Ixodidae).</a> Journal of Medical Entomology. 35, 177-179 (1998).</li><li>Almaz&#225;n, C., 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=Characterization+of+three+Ixodes+scapularis+cDNAs+protective+against+tick+infestations.">Characterization of three Ixodes scapularis cDNAs protective against tick infestations.</a> Vaccine. 23 (35), 4403-4416 (2005).</li><li>Levin, M. L., Ross, 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=Acquisition+of+different+isolates+of+Anaplasma+phagocytophilum+by+Ixodes+scapularis+from+a+model+animal.">Acquisition of different isolates of Anaplasma phagocytophilum by Ixodes scapularis from a model animal.</a> Vector Borne Zoonotic Diseases. 4 (1), 53-59 (2004).</li><li>Heinze, D. M., Wikel, S. K., Thangamani, S., Alarcon-Chaidez, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcriptional+profiling+of+the+murine+cutaneous+response+during+initial+and+subsequent+infestations+with+Ixodes+scapularis+nymphs.">Transcriptional profiling of the murine cutaneous response during initial and subsequent infestations with Ixodes scapularis nymphs.</a> Parasites &amp; Vectors. 6 (5), 26 (2012).</li><li>Nuss, A. B., Mathew, M. G., Gulia-Nuss, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearing,+Ixodes+scapularis,+the+Black-legged+Tick:+Feeding+Immature+Stages+on+Mice.">Rearing, Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice.</a> Journal of Visualized Experiments. (123), e55286 (2017).</li><li>Wada, T., 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=Selective+ablation+of+basophils+in+mice+reveals+their+nonredundant+role+in+acquired+immunity+against+ticks.">Selective ablation of basophils in mice reveals their nonredundant role in acquired immunity against ticks.</a> Journal of Clinical Investigation. 120 (8), 2867-2875 (2010).</li><li>Saito, T. B., Walker, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Tick+Vector+Transmission+Model+of+Monocytotropic+Ehrlichiosis.">A Tick Vector Transmission Model of Monocytotropic Ehrlichiosis.</a> The Journal of Infectious Diseases. 212 (6), 968-977 (2015).</li><li>Boppana, V. D., Thangamani, S., Alarcon-Chaidez, F. J., Adler, A. J., Wikel, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+feeding+by+the+Rocky+Mountain+spotted+fever+vector,+Dermacentor+andersoni,+induces+interleukin-4+expression+by+cognate+antigen+responding+CD4++T+cells.">Blood feeding by the Rocky Mountain spotted fever vector, Dermacentor andersoni, induces interleukin-4 expression by cognate antigen responding CD4+ T cells.</a> Parasites &amp; Vectors. 2 (1), 47 (2009).</li><li>Gargili, A., Thangamani, S., Bente, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+laboratory+animal+hosts+on+the+life+cycle+of+Hyalomma+marginatum+and+implications+for+an+in+vivo+transmission+model+for+Crimean-Congo+hemorrhagic+fever+virus.">Influence of laboratory animal hosts on the life cycle of Hyalomma marginatum and implications for an in vivo transmission model for Crimean-Congo hemorrhagic fever virus.</a> Frontiers in Cell and Infection Microbiology. 20 (3), 39 (2013).</li><li>Almaz&#225;n, C., 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=A+Versatile+Model+of+Hard+Tick+Infestation+on+Laboratory+Rabbits.">A Versatile Model of Hard Tick Infestation on Laboratory Rabbits.</a> Journal of Visualized Experiments. (140), e57994 (2018).</li><li>Zhijun, Y., 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=The+life+cycle+and+biological+characteristics+of+Dermacentor+silvarum+Olenev+(Acari:+Ixodidae)+under+field+conditions.">The life cycle and biological characteristics of Dermacentor silvarum Olenev (Acari: Ixodidae) under field conditions.</a> Veterinary Parasitology. 168 (3-4), 323-328 (2010).</li><li>Ahmed, B. M., Taha, K. M., El Hussein, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+cycle+of+Hyalomma+anatolicum+Koch,+1844+(Acari:+Ixodidae)+fed+on+rabbits,+sheep+and+goats.">Life cycle of Hyalomma anatolicum Koch, 1844 (Acari: Ixodidae) fed on rabbits, sheep and goats.</a> Veterinary Parasitology. 177 (3-4), 353-358 (2011).</li><li>&#352;irok&#253;, P., Erhart, J., Petr&#382;elkov&#225;, K. J., Kamler, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+cycle+of+tortoise+tick+Hyalomma+aegyptium+under+laboratory+conditions.">Life cycle of tortoise tick Hyalomma aegyptium under laboratory conditions.</a> Experimental and Applied Acarology. 54, 277-284 (2011).</li><li>Chen, X., 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=Life+cycle+of+Haemaphysalis+doenitzi+(Acari:+Ixodidae)+under+laboratory+conditions+and+its+phylogeny+based+on+mitochondrial+16S+rDNA.">Life cycle of Haemaphysalis doenitzi (Acari: Ixodidae) under laboratory conditions and its phylogeny based on mitochondrial 16S rDNA.</a> Experimental and Applied Acarology. 56, 143-150 (2012).</li><li>Jin, S. W., 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=Life+Cycle+of+Dermacentor+everestianus+Hirst,+1926+(Acari:+Ixodidae)+under+Laboratory+Conditions.">Life Cycle of Dermacentor everestianus Hirst, 1926 (Acari: Ixodidae) under Laboratory Conditions.</a> Korean Journal of Parasitology. 55 (2), 193-196 (2017).</li><li>Labruna, M. B., Fugisaki, E. Y., Pinter, A., Duarte, J. M., Szab&#243;, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+cycle+and+host+specificity+of+Amblyomma+triste+(Acari:+Ixodidae)+under+laboratory+conditions.">Life cycle and host specificity of Amblyomma triste (Acari: Ixodidae) under laboratory conditions.</a> Experimental and Applied Acarology. 30 (4), 305-316 (2003).</li><li>Breuner, N. E., 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=Failure+of+the+Asian+longhorned+tick,+Haemaphysalis+longicornis,+to+serve+as+an+experimental+vector+of+the+Lyme+disease+spirochete,+Borrelia+burgdorferi+sensu+stricto.">Failure of the Asian longhorned tick, Haemaphysalis longicornis, to serve as an experimental vector of the Lyme disease spirochete, Borrelia burgdorferi sensu stricto.</a> Ticks Tick Borne Diseases. 11 (1), 101311 (2020).</li></ol>

