Leishmaniasis in the Lowlawnds of Bolivia (Leishbol): Part VIII. Characterization and Identification of Bolivian Isolates by PFG Karyotyping

Dujardin, Jean‐Claude; Gajendran, Nadesan; Hamers, R.; Matthijsen, G.; Urjel, R.; Recacoechea, M.; Villarroel, G; Bermúdez, H.; Desjeux, P.; Doncker, Simonne De; Ray, D. Le

doi:10.1007/978-1-4613-1575-9_19

Cited by 6 publications

(4 citation statements)

References 13 publications

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“…Molecular karyotyping, hybridization and densitometry OFAGE electrophoresis using equipment described by Dujardin et al (1987) were performed on 1 . 5% agarose gels in 0 .…”

Section: Parasitesmentioning

confidence: 99%

Clonal propagation and the fast generation of karyotype diversity: anin vitro Leishmaniamodel

et al. 2006

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In the present work we studied the karyotype stability during long-term in vitro maintenance in 3 cloned strains of Leishmania (Viannia) peruviana, Leishmania (Viannia) braziliensis and a hybrid between both species. Only the L. (V.) peruviana strain showed an unstable karyotype, even after subcloning. Four chromosomes were studied in detail, each of them characterized by homologous chromosomes of different size (heteromorphy). Variations in chromosome patterns during in vitro maintenance were rapid and discrete, involving loss of heteromorphy or appearance of additional chromosome size variants. The resulting pattern was not the same according to experimental conditions (subinoculation rate or incubation temperature), and interestingly, this was associated with differences in growth behaviour of the respective parasites. No change in total ploidy of the cells was observed by flow cytometry. We discuss several mechanisms that might account for this variation of chromosome patterns, but we favour the occurrence of aneuploidy, caused by aberrant chromosome segregation during mitosis. Our results provide insight into the generation of karyotype diversity in natural conditions and highlight the relativity of the clone concept in parasitology.

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“…Molecular karyotyping, hybridization and densitometry OFAGE electrophoresis using equipment described by Dujardin et al (1987) were performed on 1 . 5% agarose gels in 0 .…”

Section: Parasitesmentioning

confidence: 99%

Clonal propagation and the fast generation of karyotype diversity: anin vitro Leishmaniamodel

et al. 2006

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“…The first one is a widespread parasite in Bolivia, [1][2][3][4] while the second one has been reported only rarely. [5][6][7] Leishmania amazonensis is better known from sylvatic lowlands, especially in Amazonia, 8,9 where all proven vectors belong to the Lutzomyia flaviscutellata complex, 9,10 but the cycle is able to survive in plantation woodland and deforested areas as in Brazil. 11 In the Venezuelan and Ecuadorian Andes, parasites related to L. amazonensis have been described (L. garnhami 12 and L. mexicana, 13 respectively).…”

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confidence: 99%

Lutzomyia nuneztovari anglesi (Le pont & Desjeux, 1984) as a vector of Leishmania amazonensis in a sub-Andean leishmaniasis focus of Bolivia.

