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Inventory of the Spanish Institutions and Scientists Involved in Alternatives to the use of Laboratory Animals (Refinement, Reduction or Replacement)+

Inventory: 18g. Inventory of Institutions

Type of Institution: University

Staff:

Maria Jose González Muñoz, Dr Farmacia, Titular , 30 % time devoted to altern. meth.
Mª Carmen Martínez Para, Dr Farmacia, Catedrática, 10 % time devoted to altern. meth.
Isabel Meseguer Soler, Dr Farmacia, 30 % time devoted to altern. meth.
Mª Victorina Aguliar Vilas, Dr Farmacia, Titular, 40 % time devoted to altern. meth.
Carmen Jose Mateos Vega, Ld Farmacia, Becaria , 80 % time devoted to altern. meth.

Total staff involved in alternative methods is 7 people.

Activities / aims: This university department is mainly involved in non-regulated applied research; it performs toxicology and nutrition testing on a routine basis, and toxicology (neurotoxicity) studies at a research level. Nutrients are routinely evaluated.

The main use of alternative methods is for replacement studies.

Model systems: The model systems used are conventional animal models.

Work lines: Bioavailability of several mineral elements in different food: Cr, Fe, Se, P in breakfast cereals, using an "in vitro" and "in vivo" method. Role of certain minerals in several neurological diseases: determination of Se in the cerebrospinal fluid of Parkinson and Alzheimer patients.

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices rules according to the FDA. The team are available to participate in EU validation programmes.

Sources of financing: Universidad de Alcalá, FIS

Type of Institution: University

Staff:

Jose María Cobo Sanz, Email: nbcobo@nutri.alcala.es, Dr Farmacia, Profesor Universidad Alcalá, 50 % time devoted to altern. meth.
Rafael García-Cañero, Email: rgcanero@hepatexp.cph.es, Dr Ciencias Químicas, Adjunto Jefe Servicio Bioquímica, 90 % time devoted to altern. meth., duties related to altern. meth.: Vocal de la Sociedad Española de Biología Cellular
Javier Pérez de Diego, Dr Farmacia, Profesor Universidad Alfonso X, 20 % time devoted to altern. meth.

Total staff involved in alternative methods is 3 people.

Activities / aims: This university department is mainly involved in basic research and method development; it performs biochemistry, toxicology (mechanisms of toxicity, basal cytotoxicity, carcinogenicity, hepatotoxicity, chronic toxicity) and culture methodology studies on a routine basis, and cell biology and molecular biology studies at a research level. Diverse chemical compounds and wastes are routinely evaluated; pharmaceuticals also being occasionally studied.

The main use of alternative methods is for complementary studies.

Model systems: The model systems used are conventional animal models and in vitro methods: primary culture of dispersed cells (rat liver cells) and cell lines culture (CHO, HL60, Hela, HepG2).

The endpoints employed in vitro are cellular proliferation (fluorescence), metabolic activity (apoptosis), nucleic acids (DNA degradation) and organ-specific indicators (ion uptake, cell membranes).

Experimental systems (examples):

1 HL60 cells, cell cycle and apoptosis, by flow cytometry, gel electrophoresis, DNA degradation, replacement test

Work lines: The most important field is to study the interrelation between different chemicals and cancer and to establish the role of apoptosis as regulator of DNA degradation. Interest in the chemicals that change Na/K across membranes and how this change might affect intracellular pH and stop mitosis.

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices. The team are available to participate in EU validation programmes in relation to biochemical methods applied to cell cultures, detection of carcinogenicity, etc.

Sources of financing: Fondo de Investigación Sanitaria, Dirección General de Investigación y Ciencia, Fondos de Investigación de la Universidad Privada, Diversas ayudas para el Desarrollo de Cooperación con otros laboratorios internacionales.

