201-207
Induction of partial protection against Leishmania major in BALB/c mice by Leishmania tropica
by Hamid Mahmoudzadeh-Niknam

Leishmaniasis is a parasitic disease of man and other mammals. Immunity against leishmaniasis appears to remain essentially species specific; however some cross-reactivity has been reported. The aim of this study was analyzing cross-protection induced by Leishmania tropica (L. tropica) against Leishmania major (L. major). BALB/c mice were infected with L. tropica in the footpad followed by a challenge infection in the contra lateral footpad by L. tropica or L. major. Footpad thickness and parasite load in the footpad, popliteal lymph node, and spleen were determined after challenge. The results demonstrate that L. tropica induces partial protection of BALB/c mice against L. tropica as well as L. major. The protection was more efficient against a homologous strain (L. tropica) than against a heterologous strain (L. major). The partial protection against L. major was detected in the footpad tissues as well as popliteal lymph node. No protection was observed against L. major in the spleen tissue. These findings have implications in vaccination strategies for Leishmaniasis based on the use of heterologous species of the parasite.

209-213
Bordetella avium cross-reacts with B. bronchiseptica by ELISA but natural B. avium infection in rats is unlikely
by R Boot L. van den Berg, M A Koedam& J L Veenema

The specificity of a Bordetella bronchiseptica antibody ELISA for the monitoring of laboratory rodents was re-evaluated by studying the serological relationship of the B. bronchiseptica antigen and antigens of B. avium, B. hinzii, B. holmesii and an unclassified Bordetella sp. Immunization of rats with B. avium strains induced antibodies to the B. bronchiseptica antigen. Bordetella antibody free rats that were experimentally infected with a B. avium strain seroconverted to the bacterium but not to B. bronchiseptica and were B. avium culture positive at one week postinfection but not at 6 weeks. Pathogen free rats exposed to the B. avium infected rats remained free from any cultural or serological evidence of B. avium infection. Lung lesions were not seen in experimentally infected and exposed rats. Natural B. avium infection in rats therefore seems unlikely.

215-219
Experimental calcification of the aorta in rabbits: Effects of chelating agents and glucagon
by C Cavallotti, F M Tranquilli Leali, M Artico, V D’Andrea& V. Malinovska

Experimental calcification of the aorta, induced in rabbits by a single dose of 8 mg of dihydro-tachysterol (AT 10), represents an optimal model for studying vascular diseases caused by lipid and calcium deposition on the artery wall. This lesion may be treated with chelating agents and/or with glucagon. The effect of chelating agents and/or of glucagon on lipid and calcium deposition involving the aortic wall was studied in 12 adult white Flemish rabbits. The animals were divided into four groups and in each of them experimental calcification of the aorta was induced by AT 10. The first group did not receive any treatment and served as control. The second group received only ethylenediamine tetraacetic acid (EDTA) (120 mg daily). The third group received only glucagon (0.1 mg subcutaneously). In the fourth group, the glucagon (0.1 mg subcutaneously) was injected with EDTA (120 mg daily). The animals were killed 30 days after suspension of therapy. The aorta was rapidly harvested in its thoracic tract. The calcium content of the sclerosed aorta was determined by flame photometer, while the cytochemical detection of calcium was performed using N,N-Naphthaloyl-hydroxyl-amine. Lipid deposition was studied by Sudan black B staining. Usual laboratory staining (hematoxyline - eosine and Toluidin blue ) were used for detection of structural details. All morphological results were submitted to the quantitative analysis of images and statistical analysis. The results of the present study suggest that glucagon may play a role in the biochemical and morphological modifications usually found in atherosclerosis. In fact, macroscopic and ultrastructural evaluation of our morphological results suggest that the pathological changes observed may be restored by administration of glucagons and/or by chelating therapy.

221-223
Case Report and Short Communication: Rectal prolapse associated with an unusual combination of pinworms and citrobacter species infection in FVB mice colony
by MJ Mahesh Kumar, P Nagarajan, R Venkatesan & Ramesh C Juyal

Spontaneous cases of rectal prolapse in a breeding colony of FVB mice were found to be due to infection with Syphacia obvelata and Citrobacter freundii. Microbiology, biochemical and parasitological examination revealed Citrobacter freundii and eggs of Syphacia obvelata. After treatment with antibiotics, antihelminthic drugs and manual reduction prevented further occurrence.

245-249
Fluctuating Asymmetry in relation to single housing versus group housing in three inbred mouse strains
by Charlotte Stub, Merel Ritskes-Hoitinga, Aage Kristian Olsen, Thomas C Krohn& Axel Kornerup Hansen

Fluctuating asymmetry (FA) reflects small, random deviations from symmetry in otherwise bilaterally symmetrical characters and has been used to detect harmful conditions such as environmental and genomic stress in growing animals and humans. The development of FA may be related to the balance between canalization (the ability of the genotype to develop a constant phenotype under changing environments) and plasticity (the ability of the genotype to change phenotype dependent on the environment) of the individual. Different mouse strains differ in coping strategies in stress situations, and these coping strategies may be related to this balance. In this study, development of FA was studied in female mice of three different inbred strains, 129s6/Sv, C57BL/6J, and BALB/c, during a 6 week period. Besides the comparison of different strains, single housing was compared to group housing conditions. Overall, FA did not differ between strains. After six weeks, single-housed mice had higher FA than those that were group housed (P<0.001), which may indicate that single housing causes a higher degree of environmental stress than group housing does.

251-267
Nordic-European Workshop on Ethical Evaluation of Animal Experiments
Workshop Report on the Cost-Benefit Principle in Hanasaari, Helsinki, Finland 7-9 November 2003
by Hanna-Marja Voipio, Eila Kaliste, Paula Hirsjärvi, Timo Nevalainen, Merel Ritskes-Hoitinga

Ethical evaluation of animal experiments is gaining an increasingly important role in the general review process of animal experiments. In order to discuss various aspects of ethical evaluation, a workshop was organised in Helsinki. Forty participants representing the scientific community, animal welfare organisations and regulators from Nordic and Baltic countries and The Netherlands took part. The workshop was organised by the Cooperation Group for Laboratory Animal Sciences within the Finnish Ministry of Education. Costs refer to the cost or harm to animals when they are used in experiments, and include aspects such as pain, suffering and distress. Benefits are defined as the benefits gained by humans or other target groups resulting from animal studies, e.g. therapies for human diseases, and/or increased knowledge from basic scientific studies. Use of animals is considered ethically justifiable if the benefits outweigh the costs. “Cost modifiers” or “means” can be used to decrease the costs for the animals, e.g. an improved or refined technique that is less distressing to the animal. This will translate to better ethical acceptability. The use of scoring systems for ethical assessment was not supported by the participants since these systems can lead to a false impression of objectivity. A classification of costs versus benefits into three degrees (low, medium and high) was considered the most suitable analysis method. Furthermore, some example protocols were evaluated by the workshop participants; the result revealed a large variation in scoring the degree of costs, the importance of the benefits, and the possibilities of modifying the means. Clearly, further and continuing interaction between all of the interest parties is necessary for the creation of precision tools for ethical assessment of animal studies necessity.