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Improvements in obtaining New World Leishmania sp from mucosal lesions: notes on isolating and stocking parasites.
Exp Parasitol. 2012 Oct; 132(2):300-3.EP

Abstract

Tegumentary leishmaniasis is an endemic protozoan disease that, in Brazil, is caused by parasites from Viannia or Leishmania complex. The clinical forms of cutaneous disease comprise localized, disseminated, mucosal or mucocutaneous, and diffuse leishmaniasis. Viannia complex parasites are not easy to isolate from patient lesions, especially from mucosal lesions, and they are difficult to culture. The aim of the present study was to compare the efficiency of ex vivo (culture) and in vivo (IFNγ-deficient mice) parasite isolation methods to improve the isolation rate and storage of stocks of New World Leishmania sp that cause cutaneous leishmaniasis (CL) or mucosal leishmaniasis (ML). Biopsy fragments from cutaneous or mucosal lesions were inoculated into culture medium or mouse footpads. We evaluated 114 samples (86 CL, 28 ML) using both methods independently. Samples from CL patients had a higher isolation rate in ex vivo cultures than in mice (34.1% vs. 18.7%, P<0.05). Nevertheless, almost twice the number of isolates from ML lesions was isolated using the mouse model compared to ex vivo cultures (mouse, 6/25; culture, 3/27). The overall rates of isolation were 40.2% for CL samples and 29.6% for ML samples. Of the 43 isolations, we successfully stocked 35 isolates (81.4%; 27 CL, 8 ML). Contaminations were more frequently detected in cultures of ML than CL lesions. For comparison, the use of both methods simultaneously was performed in 74 samples of CL and 25 samples of ML, and similar results were obtained. Of the eight ML isolates, five were isolated only in mice, indicating the advantage of using the in vivo method to obtain ML parasites. All parasites obtained from in vivo isolation were cryopreserved, whereas only 68% of ex vivo isolations from CL lesions were stocked. In conclusion, the use of genetically modified mice can improve the isolation of parasites from ML. Isolation and stocking of New World Leishmania parasites, especially those from ML that are almost absent in laboratory stocks, are critical for evaluating parasite genetic diversity as well as studying host-parasite interactions to identify biological markers of Leishmania. In this paper, we also discuss some of the difficulties associated with isolating and stocking parasites.

Authors+Show Affiliations

Institute of Tropical Pathology and Public Health, Universidade Federal de Goiás, Rua 235 S/N - Setor Universitário, 74605-050 Goiânia, Goiás, Brazil. mledorta@gmail.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22728105

Citation

Dorta, Miriam L., et al. "Improvements in Obtaining New World Leishmania Sp From Mucosal Lesions: Notes On Isolating and Stocking Parasites." Experimental Parasitology, vol. 132, no. 2, 2012, pp. 300-3.
Dorta ML, Oliveira MA, Fleuri AK, et al. Improvements in obtaining New World Leishmania sp from mucosal lesions: notes on isolating and stocking parasites. Exp Parasitol. 2012;132(2):300-3.
Dorta, M. L., Oliveira, M. A., Fleuri, A. K., Duarte, F. B., Pinto, S. A., Pereira, L. I., & Ribeiro-Dias, F. (2012). Improvements in obtaining New World Leishmania sp from mucosal lesions: notes on isolating and stocking parasites. Experimental Parasitology, 132(2), 300-3. https://doi.org/10.1016/j.exppara.2012.06.006
Dorta ML, et al. Improvements in Obtaining New World Leishmania Sp From Mucosal Lesions: Notes On Isolating and Stocking Parasites. Exp Parasitol. 2012;132(2):300-3. PubMed PMID: 22728105.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Improvements in obtaining New World Leishmania sp from mucosal lesions: notes on isolating and stocking parasites. AU - Dorta,Miriam L, AU - Oliveira,Milton A P, AU - Fleuri,Anna K A, AU - Duarte,Fernanda B, AU - Pinto,Sebastião A, AU - Pereira,Ledice I A, AU - Ribeiro-Dias,Fatima, Y1 - 2012/06/21/ PY - 2011/12/10/received PY - 2012/06/12/revised PY - 2012/06/13/accepted PY - 2012/6/26/entrez PY - 2012/6/26/pubmed PY - 2012/12/10/medline SP - 300 EP - 3 JF - Experimental parasitology JO - Exp Parasitol VL - 132 IS - 2 N2 - Tegumentary leishmaniasis is an endemic protozoan disease that, in Brazil, is caused by parasites from Viannia or Leishmania complex. The clinical forms of cutaneous disease comprise localized, disseminated, mucosal or mucocutaneous, and diffuse leishmaniasis. Viannia complex parasites are not easy to isolate from patient lesions, especially from mucosal lesions, and they are difficult to culture. The aim of the present study was to compare the efficiency of ex vivo (culture) and in vivo (IFNγ-deficient mice) parasite isolation methods to improve the isolation rate and storage of stocks of New World Leishmania sp that cause cutaneous leishmaniasis (CL) or mucosal leishmaniasis (ML). Biopsy fragments from cutaneous or mucosal lesions were inoculated into culture medium or mouse footpads. We evaluated 114 samples (86 CL, 28 ML) using both methods independently. Samples from CL patients had a higher isolation rate in ex vivo cultures than in mice (34.1% vs. 18.7%, P<0.05). Nevertheless, almost twice the number of isolates from ML lesions was isolated using the mouse model compared to ex vivo cultures (mouse, 6/25; culture, 3/27). The overall rates of isolation were 40.2% for CL samples and 29.6% for ML samples. Of the 43 isolations, we successfully stocked 35 isolates (81.4%; 27 CL, 8 ML). Contaminations were more frequently detected in cultures of ML than CL lesions. For comparison, the use of both methods simultaneously was performed in 74 samples of CL and 25 samples of ML, and similar results were obtained. Of the eight ML isolates, five were isolated only in mice, indicating the advantage of using the in vivo method to obtain ML parasites. All parasites obtained from in vivo isolation were cryopreserved, whereas only 68% of ex vivo isolations from CL lesions were stocked. In conclusion, the use of genetically modified mice can improve the isolation of parasites from ML. Isolation and stocking of New World Leishmania parasites, especially those from ML that are almost absent in laboratory stocks, are critical for evaluating parasite genetic diversity as well as studying host-parasite interactions to identify biological markers of Leishmania. In this paper, we also discuss some of the difficulties associated with isolating and stocking parasites. SN - 1090-2449 UR - https://www.unboundmedicine.com/medline/citation/22728105/Improvements_in_obtaining_New_World_Leishmania_sp_from_mucosal_lesions:_notes_on_isolating_and_stocking_parasites_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0014-4894(12)00193-2 DB - PRIME DP - Unbound Medicine ER -