scholarly journals A simple method to purify biologically and antigenically preserved bloodstream trypomastigotes of Trypanosoma cruzi using Deae-cellulose columns

1983 ◽  
Vol 78 (3) ◽  
pp. 317-333 ◽  
Author(s):  
Maria Auxiliadora de Sousa
Keyword(s):  
Trypanosoma Cruzi ◽  
Deae Cellulose ◽  
Mouse Blood ◽  
Simple Method ◽  
Infected Blood ◽  
Normal Morphology ◽  
Mean Survival Time ◽  
Triatomine Bugs ◽  
Forms Processing ◽  
Polymorphism Pattern

A method to purify trypanosomastigotes of some strains of Trypanosoma cruzi (Y, CL, FL, F, "Berenice", "Colombiana" and "São Felipe") from mouse blood by using DEAE-cellulose columns was standardized. This procedure is a modification of the Lanham & Godfrey methods and differs in some aspects from others described to purify T. cruzi bloodstream trypomastigotes, mainly by avoidance of prior purifications of parasites. By this method, the broad trypomastigotes were mainly isolated, accounting for higher recoveries obtained with strains having higher percentages of these forms: processing of infected blood from irradiated mice could be advantageous by increasing the recovery of parasites (percentage and/or total number) and elution of more slender trypomastigotes. Trypomastigotes purified by this method presented normal morphology and motility, remained infective to triatomine bugs and mice, showing in the latter prepatent periods and courses parasitemia similar to those of control parasites, and also reproducing the polymorphism pattern of each strain. Their virulence and pathogenicity also remained considerably preserved, the latter property being evaluated by LD 50 tests, mortality rates and mean survival time of inoculated mice. Moreover, these parasites presented positive, clear and peripheral immunofluorescence reaction at titres similar to those of control organisms, thus suggesting important preservation of their surface antigens.

scholarly journals Triatomine bugs, their microbiota and Trypanosoma cruzi: asymmetric responses of bacteria to an infected blood meal

2016 ◽  
Vol 9 (1) ◽  
Author(s):  
Sebastián Díaz ◽  
Bianca Villavicencio ◽  
Nathália Correia ◽  
Jane Costa ◽  
Karen L. Haag
Keyword(s):  
Trypanosoma Cruzi ◽  
Blood Meal ◽  
Infected Blood ◽  
Triatomine Bugs ◽  
Asymmetric Responses

Isolation of blood and intracellular forms of Trypanosoma cruzi from rats and other rodents and preliminary studies of their metabolism

Parasitology ◽  
1978 ◽  
Vol 76 (2) ◽  
pp. 159-176 ◽  
Author(s):  
W. E. Gutteridge ◽  
B. Cover ◽  
Maria Gaborak
Keyword(s):  
Trypanosoma Cruzi ◽  
Blood Cells ◽  
Light And Electron Microscopy ◽  
Deae Cellulose ◽  
Whole Body ◽  
Differential Centrifugation ◽  
Γ Irradiation ◽  
Hind Limb Muscle ◽  
Radio Tracer

SummaryIsolation of blood and intracellular forms of Trypanosoma cruzi was made mainly from rats (90–110 g) which had received 580 rad of whole-body γ-irradiation not more than 24 h before subcutaneous inoculation with 107 trypomastigotes of the Sonya strain of T. cruzi. Unirradiated chinchillas (250–350 g) were, however, used for some experiments. Blood forms were isolated using a technique involving differential centrifugation to remove most of the erythrocytes and DEAE–cellulose chromatography to remove the remaining blood cells. Overall recoveries were usually in the range 30–70%. Parasites were mainly (approximately 98%) broad forms and were motile, metabolically active (as judged by respiratory and radio-tracer incorporation studies) and had lost none of their infectivity for mice. Intracellular forms were isolated from hind-limb muscle tissue. This was disrupted in an MSE tissue homogenizer and the homogenate incubated with DNase, collagenase and trypsin. Parasites, contaminated only by a few blood cells, were then obtained by differential centrifugation. For purer preparations, a terminal sucrose gradient step was used. Recoveries ranged between 40 and 70%. About 1–3% of the parasites isolated were epimastigotes and trypomastigotes; the remainder are probably best collectively termed ‘amastigotes’, though they were pointed and most had a short, free flagellum. They were undamaged as judged by light and electron microscopy and metabolically active as judged by respiratory and radio-tracer incorporation studies. However, the infectivity for mice of both these purified preparations and the initial cell homogenates could be accounted for by the epimastigotes and trypomastigotes present in them. Preliminary biochemical studies with isolated parasites have shown that blood, intracellular and culture forms of T. cruzi have a respiratory system which is in part sensitive to CN- and that all forms synthesize nucleic acids and proteins when incubated in vitro. There appears, however, to be a lack of DNA synthesis in blood stages, and thus it is not surprising that these forms do not divide.

