Continuous Cultivation of Trypanosoma brucei Blood Stream Forms in a Medium Containing a Low Concentration of Serum Protein without Feeder Cell Layers

1989 ◽  
Vol 75 (6) ◽  
pp. 985 ◽  
Author(s):  
Hiroyuki Hirumi ◽  
Kazuko Hirumi
Keyword(s):  
Trypanosoma Brucei ◽  
Serum Protein ◽  
Continuous Cultivation ◽  
Blood Stream ◽  
Feeder Cell ◽  
Low Concentration ◽  
Cell Layers ◽  
Feeder Cell Layers

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with γ-Irradiation

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4400 ◽  
Author(s):  
Andras Nagy ◽  
Marina Gertsenstein ◽  
Kristina Vintersten ◽  
Richard Behringer
Keyword(s):  
Mouse Embryo ◽  
Feeder Cell ◽  
Γ Irradiation ◽  
Mouse Embryo Fibroblast ◽  
Cell Layers ◽  
Feeder Cell Layers

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomycin C

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4399 ◽  
Author(s):  
Andras Nagy ◽  
Marina Gertsenstein ◽  
Kristina Vintersten ◽  
Richard Behringer
Keyword(s):  
Mitomycin C ◽  
Mouse Embryo ◽  
Feeder Cell ◽  
Mouse Embryo Fibroblast ◽  
Cell Layers ◽  
Feeder Cell Layers

In vitrocultivation ofTrypanosoma congolensebloodstream forms in the absence of feeder cell layers

Parasitology ◽  
1991 ◽  
Vol 102 (2) ◽  
pp. 225-236 ◽  
Author(s):  
H. Hirumi ◽  
K. Hirumi
Keyword(s):  
Infected Mouse ◽  
Primary Cultures ◽  
Population Doubling ◽  
Surface Coat ◽  
Feeder Cell ◽  
Mouse Blood ◽  
Diethylaminoethyl Cellulose ◽  
Cell Layers ◽  
Feeder Cell Layers

SUMMARYBloodstream forms ofTrypanosoma congolense(2 clones: ILNat3·1 and IL3000, and 4 stocks: IL2079, IL2466, IL3266 and CP-81) were continuously cultivatedin vitroat 34–36 °C in the absence of feeder cell layers, using HMI-93 medium which was modified from Iscove's modified Dulbecco's MEM (Flow Laboratories, Irvine, Scotland). The modification was done by supplementing the medium with 0·05 mM bathocuproine sulphonate, 1·5 mM L-cysteine, 0·5 mM hypoxanthine, 0·12 mM 2-mercaptoethanol, 1 mM sodium pyruvate, 0·16 mM thymidine, 20% (v/v) heat-inactivated young goat serum and 5% (v/v) Serum Plus™ (Hazleton Biologics, Lenaxa, KS, USA). Trypanosomes obtained from two different sources were used to initiate primary cultures: (1) metacyclic forms which were producedin vitroat 27 °C, and (2) bloodstream forms obtained from Balb/c mice which had been infected with the bloodstream forms transformedin vitrofrom the metacyclic forms. Metacyclic forms placed in 25 cm2T-type (T-25) flasks rapidly attached to the bottom of the flasks and transformed to bloodstream forms during the initial 24 h and continued to proliferate. The bloodstream forms isolated from the infected mouse blood by means of diethylaminoethyl cellulose (DE52) column chromatography also continued to proliferate in the flasks. Cultures were maintained by replacing the medium every 24 h. Every 4–5 days, the attached bloodstream forms were resuspended in fresh medium by gentle pipetting and then were subcultured. The method was further simplified by initiating primary cultures directly with 10 μl of the tail blood of infected mice in 24-well culture plates and then by subcultivating either in wells or in T-25 flasks. The shortest population doubling time, 9 h, was achieved by seeding subcultures with 106bloodstream forms/ml. The bloodstream forms propagated in this system were morphologically similar to those seen in infected mouse blood, they were covered with a surface coat as examined by electron microscopy and they were infective to mice.

scholarly journals Comparative evaluation of the chemotherapeutic efficacies of two salts of diminazene aceturate in Trypanosoma brucei brucei infected dogs

