European Union research and innovation perspectives on biotechnology

"Food, Agriculture and Fisheries and Biotechnology" is one of 10 thematic areas in the Cooperation programme of the European Union's 7th Framework Programme for Research, Technological Development and Demonstration Activities (FP7). With a budget of nearly €2 billion for the period 2007-2013, its objective is to foster the development of a European Knowledge-Based Bio-Economy (KBBE) by bringing together science, industry and other stakeholders that produce, manage or otherwise exploit biological resources. Biotechnology plays an important role in addressing social, environmental and economic challenges and it is recognised as a key enabling technology in the transition to a green, low carbon and resource-efficient economy. Biotechnologies for non-health applications have received a considerable attention in FP7 and to date 61 projects on industrial, marine, plant, environmental and emerging biotechnologies have been supported with a contribution of €262.8 million from the European Commission (EC). This article presents an outlook of the research, technological development and demonstration activities in biotechnology currently supported in FP7 within the Cooperation programme, including a brief overview of the policy context.     

Biotechnology as the engine for the Knowledge-Based Bio-Economy

Abstract

The European Commission has defined the Knowledge-Based Bio-Economy (KBBE) as the process of transforming life science knowledge into new, sustainable, eco-efficient and competitive products. The term “Bio-Economy” encompasses all industries and economic sectors that produce, manage and otherwise exploit biological resources and related services. Over the last decades biotechnologies have led to innovations in many agricultural, industrial, medical sectors and societal activities. Biotechnology will continue to be a major contributor to the Bio-Economy, playing an essential role in support of economic growth, employment, energy supply and a new generation

FP7 provides the research community with funding certainty over the next few years. One of the FP7 thematic priorities is dedicated to the strengthening the European knowledge-based bio-economy bringing together science, industry and relevant stakeholders from Europe and the rest of the world. The conditions are, therefore, favourable towards the sustainable development and deployment of biotechnologies as an engine for the knowledge-based bio-economy.     

Comparative development of knowledge-based bioeconomy in the European Union and Turkey

Abstract

Biotechnology, defined as the technological application that uses biological systems and living organisms, or their derivatives, to create or modify diverse products or processes, is widely used for healthcare, agricultural and environmental applications. The continuity in industrial applications of biotechnology enables the rise and development of the bioeconomy concept. Bioeconomy, including all applications of biotechnology, is defined as translation of knowledge received from life sciences into new, sustainable, environment friendly and competitive products. With the advanced research and eco-efficient processes in the scope of bioeconomy, more healthy and sustainable life is promised. Knowledge-based bioeconomy with its economic, social and environmental potential has already been brought to the research agendas of European Union (EU) countries. The aim of this study is to summarize the development of knowledge-based bioeconomy in EU countries and to evaluate Turkey’s current situation compared to them. EU-funded biotechnology research projects under FP6 and FP7 and nationally-funded biotechnology projects under The Scientific and Technological Research Council of Turkey (TUBITAK) Academic Research Funding Program Directorate (ARDEB) and Technology and Innovation Funding Programs Directorate (TEYDEB) were examined. In the context of this study, the main research areas and subfields which have been funded, the budget spent and the number of projects funded since 2003 both nationally and EU-wide and the gaps and overlapping topics were analyzed. In consideration of the results, detailed suggestions for Turkey have been proposed. The research results are expected to be used as a roadmap for coordinating the stakeholders of bioeconomy and integrating Turkish Research Areas into European Research Areas.     

