Name: cutaneous leishmaniasis in the dorsal skin of hamsters: a useful model for the screening of antileishmanial drugs
Uploaded: 2012-04-21T14:00:00
Description: Watch this Scientific Journal Video about Cutaneous Leishmaniasis in the Dorsal Skin of Hamsters: a Useful Model for the Screening of Antileishmanial Drugs at JoVE.com

This study was supported by grants from The Committee for Research at the University of Antioquia (CODI), The Colombian Institute for Development of Science and Technology &#x2013; COLCIENCIAS and The Center for Development of Products against Tropical Diseases - CIDEPRO.

1. Infection of Hamsters
1. Animals
Inbred female and male golden hamsters (Mesocricetus auratus), 6-8 weeks, weighing 140-160 g are used. They are housed at the animal facility, in temperature-controlled accommodation, fed with standard rodent dried food and provided with water ad libitum. All the procedures involving animals are approved by the institutional Ethical Committee for Experimental Animal Use. Before experimental infection with dermotropic ...

<ol>
	<li>WHO. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+the+leishmaniasis.">Control of the leishmaniasis.</a> Report of a Meeting of the WHO Expert Committee on the Control of Leishmaniasis WHO. 949, (2010).</li><li>Titus, R. G., Marchand, M., Boon, T., Louis, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+limiting+dilution+assay+for+quantifying+Leishmania+major+in+tissues+of+infected+mice.">A limiting dilution assay for quantifying Leishmania major in tissues of infected mice.</a> Parasite Immunol. 7, 545-555 (1985).</li><li>Reynolds, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Kodak+Ektachem+dry+layer+technology+for+clinical+chemistry.">The Kodak Ektachem dry layer technology for clinical chemistry.</a> Upsala. J. Med. Sci. 91, 143-146 (1986).</li><li>Handman, E., Ceredig, R., Mitchell, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+cutaneous+leishmaniasis:+disease+patterns+in+intact+and+nude+mice+of+various+genotypes+and+examination+of+some+differences+between+normal+and+infected+macrophages.">Murine cutaneous leishmaniasis: disease patterns in intact and nude mice of various genotypes and examination of some differences between normal and infected macrophages.</a> Australian J. Exp. Biol. Med. Sci. 57, 9-29 (1979).</li><li>Hanson, W., Chapman, W., Waits, V., Lovelace, 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+Leishmania+(Viannia)+panamensis+lesions+and+relationship+of+numbers+of+amastigotes+to+lesion+area+antimony-treated+and+untreated+hamsters.">Development of Leishmania (Viannia) panamensis lesions and relationship of numbers of amastigotes to lesion area antimony-treated and untreated hamsters.</a> J. Parasitol. 77, 780-783 (1991).</li><li>Henao, H. H., Osorio, Y., Saravia, N. G., G&#243;mez, A., Travi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+and+toxicity+of+pentavalentantimonials+(Glucantime+and+Pentostam)+in+an+American+cutaneous+leishmaniasis+animal+model:+luminometry+application.">Efficacy and toxicity of pentavalentantimonials (Glucantime and Pentostam) in an American cutaneous leishmaniasis animal model: luminometry application.</a> Biom&#233;dica. 24, 393-402 (2004).</li><li>Travi, B. L., Martinez, J. E., Zea, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimonial+treatment+of+hamsters+infected+with+Leishmania+(Viannia)+panamensis:+assessment+of+parasitological+cure+with+different+therapeutic+schedules.">Antimonial treatment of hamsters infected with Leishmania (Viannia) panamensis: assessment of parasitological cure with different therapeutic schedules.</a> Trans. R. Soc. Trop. Med. Hyg. 87, 567-569 (1993).</li><li>Hommel, M., jaffe, C. L., Travi, B., Milon, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+for+leishmaniasis+and+for+testing+anti-leishmanial+vaccines.">Experimental models for leishmaniasis and for testing anti-leishmanial vaccines.</a> Ann. Trop. Med. Parasitol. 89, 55-73 (1995).</li><li>Travi, B. L., Osorio, Y., Saravia, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+inflammatory+response+promotes+cutaneous+metastasis+in+hamsters+infected+with+Leishmania+(Viannia)+panamensis.">The inflammatory response promotes cutaneous metastasis in hamsters infected with Leishmania (Viannia) panamensis.</a> J. Parasitol. 82, 454-457 (1996).</li><li>Travi, B. L., Osorio, Y. <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+Albendazole+as+an+alternative+treatment+of+cutaneous+Leishmaniasis+in+the+hamster+model.">Failure of Albendazole as an alternative treatment of cutaneous Leishmaniasis in the hamster model.</a> Memorias Instituto Oswaldo. 93, 515 (1998).</li><li>Travi, B. L., Osorio, Y., Melby, P. C., Chandrasekar, B., Arteaga, L., Saravia, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+is+a+major+determinant+of+the+clinical+evolution+and+immune+response+in+hamsters+infected+with+Leishmania+spp.">Gender is a major determinant of the clinical evolution and immune response in hamsters infected with Leishmania spp.</a> Infect Immun. 70, 2288-2296 (2002).</li><li>Osorio, Y., Melby, P. C., Pirmez, C., Chandrasekar, B., Guar&#237;n, N., Travi, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+site+of+cutaneous+infection+influences+the+immunological+response+and+clinical+outcome+of+hamsters+infected+with+Leishmania+panamensis.">The site of cutaneous infection influences the immunological response and clinical outcome of hamsters infected with Leishmania panamensis.</a> Parasite Immunol. 25, 139-148 (2003).</li><li>Osorio, Y., Bonilla, D. L., Peniche, A. G., Melby, P. C., Travi, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pregnancy+enhances+the+innate+immune+response+in+experimental+cutaneous+leishmaniasis+through+hormone-modulated+nitric+oxide+production.">Pregnancy enhances the innate immune response in experimental cutaneous leishmaniasis through hormone-modulated nitric oxide production.</a> J. Leukoc. Biol. 83, 1413-1422 (2008).</li><li>Espitia, C. M., Zhao, W., Saldarriaga, O., Osorio, Y., Harrison, L. M., Cappello, M., Travi, B. L., Melby, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Duplex+real-time+reverse+transcriptase+PCR+to+determine+cytokine+mRNA+expression+in+a+hamster+model+of+New+World+cutaneous+leishmaniasis.">Duplex real-time reverse transcriptase PCR to determine cytokine mRNA expression in a hamster model of New World cutaneous leishmaniasis.</a> BMC Immunol. 22, 11-31 (2010).</li></ol>

