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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: 18e. Inventory of Institutions

Type of Institution: Governmental facility for service (diagnostic, control, monitoring)

Staff:

Jesus Salas Zapatero, Email: istzo350@tsai.es, Ld Veterinaria, Jefe de Servicio de Productos Alimenticios.
Pilar Moya Esteve, Dr Biología, Unidad de Productos de Origen Animal
Angel Blazquez de los Riscos, Ld Veterinaria, Unidad Electroquímica y PCR
Mª Rosa Sarazá Linares, Ld Veterinaria, Unidad Microbiología.
Mª Teresa González Villa, Ld Biología, Unidad Microbiología.
Mª del Pilar Clemente Belmonte, Ld Biología, Unidad de Parasitología.
Raquel Fernández Peiteado, Ld Biología, Unidad de Productos de Origen Animal y Vegetal
Carlos Arnaiz Ronda, Dr Biología, Director Técnico del Centro de Investigación y Desarrollo

Total staff involved in alternative methods is 8 people.

Activities / aims: This governmental service facility (diagnostic, control, monitoring) department is mainly involved in regulatory testing, method validation, quality control of food and industrial products; it performs toxicology (mechanisms of toxicity, basal cytotoxicity, acute systemic toxicity, ocular cytotoxicity, dermal irritation and corrosivity, nephrotoxicity, respiratory cytotoxicity, reproductive cytotoxicity, neurotoxicity, immunotoxicity and sensitisation, haematoxicity, genotoxicity / mutagenicity, carcinogenicity, hepatotoxicity, endocrine disruption, chronic toxicity, integrated test strategies), nutrition studies and quality control testing on a routine basis. Cosmetics, diverse chemical compounds, pesticides, food additives, colourings and wastes are routinely evaluated.

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

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

Work lines: Quality control of food and related products of human use. Evaluation of toxicological tests for the evaluation of pesticides, food additives and food traces. The use of mutagenicity in vitro tests applied to quality control activities.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes, in relation to in vitro methods for detecting toxic substances in food products

Sources of financing: Self financing

Type of Institution: Governmental research facility

Staff:

María Argelia Castaño Calvo, Email: castano@inia.es, Dr Ciencias Biológicas, Investigador A3, 90 % time devoted to altern. meth.
María Jesús Muñoz Reoyo, Email: reoyo@inia.es, Dr Ciencias Biológicas, Investigador A3, 80 % time devoted to altern. meth.
Matilde Carballo Santaolaya, Email: carballo@inia.es, Dr Ciencias Biológicas, Investigador A3, 40 % time devoted to altern. meth.
María Teresa Llorente Rodríguez, Email: mteresa@inia.es, Ld Ciencias Biológicas, Becaria Predoctoral INIA, 100 % time devoted to altern. meth.
Apolonia Novillo Villajo, Email: novillo@inia.es, Dr Ciencias Biológicas, Contratada Postdoctoral, 100 % time devoted to altern. meth.
Ana Isabel de la Torre Reoyo, Email: torre@inia.es, Dr Ciencias Biológicas, Becaria Postdoctoral INIA, 50 % time devoted to altern. meth.
Jaime Roset Álvarez, Email: roset@inia.es, Ld Ciencias Biológicas, Becario Predoctoral INIA, 50 % time devoted to altern. meth.
María Luisa Cuéllar Cuéllar, FPII, Ayudante de Laboratorio de Plantilla, 70 % time devoted to altern. meth.

Total staff involved in alternative methods is 8 people.

Activities / aims: This governmental research facility department is mainly involved in non-regulated applied research, method development and method validation; it performs toxicology testing (basal cytotoxicity, reproductive cytotoxicity, genotoxicity / mutagenicity, ecotoxicity), monitoring -chemical, biological studies on a routine basis. Diverse chemical compounds, pesticides, environmental pollutants and wastes are routinely evaluated.

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

Model systems: The model systems used are conventional animal models (sea and freshwater fishes), invertebrates (Daphnia magna, Folsomia candida, Enchitraeus albidus, Eisenia foetida), embryos (medaka Oryzias latipes), vegetables (seed germination and unicellular algae), and in vitro methods - primary culture of dispersed cells (fish O mykiss macrophages), and cell lines culture (RTG-2, CHSE-214, EPC).