Type of Institution: University

Staff:

Francisco Javier Moyano López, Email: fjmoyano@ualm.es, Dr Ciencias Biológicas, Titular de Universidad, 50 % time devoted to altern. meth.
Manuel Díaz López, Email: mdiaz@ualm.es, Dr Farmacia, Titular de Universidad, 50 % time devoted to altern. meth.
Francisco Javier Alarcón López, Email: falarcon@ualm.es, Dr Ciencias Biológicas, Becario Postdoctoral, 75 % time devoted to altern. meth.
Tomás Martínez Moya, Email: tomas@ualm.es, Ld Veterinaria, Becario Predoctoral, 100 % time devoted to altern. meth.
Inmaculada Martínez Díaz, Email: imdiaz@ualm.es, Ld Ciencias del Mar, Becaria de Proyecto, 100 % time devoted to altern. meth.

Total staff involved in alternative methods is 5 people.

Activities / aims: This university department is mainly involved in non-regulated applied research, method development and alternatives to animals in education; it performs physiology and nutrition testing on a routine basis. Protease inhibitors and nutrient digestibility are routinely evaluated.

Model systems: The model systems used are in vitro methods -- cell-free systems (digestive enzymes) and education models (computer simulations)

The endpoints employed are in vitro metabolic activity (controlled proteins and fatty acids hydrolysis).

Experimental systems (examples):

1 digestive enzyme from fish, protein hydrolysis, digestibility, replacement test
2 ruminant liquid extract, changes in nutrient content, digestibility, replacement test
3 digestive enzyme from fishes, protein degradation studied by electrophoresis, digestibility studies, replacement test
4 digestive enzyme from fish, protease inhibition studied by zymogramme, protease inhibition, complementary test

Work lines: Design and nutritive evaluation of feed sources for marine fish. Design and nutritive evaluation of artificial feed for larval marine fish. Nutritive evaluation of feed for ruminants

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes in relation to digestibility techniques in vitro.

Sources of financing: 3 CICYT projects on water culture.

Type of Institution: University

Staff:

Ricardo Marcos Dauder, Email: rmd@cc.uab.es, Biólogo, Catedrático, 50 % time devoted to altern. meth.
Amadeu Creus Capdevila, Email: a.creus@uab.es, Biólogo, Prof. Titular, 50 % time devoted to altern. meth.
Noel Xamena López, Email: n.xamena@uab.es, Biólogo, Prof. Titular, 30 % time devoted to altern. meth.
Antonia Velázquez Henar, Email: a.velazquez@uab.es, Biólogo, Prof. Titular, 30 % time devoted to altern. meth.
Jordi Surralles Calonge, Email: j.surralles@uab.es, Biólogo, Post. Doct. Contratado,30 % time devoted to altern. meth.

Total staff involved in alternative methods is 10 people.

Activities / aims: This University department is mainly involved in basic research, non-regulated applied research, method development and method validation; it performs toxicology (mechanisms of toxicity, genotoxicity / mutagenicity, carcinogenicity) testing, genetics and monitoring --chemical, biological- studies on a routine basis. Pharmaceuticals, diverse chemical compounds and pesticides are routinely evaluated; environmental pollutants and physical agents also being occasionally studied.

The main use of alternative methods is for screening and complementary studies.

Model systems: The model systems used are invertebrate animal models (Drosophila), in vitro methods -- primary culture of dispersed cells (human lymphocytes), cell lines culture (human lymphoblast cell lines)--, and human volunteers (biomonitoring) studies.

The endpoints employed are in vivo genetic and cytogenetic lesions and in vitro nucleic acids.

Experimental systems (examples):

1 Drosophila, germinal and somatic mutations, genotoxicity, screening and complementary test
2 human lymphocytes, chromosomal mutations, genotoxicity, screening and complementary test
3 human volunteers, chromosomal mutations, genotoxicity, screening and complementary test

Work lines: Genotoxicity testing of different chemicals, mainly pesticides; biomonitoring of people environmentally, occupationally or therapeutically exposed to genotoxicants; molecular basis of chemically induced mutants; improvement of different tests used in genotoxicity testing; molecular epidemiology.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes in relation to genotoxic evaluation

Sources of financing: CICyT, CIRIT (Generalitat de Catalunya), Environment Programme (EU) and pharmaceutical industries.