The most critical step in the protocol is firm gluing of the capsule to the mouse skin. Therefore, the latex glue should be homogenously applied to the entire EVA-foam surface of the capsule and constant pressure for 3 minutes should be applied, especially to the left and right side of the capsule. We also recommend placement of the capsule as far forward on the back as possible to avoid its removal by the mouse using its rear paws. In our experiments, only the adhesion of the EVA-foam and latex glue to the mouse skin ha...

Ticks are important vectors of several pathogens and represent a serious risk to animal and human health1. Setting up an effective feeding system is crucial when studying their biology, tick-host-pathogen interactions, or establishing effective control measures. Currently, several artificial feeding systems, which avoid the use of live animals are available for ticks2,3,4 and these should be utilized whenever experimental conditions allow. However, in various experimental settings these systems fail to appropriately mimic the specific physiologic features and the use of live animals is necessary to achieve relevant results.
Laboratory mice are commonly used for the study of many biological systems and are routinely utilized as hosts for feeding ticks5,6,7,8,9. The two most common methods of feeding immature ticks on mice include free infestations and the use of confinement chambers attached to the mouse. Free infestations are primarily used for larval stages and engorged ticks can drop to an area where they can be recovered. Confinement chambers are usually composed of acrylic or polypropylene caps which are glued to the mouse&#8217;s back. The first technique is an effective natural system for tick feeding but does not allow close monitoring during the experiment because the individual ticks are dispersed in different parts of the host body. Additionally, engorged ticks that drop to a recovery area can become contaminated with feces and urine10,11,12,13,14 that may severely affect the tick fitness or they can be damaged or eaten by the mouse if there is no separation between the animal and the recovery area15. Chamber-based systems allow the confinement of ticks to a defined area, however, the gluing process is laborious and the caps are often weakly adherent to the glue and thus they often detach during the experiment16,17,18,19. The caps are also stiff, uncomfortable, and lead to skin reactions, which prevent the re-use of the mice and necessitates their euthanasia after the experiment.
In our previous study, we successfully developed an effective system using chambers made of ethylene-vinyl acetate (EVA) foam for feeding ticks on laboratory rabbits20. Herein, we adapted this system to a mouse model and propose a simple and clean method to feed immature hard tick stages in closed capsules made from EVA-foam. Specifically, our system uses elastic EVA-foam capsules glued to the shaved mice back with fast drying (3 min), non-irritating latex glue. This technique allows firm and long-lasting attachment of capsules to the experimental mouse, as well as effective tick infestation/collection during the entire course of the experiment. The flat capsule is made from flexible materials and does not impede manipulation of the mouse for blood collection or other purposes. The system is suitable mainly for the nymphal tick stages, but with slight modification it can be used for feeding larvae as well. The method can be completed by one single experienced person and extensive training is not required.

