Rapid isolation of metacyclic trypomastigotes of Trypanosoma cruzi from feces and urine of the vector

Schaub, Günter A.

doi:10.1016/0001-706x(91)90072-r

Cited by 5 publications

(11 citation statements)

References 15 publications

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“…The overall biological role of this coat is unknown but it seems to be associated with stabilization of the plasma-membrane or protection of antigenic determinants. The binding of albumin was clearly indicated by the observation that adherence of eluted, vector-derived metacyclic trypomastigotes of T. cruzi onto glass surfaces after isolation via a DEAE-Sephacel column is excluded if the parasites are collected in an albumin-supplemented buffer (Schaub, 1991). The same phenomenon occurred using blood trypomastigotes (Schaub, unpublished data).…”

Section: Discussionmentioning

confidence: 96%

“…The pellet was resuspended in 2-3 ml of a phosphatebuffered saline solution supplemented with 0 . 5% glucose (PBSG) (Schaub, 1991) and cells were transferred onto a pre-equilibrated column filled with 1 ml of cold DEAE-Sephacel (Pharmacia , Uppsala, Sweden) (Stiles & Kierszenbaum, 1986 ;Schaub, 1991). Trypomastigotes were eluted with PBSG and the eluate was collected in fractions of approximately 5 ml each into tubes containing 1 .…”

Section: Isolation and Purification Of Blood Trypomastigotesmentioning

confidence: 99%

“…Isolation procedures for trypomastigotes from either mammalian or insect hosts usually employ a DEAEcellulose column followed by concentration of cells using centrifugation (Lanham, 1968). Isolation protocols differ in the composition of buffers and centrifugal forces (Colli, Andrews & Zingales, 1980 ;Araujo & Remington, 1981 ;Zingales et al 1982 ;Katzin & Colli, 1983 ;Lanar & Manning, 1984 ;Stiles & Kierszenbaum, 1986 ;Piras, Henriquez & Piras, 1987 ;Andrews et al 1988 ;Couto et al 1990 ;Schaub, 1991). The observation that trypomastigotes isolated using DEAE columns adhere to glass-surfaces if they are not collected in a protein-supplemented buffer indicates that column-induced changes of the surface of the parasites have occurred (Schaub, 1991).…”

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confidence: 99%

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Effects of the isolation methodology on protein profiles of blood trypomastigotes of Trypanosoma cruzi

Hölscher

Hartmann²,

Mossmann

et al. 2003

Parasitology

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Blood trypomastigotes of Trypanosoma cruzi were isolated from infected athymic rnu/rnu rats and purified by an improved procedure of DEAE-Sephacel ion-exchange chromatography. Elution into a buffer supplemented with bovine serum albumin avoided column-induced changes on the surface of the parasites. Biotin-labelled bovine serum albumin, fluorescence microscopy, flow cytometry and Western blot analysis revealed a very intense binding of albumin to the parasite. Incubation and washing of cells without protein supplementation did not result in any damage or lysis of parasites but it did cause extensive shedding of cellular and surface proteins into the supernatant which could be prevented by using the protein-supplemented buffer. A decreasing yield of high molecular weight cellular proteins in relation to centrifugal force was a general phenomenon observed in scanning densitometry of SDS gels after isolation in either protein-supplemented buffer or protein-free buffer. The quantity of shed cellular components increased with increasing centrifugal force. In contrast, quantities of high molecular weight, biotin-labelled surface proteins increased with greater centrifugal force, indicating labelling of otherwise inaccessible residues. These data emphasize the importance of protein supplementation of buffers with proteins and of choosing low centrifugation forces (<400 g) during investigations of T. cruzi.