The endpoints employed are in vivo stress parameters, micronuclei induction, metahaemoglobin, retinol, acetylcholinesterase, P-450 induction, macroscopic alterations of development, larva viability, histopathologic alterations in fish species and in vitro cell viability (neutral red uptake, exclusion of dyes (trypan blue, ethidium bromide)), cellular proliferation (total protein content (kenacide blue, Bradforf method)) and metabolic activity (ATP intracellular content).

Experimental systems (examples):

1 RTG-2 rainbow trout fibroblast cell line, neutral red uptake, total protein content, ATP, acute cytotoxicity, replacement test
2 RTG-2 rainbow trout fibroblast cell line, micronuclei frequency and cell cycle alterations, by flow cytometry, detection of genotoxicity
alterations, replacement test
3 Daphnia magna and Dahpnia pulex, immobilization and reproduction, acute and chronic toxicity
4 Chlorella vulgaris, growth, by spectrophotometry on 96-well, sub-acute toxicity, replacement test
5 eggs and larva of medaka (Oryzias latipes), alterations in development, viability. Stereomicroscopic observation, lethality, teratogenesis, chronic toxicity, replacement test
6 Folsomia candida and Enchitraeus albidus, alterations in reproduction, acute toxicity, by count and uptake, lethality and reproduction, replacement test

Work lines: Diagnosis and risk assessment of environmental toxicological processes by the development of new miniaturized short-term tests for soil and water (on fish cell lines, algae, invertebrates, multispecies, microcosms test). Invertebrates and fish biomarkers: stress, cytochrome P-450, ACTH, micronuclei frequency by flow cytometry and mutations detection by DNA fingerprinting.

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices rules according to the EU, OECD and the FDA The team have previously been involved in the following alternative method validation programmes: ring test on reproduction of Daphnia magna, MEIC and EDIT (RTG2 cytotoxicity test, and ovoalbumin denaturalization test). They are available to participate in EU validation programmes.

Sources of financing: Project AIR1-AZ92-0036 New in vitro approaches for fish protection in aquaculture: application of cell and tissue cultures for pathogen detection and for vaccines development. --Commission of the European Communities 1992-1996; Proyecto CAM AMB COR 0010/94 Valoración ecotoxicológica del Parque Sudeste de la Comunidad de Madrid. Incorporación de bioensayos de toxicidad en programas de seguimiento y control. Comunidad de Madrid 1994-1997; Proyecto AMB 94-0655-C02-01 Desarrollo y validación en condiciones de campo de sistemas de detección y biomarcadores de genotoxicidad en organismos acuáticos CICYT 1994-1997; Proyecto SC98-098-C2 Identificación y valoración de impactos medioambientales de las explotaciones porcinas. Adecuación a las nuevas directivas de la Unión Europea. INIA 1998-2001; Proyecto SC95-091. Desarrollo y validación de sistemas de detección de efluentes y vertidos. INIA 1995-1997; Aspectos toxicológicos y medioambientales relacionados con el regadio, la utilización de aguas residuales y el estudio de sistemas de depuración por lagunaje. IRYDA 1994-1997 (15750); Proyecto AMB 97-0431-C02-01 Valoración ecotoxicológica de efluentes industriales sobre organismos acuáticos. Validación de sistemas alternativos y biomarcadores de exposición de genotoxicidad CICYT 1997-2000 (5006); Caracterización toxicológica de vertidos y aguas residuales de la industria química. Infraestructura y Ecología 1996-1997 (7000)

Type of Institution: Governmental service facility

Staff:

Françoise Lhoest Mathijsen, Ld Biología, Facultativo, 10 % time devoted to altern. meth.


Total staff involved in alternative methods is 1 person.

Activities / aims: This governmental service facility department (diagnostic, control, monitoring, expert work) is mainly involved in regulatory testing; it performs toxicology (ecotoxicity) testing on a routine basis. Environmental pollutants and wastes are routinely evaluated; pesticides also being occasionally studied.

The main use of alternative methods is for complementary studies.

Model systems: The model systems used are animal models -- invertebrates (Daphnia magna) and in vitro methods -- micro-organisms (Photobacterium phosphoreum).

The endpoints employed are in vivo (immobilization) and in vitro metabolic activity (bioluminescence inhibition).

Experimental systems (examples):

1 Daphnia magna , immobilization, characterisation of wastes, replacement test
2 Photobacterium phosphoreum, bioluminescence inhibition, characterization of wastes, replacement test
3 cell culture, cell viability, cytotoxicity, complementary test

Work lines: Toxicological evaluation of wastes using ecotoxicity assays.Determination and evaluation of acute and chronic toxicity of pharmaceuticals using cell lines

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes.