<table>
	<tbody>
		<tr>
			<td>EVA-foam 2 mm thick, (low density)</td>
			<td>Cosplay Shop</td>
			<td>EVA-45kg (950/450/2 mm)</td>
			<td>It can be ordered also via Amazon (ref. no. B07BLMJDXD)</td>
		</tr>
		<tr>
			<td>Heat Shrink Tubing Electric Wire Wrap Sleeve</td>
			<td>Amazon</td>
			<td>B014GMT1AM</td>
			<td>Different diameters of Heat Shrink Tubing are available via Amazon.</td>
		</tr>
		<tr>
			<td>Mice BALB/cByJ</td>
			<td>Charles River</td>
			<td>Strain code 627</td>
			<td></td>
		</tr>
		<tr>
			<td>Mice C57BL/6</td>
			<td>Charles River</td>
			<td>Strain code 664</td>
			<td></td>
		</tr>
		<tr>
			<td>No-toxic Latex Glue</td>
			<td>Tear mender</td>
			<td>Fabric &#38; Leather Adhesive</td>
			<td>Also available also via Amazon (ref. no. B001RQCTUU)</td>
		</tr>
		<tr>
			<td>Punch Tool Hand Art Tool</td>
			<td>Amazon</td>
			<td>B07QPWNGBF</td>
			<td>Saled by amazon as Leather Working Tools 1-25mm Round Steel Leather Craft Cutter Working for Belt Strap</td>
		</tr>
		<tr>
			<td>PVC Binding Covers Transparent</td>
			<td>Amazon</td>
			<td>B078BNLSNP</td>
			<td>Any transparent PVC sheet of ticknes between 0.150 mm to 0.180 mm is suitable</td>
		</tr>
		<tr>
			<td>Self Adhesive Pad Sponge Double Coated Foam Tape</td>
			<td>Amazon</td>
			<td>B07RHDZ35J</td>
			<td>Saled by amazon as 2 Rolls Double Sided Foam Tape, Super Strong White Mounting Tape Foam</td>
		</tr>
		<tr>
			<td>Transparent seal stickers (20 mm diameter circles)</td>
			<td>Amazon</td>
			<td>B01DAA6X66</td>
			<td></td>
		</tr>
	</tbody>
</table>

a capsule-based model for immature hard tick stages infestation on laboratory mice

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of mouse models in tick research is critical for understanding their biology and tick-host-pathogen interactions. Here we demonstrate a non-laborious technique for the feeding of immature stages of hard ticks on laboratory mice. The benefit of the method is its simplicity, short duration, and the ability to monitor or collect ticks at different time points of an experiment. In addition, the technique allows attachment of two individual capsules on the same mouse, which is beneficial for a variety of experiments where two different groups of ticks are required to feed on the same animal. The non-irritating and flexible capsule is made from easily accessible materials and minimizes the discomfort of the experimental animals. Furthermore, euthanasia is not necessary, mice recover completely after the experiment and are available for re-use.

We propose the detailed step-by step method for feeding immature hard tick stages in EVA-foam capsules applied to a mouse&#8217;s back (Figure 2). This non-laborious protocol is suitable for various types of experiments when precise tick monitoring and collection is required. The main advantages of this method are its simplicity, easily accessible cost-effective materials, and short duration. In addition, we succeeded in attaching two capsules to one mouse individual (Fi...

Watch this Scientific Journal Video about A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice at JoVE.com

A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice

In this study, a feeding system for nymphal and larval stages of hard tick was developed using a capsule attached to laboratory mouse. The feeding capsule is made from flexible materials and remains firmly attached to the mouse for at least one week and allows comfortable monitoring of tick feeding.

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of ...