Martínez¹,

Pont²,

Torrez³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. Recently, a new Leishmania amazonensis focus was described in a sub-Andean region (1,450-2,100 meters above sea level) of Bolivia. In this area, three anthropophilic sandfly species were identified: Lutzomyia nuneztovari anglesi Le Pont & Desjeux, 1984, which represented 86-99% of the captures, Lu. galatiae Lu. shannoni Dyar 1929. Only Lu. nuneztovari anglesi was found naturally infected by flagellates (16 of 1,715 females). Three Leishmania stocks were isolated and analyzed by isoenzyme electrophoresis at 11 loci. No significant isoenzymatic differences were demonstrated between them and 7 stocks isolated from patients from the same area, and previously characterized as L. amazonensis. Moreover, in a simplified protocol, the experimental infection of Lu. nuneztovari anglesi by L. amazonensis was successful in 92% of the surviving specimens. These data are discussed in relation to the Killick-Kendrick criteria. These results strongly suggest that Lu. nuneztovari anglesi is the vector of L. amazonensis at Cajuata, Inquisivi, La Paz, Bolivia.In Bolivia, only two Leishmania species have been identified as agents of human cutaneous leishmaniasis:The first one is a widespread parasite in Bolivia, 1-4 while the second one has been reported only rarely. 5-7 Leishmania amazonensis is better known from sylvatic lowlands, especially in Amazonia, 8,9 where all proven vectors belong to the Lutzomyia flaviscutellata complex, 9,10 but the cycle is able to survive in plantation woodland and deforested areas as in Brazil. 11 In the Venezuelan and Ecuadorian Andes, parasites related to L. amazonensis have been described (L. garnhami 12 and L. mexicana, 13 respectively). Leishmania amazonensis is potentially very dangerous and occasionally induces a chronic and eventually fatal disease known as cutaneous diffuse leishmaniasis (CDL). The first Bolivian case reported by Prado Barrientos from the neighboring region of the Yungas was a typical case of CDL probably due to L. amazonensis. 14 Recently, we described an outbreak of cutaneous leishmaniasis in the province of Inquisivi, La Paz, 15 where the parasite was identified as L. amazonensis on the basis of biologic and molecular data. The present study provides evidence indicating that the vector of L. amazonensis is Lu. nuneztovari anglesi Le Pont and Desjeux, 1984. MATERIALS AND METHODSStudy area. The L. amazonensis focus is located at Cajuata and surrounding communities in the province of Inquisivi in southeastern region (67º15ЈW, 16º42ЈS) of the Department of La Paz, Bolivia. The study area is at an altitude ranging from 1,450 to 2,100 meters above sea level. 15 It is a deforested valley with very steep slopes such that the bottom of the valley is shaded early in the afternoon. The human population lives in scattered adobe brick houses with corrugated iron roofs. Around the settlements, land is cultivated with coca plantations, root vegetables, and papaya crops, while residual deciduous forests with xerophytes and epiphytes cover the steepest places. The cumulative pr...

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“…Agarose blocks containing chromosomal DNA were prepared as described by Van Der Ploeg et al (1984) for trypanosomes. OFAGE using equipment described by Dujardin et al (1987) were performed on 1.5% agarose gels in 0.4 × TBE running buffer at 7-12°C for 23 hours. Three distinct runs with 45-, 65-, and 125-s pulses achieved resolution of the whole karyotype.…”

Section: Methodsmentioning

confidence: 99%

Virulent and attenuated lines of Leishmania major: DNA karyotypes and differences in metalloproteinase GP63

Sádlová

Volf²,

Victoir

et al. 2006

FOLIA PARASIT

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Abstract. The Leishmania metalloproteinase GP63 has been reported to play important roles mainly in resistance of promastigotes to complement-mediated lysis and in interaction with macrophage receptors. On the other hand, its function in insect vectors is still unclear. We compared the structure and dosage of gp63 genes and the activity of GP63 in Leishmania major Yakimoff et Schokhor strains and lines differing in virulence for mice and ability to develop in sand flies. The results demonstrate considerable variability in amount and proteolytical activity of GP63 among L. major strains although genomic changes in the gp63 locus were not found. Attenuated LV561/AV line showed low amount and low enzymatic activity of GP63. Serial passages of attenuated parasites through either Phlebotomus duboscqi Neveu-Lemaire or through mice led to a recovery of GP63 proteolytical activity to the level present in virulent LV561/V line. Overexpression of GP63 was found in two L. major strains (L119, Neal) with defective lipophosphoglycan (LPG); both these strains were capable to cause mice infection but unable to survive and multiply in sand flies. Differences were found also in karyotypes and in amount of minichromosomes amplified in some lines of the LV561 strain. The results suggest that parasite virulence is not simply correlated with the activity of GP63; however, this enzyme plays a significant role in association with other surface molecules, especially LPG. Overexpression of GP63 can compensate LPG defect in the vertebrate host but in sand flies both molecules fulfil quite different functions and the defect in LPG is lethal for the parasite. On the other hand, linear minichromosomes of about 200 kb found in some lines of the LV561 strain are associated with development in vitro and in the vector but they are not essential for the infection of the vertebrate host.

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Leishmaniasis in the Lowlawnds of Bolivia (Leishbol): Part VIII. Characterization and Identification of Bolivian Isolates by PFG Karyotyping

Cited by 6 publications

References 13 publications

Clonal propagation and the fast generation of karyotype diversity: anin vitro Leishmaniamodel

Clonal propagation and the fast generation of karyotype diversity: anin vitro Leishmaniamodel

Lutzomyia nuneztovari anglesi (Le pont & Desjeux, 1984) as a vector of Leishmania amazonensis in a sub-Andean leishmaniasis focus of Bolivia.

Virulent and attenuated lines of Leishmania major: DNA karyotypes and differences in metalloproteinase GP63

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