In vitro isolation and cultivation of Cowdria ruminantium under serum-free culture conditions

Zweygarth, E.; Vogel, Susanne; Josemans, Antoinette I.; Horn, Ernst

doi:10.1016/s0034-5288(97)90011-4

Cited by 12 publications

(17 citation statements)

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“…The Welgevonden isolate has been shown, as discussed above, to provide cross-protection against a needle challenge with a range of other isolates (Collins et al, 2003). This could make the Welgevonden isolate a good candidate to use as an attenuated vaccine, but unfortunately it did not attenuate while being grown in culture through hundreds of passages over several years (Gueye et al, 1994;Zweygarth et al, 1997). Recently, however, the Welgevonden isolate was attenuated by being cultured in a canine macrophage-monocyte cell line, after which it was by re-adapted to grow in bovine endothelial cells (Zweygarth et al, 2005).…”

Section: Attenuated Vaccinesmentioning

confidence: 99%

Natural history of Ehrlichia ruminantium

Allsopp

2010

Veterinary Parasitology

121

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Ehrlichia ruminantium is an obligately intra-cellular "-proteobacterium which causes a disease known as heartwater or cowdriosis in some wild, and all domestic, ruminants.The organism is transmitted by ticks of the genus Amblyomma, and it is of serious economic importance wherever the natural vectors occur, an area which includes all of sub-Saharan Africa, and several islands in the Caribbean. The disease was first

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Section: Attenuated Vaccinesmentioning

confidence: 99%

Natural history of Ehrlichia ruminantium

Allsopp

2010

Veterinary Parasitology

121

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“…The in vitro isolation and cultivation of C. ruminantium under serum‐free culture conditions has been reported previously. 10 Three stocks of C. ruminantium were propagated in a serum‐free medium based on a modified HL‐1 medium, consisting of a DME/F‐12‐based medium and compound solution A (see under Materials and Methods). Although HL‐1 medium is chemically defined, the supplier did not publish the complete formulation and only limited information is made available.…”

Section: Resultsmentioning

confidence: 99%

“…Various synthetic culture media were used: HL‐1 (Hycor Biomedical Inc., Portland, Maine, USA), Dulbecco's modified Eagle's medium nutrient mixture Ham F‐12 (DME/F‐12) (Sigma, St. Louis, MO, USA; D 0547), SFRE 199‐1 (Sigma; S 2013), Iscove's modified Dulbecco's medium (IMDM) (Sigma; 17633), DME (Highveld Biologicals), Leibovitz L‐15 (Highveld Biologicals; CN 1225‐1) and RPMI 1640 (Highveld Biologicals; CN 1735). The complete, serum‐free media prepared from these synthetic culture media are referred to as SFMC‐1 (HL‐1), 10 SFMC‐3 (DME/F‐12), SFMC‐9 (SFRE 199‐1), SFMC‐10 (IMDM), SFMC‐13 (Leibovitz L‐15) and SFMC‐14 (RPMI 1640) ( s erum‐ f ree m edium for C owdria No. nn).…”

Section: Methodsmentioning

confidence: 99%

“…Propagation of cultures was carried out as described previously 10 with minor modifications. Briefly, infected cultures were harvested for subcultivation by scraping off the cell monolayer into the medium using a Pasteur pipette bent at the tip to form a small hook.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, Zweygarth et al 10 succeeded in propagating three stocks of C. ruminantium in a serum‐free medium, two of which were also initiated under these conditions. A commercial medium kit was used and although the complete culture medium was chemically defined, the exact formulation is not completely published.…”

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Serum‐free Media for the in Vitro Cultivation of Cowdria ruminantium

Zweygarth

Josemans

Horn

1998

Annals of the New York Academy of Sciences

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The in vitro culture of Cowdria ruminantium, the causative agent of heartwater in domestic ruminants, was first achieved in 1985; since then, most groups working with this culture system have used media which were supplemented with serum and, in addition, most of them contained tryptose phosphate broth. These undefined products vary from batch to batch and often fail to support the growth of C. ruminantium. We are therefore working towards the development of a completely chemically defined medium for Cowdria culture. We attempted the propagation of the Welgevonden stock of C. ruminantium in bovine endothelial cell cultures in a variety of serum-free culture media. Four synthetic media gave unsatisfactory results, these were: SFRE-199, Iscove's modified Dulbecco's medium, Dulbecco's modified Eagle's medium, and Leibovitz L-15. These media were all supplemented with a proprietary solution A (components solution A of the HL-1 medium kit, containing transferrin, testosterone, sodium selenite, ethanolamine, saturated and unsaturated fatty acids, and stabilizing proteins). Three other serum-free media did support the growth of C. ruminantium: a modified HL-1 medium, Dulbecco's modified Eagle's medium nutrient mixture Ham F-12 (DME/F-12), and RPMI 1640. The chemical composition of DME/F-12 and RPMI 1640 are published, but not that of the HL-1 medium. Each of these media was supplemented with proprietary solution A. Various supplements were investigated as alternative to the incompletely specified solution A; bovine lipoproteins and bovine transferrin were identified as essential supplements which effectively replaced compound solution A. C. ruminantium was propagated in the three growth-supportive media for at least 10 passages.

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Ehrlichia

Allsopp¹,

McBride²

Genome Mapping and Genomics in Animal-Associated Microbes

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In vitro isolation and cultivation of Cowdria ruminantium under serum-free culture conditions

Cited by 12 publications

References 9 publications

Natural history of Ehrlichia ruminantium

Natural history of Ehrlichia ruminantium

Serum‐free Media for the in Vitro Cultivation of Cowdria ruminantium

Ehrlichia

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