Sources of financing: Self financing

Type of Institution: Governmental service facility

Staff:

Pilar Gascó Alberich, Email: extox@mad.inaltox.es, Dr Farmacia, Jefe Sec Experimentación Toxicológica. Prof Asociado, 20 % time devoted to altern. meth.
Javier Piga de Lariba, Ld Ciencias Biológicas, Facultativo Sección Experimentación Toxicológica, 20 % time devoted to altern. meth.

Total staff involved in alternative methods is 2 people.

Activities / aims: This governmental service facility department (diagnostic, control, monitoring, expert work) is mainly involved in non-regulated applied research, regulatory testing and alternatives to animals in education; it performs toxicology (ecotoxicity) testing on a routine basis and toxicology (dermal irritation and corrosivity, hepatotoxicity) studies at a research level. Environmental pollutants and wastes are routinely evaluated; diverse chemical compounds also being occasionally studied.

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

Model systems: The model systems used are conventional animal models (rat, mouse, rabbit), invertebrates (Daphnia magna), vegetables (algae), in vitro methods -- micro-organisms (Photobacterium phosphoreum) and education models (hepatocytes).

The endpoints employed are in vivo (toxicity/ lethality, immobilization, growth) and in vitro cell viability (dyes) and

metabolic activity (inhibition of bioluminescence).

Experimental systems (examples):

1 Daphnia magna, immobilization, visual, CE50, ecotoxicity
2 Chlorella vulgaris, inhibition of growth, by spectrophotometry, CI50, ecotoxicity
3 Photobacterium phosphoreum, bioluminescence inhibition, CE50, ecotoxicity

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes related to ecotoxicity and toxicology.

Type of Institution: Governmental service facility

Staff:

Carmen Rodríguez Vicente, Dr Ciencias Químicas y Farmacia, Jefe de Servicio, 50 % time devoted to altern. meth.
Patrocinio Villar López, Ld Ciencias y Farmacia, Jefe Servicio Criminalística, 20 % time devoted to altern. meth.
Carmen Martínez Sánchez, Dr Medicina, Especialista en Anatomía Patológica, Facultativo, 10 % time devoted to altern. meth.
Josefa Valenzuela Moreno, Capacitación para Animales de Laboratorio, Auxiliar de Laboratorio, 80 % time devoted to altern. meth.
Josefina Barba Quintero, Grado Medio, Auxiliar de Laboratorio, 20 % time devoted to altern. meth.

Total staff involved in alternative methods is 5 people.

Activities / aims: This governmental service facility department (diagnostic, control, monitoring, expert work) is mainly involved in basic research, regulatory testing and method development; it performs biochemistry and toxicology (mechanisms of toxicity, acute systemic toxicity, hepatotoxicity, chronic toxicity, ecotoxicity) testing on a routine basis. Pharmaceuticals, cosmetics, pesticides, drugs of abuse, environmental pollutants and wastes are routinely evaluated; toxins, food additives and colourings also being occasionally studied.

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

Model systems: The model systems used are conventional animal models (BrSelf Norway: nefrotoxicity studies), invertebrates (Daphnia magna), vegetables (Algae: Chlorella vulgaris), in vitro methods - micro-organisms (Photobacterium phosphoreum) and human volunteers.

The endpoints employed are in vivo (enzyme activities, haematology, biochemical parameters, histological observations, behavioural tests) and in vitro metabolic activity (bioluminescence).

Experimental systems (examples):

1 Photobacterium phosphoreum, bioluminescence inhibition, ecotoxicological evaluation, replacement test
2 brSelf Norway rats, biochemical evaluation: NAG creatinine, urea, bilirrubin, nephrotoxicity studies, refinement
3 rabbits, ADH, hair accumulation of ethanol, chronic alcoholism refinement
4 fauve Bourgogne red-haired rabbits, hair and cannabis accumulation, toxicokinetics of abused drugs, refinement
5 wistar rats, enzymes. GOT, GPT, transaminases, bilirrubin, pharmaceutical toxicity, refinement
6 rat, rabbit, human, Ache, Che inhibition, interference of pharmaceuticals, refinement

Work lines: The ecotoxicological evaluation of wastes and waters by means of Photobacterium phosphoreum, Daphnia magna and Chlorella vulgaris tests. Toxicokinetic studies of abused drugs on fauve Bourgogne red-haired rabbits and their application for forensic purposes. Nephrotoxicity studies using sensitive models such as the brSelf Norway rats. The hepatotoxic effects of pharmaceuticals on Wistar rats. The neurotoxicity of pesticides.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes.