Our technique is suitable for various types of experiments but our mouse model is required for feeding ticks and is necessary to keep the ticks in enclosed area for easy collection and monitoring. The benefit of this simple method is it efficiency, short duration, and the ability to monitor or collect ticks at different time points of an experiment. Our system is suitable for studying tick biology, host-vector pathogen interactions or evaluating different control measure of these medically important arthropods.This method requires accuracy and attention to detail as immature tick stages are small and feeding them on mice necessitates the enclosed capsule to be be firmly attached to the mouse for several days. Begin by sticking together a two millimeter EVA foam, an adhesive double sticky foam. Use a 20 millimeter diameter leather hole punch to cut a circle from the foam pieces.Then, use a 12 millimeter hole punch to cut the interior. Peel the protective paper strip from the adhesive double sticky foam and attach a 20 millimeter transparent plastic circle to the foam. If feeding larvae, do not remove the protective paper strip from the adhesive foam and proceed directly with capsule attaching.Make a one centimeter slit in the transparent plastic then use an entomological pin to create at least 10 small holes for moisture evaporation during the experiment. After the mouse has been anesthetized place it on the manipulation pad and attach a nose cone for continuous isoflurane supply. Ensure that the breathing rate is less than 80 breaths per minute by adjusting the isoflurane level.Shave the anterior part of the mouse from behind the shoulder blades up to the area just behind the ears creating a shaved area that is greater than the capsule surface. Apply non irritating latex glue to the entire EVA foam site of the capsule and wait for one minute. Place the capsule on the mouse's back and apply constant pressure for three minutes especially on the left and right side of the capsule.Slightly lift the capsule to visually check its attachment to the skin. To ensure the firm attachment of the capsule apply a slight three minute constant pressure with the fingers especially on the left and right side of the capsule. If non-attached regions are found, apply more glue with a spatula and press for another three minutes.If a non-attachment region forms during the experiment fix it in the same way. Introduce individual nymphs into the capsule via the one centimeter slit. Slightly squeeze the capsule from two sides to allow the transparent plastic to bend then push the nymph inside the capsule.Turn the forceps 90 degrees and pull them out. For larvae infestation, remove the paper slip from the attached capsule and place the syringe with the larvae directly inside. Deposit the ticks by pushing the syringe plunger then gently turn the plunger towards the skin to remove the remaining attached larvae.Once the larvae are deposited onto the skin close the capsule by attaching the transparent plastic circle. Cut a 3.5 millimeter strip of the protective plastic band and apply the protective plastic band around the capsule then return the mouse to the cage. To collect the ticks, create a cross shaped cut in the plastic with a scalpel and use forceps to collect the ticks.If necessary, reclose the capsule by sticking an adhesive plastic patch on the transparent plastic. the same plastic patch can be used if collection of ticks at multiple time points is required. Alternatively, the mouse can be euthanized and the ticks collected or manually detached after capsule removal.For successful recovery of the mice, keep them in a cage for one additional week and let the capsules detach naturally. Once the capsule is off, check for abnormal reactions on the skin. Apply any emollient lotion if there is any irritation.This protocol makes it possible to feed immature hard ticks in EVA foam capsules applied to a mouse's back. The use of the fast drying and non irritating latex glue ensures that the capsule is firmly attached within three minutes and remains attached for at least seven days. Attaching two capsules makes it possible to feed two different groups of ticks on the same animal.Furthermore, complete recovery of the mice after the experiments, makes it possible to reuse the animals and avoid euthanasia. This system has been successfully used to feed Ixodes ricinus nymphs. A moderate to high engorgement success rate was achieved in two different mouse strains.In both strains, all nymphs finished the feeding within four to five days while the majority dropped off on the fourth day. When attempting this protocol, it's important to shave a sufficient area of mouse's skin and effectively attach the capsule to avoid it's accidental detachment during the experiment. Besides the study of basic tick biology, this method can be used to explore pathogen transmission from tick to host or host to ticks.Additionally, the option to reuse the mouse offers a possibility for multiple re-infestations that can be beneficial for immunological studies or investigating different tick-borne pathogen transmissions.

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Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human ...

Isolation of microRNAs from Tick Ex Vivo Salivary Gland Cultures and Extracellular Vesicles

Isolation of microRNAs from Tick Ex Vivo Salivary Gland Cultures and Extracellular Vesicles

Ticks are important ectoparasites that can vector multiple pathogens. The salivary glands of ticks are essential for feeding as their saliva contains many ...

A Simple and Inexpensive Running Wheel Model for Progressive Resistance Training in Mice

Previously developed rodent resistance-based exercise models, including synergistic ablation, electrical stimulation, weighted-ladder climbing, and most ...

Embryo Injection Technique for Gene Editing in the Black-Legged Tick, Ixodes scapularis

Embryo Injection Technique for Gene Editing in the Black-Legged Tick, Ixodes scapularis

Ticks can transmit various viral, bacterial, and protozoan pathogens and are therefore considered vectors of medical and veterinary importance. Despite ...