The in vitro cultivation of the filarial nematode Brugia malayi from the infective stage to the fourth and the young adult stage is described. Different culture conditions including cell-free systems and coculture with different human lymphatic cell lines were compared. Cocultivation with each of the cell lines used enhanced the survival of the parasites. The best results were obtained employing the human T cell leukemia line Jurkat and human dermal fibroblasts as feeder cells in RPMI 1640 supplemented with 10% heat-inactivated human serum. This culture system allowed the cultivation of B. malayi for more than 7 weeks with an average growth of the larvae by factor 6.4 (0.77 ± 0.035 cm) and a maximum growth by factor 10 (1.2 cm). 100% of the parasites reached the fourth larval stage after 14 days. By day 37 of culture, 17% of the larvae had completed their final moult. Parasites remained alive up to 52 days. During the first four weeks of culture, both the length and the periods of moulting of the in vitro cultivated filariae closely resembled those observed with B. malayi in vivo in rodent hosts. This system provides a new approach for successful cultivation of B. malayi, that should help to overcome the difficulties of reproduction by other laboratories encountered with previous culture systems. It appeared highly reproducible in our hands and possibly further improvement can be obtained on its basis. This cultivation system represents a further step towards the complete replacement of the vertebrate host by means of in vitro culture systems in parasitological research.

An in vitro cytotoxicity assay with fish cell lines was established in order to reduce the number of animals used in fish tests with the golden orfe (leuciscus idus melanotus). In three co-ordinated working laboratories the toxicity of seven chemicals was determined according to a standardised protocol using the permanent cell lines D11 and RTG-2. There was an almost linear correlation (r=0.99) between the IC50 values after neutral red uptake and crystal violet staining of the chemically treated RTG-2 cells. These cells were more sensitive than D11 cells (r=0.93) with regard to chemically induced cytotoxicity. The test protocol was adapted for the testing of waste water (ten samples). Two samples induced cytotoxic effects in RTG-2 cells while in none of the three laboratories the same samples were found cytotoxic in D11 cells. The data with the seven chemicals and the still limited number of waste water samples (10) lend strong support to the assumption that the fish cell line RTG-2 can be regarded as well suited for detecting toxic properties of waste water.

The isolated perfused Bovine Udder Skin (BUS) model from slaughter house material is an ex vivo skin model that can easily be maintained in culture at high vitality for at least 8 hours. The BUS model allows to study skin penetration and absorption as well as skin irritation almost under in vivo conditions. Moreover, the skin of the bovine udder is morphologically resembling human skin to a higher extent than the skin of rodents. Pharmacological and toxicological studies on raw materials and formulated products of interest to cosmetics and skin products industry reveal a high correlation to data obtained in humans. Therefore, the BUS model holds great promise since it allows to reduce the number of test animals in skin pharmacology and toxicology, since it provides information which cannot be obtained in other in vitro models and not even in human studies, e.g. pharmacokinetics and drug metabolism.

In 1994 ZEBET, the German Center for Documentation and Evaluation of Alternatives to Animal Experi­ments, at the Federal Institute for Health Protection of Consumers and Veterinary Medicine, has asked 32 government institutions in the Federal Republic of Germany about animal experiments conductedfor diagnosis of microbiological diseases and diseases caused by parasites in veterinary medicine in Germany in 1988-1993. Among the 22 institutions answering some had totally replaced animal experiments for the diagnosis of rabies and psittacosis by cell culture methods and of tuberculosis using appropriate culture media. In some institutions diagnosis of tuberculosis is still performed in animals despite the general statement  in the standard for diagnosis of tuberculosis DIN 58943 part 31 (1989) "due to improved diagnostic culture methods ... animaI experiments need not be carried out routinely for the diagnosis of tuberculosis H."  The result of the inquiry shows that not all of the institutes have replaced animal experiments by in vitro alternatives even though they are recommended  in standards, guideli­nes or other regulatory documents. This particularly holds for the diagnosis of rabies, psittacosis and tuberculosis. In some institutions lack of funding may not have allo­wed to establish cell culture methods.

NEWS
SIAT Newsletter 7, Schliessung des IVT-Labors
Gründung eines Steinbeis Technologietransfer-Zentrums für in vitro Pharmakologie und Toxikologie Jahresbericht der Stiftung set
ECVAM Corner
Verleihung des BMG-Forschungspreises 94
Verleihung des "Herbert-Stiller-Preises" 1995
BMG-Ausschreibung Forschungspreis
Stiftung Fo rschung 3R: Call for 3R-Research-Proposals
Ausschreibung Felix-Wankel-Tierschutz-Forschungspreis 1996
Novellierung des deutschen Tierschutzgesetzes
Teilrevision der Schweizer Tierschutzverordnung

COMMENTS
Stellungnahme des DTB zur Novellierung
Stellungnahme der TVT zur Novellierung
Kommentar zur Teilrevision der TSchV

CONFERENCE REPORT
Linz '95

BOOK REVIEW
Matthias Kaiser and Stellan Welin (eds.): Ethical Aspects of Modern Biology

CALENDAR

INDEX 

INSTRUCTIONS TO AUTHORS