Abstract
Since large animal transgenesis has been successfully attempted for the first time about 25 years ago, the technology has been applied in various lines of transgenic pigs. Nevertheless one of the concerns with the technology—animal welfare—has not been approached through systematic assessment and statements regarding the welfare of transgenic pigs have been based on anecdotal observations during early stages of transgenic programs. The main aim of the present study was therefore to perform an extensive welfare assessment comparing heterozygous transgenic animals expressing GFP with wildtype animals along various stages of post natal development. The protocol used covered reproductory performance and behaviour in GFP and wildtype sows and general health and development, social behaviour, exploratory behaviour and emotionality in GFP and wildtype littermates from birth until an age of roughly 4 months. The absence of significant differences between GFP and wildtype animals in the parameters observed suggests that the transgenic animals in question are unlikely to suffer from deleterious effects of transgene expression on their welfare and thus support existing anecdotal observations of pigs expressing GFP as healthy. Although the results are not surprising in the light of previous experience, they give a more solid fundament to the evaluation of GFP expression as being relatively non-invasive in pigs. The present study may furthermore serve as starting point for researchers aiming at a systematic characterization of welfare relevant effects in the line of transgenic pigs they are working with.
Similar content being viewed by others
References
Brunetti D, Perota A, Lagutina I, Colleoni S, Duchi R, Calabrese F, Seveso M, Cozzi E, Lazzari G, Lucchini F, Galli C (2008) Transgene expression of green fluorescent protein and germ line transmission in cloned pigs derived from in vitro transfected adult fibroblasts. Cloning Stem Cells 10(4):409–419. doi:10.1089/clo.2008.0036
Cabot RA, Kuhholzer B, Chan AWS, Lai L, Park KW, Chong KY, Schatten G, Murphy CN, Abeydeera LR, Day BN, Prather RS (2001) Transgenic pigs produced using in vitro matured oocytes infected with a retroviral vector. Anim Biotechnol 12(2):205
Chaloupkova H, Illmann G, Neuhauserova K, Tomanek M, Valis L (2007) Preweaning housing effects on behavior and physiological measures in pigs during the suckling and fattening periods. J Anim Sci 85(7):1741–1749. doi:10.2527/jas.2006-504
Clark J, Whitelaw B (2003) A future for transgenic livestock. Nat Rev Genet 4(10):825–833. doi:10.1038/nrg1183
Crawley JN (2007) What’s wrong with my mouse? behavioral phenotyping of transgenic and knockout mice, 2nd edn. Wiley-Liss, New York
D’Eath RB (2005) Socialising piglets before weaning improves social hierarchy formation when pigs are mixed post-weaning. Appl Anim Behav Sci 93(3–4):199–211. doi:10.1016/j.applanim.2004.11.019
Duncan IJH (2005) Science-based assessment of animal welfare: farm animals. Rev Sci Tech Off Int Epizoot 24(2):483–492
Einsiedel EF (2005) Public perceptions of transgenic animals. Rev Sci Tech Off Int Epizoot 24(1):149–157
Hadjantonakis AK, Nagy A (2001) The color of mice: in the light of GFP-variant reporters. Histochem Cell Biol 115(1):49–58
Hofmann A, Kessler B, Ewerling S, Weppert M, Vogg B, Ludwig H, Stojkovic M, Boelhauve M, Brem G, Wolf E, Pfeifer A (2003) Efficient transgenesis in farm animals by lentiviral vectors. EMBO Rep 4(11):1054–1060. doi:10.1038/sj.embor.7400007
Kaiser M (2005) Assessing ethics and animal welfare in animal biotechnology for farm production. Rev Sci Tech Off Int Epizoot 24(1):75–87
Kurome M, Ueda H, Tomii R, Naruse K, Nagashima H (2006) Production of transgenic-clone pigs by the combination of ICSI-mediated gene transfer with somatic cell nuclear transfer. Transgenic Res 15(2):229–240. doi:10.1007/s11248-006-0004-5
Laible G (2009) Enhancing livestock through genetic engineering-recent advances and future prospects. Comp Immunol Microbiol Infect Dis 32(2):123–137. doi:10.1016/j.cimid.2007.11.012
Lassen J, Gjerris M, Sandoe P (2005) After Dolly—ethical limits to the use of biotechnology on farm animals. In: Symposium on agricultural and societal implications of contemporary Embry-technologies in farm animals, Copenhagen, Denmark, 12 Jan 2005. Elsevier Science Inc, New York, pp 992–1004. doi:10.1016/j.theriogenology.2005.09.012
Li L, Pang DX, Wang TD, Li ZJ, Chen LM, Zhang MJ, Song N, Nie DB, Chen ZW, Lai LX, Ouyang HS (2009) Production of a reporter transgenic pig for monitoring Cre recombinase activity. Biochem Biophys Res Commun 382(2):232–235. doi:10.1016/j.bbrc.2009.02.146
Litten JC, Drury PC, Corson AM, Lean IJ, Clarke L (2003) The influence of piglet birth weight on physical and behavioural development in early life. Biol Neonate 84(4):311–318. doi:10.1159/000073640
Martin P, Bateson P (1993) Measuring behaviour. Cambridge University Press, UK