TRAIN: Training through Research Application Italian iNitiative

Training through Research Application Italian iNitiative (TRAIN) is a mobility program financed under the EU action called "Cofinancing of regional, national and international programs" (COFUND) of the European Commission Seventh Framework Program (FP7) - People, and has been designed to encourage the promotion and development of international programs of research through mobility at various stages of research careers. The aim of TRAIN is to improve translational skills in the field of cancer by promoting a three-year international mobility program assigning a total of 51 fellowships subdivided into incoming, outgoing and reintegration fellowships.?The TRAIN proposal has been submitted in February 2009 to the European Commission in reply to the 2008 FP7-PEOPLE-COFUND call and has been successfully evaluated. TRAIN is addressed to postdoctoral scientists or scientists who have at least four years' full-time equivalent research experience and who wish to improve their careers spending one year abroad. The mobility program is open also to non-Italian experienced scientists wishing to spend one year in an Italian research center or private company. Part of the scheme is targeted to experienced Italian scientists who have completed at least three years of research in a foreign country and are interested in returning to Italy.?TRAIN is part of an overall Italian strategy outlined by the International Program of the Italian Cancer Network "Alleanza Contro il Cancro" to promote Italian participation in the building of the European Area for translational cancer research and to enhance the interaction between academy and industry.     

Economic valuation of air pollution mortality: A 9-country contingent valuation survey of value of a life year (VOLY)

This paper provides a key element for the calculation of the damage costs of air pollution, namely the valuation of mortality, important because premature mortality makes by far the largest contribution. Whereas several studies have tried to quantify the cost of air pollution mortality by multiplying a number of deaths by the ‘value of prevented fatality’ (also known as ‘value of statistical life’), we explain why such an approach is not correct and why one needs to evaluate the change in life expectancy due to air pollution. Therefore, an estimate for the monetary value of a life year (VOLY) is needed. The most appropriate method for determining VOLY is contingent valuation (CV). To determine VOLY for the EU, we have conducted a CV survey in 9 European countries: France, Spain, UK, Denmark, Germany, Switzerland, Czech Republic, Hungary, and Poland with a total sample size of 1463 persons. Based on the results from this 9-country CV survey we recommend a VOLY estimate of 40,000 € for cost–benefit analysis of air pollution policies for the European Union. As for confidence intervals, we argue that VOLY is at least 25,000 € and at the most 100,000 €.     

Modelling interactions between farmer practices and fattening pig performances with an individual-based model

European pig production continues to encounter economic and environmental challenges. To address these issues, methods have been developed to assess performances of pig production systems. Recent studies indicate that considering variability in performances among pigs improves the accuracy and reliability of results compared with modelling an average animal. Our objective was to develop a pig fattening unit model able to (i) simulate individual pig performances, including their variability in interaction with farmers’ practices and management, and (ii) assess their effects on technical, economic and environmental performances. Farmer practices included in the model were chosen from a typology generated from on-farm surveys focused on batch management, pig allocation to pens, pig feeding practices, practices of shipping to the slaughterhouse, and management of the remaining pigs. Pigs are represented using an individual-based model adapted from the InraPorc® model. To illustrate the model’s abilities, four scenarios were simulated that combine two feed rationing plans (ad libitum, restricted to 2.5 kg/day) and two feed sequence plans (two-phase, 10-phase). Analysis of variance was performed on the simulated technical, economic and environmental indicators (calculated via Life Cycle Assessment). The feed rationing plan and feed sequence plan significantly affected all indicators except for the premium per pig, for which the feed sequence plan did not have a significant effect. The ‘restricted 10-phase’ scenario maximised gross margin of the fattening unit (14.2 €/pig) and minimised environmental impacts per kg of pig produced. In contrast, the ‘ad libitum two-phase’ scenario generated the lowest margin (8.20 €/pig) and the highest environmental impacts. The model appears to be a promising tool to assess effects of farmers’ practices, pig characteristics and farm infrastructure on technical, economic and environmental performances of the fattening unit, and to investigate the potential of improvement. However, further work is needed, based on virtual experiments, in order to evaluate the effects of a larger diversity of practices.     

Cost-utility of an 8-month aquatic training for women with fibromyalgia: a randomized controlled trial

Introduction

Physical therapy in warm water has been effective and highly recommended for persons with fibromyalgia, but its efficiency remains largely unknown. Should patients or health care managers invest in this therapy? The aim of the current study was to assess the cost-utility of adding an aquatic exercise programme to the usual care of women with fibromyalgia.

