Cytological and immunological responses toBabesia divergens in different hosts: Ox, gerbil, man

Gorenflot, A.; Brasseur, Philippe; Précigout, Éric; L’Hostis, Monique; Marchand, A.; Schrével, Joseph

doi:10.1007/bf00934377

Cited by 98 publications

(99 citation statements)

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“…The in vitro culture of B. bovis has been described in sheep, goat, horse and rabbit erythrocytes [11]. Similarly, Ben Musa and Phillips [1] and Gorenflot et al [5] described continuous culture of B. divergens in rat erythrocytes and human erythrocytes respectively; they both used RPMI 1640 supplemented with 20% FCS and 10% human serum respectively. The culture in sheep erythrocytes described in this paper is closely related to these protocols.…”

Section: Discussionmentioning

confidence: 99%

“…divergens Rouen 1987 was obtained from a man who had recovered from acute babesiosis [5]. Additionally, five B. divergens isolates were obtained from infected cattle in 1988: 1903B (Corrèze), 2212B (Côtes d'Armor), 2704A (Eure), 3606A (Indre), 5601A (Morbihan).…”

Section: B Divergens Isolatesmentioning

confidence: 99%

“…Splenectomized rats could be experimentally infected by B. divergens [10], but the main experimental model for B. divergens babesiosis remains the Gerbil (Meriones unguiculatus), which develops an acute and fatal babesiosis [9]. The in vitro culture of B. divergens has also been described in erythrocytes from different origins: rat [1] and humans [5]. This paper presents the in vitro culture of B. divergens in sheep erythrocytes and the first results on experimental infections of splenectomized sheep with B. divergens.…”

Section: Introductionmentioning

confidence: 99%

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Sheep as a new experimental host for Babesia divergens

et al. 2002

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-Babesia divergens was cultivated in sheep erythrocytes in RPMI 1640 supplemented with 10% Fetal Calf Serum (FCS) or sheep serum. In vitro cultures in sheep red blood cells were initiated with human erythrocytes infected in vitro with B. divergens Rouen 1987 or with gerbil blood infected with several isolates from bovine origin. After the first subcultures on sheep erythrocytes, a ten-fold multiplication of the parasites was obtained within 48 h. Erythrocytes from three splenectomized sheep were infected in vitro with B. divergens; when parasitaemia reached 10%, the animals were inoculated with homologous parasitized erythrocytes. All sheep expressed hyperthermia with a peak between the 6th and the 9th day post-infection (p-i) and a transitory parasitaemia 10 days p-i. In vitro primary cultures were performed on two of these sheep, demonstrating the parasite persistence at very low parasitaemia in the infected animals. Splenectomized sheep can be used as a new model for B. divergens chronic infection.Babesia divergens / sheep / in vitro culture / experimental infection Résumé -Le Mouton : un nouvel hôte expérimental de Babesia divergens. Babesia divergens a été cultivé in vitro dans des hématies de Mouton suspendues dans du RPMI 1640 supplémenté par 10 % de sérum de veau foetal ou de sérum de Mouton. Les cultures en hématies de mouton ont été initiées avec des hématies humaines infectées in vitro par B. divergens Rouen 1987 ou avec des hématies de gerbilles infectées par différents isolats d'origine bovine. Après les premiers passages sur héma-ties de mouton, les parasites se multiplient 10 fois en 48 h. Les hématies de trois moutons splénecto-misés ont été infectées in vitro par B. divergens ; lorsque la parasitémie in vitro a atteint 10 %, les animaux ont été infectés expérimentalement avec les hématies homologues. Tous les moutons ont présenté un pic d'hyperthermie entre le 6 e et le 9 e jour post-infection (p-i) et une parasitémie