Sources of financing: Self financing, Junta de Andalucía, food additives and pharmaceutical industries.

Type of Institution: Governmental service facility (diagnostic, control, monitoring, expert work)

Staff:

Guillermo Repetto Kuhn, Email: repetto@sev.inaltox.es, Dr Medicina, Facultativo, 50 % time devoted to altern. meth., duties related to altern. meth.: Coordinador del GTEMA, Representante español en ERGATT
Pilar Sanz Nicolás, Email: biol@sev.inaltox.es, Dr Biología, Jefe Servicio Biología, 10 % time devoted to altern. meth.
Antonio Garfia, Dr Medicina, Jefe Servicio Anatomía Patológica
Ana del Peso Bejarano, Ld Farmacia, Facultativo Servicio Química, 30 % time devoted to altern. meth.
Inmaculada Flores García, Ld Farmacia, Facultativo Servicio Biología, 10 % time devoted to altern. meth.
Susana Jiménez Fuentes, Técnico Auxiliar de Farmacia, Agente de Laboratorio Servicio Biología, 50 % time devoted to altern.meth.
Dolores Osuna Guillén, Grado Medio, Auxiliar de Laboratorio Servicio Biología, 50 % time devoted to altern. meth.
Manuel Salguero Villadiego, Dr Medicina, Especialista Anatomía Patológica, Profesor Universidad, 40 % time devoted to altern. meth.
Manuel López Soto, Ld Biología, Facultativo de Biología, 50 % time devoted to altern. meth.
M Rosario Repetto Kuhn, Dr Medicina, Servicio de Información Toxicológica, 10 % time devoted to altern. meth.
Manuel Repetto Jiménez, Dr Ciencias y Medicina, Director INT-SE, 10 % time devoted to altern. meth.

Total staff involved in alternative methods is 10 people.

Activities / aims: This governmental service facility department (diagnostic, control, monitoring and expert work) is mainly involved in basic research, non-regulated applied research, regulatory testing, method development, method validation and alternatives to animals in education; it performs biochemistry and toxicology (mechanisms of toxicity, basal cytotoxicity, acute systemic toxicity, neurotoxicity, ecotoxicity) testing on a routine basis, and cell biology, molecular biology, biokinetics and biotransformation, genetics, pathology, nutrition, diagnostics and monitoring --chemical, biological-- studies at a research level. Pharmaceuticals, cosmetics, diverse chemical compounds, pesticides, drugs of abuse, environmental pollutants and wastes are routinely evaluated; toxins, food additives and colourings also being occasionally studied.

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

Model systems: The model systems used are animal models -- invertebrates (crayfish Procambarus clarkii, medaka), embryos (chick embryo), vegetables (chloroplasts), in vitro methods -- micro-organisms (E coli), organ culture (skin), primary culture of dispersed cells (human lymphocytes), cell lines culture (Neuro 2a, SH-5Y5Y, Vero, RTG-2), cell-free systems (microsomal), human volunteers (monitoring), education models (computer models) and mathematical modelling (toxic effects, education).

The endpoints employed in vitro are morphology (optical microscopy, immunohistochemistry), cell viability (neutral red uptake, propidium iodide by spectrophotometry), cytoskeleton/membranes/enzyme release (LDH leakage, hexosaminidase release, neurofilaments), cellular proliferation (total protein content, DNA), metabolic activity (PFK, LDH, ATPase, SDH), nucleic acids (apoptosis / necrosis by TUNEL and gel electrophoresis), biotransformation systems (P450), defence systems (GOR, GST, G6P-DH, SOD) and organ-specific indicators (acetylcholinesterase, enolase, galactosidase, TAU protein).