Methods

Costs to the health care system and to society were considered in this study that included 33 participants, randomly assigned to the experimental group (n = 17) or a control group (n = 16). The intervention in the experimental group consisted of a 1-h, supervised, water-based exercise sessions, three times per week for 8 months. The main outcome measures were the health care costs and the number of quality-adjusted life-years (QALYs) using the time trade-off elicitation technique from the EuroQol EQ-5D instrument. Sensitivity analyses were performed for variations in staff salary, number of women attending sessions and time spent going to the pool. The cost effectiveness acceptability curves were created using a non-parametric bootstrap technique.

Results

The mean incremental treatment costs exceeded those for usual care per patient by € 517 for health care costs and € 1,032 for societal costs. The mean incremental QALY associated with the intervention was 0.131 (95% CI: 0.011 to 0.290). Each QALY gained in association with the exercise programme cost an additional € 3,947/QALY (95% CI: 1,782 to 47,000) for a health care perspective and € 7,878/QALY (3,559 to 93,818) from a societal perspective. The curves showed a 95% probability that the addition of the water-based programme is a cost-effective strategy if the ceiling of inversion is € 14,200/QALY from a health care perspective and € 28,300/QALY from a societal perspective.

Conclusion

The addition of an aquatic exercise programme to the usual care regime for fibromyalgia in women is cost effective in terms of both health care costs and societal costs. However, the characteristics of facilities (distance from the patients' homes and number of patients that can be accommodated per session) are major determinants to consider before investing in such a programme.

Trial registration

Current controlled trials ISRCTN53367487.

     

The effect of colony age on PCR fingerprinting

The pattern of DNA fragments produced by the polymerase chain reaction using arbitrary primers is strongly influenced by the age of the colonies sampled. We show that the number, clarity and reproducibility of the bands produced is optimal when very young colonies are sampled.
This work was supported by European Community grants (EEC CI1* 0545-UK, JEB and HC; BIOT-CT91-0283, JEB and HK), the AFRC (PG7 586, JEB and BH) and the SERC Biotechnology Directorate and Zeneca Ltd through a CASE studentship to LS.     

European research infrastructures for the development of nanobiotechnologies

Research infrastructures are essential for top-level academic and industrial research activities. Throughout the successive framework programmes (FPs) of the EU, actions have been gradually developed to support researchers in accessing top-level European research infrastructures located outside their own country and also to better coordinate and integrate these infrastructures Europe-wide, enabling better research services. At the same time, research infrastructures pave the way for the development of scientific and technological advances. Under the sixth Framework Programme (FP6; 2002-2006), for example, nanobiotechnologies have benefited from these European actions through three approaches: the support of multi-disciplinary pan-European infrastructures; the support of pan-European infrastructures dedicated to biology but with usage in multiple domains of biology; and the funding of integrated centers for nanobiotechnologies. The seventh Framework Programme (FP7; 2007-2013) will reinforce these actions toward research infrastructures, with particular attention to the emergence of new ones as well as to the provision of important strategic research services in fields such as nanobiotechnologies.     

Current state and prospects of biotechnology in Central and Eastern European countries. Part I: Visegrad countries (CZ,H, PL,SK)

Innovation is a key determinant of sustainable growth. Biotechnology (BT) is one such industry that has witnessed a revolution in innovative ideas leading to the founding of many new companies based on providing products, solutions and services, stretching from the food industry to environmental remediation, and new medicines. BT holds much promise for the development of national and local economies, however, this requires a strategic approach involving actors within government, industry, and academia working in concert to maximize this potential. This first article reviews the current “state of play” in the field of BT within the Central Eastern European (CEE) countries. For the purposes of this article, CEE refers to the countries of Czech Republic, Hungary, Poland, and Slovakia (the so-called Visegrad – V4 countries). We examine the components that support the creation and development of a BT sector in CEE and also highlight the barriers to these objectives. Clearly setting priorities for the countries’ policy agenda, as well as the alignment of Smart Specialization Strategy will help to focus efforts. Recent investments in R&D infrastructure within CEE have been substantial, but conditions will need to be optimized to harness these largely European investments for effective use towards SME high-tech development.     