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Section: Discussionmentioning

confidence: 96%

Section: Isolation and Purification Of Blood Trypomastigotesmentioning

confidence: 99%

mentioning

confidence: 99%

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Effects of the isolation methodology on protein profiles of blood trypomastigotes of Trypanosoma cruzi

Hölscher

Hartmann²,

Mossmann

et al. 2003

Parasitology

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“…Isolated from Triatoma infestans, it is cyclically maintained between this vector and mice or stored at )80°C (Schaub 1988). Since bacterial contamination of the vectorderived metacyclic trypomastigotes could aect the parasitemia in the athymic nude rats, metacyclic trypomastigotes were isolated from faeces and urine of T. infestans (Schaub 1992) and subcutaneously injected into immunocompetent Balb/c mice. Three weeks p.i., at the peak of parasitemia, nude rats were infected.…”

Section: Methodsmentioning

confidence: 99%

“…(Ebert and Schaub 1983;Schaub and Schottelius 1984;Schaub and LoÈ sch 1988;Schaub 1988Schaub , 1989. For a characterization of vector-and mammal-derived trypomastigotes of low virulent strains, not only high numbers of vector-derived trypomastigotes (Schaub 1992) but also of blood trypomastigotes were needed. Parasitemia (and thereby the amount of obtainable blood trypomastigotes) could be increased by using strains of mammals which are highly susceptible, e.g.…”

Section: Introductionmentioning

confidence: 99%

Development of high numbers of blood trypomastigotes of Trypanosoma cruzi in nude rats

Schaub

Hölscher

Mossmann

2001

Parasitology Research

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The development and pathogenicity of a Trypanosoma cruzi strain ("Chile 5") of low virulence were studied after infection of nude rats with different doses of blood trypomastigotes (10-10(7) parasites/rat). Peak parasitemias were correlated with the infection dose, which also influenced the mean survival times (26-36 days post-infection). Within 26 or 27 days, a subcutaneous injection of 10(7) blood trypomastigotes developed to about 8-20 x 10(7) parasites/ml.

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“Natural infections” with Trypanosoma cruzi via the skin of mice: size of mouthparts of vectors and numbers of invading parasites

Waldeck

Schaub

2022

Parasitol Res

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Investigating parameters influencing natural infections with Trypanosoma cruzi via the skin, the diameters of mouthparts of different stages of triatomines vectors were measured to determine the size of the channel accessible for T. cruzi during cutaneous infection. The mean diameters of the skin-penetrating mandibles of first to fifth instar nymphs of the vector Triatoma infestans increased from 18 to 65 µm. The mean diameter in fourth instar nymphs of Dipetalogaster maxima was 86 µm. Different numbers of isolated vector-derived metacyclic trypomastigotes (10–10,000) were injected intradermally into mice. Prepatent periods, parasitemia and mortality rates were compared with those of mice obtaining 10,000 metacyclic trypomastigotes that are usually present in the first drop of faeces onto the feeding wounds of fifth and fourth instar nymphs of T. infestans and D. maxima, respectively. After injection of 50–10,000 T. cruzi, in all 42 mice the infection developed. An injection of 10 parasites induced an infection in 8 out of 15 mice. With increasing doses of parasites, prepatent periods tended to decrease. The level of parasitemia was higher after injection of the lowest dose. Except for one mouse all infected mice died. After placement of 10,000 metacyclic trypomastigotes onto the feeding wound of fifth or fourth instar nymphs of T. infestans and D. maxima, respectively, the infection rates of the groups, prepatent periods and the levels of parasitemia of T. cruzi in mice indicated that about 10–1,000 metacyclic trypomastigotes entered the skin via this route. For the first time, the present data emphasise the risk of an infection by infectious excreta of triatomines deposited near the feeding wound and the low number of invading parasites.

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Rapid isolation of metacyclic trypomastigotes of Trypanosoma cruzi from feces and urine of the vector

Cited by 5 publications

References 15 publications

Effects of the isolation methodology on protein profiles of blood trypomastigotes of Trypanosoma cruzi

Effects of the isolation methodology on protein profiles of blood trypomastigotes of Trypanosoma cruzi

Development of high numbers of blood trypomastigotes of Trypanosoma cruzi in nude rats

“Natural infections” with Trypanosoma cruzi via the skin of mice: size of mouthparts of vectors and numbers of invading parasites

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