Experimental systems (examples):

1 SH-SY5Y human differentiated neuroblastoma cells, apoptosis by TUNEL and gel electrophoresis differentiation, cell death mechanisms, replacement test
2 SH-SY5Y human differentiated neuroblastoma cells, acetylcholinesterase, enolase, galactosidase Tau protein, neurofilaments by immunocytochemistry, neurotoxicity mechanisms, replacement test
3 Neuro-2a mouse neuroblastoma cell culture, ATPase, phosphofructokinase, LDH, SDH, metabolic alterations, replacement test
4 Neuro-2a mouse neuroblastoma cell culture, neutral red uptake, MTT reduction, cell proliferation, basal cytotoxicity, replacement test
5 Monkey kidney Vero cell culture, glutathione reductase, glutathione transferase, glucose-6P-DH, SOD, antioxidative mechanisms, replacement test
6 RTG-2 rainbow trout fibroblasts cell proliferation by total protein content, ecotoxicity tests, replacement test
7 rat microsomal fraction, pyrrole adduct formation, by spectrofluorimetry, metabolic activation of diketone neurotoxic solvents, replacement test
8 muscle microsomal fractions of the crayfish Procambarus clarkii, acetylcholinesterase, neuropathy-target sterase, ecotoxicity effects of environmental pollutants in invertebrates, replacement test

Work lines: The development and use of in vitro methods applied to different materials and areas. The development of models of human neuronal differentiation and the interaction produced by drugs of abuse. The neurotoxic effects of environmental pollutants on mouse and human neuroblastoma cells, as alternative procedures to the use of animals. Interference in chick embryo development is studied. The investigation of chemical induced oxidative stress using Vero monkey kidney cells. The effects produced on very simple vegetal systems, including chloroplasts. As ecotoxicological tests, in vivo and in vitro cell fractions from the crayfish Procambarus clarkii are used in parallel to cultures of fish cell lines (RTG-2).

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes.

Sources of financing: Aid for the consolidation of research groups. Junta de Andalucia. Desarrollo de un modelo in vitro con celulas de neuroblastoma para la evaluación de la neurotoxicidad de compuestos orgánicos. Desarrollo de modelos experimentales in vitro para la evaluacion de la toxicidad de compuestos metalicos. CICYT. Evaluacion de los efectos de la cocaina sobre celulas de neuroblastoma. Comunidad de Madrid. Studies for the pharmaceutical, cosmetic, and food additive industries. Inventory of Spanish institutions and Scientists interested in Alternative Methods. European Union.

Others: The team provides help for activities organized by GTEMA- Grupo de Trabajo Especializado en métodos Alternativos (http://tox.umh.es/aet/gtema) and REMA- Red Española de métodos Alternativos (http://tox.umh.es/rema/).

Type of Institution: Governmental research facility

Staff:

Ana I. Ortiz, Dr Veterinaria, Jefe de Servicio, 50 % time devoted to altern. meth.
Ana Santa Maria, Ingeniero T. Agrícola, Jefe de Sección, 80 % time devoted to altern. meth.
Alicia Galindo, Ld. Ciencias Bilógicas, Facultativo, 50 % time devoted to altern. meth.
Maria del Mar Díaz Llorente, Ld. Farmacia, Ax Investigación, 100 % time devoted to altern. meth.
Ana López Jiménez, Ld. Farmacia, Ax. Investigación, 100 % time devoted to altern. meth.

Total staff involved in alternative methods is 5 people.

Activities / aims: This governmental research facility department is mainly involved in basic research and non-regulated applied research; it performs toxicology (basal cytotoxicity, genotoxicity / mutagenicity) testing on a routine basis. Diverse chemical compounds, food additives and colourings are routinely evaluated; pharmaceuticals, pesticides and wastes also being occasionally studied.

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

Model systems: The model systems used are in vitro methods -- cell lines culture (CHO-K1, Vero, A-549, Hep-2, NRK, BGM, RD).

The endpoints employed are in vitro cell viability, cellular proliferation and metabolic activity.

Experimental systems (examples):

1 Chinese hamster ovary cell line (CHO), neutral red uptake, cytotoxicity, replacement test
2 Chinese hamster ovary cell line (CHO), total protein content, cytotoxicity, replacement test
3 Chinese hamster ovary cell line (CHO), MTT reduction, cytotoxicity, replacement test
4 Chinese hamster ovary cell line (CHO), gene mutation, micronuclei, genotoxicity, replacement test

Work lines: Cytotoxicity assays: neutral red (NR) uptake test; total protein content (kenacide blue (KB)) test, MTT assay; genotoxicity-mutagenicity: gene mutation assay, micronucleus assay.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The team are available to participate in EU validation programmes.