Assessing the cost-effectiveness of the water purification function of wetlands for environmental planning

A reduction of nutrient input into freshwater and marine water bodies is a prerequisite for the restoration of healthy aquatic ecosystems and for the achievement of the ambitious goals of the European Water Framework Directive and the HELCOM and OSPAR convention. In Northern Germany, phosphorus inputs from point sources were reduced by nearly two-thirds compared to the phosphorus load in the period 1980–1985. This success was made possible by the investment of 2.1 billion € since 1985 in the development and enhancement of waste water treatment plants. However, these investments had only little effect on nitrogen inputs, which enter water bodies mainly from non-point sources. In 2002, the federal state Schleswig-Holstein implemented a peatland rehabilitation program aiming to restore the water purification function of this wetland type. The rationale of this plan is based on an economical analysis of the effects of the investments in the waste water action plan. A reduction of the P load by 1 kg has mean costs between 70 and 100 €. Based on model calculation, restoring wetlands for water quality purification will cost between 1 and 50 € per retained kg of nitrogen when considering a 10-year time span. Compared to the waste water action plan, wetland restoration is also a cost-efficient strategy for nutrient load reduction. For 10% of the investments of the waste water action plan, the nitrogen load from Schleswig-Holstein can be reduced by 10%; this will require rewetting of 20,000 ha of wetlands. The implementation of this strategy as well as the application of the concept of ecosystem services in environmental management is currently limited by a lack of political support and funding for such projects from higher political bodies such as the European Union.     

Safe biotechnology (4). Recommendations for safety levels for biotechnological operations with microorganisms that cause diseases in plants

The Working Party on Safety in Biotechnology of the European Federation of Biotechnology has proposed a classification of microorganisms that cause diseases in plants. In this paper appropriate safety levels are proposed for these classes of microorganisms in order to ensure that research, development and industrial fermentation work with plant pathogens will limit the risk of outbreaks of diseases in crops that could result from work with such microorganisms when they are cultivated in laboratories, glasshouses and biotechnology installations.Co-opted: J. Dähne, J. Drozd, M. Lemattre, I. M. Smith , E. M. A. WaterschootA Report prepared by the Working Party on Safety in Biotechnology of the European Federation of Biotechnology (EFB)

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Odense Pilot River Basin: implementation of the EU Water Framework Directive in a shallow eutrophic estuary (Odense Fjord,Denmark) and its upstream catchment

Implementation of the EU Water Framework Directive (WFD) is a huge environmental management challenge for Europe, demanding an integrated sustainable approach to water management and a common objective of obtaining ‘good status’ for all water bodies before 2015. The main task is the preparation of a river basin management plan for each of the 96 European river basin districts before the end of 2009. In Odense River Basin (island of Fyn, Denmark), one of 14 appointed European Pilot River Basins, the implementation of the WFD has been developed and tested in practice. Reference conditions and ecological status classification for Odense Fjord, based on eelgrass (Zostera marina) depth limit and nutrient concentrations, have been drawn up through a combination of historical data and modelling tools. A subsequent quantitative linking of pressures and impact, in casu between land-based nitrogen (N) loading of the fjord and resulting nutrient concentrations and eelgrass appearance, provided an estimate of the needed nitrogen load reduction of the fjord. This amounted to approx. 1,200 tonnes N per year (an annual load reduction of ca. 11 kg N ha^?1 of catchment area or ca. 19.5 g N m^?2 of fjord surface)—a load reduction of ca. 60% from the present level—to obtain at least ‘good’ ecological status sensu WFD. It is presently not possible to quantify a target load for phosphorus (P) in relation to marine environmental objectives. An economically feasible programme of measures to obtain ‘good’ status in all surface water and groundwater bodies in Odense River Basin, using an integrated cost-effectiveness analysis, showed that re-establishment of wetlands, catchcrops, and reduced fertilisation norms are the most effective measures if large reductions in N loads to the aquatic environment are to be achieved. The total socio-economic cost of implementing the WFD in the river basin amounts to about 13 million €/year, which will increase the expense for water services by only 0.5–0.6% of the total income and production value in the basin (15,650 million €/year). Investments to obtain the needed nitrogen load reductions from agriculture are thus economically feasible. Further, it is not an impossible task, either economically or technically, to reach the objectives of the WFD while still retaining the possibility of keeping a high agricultural production in the catchment (maintaining livestock production but decreasing crop production in the case of Odense River Basin). The future conditions in Odense Fjord will not only depend on the success in reducing the load from the river basin area, but will also be affected by the trend in the nutrient loss from the whole Baltic catchment area. The high growth rates in the new EU Member States thus pose an important challenge to water managers, and decoupling of economic growth from pressure on water bodies will be necessary. Finally, a number of challenges facing water managers around the Baltic and within the EU, namely preconditions required to successfully implement the WFD, are presented.     

