Brenière, Simone Frédérique

Desjeux, P.

Le Pont, François

Valette, E.

Horizon / Pleins textes

Mem. Inst. Osrvaldo Cncz, Rio de Janeiro 
Vol. 83(1): 139-140, jan./mar. 1988 RESEARCH NOTE 
ZYMODEMES OF TRYPANOSOMA CRUZ1 ISOLATED. 
FROM WILD MAMMALS IN BOLIVIA 
E. VALETTE, S. F. BRENIERE, F. LE PONT & P. DESJEUX 
IBBA - ORSTOM, Embajada de Francia, Casilla 824, La Paz, Bolivia 
Trypanosoma cruzi, the etiological agent of 
Chagas’ disease, is a very heterogeneous popula- 
tion of parasites circulating among man, vectors 
and wild and domestic reservoirs. Until now,. 
few data have been reported about the T. cnizi 
isoenzymatic strains circulating in the Bolivian 
sylvatic reservoirs. In the present study we 
reported the isoenzyme typification of 9 
Trypanosomatidae stocks isolated from wild 
Bolivian mammals. 
The stocks were obtained from wild mam- 
mals caught in the Yungas and Alto Beni 
regions of Bolivia (North East from La Paz) by 
means of blood culture: 2 ml of sterile heart 
blood were introduced into 3 ml of LIT medi- 
um and the cultures amplified to a good con- 
centration of parasites for the isoenzyme 
typification. These stocks were analyzed by 
isoenzyme electrophoresis carried out in cel- 
lulose acetate plates. The eleven enzyme 
systems assayed were as follows: glucose-6- 
phosphate dehydrogenase (EC 1.1.1.49XG6P), 
glucose-6-phosphate isomerase (EC 5.3.1.9) 
(GPI), glutamate dehydrogenase (EC 1.4.1.2) 
(GDl), glutamate dehydrogenase (NADP+) 
(EC 1.4.1.4)(GD2), isocitrate dehydrogenase 
(NADP+) (EC 1.1.1.42) (ICD), malate dehydro- 
genase (EC 1.1.1.37)(MDH), malate dehydroge- 
nase (oxaloacetate-decarboxylating) (NADP+) 
or malic enzyme (EC 1.1.1.40)(ME), peptidase 
1 (ficin, EC 3.4.22.3, formely EC 3.4.4.12, 
substrate: leucylleucylleucine) (PEPl), peptidase 
2 (bromelain, EC 3.4.22.4, formely EC 3.4.4.24, 
substrate: leucyl-L-alanine) (PEP2), phospho- 
glucomulatase (EC 5.4.2.2, formely EC 2.7.5.1) 
(PGM), and phosphogluconate dehydrogenase 
(EC 1.1.1.44) (6PGD). The stocks were 
classified according to their genetic distances 
(average number of codon differences between 
populations) (Nei, 1972, Am Nat., 106: 
283-292). We used as reference samples: (a) the 
three zymodemes previously described by Miles 
Received September 28, 1987. 
Accepted January 18, 1988. 
et al. (1980, Trans. R. Soc. Trop. Med. Hyg.., 
74: 221-237) referred to  as Z1, 22  and 23; 
(b) the Chilean Tulahuen strain. 
Out of 9 stocks, 5 were identified as Trypa- 
nosoma cruzi (2 of them already reported, 3 of 
them close to two variants already reported). 
These 5 T. cmzi stocks were isolated from 4 
Didelphis marsupialis (opossum) and 1 Dasypus 
novemcinctus (armadillo). Two interesting 
features should be emphasized regarding the 
enzyme profiles of the T. cruzi stocks: (a) 1 of 
them appeared identical to the Tulahuen strain, 
and also exhibited typical heterozygous pattern 
for the GPI, like the Tulahuen strain (Tibayrenc 
& Desjeux, 1983, Trans. R. Soc. Trop. Med 
Hyg., 77: 73-75). Until now such zymodeme, 
with typical heterozygous patterns, had been 
reported only in domestic transmission cycles 
(Tibayrenc & Miles, 1983, Trans. R. Soc. Trop. 
Med. Hyg., 7 %  76-83; Tibayrenc et al., 1984, 
TABLE 
Genetic distances among sylvatic Bolivian stocks and 
reference Brazilian zymodemes 
Reference 
strains Sylvatic strains* Sylvatic 
strains* 
N ? 2 0 N ? 4 3  NP1 N P 2  Z1 22 23  
N? 20 - 
N? 43 1.58 - 
NP 1 0.24 1.60 - 
NP 2 0.78 0.69 0.85 - 
References 
z1 0.64 1.03 0.37, 0.85 - 
22  1.34 0.84 1.36 1.26 1.65 - 
23 1.77 1,24 1.79 1.07 2.26 0.85 - 
* Sylvatic strains: NP 20 and NO 43, reference num- 
ber from Tibayrenc classification (Tibayrenc e t  al., 
1986, Proc. Nat1 Acad. Scì. USA, 83: 115-1191, 
NP =New profiles. ORSTOM Fonds Documentaire 
E. VALETTE ET AL. 140 
ISOENZYMIC STRAINS" 
22 Enzymes W.S Tula Z1 
œ - -  I I -  
ME œ - - - -  - - -  
MDH U 
PEP 2 9 
6PGD 
* w. s =Wild stocks unrelated to T. cruzi, Tula =Tu- 
lahuen T. cruzi reference strain, Z1 and 22 =refer- 
ence Brazilian T. cruzi strains. 
Trans. R. Soc. Trop. Med. Hyg., 78: 519-525). 
(b) Even for this very limited sample, the 
distance values are spread over a continuum and 
show that the zymodemes recorded here are 
impossible to be clustered into few well-defined 
groups or "principal zymodemes" : they exhibit 
a non-hierarchized structure (see Table), a 
result already noted for a more extensive set of 
stocks by Tibayrenc et al. (1986, Proc. Nat2 
A cad Sci. USA, 83: 1 15-1 1 7) .  
The 4 other stocks were not identified as 
T. cruzi. They shared the same isoenzyme 
profiles and were isolated from 1 Bradypus tri- 
dactylus (sloth) and 3 Coendou preizensilis 
(porcupine). The malic enzyme and the malate 
dehydrogenase enzyme systems did not reveal 
2 loci as in i? cruzi and three other enzyme 
systems (PGM, 6PGD and PEP 2) presented 
new patterns never reported for the extensive 
set of stocks of i? cruzi studied by Tibayrenc 
et al. (1986, &oc. Nat1 Acad. Sci. USA, 83: 
115-119) (see Fig.). The taxonomic status 
of these stocks is presently under analysis. 
Further details will be included in a more 
elaborated paper. 
Acknowlegments - We are indebted to 
Isoenzymic patterns of wild Trypanosomatidae stocks Dra Michel Tibayrenc (ORSTOM) who revised 
and Trypanosoma cruzi reference strains. the manuscript. 
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Zymodemes of Trypanosoma cruzi isolated from wild mammals in Bolivia

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