scholarly journals Elucidating the Mechanism of Trypanosoma cruzi Acquisition by Triatomine Insects: Evidence from a Large Field Survey of Triatoma infestans

2020 ◽  
Vol 5 (2) ◽  
pp. 87
Author(s):  
Aaron W. Tustin ◽  
Ricardo Castillo-Neyra ◽  
Laura D. Tamayo ◽  
Renzo Salazar ◽  
Katty Borini-Mayorí ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Regression Models ◽  
Control Strategies ◽  
Protozoan Parasite ◽  
Etiologic Agent ◽  
Triatoma Infestans ◽  
Large Field ◽  
Blood Sucking ◽  
Triatomine Bugs ◽  
Cruzi Infection

Blood-sucking triatomine bugs transmit the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease. We measured the prevalence of T. cruzi infection in 58,519 Triatoma infestans captured in residences in and near Arequipa, Peru. Among bugs from infected colonies, T. cruzi prevalence increased with stage from 12% in second instars to 36% in adults. Regression models demonstrated that the probability of parasite acquisition was roughly the same for each developmental stage. Prevalence increased by 5.9% with each additional stage. We postulate that the probability of acquiring the parasite may be related to the number of feeding events. Transmission of the parasite does not appear to be correlated with the amount of blood ingested during feeding. Similarly, other hypothesized transmission routes such as coprophagy fail to explain the observed pattern of prevalence. Our results could have implications for the feasibility of late-acting control strategies that preferentially kill older insects.

scholarly journals Surface electrical charge of bloodstream trypomastigotes of Trypanosoma cruzi strains

1983 ◽  
Vol 78 (4) ◽  
pp. 497-500 ◽  
Author(s):  
Maria Auxiliadora de Sousa
Keyword(s):  
Trypanosoma Cruzi ◽  
Deae Cellulose ◽  
Electrical Charge ◽  
Surface Charges

Bloodstream trypomastigotes of some Trypanosoma cruzi strains were processed through DEAE-cellulose columns under standardized conditions. The results obtained suggest mainly that these strains present different surface charges, that there are subpopulations of bloodstream trypomastigotes as regards electrical charges and that the broad forms are less negative than the slender ones.

Plasmodium berghei-infected red cells sorted according to DNA content

Parasitology ◽  
1979 ◽  
Vol 78 (3) ◽  
pp. 263-270 ◽  
Author(s):  
R. J. Howard ◽  
F. L. Battye
Keyword(s):  
Red Blood Cells ◽  
Dna Content ◽  
Cell Sorting ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Dye Uptake ◽  
Direct Proportion ◽  
Mouse Blood ◽  
Infected Blood ◽  
A Cell

SUMMARYA cell-sorting method is described for the analysis and separation of red blood cells in Plasmodium berghei-infected mouse blood based on their DNA content. This method involves a selective uptake of the bis-benzimidazole dye 33258 Hoechst, a DNA-binding dye, by red blood cells containing parasites. Infected blood is incubated at 37 °C with the dye then washed at 4 °C to remove unbound dye. Uninfected cells are then non-fluorescent at the characteristic wavelengths for 33258 Hoechst excitation and emission, whereas parasitized cells display fluorescence intensities in approximately direct proportion to the number of parasite nuclei (i.e. amount of parasite DNA) within the cell and can be sorted accordingly. Providing cells were incubated in a complex nutrient medium during dye uptake at 37°C, the sorted parasite cells produced lethal P. berghei infections when injected into BALB/c mice. The dyelabelling technique is simple and sufficient red blood cells at various stages of infection can be collected for biochemical or immunochemical studies by cell sorting.

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