The conditions required for the maintenance of Onchocerca lienalis microfilariae in vitro. 1991

T M Litchfield, and C D Mackenzie, and M G Varma
Department of Allergy and Allied Respiratory Diseases, Guy's Hospital, London, U.K.

Microfilariae of the bovine parasite Onchocerca lienalis were maintained in vitro using a tick cell line as a feeder layer, and under the conditions provided would develop to the sausage stage. A number of media supported this achievement with CMRL 1066 producing the highest yields, particularly when used in conjunction with either medium 199, HAM's F-12, RPMI 1640 or Mark's M20. The addition of inactivated foetal calf serum (iFCS) suppressed development; on the other hand, the addition of tryptose phosphate broth (TPB) enhanced it. The pH range of the medium preferred by the developing worms was 7.35-7.85, and the most favourable osmolality lay in the range 360 to 390 mosM/kg. Insect-derived hormones did not improve yields of developed larvae nor promote moulting to the second larval stage. The culture conditions described in this work, which favour parasite survival and development, provide further insights into the physiological requirements of filariae as well as guide lines to achieving successful in vitro maintenance of Onchocerca sp. A morphological examination of the developing larvae was carried out at electron microscopical level.

UI MeSH Term Description Entries
D008842 Microfilariae The prelarval stage of Filarioidea found in the blood and tissues of mammals, birds and intermediate hosts (vector). Microfilaria
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D009854 Onchocerca A genus of parasitic nematodes whose organisms live and breed in skin and subcutaneous tissues. Onchocercal microfilariae may also be found in the urine, blood, or sputum. Onchocercas
D009994 Osmolar Concentration The concentration of osmotically active particles in solution expressed in terms of osmoles of solute per liter of solution. Osmolality is expressed in terms of osmoles of solute per kilogram of solvent. Ionic Strength,Osmolality,Osmolarity,Concentration, Osmolar,Concentrations, Osmolar,Ionic Strengths,Osmolalities,Osmolar Concentrations,Osmolarities,Strength, Ionic,Strengths, Ionic
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013696 Temperature The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms. Temperatures
D013987 Ticks Blood-sucking acarid parasites of the order Ixodida comprising two families: the softbacked ticks (ARGASIDAE) and hardbacked ticks (IXODIDAE). Ticks are larger than their relatives, the MITES. They penetrate the skin of their host by means of highly specialized, hooked mouth parts and feed on its blood. Ticks attack all groups of terrestrial vertebrates. In humans they are responsible for many TICK-BORNE DISEASES, including the transmission of ROCKY MOUNTAIN SPOTTED FEVER; TULAREMIA; BABESIOSIS; AFRICAN SWINE FEVER; and RELAPSING FEVER. (From Barnes, Invertebrate Zoology, 5th ed, pp543-44) Ixodida,Ixodidas,Tick

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