Munsterhjelm C, Valros A, Heinonen M, Halli O, Peltoniemi OAT (2008) Housing during early pregnancy affects fertility and behaviour of sows. Reprod Domest Anim 43(5):584–591. doi:10.1111/j.1439-0531.2007.00956.x
Niemann H, Kues W, Carnwath JW (2005) Transgenic farm animals: present and future. Rev Sci Tech Off Int Epizoot 24(1):285–298
Olsson IAS, Sandoe P (2003) Ethical decisions concerning animal biotechnology: what is the role of animal welfare science? In: UFAW International symposium on science in the service of animal welfare, Edinburgh, Scotland, 2–4 Apr 2003. Universities Federation Animal Welfare, pp S139–S144
Pearce GP, Paterson AM (1993) The effect of space restriction and provision of toys during rearing on the behavior, productivity and physiology of male pigs. Appl Anim Behav Sci 36(1):11–28
Puppe B, Tuchscherer A (1999) Developmental and territorial aspects of suckling behaviour in the domestic pig (Sus scrofa f. domestica). J Zool 249:307–313
Rogers DC, Fisher EMC, Brown SDM, Peters J, Hunter AJ, Martin JE (1997) Behavioral and functional analysis of mouse phenotype: SHIRPA, a proposed protocol for comprehensive phenotype assessment. Mamm Genome 8(10):711–713
Smulders D, Verbeke G, Mormede P, Geers R (2006) Validation of a behavioral observation tool to assess pig welfare. Physiol Behav 89(3):438–447. doi:10.1016/j.physbeh.2006.07.002
van der Kooij EV, Kuijpers AH, Schrama JW, van Eerdenburg F, Schouten WGP, Tielen MJM (2002) Can we predict behaviour in pigs? Searching for consistency in behaviour over time and across situations. Appl Anim Behav Sci 75(4):293–305
van der Meer M, Rolls A, Baumans V, Olivier B, van Zutphen LFM (2001) Use of score sheets for welfare assessment of transgenic mice. Lab Anim 35(4):379–389
van Dijk AJ, van Rens BTTM, van der Lende T, Taverne MAM (2005) Factors affecting duration of the expulsive stage of parturition and piglet birth intervals in sows with uncomplicated, spontaneous farrowings. Theriogenology 64(7):1573–1590
Van Reenen CG (2007) Assessing the welfare of transgenic farm animals. In: Engelhard M, Hagen K, Boysen M (eds) Symposium on new application of livestock genetic engineering, Berlin, Germany, 21–22 Sep 2007. Springer, Berlin, pp 119–143
Van Reenen CG, Meuwissen THE, Hopster H, Oldenbroek K, Kruip TAM, Blokhuis HJ (2001) Transgenesis may affect farm animal welfare: a case for systematic risk assessment. J Anim Sci 79(7):1763–1779
Vasey DB, Lillico SG, Sang HM, King TJ, Whitelaw CBA (2009) CMV enhancer-promoter is preferentially active in exocrine cells in vivo. Transgenic Res 18(2):309–314. doi:10.1007/s11248-008-9235-y
Velarde A, Geers R (eds) (2007) On farm monitoring of pig welfare. Wageningen Academic Publishers, Wageningen
Webster J (2005) The assessment and implementation of animal welfare: theory into practice. Rev Sci Tech Off Int Epizoot 24(2):723–734
Webster NL, Forni M, Bacci ML, Giovannoni R, Razzini R, Fantinati P, Zannoni A, Fusetti L, Dalpra L, Bianco MR, Papa M, Seren E, Sandrin MS, Mc Kenzie IFC, Lavitrano M (2005) Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer. Mol Reprod Dev 72(1):68–76. doi:10.1002/mrd.20316
Whitelaw CBA, Radcliffe PA, Ritchie WA, Carlisle A, Ellard FM, Pena RN, Rowe J, Clark AJ, King TJ, Mitrophanous KA (2004) Efficient generation of transgenic pigs using equine infectious anaemia virus (EIAV) derived vector. FEBS Lett 571(1–3):233–236. doi:10.1016/j.febslet.2004.06.076
Zoric M, Nilsson E, Mattsson S, Lundeheim N, Wallgren P (2008) Abrasions and lameness in piglets born in different farrowing systems with different types of floor. Acta Vet Scand 50(1):37
Acknowledgments
This work was performed as part of and partly financed through the EC NEST029025 project INTEGRA, with additional support from The Roslin Institute Strategic Grant funding from the BBSRC. R Huber received a PhD grant (SFRH/BD/36682/2007) and Liliana Remuge a Bolsa de Iniciação Científica grant (C2008-LA770224-BII). by Fundação para a Ciência e a Tecnologia (FCT). Thanks to the Named Veterinary Surgeon and Pig Unit Manager at The Roslin Institute for valuable input, parameter choice and overall support; to farm staff for essential assistance during data collection; to Norrie Russell for photography and to Douglas Vasey for assistance in the Human Approach test. Statistical analysis was performed with help of Armando Teixeira Pinto from the Department of Biostatistics and Medical Informatics, Faculty of Medicine of the University of Porto.
Author information
Authors and Affiliations
Corresponding authors
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Huber, R.C., Remuge, L., Carlisle, A. et al. Welfare assessment in transgenic pigs expressing green fluorescent protein (GFP). Transgenic Res 21, 773–784 (2012). https://doi.org/10.1007/s11248-011-9571-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-011-9571-1