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Section: Discussionmentioning

confidence: 99%

Section: B Divergens Isolatesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Sheep as a new experimental host for Babesia divergens

et al. 2002

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“…Later, erythrocytes from various animal origins corresponding to natural (bovine, human) or experimental hosts (rat, sheep) were proved to support B. divergens continuous in vitro growth [2,4,10,21]. The culture system developed in our laboratory [4] with sheep erythrocytes allowed high in vitro parasitemia (50%) with only 10% serum (fetal calf or sheep).…”

Section: Introductionmentioning

confidence: 99%

Isolation of Babesia divergens from carrier cattle blood using in vitro culture

2004

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-Babesia divergens, the main causative agent of bovine babesiosis in Western Europe, was isolated from naturally infected cattle. Ninety-six blood samples were examined by means of an in vitro culture technique in sheep erythrocytes: 19 of them were collected from animals in the acute phase of the disease with visible parasitemia on blood smears, while the 77 remaining animals showed no microscopically detectable parasites. B. divergens was cultured from the 19 first blood samples as well as from 31 samples collected from asymptomatic animals. The time period before parasites could be detected in the culture varied in the latter samples from 6 to 20 days. The effects of sampling condition (anticoagulant used) and storage length were tested. A good correlation was obtained between immunofluorescent antibody test and culture, with identical results (positive or negative) for 89.6% of the samples collected from asymptomatic animals. The sensitivity of the in vitro culture method was determined and was about 10 parasites/mL of whole blood from three independent experiments performed with three different isolates, confirming its suitability to detect and culture diverse B. divergens isolates from carrier cattle. The parasites could indeed be isolated 9 months after the acute babesiosis phase in the blood of naturally infected animals. The 50 isolates collected in this study were successfully subcultured, cryopreserved and resuscitated using the same culture medium. The in vitro isolation of B. divergens from asymptomatic carrier cattle was achieved and will allow the analysis of parasite diversity within cattle herds. Babesia divergens / carrier cattle / in vitro culture / isolation

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“…Routine diagnosis of Babesia bovis babesiosis with an enzyme linked immunosorbent assay (ELISA) using a defined parasite antigenic fraction has been described (Waltisbuhl et al, 1987). For B. divergens, ELISA requires parasitized erythrocytes obtained from live animals (Purnell et al, 1976;Gray and Kaye, 1991) The previous description of a long term in vitro culture for B. divergens has allowed us to produce purified parasites (Gorenflot et al, 1991;Becuwe et al, 1992). The purpose of the present study was to develop an ELISA using these purified parasites as a source of antigen.…”

Section: Introductionmentioning

confidence: 99%

Babesia divergens: an Elisa with soluble parasite antigen for monitoring the epidemiology of bovine babesiosis

Chauvin

L’Hostis²,

Valentin

et al. 1995

Parasite

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Summary :An enzyme linked immunosorbent assay (ELISA) for bovine babesiosis caused by Babesia divergens was developed to analyse the evolution of the serological status of cattle living in an enzootic area. The antigen used was a soluble extract of B. divergens obtained from in vitro culture. Specificity, evaluated with negative seta, was 96,6%. The ELISA was compared to indirect immunofluorescence analysis (IFA) on naturally or experimentally infected animals. It appeared that IFA was positive for at least seven or eight weeks; on the contrary, B. divergens-specific antibodies were only transitorily detected by ELISA. Furthermore, the ELISA was more sensitive than the IFA for the detection of post-infection antibody increases, particularly when infection-pressure was low.These results suggest that IFA and ELISA could be complementary tools in epidemiological surveys; indeed, this ELISA could be the most efficient tool to detect new infections in cohort monitoring.

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Cytological and immunological responses toBabesia divergens in different hosts: Ox, gerbil, man

Cited by 98 publications

References 23 publications

Sheep as a new experimental host for Babesia divergens

Sheep as a new experimental host for Babesia divergens

Isolation of Babesia divergens from carrier cattle blood using in vitro culture

Babesia divergens: an Elisa with soluble parasite antigen for monitoring the epidemiology of bovine babesiosis

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