Type of Institution: University

Staff:

Montserratt Llagostera Casas, Email: ibm4@cc.uab.es, Dr Biología, Prof. Titular, 20 % time devoted to altern. meth.
J. Antonio Moreno, Ld Biología, Prof. Ayudante, 20 % time devoted to altern. meth.
Maribel Cárdenas, Ld Biología, Estudiante Tercer Ciclo, 70 % time devoted to altern. meth.
Mirle Ferrer, Ld, Becaria ICI, 100 % time devoted to altern. meth.

Total staff involved in alternative methods is 4 people.

Activities / aims: This university department is mainly involved in basic research and non-regulated applied research; it performs toxicology (genotoxicity / mutagenicity, ) testing on a routine basis. Diverse chemical compounds, pesticides, biomaterials, environmental pollutants and wastes are routinely evaluated.

Model systems: The model systems used are in vitro methods: micro-organisms.

The endpoints employed in vitro are biotransformation systems (vegetable enzymes: peroxidase, cit P450).

Experimental systems (examples):

1 Ames assay, reversion of mutation, vegetal promutagens activation, complementary test
2 micro-organisms, SOS gene induction, by betagalactosidase activity, comparison with Ames test, complementary test

Work lines: The study of the capability of plants to metabolize environmental pollutants to mutagenic metabolites and the identification of plant enzymatic activities involved in this process.

Quality assurance / Validation programmes: The team have previously been involved in alternative method validation programmes. They are available to participate in EU validation programmes in relation to genotoxicity and environmental mutagenesis

Sources of financing: CE, DGCYT, Generalitat de Catalunya

Type of Institution: University

Staff:

María José Hazen de San Juan, Email: mariajose.hazen@uam.es, Dra Biología, Profesor Titular, 80 % time devoted to altern. meth.
María Luisa Molero Vergara, Dr Biología, Profesor Titular, 80 % time devoted to altern. meth.
Ana Isabel Pérez Gorroño, Ld Biológia, 40 % time devoted to altern. meth.
Nuria Ortiz Movilla, Ld Biología, 40 % time devoted to altern. meth.
Miguel Angel Fernández, Técnico, 25 % time devoted to altern. meth.

Total staff involved in alternative methods is 5 people.

Activities / aims: This university department is mainly involved in non-regulated applied research and method development; it performs cell biology, toxicology (mechanisms of toxicity, basal cytotoxicity, phototoxicity) testing on a routine basis and vegetal biology studies at a research level. Diverse chemical compounds and colourings are routinely evaluated; physical agents also being occasionally studied.

The main use of alternative methods is for replacement studies.

Model systems: The model systems used are in vitro methods - cell lines culture (Hela, 3T3 mouse fibroblasts, Vero).

The endpoints employed in vitro are cell morphology (scanning and transmission electronic microscopy) and cell viability (MTT reduction, neutral red uptake, total protein content, LDH).

Experimental systems (examples):

1 Hela tumour human cells, basal cytotoxicity by spectrophotometry, photodynamics, replacement test
2 3T3, Vero, basal cytotoxicity by spectrophotometry, photodynamic compounds analysis, replacement test
3 3T3, Vero, basal cytotoxicity by spectrophotometry, participation of oxygen free-radicals, replacement test

Work lines: The major interest of this lab is focused on the development of in vitro protocols which allow a rapid and reliable analysis of the photosensitizing properties of chemical compounds. The applications of the work are both to define potential photochemotherapeutic agents and to search for new phototoxic agents.

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes.

Fuente / Source: Guillermo Repetto, Ana del Peso, Manuel Salguero, Manuel Repetto (1999) Inventory of the Spanish Institutions and Scientists Involved in Alternatives to the use of Laboratory Animals (Refinement, Reduction or Replacement) Revista de Toxicología 16: 50-127.