Besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates

Frey, Caroline; Regidor-Cerrillo, J; Marreros Canales, Nelson Antonio; García-Lunar, P; Gutiérrez-Expósito, D; Schares, G; Dubey, JP; Gentile, A; Jacquiet, P; Shkap, V; Cortes, H; Ortega-Mora, LM; Álvarez-García, G (2016). Besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates. Parasites & Vectors, 9(1), p. 115. BioMed Central 10.1186/s13071-016-1405-9

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Background Bovine besnoitiosis, caused by the protozoan Besnoitia besnoiti, reduces productivity and fertility of affected herds. Besnoitiosis continues to expand in Europe and no effective control tools are currently available. Experimental models are urgently needed. Herein, we describe for the first time the kinetics of standardised in vitro models for the B. besnoiti lytic cycle. This will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful for the treatment of besnoitiosis. Methods We compared invasion and proliferation of one B. tarandi (from Finland) and seven B. besnoiti isolates (Bb-Spain1, Bb-Spain2, Bb-Israel, Bb-Evora03, Bb-Ger1, Bb-France, Bb-Italy2) in MARC-145 cell culture. Host cell invasion was studied at 4, 6, 8 and 24 h post infection (hpi), and proliferation characteristics were compared at 24, 48, 72, 96, 120, and 144 hpi. Results In Besnoitia spp., the key parameters that determine the sequential adhesion-invasion, proliferation and egress steps are clearly distinct from those in the related apicomplexans Toxoplasma gondii and Neospora caninum. Besnoitia spp. host cell invasion is a rather slow process, since only 50 % of parasites were found intracellular after 3–6 h of exposure to host cells, and invasion still took place after 24 h. Invasion efficacy was significantly higher for Bb-France, Bb-Evora03 and Bb-Israel. In addition, the time span for endodyogeny to take place was as long as 18–35 h. Bb-Israel and B. tarandi isolates were most prolific, as determined by the tachyzoite yield at 72 hpi. The total tachyzoite yield could not be predicted neither by invasion-related parameters (velocity and half time invasion) nor by proliferation parameters (lag phase and doubling time (dT)). The lytic cycle of Besnoitia was asynchronous as evidenced by the presence of three different plaque-forming tachyzoite categories (lysis plaques, large and small parasitophorous vacuoles). Conclusions This study provides first insights into the lytic cycle of B. besnoiti isolates and a standardised in vitro model that allows screening of drug candidates for the treatment of besnoitiosis.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Center for Fish and Wildlife Health (FIWI)

UniBE Contributor:

Frey, Caroline and Marreros Canales, Nelson Antonio

Subjects:

600 Technology > 630 Agriculture

ISSN:

1756-3305

Publisher:

BioMed Central

Funders:

[4] Swiss National Science Foundation

Language:

English

Submitter:

Caroline Franziska Frey Marreros Canales

Date Deposited:

22 Mar 2016 11:34

Last Modified:

22 Mar 2016 11:34

Publisher DOI:

10.1186/s13071-016-1405-9

PubMed ID:

26928430

Uncontrolled Keywords:

Besnoitia besnoiti, Besnoitia tarandi, Lytic cycle, In vitro model, Isolates, Invasion, Proliferation

BORIS DOI:

10.7892/boris.79072

URI:

https://boris.unibe.ch/id/eprint/79072

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