Sources of financing: Comunidad Autónoma de Madrid, Fondo de Investigación Sanitaria (FIS)

Type of Institution: Governmental research facility

Staff:

Ingrid Maive Outschoorn, Email: ioutscho@isciii.es, Dr BSc (St Andrew´s), DEA (Montpellier), Mphill (Columbia), Dr Ciencias , J. Sección Respuesta Inmune, 100 % time devoted to altern. meth., duties related to altern. meth.: FELASA Workshop Participant Utrecht ND
José Díaz Romero. Dr., Novartis, Suiza.
Jesus Colino Gutierrez, Dr, Univ Health Sciences, Jones Br, Rd, Bethesda, US

Total staff involved in alternative methods is 3 people.

Activities / aims: This governmental research facility department is mainly involved in basic research, regulatory testing, method development, method validation and alternatives to animals in education; it performs cell biology, molecular biology, biochemistry, physiology, biokinetics and biotransformation, toxicology (basal cytotoxicity, reproductive, cytotoxicity, immunotoxicity and sensitisation, haematoxicity, hepatotoxicity, endocrine disruption, chronic toxicity, ecotoxicity, integrated test strategies), nutrition, quality control, culture methodology testing studies on a routine basis, and vegetal biology, pathology, diagnostics (autoimmune infections), monitoring --chemical, biological-- studies at a research level. Vaccines, hormones, pharmaceuticals and environmental pollutants are routinely evaluated.

The main use of alternative methods is for screening.

Model systems: The model systems used are conventional animal models (hamster, rabbit, rat, mice, monkey), embryos (mouse, hamster), vegetables (fungi) and in vitro methods, including micro-organisms (Candida albicans, yeast, neisseria, coxiella, ricketsia), primary culture of dispersed cells (thymus, thyroid, spleen [mice, human]) and cell-free systems (lymphoid).

The endpoints employed are in vitro cellular proliferation (Tyd incorporation), cell signalling (surface marker expression), nucleic acids (DNA proliferation, mitochondrial DNA in ageing models), defence systems (antibodies, cytokines) and organ-specific indicators (tyroglobulin, microsomal).

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

Sources of financing: FIS, Instituto Universitario de Estudios de la Mujer, Universidad Autónoma de Madrid

Type of Institution: Governmental research facility

Staff:

Bartolomé Ribas Ozonas, Email: bribas@isciii.es, Dr Medicina y Dr Farmacia, Jefe de Area, 55 % time devoted to altern. meth., duties related to altern. meth.: Director de Estudio
Mercedes Nuñez García, Email: mnuñez@isciii.es, Ld. Ciencias Biológicas, Facultativo Especialista, 80 % time devoted to altern. meth., duties related to altern. meth.: Responsable Unidad Laboratorio
Macarena Gamoneda López-Ibor, Email: gamoneda@isciii.es, Ld. Ciencias Químicas, Becaria, 25 % time devoted to altern. meth.
Miguel González-Doncel, Ld. Ciencias Biológicas, Becario, 40 % time devoted to altern. meth.
Paloma Mendez Ureña, Auxiliar de Investigación, 30 % time devoted to altern. meth.
Catalina Andrés Ratón, Auxiliar de Investigación, 40 % time devoted to altern. meth.
Mercedes Fernández de la Puebla, T. E. S. A., Becaria.
Isabel Orgaz Ariza, Email: iorgaz@isciii.es, Ld Ciencias Químicas, Jefe de Sección, 80 % time devoted to altern. meth., duties related to altern. meth.: Responsable Unidad Laboratorio
Javier Méndez González, Email: jmendez@isciii.es, Dr Ciencias Químicas, Jefe de Servicio, 100 % time devoted to altern. meth., duties related to altern. meth.: Responsable Técnico
Rosario Mancha López-Jurado, Email: rmancha@isciii.es Ld, Jefe de Sección., duties related to altern. meth.: Responsable Unidad Laboratorio
Gema Díaz López, Email: gdiaz@isciii.es, Facultativo Especialista, duties related to altern. meth.: Responsable Unidad Laboratorio

Total staff involved in alternative methods is 15 people.

Activities / aims: This governmental research facility department is mainly involved in basic research, non-regulated applied research, regulatory testing, method development, method validation and alternatives to animals in education; it performs pharmacodynamics (dose-response relationships), toxicology (mechanisms of toxicity, basal cytotoxicity, acute systemic toxicity, reproductive cytotoxicity, haematoxicity, genotoxicity / mutagenicity, carcinogenicity, hepatotoxicity and ecotoxicity) studies, monitoring --chemical, biological--, quality control and risk assessment testing on a routine basis, and cell biology and molecular biology studies at a research level. Diverse chemical compounds, environmental pollutants and wastes are routinely evaluated; pesticides and toxins also being occasionally studied.