2021 ◽  
Vol 18 (9) ◽  
pp. 1961-1967
Author(s):  
Ukamaka U. Eze ◽  
Ifeanyi G. Eke ◽  
Ikenna O. Ezeh ◽  
Terry A. Nzeakor ◽  
Callistus Owube ◽  
...  
Keyword(s):  
Drug Treatment ◽  
Trypanosoma Brucei ◽  
Clinical Management ◽  
Parasite Clearance ◽  
Treated Group ◽  
Blood Stream ◽  
Post Treatment ◽  
Trypanosoma Brucei Brucei ◽  
Diminazene Aceturate ◽  
Experimental Canine

Purpose: To compare the anti-trypanosomal efficacies of 4,4-(diazoaminedibenzamidinetrihydrate) diacetate (4,4-DDBT) and 4,4-(diazoamino) benzamidine (4,4-DB) in experimental canine trypanosomosis. Methods: The efficacies of 4,4-DDBT and 4,4-DB were evaluated in 4 groups of dogs (n = 3) designated A-D. Group A was normal control without infection or drug treatment, group B did not receive any drug treatment but was infected with Trypanosoma brucei brucei, while groups C and D were infected with T. b. brucei and treated with 4,4-DDBT(3.5 mg/kg) and 4,4-DB (3.5 mg/kg), respectively. Results: The incubation period of the infection was 6 - 9 days post-infection. Treatment of the dogs with 4,4-DDBT led to zero parasitaemia 48 h post-treatment, while there was only a decrease in parasitemia to log 6 in 4,4-DB-treated dogs. Resurgence of parasite into the blood stream occurred in 4,4-DDBTtreated dogs 6 days after initial parasite clearance. Blood analyses post-treatment revealed elevated leucocytes and lymphocytes in 4,4-DB-treated dogs (p < 0.05). Packed cell volume was also observed to be higher in 4,4-DDBT-treated group when compared to 4,4-DB group (p < 0.05). Conclusion: These findings suggest that 4,4-DDBT is more efficacious in the clinical management of canine trypanosomosis caused by T. b. brucei. However, it does not prevent relapse of infection. Based on these findings, therefore, 4,4-DDBT should be the diminazene salt of choice when indicated in the clinical management of T. b. brucei infection in dogs.

scholarly journals Clinicopathological and Microscopic Features of Trypanosoma brucei and Trypanosoma evansi Induced Infections in Sheep II

2021 ◽  
Vol 41 (2) ◽  
pp. 144-160
Author(s):  
Y.A. Wada ◽  
P.I. Rekwot ◽  
O.O. Okubanjo ◽  
B. Mohammed ◽  
S.J. Oniye
Keyword(s):  
Trypanosoma Brucei ◽  
Trypanosoma Evansi ◽  
Carcass Composition ◽  
Mixed Infections ◽  
Mucus Membrane ◽  
Microscopic Features ◽  
Cell Layers ◽  
Gross Lesions ◽  
Proliferation Of Lymphocytes ◽  
Renal Tubular

The present study elucidates further on clinical, gross, and microscopic pathologies induced by single or mixed infections with  Trypanosoma evansi and Trypanosoma brucei in sheep. Briefly, the experimental animals were divided into four groups of three  animals each. Animals in each group were either infected with T. brucei, T. evansi, mixed (T. brucei and T. evansi), or noninfected. Animals were observed for clinical, gross, and microscopic pathologies for 98 days (14 weeks). The clinical pathologies observed included loss of body condition, pale ocular mucus membrane, rough hair coat, scrotal oedema, scrotal degeneration, emaciation, and death. At necropsy, macroscopic or gross lesions included very pale and anaemic carcass composition, congested and pneumonic lungs with severe haemorrhages, serous atrophy of intestinal and body fats, lymphadenopathy, splenomegaly, and hepatomegaly. Microscopic lesions observed in the testes, spleen, liver, lungs, lymphoid, heart, and brain tissues of infected sheep were varied and included swollen kidney with renal tubular degeneration, the proliferation of lymphocytes at the germinal centers of the spleen, degeneration of the bronchioles, severe testicular degeneration with a reduction in the number of spermatogenic cell layers,  degenerated Leydig and Sertoli cells with loss of sperm reserves in the seminiferous lumen, congested liver with sinusoidal spaces and the proliferation of monocytes and lymphocytes. The results indicate that trypanosomosis due to experimental T. brucei, T. evansi, or mixed infections may be an important cause of various grades of tissue and organ pathologies in sheep in trypanosome-endemic areas. Keywords: Trypanosomosis; Clinico-pathological and microscopic features; Trypanosoma brucei; Trypanosoma evansi; Mixed  infections; Sheep

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