Growth characteristics and chemical composition of Trypanosoma cruzi

Fernandes, J.F.; Castellani, Olga

doi:10.1016/0014-4894(66)90016-6

Cited by 77 publications

(22 citation statements)

References 12 publications

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“…Previous reports of the amount of DNA in T. cruzi have indicated ca. 1.7 × 10 -13 g of DNA per cell [5,23,24]. Our results, however, indicate that the total DNA content is about 3.3 X 10 -13 g, or about two times the above amount.…”

Section: Discussioncontrasting

confidence: 59%

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Lanar

Lévy

Manning

1981

Molecular and Biochemical Parasitology

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The content and sequence complexity of the nuclear DNA and messenger RNA for epimastigotes of Trypanosoma cruzi were determined. From analysis of nuclear DNA reassociation studies and microspectrofluorometric measurements of laser induced fluorescence of cellular DNA, T. cruzi is found to be a diploid organism with a nuclear DNA content of 2.5 x 108 nucleotide pairs (2.8 X 10 -~3 g) and a kinetoplast DNA content of 4.9 X 107 nucleotide pairs (5.4 X 10 -t4 g). Reassoeiation kinetics of nuclear DNA of average length 0.4 kb reveals three kinetic components: a moderately repetitive component with a reiteration frequency of 5.1 X 103 present in 9% of the fragments, a lowly repetitive component with a reiteration frequency of 32 present in 51% of the fragments, and a single-copy component present in 23% of the fragments.By saturation hybridization of total polysomal RNA to 3H-labeled single-copy DNA, it was determined that 68% of the single-copy DNA was represented in the epimastigote polysomal RNA. This corresponds to ca. 12 000 different mRNA species. Of these, ca. 9000 are present as poly(A)÷-RNA, while the remaining 3000 appear not to be polyadenylated. Kinetic analysis of the poly(A)+-RNA population indicates it is composed of at least three classes of RNAs of different abundancy levels: two sequences which occur ca. 3000 times per cell, ca. 750 sequences which occur about 20 times per cell, and ca. 15 500 sequences which occur 1-2 times per cell.

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Section: Discussioncontrasting

confidence: 59%

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Lanar

Lévy

Manning

1981

Molecular and Biochemical Parasitology

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“…Epimastigote forms of T. cruzi (Y strain) were cultivated in LIT medium [7]. After 2 days of growth (8 × 107 cells/ml) the epimastigotes were collected by centrifugation and washed twice with 150 mM NaC1 containing 50 mM sodium phosphate (pH 7.2, PBS).…”

Section: Methodsmentioning

confidence: 99%

Endogenous and surface labeling of glycoconjugates from the three differentiation stages of Trypanosoma cruzi

Zingales¹,

Martin²,

Lederkremer³

et al. 1982

FEBS Letters

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“…The worms were observed daily for motility, moulting and survival. The culture medium (GLIT) was 1 volume of modified Yaeger's LIT (Liver Infusion Tryptose) medium (Fernandes & Castellani 1966) and 1 volume of modified Grace's medium (Grace's insect medium added with 2% v/v Basal Medium Eagle vitamins [100×] solution + 1 mM L-cysteine + 1 g l -1 glucose) plus 20% (v/v) heat-inactivated foetal bovine serum and 1% (w/v) commercial pepsin, at pH 4.0, 13°C and 5% CO 2 in air. The media, sera and reagents were purchased from Sigma Chemical Co. (USA) and Difco Laboratories (USA).…”

Section: Resale or Republication Not Permitted Without Written Consenmentioning

confidence: 99%

In vitro development of the fish parasite Hysterothylacium aduncum from the third larval stage recovered from a host to the third larval stage hatched from the egg

Adroher¹,

Malagón²,

Valero³

et al. 2004

Dis. Aquat. Org.

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Anisakids are parasitic nematodes of fish worldwide, producing economic and human health concerns. It is thus important to ascertain their in vitro life cycle in laboratory studies. Here we describe the in vitro development of third-stage larvae (L3) of Hysterothylacium aduncum isolated from blue whiting Micromesistius poutassou, to the hatching of L3 from eggs obtained from H. aduncum worms grown in GLIT medium (a modified mixture of Yaeger's LIT [Liver Infusion Tryptose] and Grace's media) at pH 4.0, 13°C and with 5% CO 2 in air. Under these conditions, L3 recovered from fish developed to mature adults (3.4 to 6.2 cm in length), with oviposition starting from Day 26 in culture. Fertilized eggs (mean size 64 × 52 µm) had a thick, rugose eggshell and were larger than unfertilized ones (mean size 49 × 42 µm), whose eggshells were thin and smooth. Eggs laid during the first and second week of oviposition, and maintained in 2.8% NaCl solution at 13°C, developed to L3. Under these maintenance conditions, between 20.6 and 52.5% of the eggs laid during the first week developed into larvae. Motile larvae, enclosed in a sheath, hatched from between 2 and 11% of these eggs. The larvae started to hatch 23 d after deposition. These larvae were 144 to 215 µm in length, enclosed in a 237 to 305 µm-long sheath. This GLIT culture medium may help to study the biology of this and other anisakids.KEY WORDS: Fish parasite · Hysterothylacium aduncum · Nematoda · Anisakidae · In vitro culture · Egg development · Hatching Resale or republication not permitted without written consent of the publisherDis Aquat Org 58: [41][42][43][44][45] 2004 mesistius poutassou (blue whiting), family Gadidae, purchased from the fish market of Granada (southern Spain). Blue whiting, on the Atlantic and Mediterranean Spanish coasts, are frequently parasitized by H. aduncum (Ruiz-Valero et al. 1992, Valero et al. 2000. The worms, found free in the host body cavity, were collected with the help of a needle with a blunt tip, placed on a Petri dish and washed in 0.9% NaCl solution several times. The worms were observed individually under an inverted microscope, and those that showed any kind of internal or external damage were discarded. They were then identified according to morphological features (Hartwich 1975, Petter & Maillard 1988, Petter & Cabaret 1995.Prior to cultivation, each larva was placed in an antibiotic-antifungal solution (80 mg gentamicin sulfate, 0.625 mg amphotericin B, 10 000 IU sodium penicillin G, 10 mg streptomycin sulfate and 4.5 ml Hanks' solution, for a final solution volume of 10 ml) and axenized as described elsewhere (Iglesias et al. 1997, Iglesias et al. 2001. Worms were cultured in sterile polystyrene 30 ml flasks. The culture medium (10 ml) was placed into each flask with 8 to 10 parasites. The culture flasks were then placed in an incubator at 13°C and 5% CO 2 in humid air, and the culture medium was renewed once a week. The worms were observed daily for motility, moulting and survival. The culture medium (GLI...

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Growth characteristics and chemical composition of Trypanosoma cruzi

Cited by 77 publications

References 12 publications

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Endogenous and surface labeling of glycoconjugates from the three differentiation stages of Trypanosoma cruzi

In vitro development of the fish parasite Hysterothylacium aduncum from the third larval stage recovered from a host to the third larval stage hatched from the egg

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