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

Model systems: The model systems used are animal models --conventional, invertebrates (Parasarcoph. argyrostoma, fish, Daphnia, algae, earthworm,vegetables, aerobic micro-organisms, fish embryo, microcrustaceans, -- vegetables (growth and development of seeds) and in vitro methods, including micro-organisms (mixed inoculum), primary culture of dispersed cells (human lymphocytes) and cell lines culture (HV-60, neurones).

The endpoints employed are in vivo (metallothioneins, G6PDH) and in vitro cellular proliferation (J Immunol Methods 65 (1983),55).

Experimental systems (examples):

1 microbial mixed inoculum, respiration inhibition, toxicity evaluation, complementary test
2 rotifers, mortality (CE50), toxicity evaluation, screening
3 microcrustaceans(Thaminocephalus platyurus), mortality (CE50), toxicity evaluation,screening
4 medaka fish (Oryzius latypes), sublethal effects, embryotoxicity evaluation
5 rats, rabbits, mice, metallothioneine metals in each isoform, by HPLC, capillary electrophoresis, specific bioindicators
6 rat tissue culture and treatments, glutathione, metallothioneine, redox state evaluation in other sulphur compounds by HPLC, capillary electrophoresis

Work lines: Biomedical effects of electromagnetic fields in extremely low frequency and in radiofrequency under the auspices of two projects and the collaboration of the Department of Physics University Complutense of Madrid, CSIC, and the Clinical Hospital of Madrid. Evaluation and validation of the new proposed technique on a Diptera larvae as alternative method. The evaluation of redox state by different sulphur compounds in degenerative diseases. Risk assessment.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU. The team have previously been involved in the following alternative method validation programmes: ring tests of a method for ready biodegradability (OECD). They are available to participate in EU validation programmes, in relation to validation of the proposed diptera larvae.

Sources of financing: CAM, FIS, CAICYT, Autonomic Community of Madrid and collaboration with ENDESA, CONAMA

Others: The validation of the diptera larvae method is proposed.

Type of Institution: Governmental research facility

Staff:

Carmen Barrueco Fdez-Cuervo, Email: cbarrue@isciii.es, Dra Ciencias Biológicas, Técnico de Gestión de Organismos Autónomos, 70 % time devoted to altern. meth., duties related to altern. meth.: Miembro del GTEMA

Total staff involved in alternative methods is 1 person.

Activities / aims: This governmental research facility department is mainly involved in risk assessment; it performs toxicology (genotoxicity / mutagenicity) testing on a routine basis. Pesticides are routinely evaluated.

The main use of alternative methods is for replacement studies.

Model systems: The model systems used are in vitro methods --micro-organisms (Salmonella typhimurium), primary culture of dispersed cells (human lymphocytes), cell lines culture (CHO), cell-free systems (microsomal fraction)-- and human volunteers (workers exposed to pesticides).

The endpoints employed are in vivo induction of chromosomal aberrations, SCE and micronuclei in human lymphocytes from exposed workers and in vitro nucleic acids reversion of mutation, gene mutations, structural mutations, SCE and micronuclei and biotransformation systems (rat liver microsomal fractions).

Experimental systems (examples):

1 Salmonella typhimurium, reversion of mutation, by count on solid medium, detection of gene mutation as genotoxicity, screening
2 human lymphocytes and CHO cell line, structural chromosomal aberrations, cytogenetic analysis of metaphases, in vitro genotoxicity, screening
3 human lymphocytes, micronuclei by cytogenetic study of cells or interphase, detection of clastogens and aneugens, screening
4 human lymphocytes and CHO cell line, SCE- sister chromatid exchange, cytogenic lesions to DNA, screening

Work lines: Genotoxicity evaluation of pesticides using in vitro and in vivo assays for determining different endpoints (gene mutation, chromosomal aberration and DNA effects). Evaluation of the report for new pesticides registration according to Annex I of Directive 91/414/EEC.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU and OECD. The team are available to participate in EU validation programmes related to genotoxicity assays.

Sources of financing: Instituto de Salud Carlos III, INIA

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.