Cutaneous leishmaniasis is endemic in the tropics and neotropics. It is often referred to as a group of diseases because of the varied spectrum of clinical manifestations, which range from small cutaneous nodules to gross mucosal tissue destruction. Most available drugs are costly, require long treatment regimens and are becoming increasingly ineffective, necessitating the discovery of new drugs.
The mouse model is widely used but has some disadvantages. Thus for example, on the basis of the l...

<table border="1">
	<tbody>
		<tr>
			<td>Name of the reagent</td>
			<td>Type</td>
			<td>Company</td>
			<td>Catalogue number</td>
		</tr>
		<tr>
			<td>Ketamine </td>
			<td>Reagent</td>
			<td>Holliday &#x2013;Scott S.A</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Xilacine</td>
			<td>Reagent</td>
			<td>Synthesis</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Reagent</td>
			<td>GIBCO</td>
			<td>14190-136</td>
		</tr>
		<tr>
			<td>Digital caliper</td>
			<td>Equipment</td>
			<td>Fisher</td>
			<td>15-077-958</td>
		</tr>
		<tr>
			<td>Giemsa</td>
			<td>Reagent</td>
			<td>SIGMA</td>
			<td>GS1L-1L</td>
		</tr>
		<tr>
			<td>Formalin</td>
			<td>Reagent</td>
			<td>Nova lab</td>
			<td>11273</td>
		</tr>
		<tr>
			<td>Paraffin</td>
			<td>Reagent</td>
			<td>Pechiney Plastic Packaging</td>
			<td>PM-996</td>
		</tr>
		<tr>
			<td>Haemotoxilin</td>
			<td>Reagent</td>
			<td>Nova lab</td>
			<td>10870103</td>
		</tr>
		<tr>
			<td>Eosin</td>
			<td>&nbsp;</td>
			<td>Nova lab</td>
			<td>10870203</td>
		</tr>
		<tr>
			<td>Kodak Ektachem dry chemistry</td>
			<td>Equipment</td>
			<td>Kodak Ektachem </td>
			<td>DT-60</td>
		</tr>
		<tr>
			<td>meglumine antimoniate</td>
			<td>Reagent</td>
			<td>Aventis</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Microscopy</td>
			<td>Equipment</td>
			<td>Nikon</td>
			<td>YS2-T</td>
		</tr>
	</tbody>
</table>

cutaneous leishmaniasis in the dorsal skin of hamsters: a useful model for the screening of antileishmanial drugs

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of lesion size is a hard procedure and animals show difficulty to eat, breathe and move because of the lesion. In order to optimize the hamster model for cutaneous leishmaniasis, young adult male and female golden hamsters (Mesocricetus auratus) were injected intradermally at the dorsal skin with 1 to 1.5 x l07 promastigotes of Leishmania species and progression of subsequent lesions were evaluated for up to 16 weeks post infection. The golden hamster was selected because it is considered the adequate bio-model to evaluate drugs against Leishmania as they are susceptible to infection by different species. Cutaneous infection of hamsters results in chronic but controlled lesions, and a clinical evolution with signs similar to those observed in humans. Therefore, the establishment of the extent of infection by measuring the size of the lesion according to the area of indurations and ulcers is feasible. This approach has proven its versatility and easy management during inoculation, follow up and characterization of typical lesions (ulcers), application of treatments through different ways and obtaining of clinical samples after different treatments. By using this method the quality of animal life regarding locomotion, search for food and water, play and social activities is also preserved.

Watch this Scientific Journal Video about Cutaneous Leishmaniasis in the Dorsal Skin of Hamsters: a Useful Model for the Screening of Antileishmanial Drugs at JoVE.com

Cutaneous Leishmaniasis in the Dorsal Skin of Hamsters: a Useful Model for the Screening of Antileishmanial Drugs

Optimization of the experimental hamster model for cutaneous leishmaniasis by intradermal injection of Leishmania promastigotes at the dorsal skin. This approach is useful during inoculation, follow-up, characterization of lesions, application of treatments and obtaining of clinical samples. Locomotion, search for food and water, play and social activities are preserved.

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of ...

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