OECD Best Practice Guidelines for Biological Resource Centers. OECD, 2007, 115 p.

The book is the first published collection of guidelines regulating operational activities of biological resource centers, approved by representatives of the 30 member nations of the Organization for Economic Cooperation and Development (OECD). The published standards were prepared as a result of multiple discussions held by expert groups of the OECD members, together with key experts of partner nations, under the auspices of a specialist task force established by the OECD Working Party on Biotechnology.     

Breeding replacement gilts for organic pig herds

In this study, breeding structures and commercial sow lines were evaluated by economic and genetic simulation studies for their suitability to provide the Dutch organic pig sector with replacement gilts. Sow and litter performance from over 2000 crossbred sows from 2006 to 2007 were collected on 11 to 14 Dutch organic pig herds, respectively, and compared with conventional herds. Results showed that organic herds had lower farrowing rates (3.6% to 7.5%), more live born piglets per litter (0.4% to 1.2%) and higher preweaning mortality rates (7% to 13%) compared to conventional herds. These results were used to simulate economic performance of various combinations of breeding structures and sow lines under organic conditions, under the assumption of absence of genotype-environment interactions. Sow and litter performance data under organic conditions (total piglets born/litter, stillborn piglets/litter, mortality until weaning, lactation length, interval weaning-oestrus and sow culling rate) and the costprice calculation for the Dutch organic pig sector were used as input for the economic simulation studies. The expected genetic progress was simulated for three potential breeding structures of the organic sector: organic breeding herds producing F1 gilts (OrgBS), a flower breeding system (FlowerBS) and a two-line rotation breeding system (RotBS). In FlowerBS, an organic purebred sow line is bred, using on-farm gilt replacement. The OrgBS with a Yorkshire × Landrace cross had the highest margin per sow place (€779), followed by RotBS with Yorkshire × Landrace cross (€706) and FlowerBS with Yorkshire sow line (€677). In case that an organic purebred sow population of 5000 sows would be available, FlowerBS gave the highest genetic progress in terms of cost price reduction (€3.72/slaughter pig per generation), followed by RotBS and OrgBS (€3.60/slaughter pig per generation). For FlowerBS, additional costs will be involved for maintaining a dedicated breeding programme. In conclusion, OrgBS using conventional genetics is economically the most viable option for the organic pig sector. However, this structure has clear disadvantages in terms of risks with regard to disease transmission and market demand. FlowerBS using a dedicated purebred organic line will only be cost-effective if sow population size is sufficiently large. RotBS might be a viable alternative, especially in combination with artificial insemination (AI) boars that are ranked according to an organic selection index. Regardless of breeding structure, the Yorkshire sow line gave the highest prolificacy and the highest economic returns on organic herds.     

Safe biotechnology

Summary The Working Party on Safety in Biotechnology of the European Federation of Biotechnology reported [Künzi M, et al. Safe Biotechnology (1) — General considerations. Appl Microbiol Biotechnol 21:1–6] on the classification of human pathogens and other microorganisms. It is proposed to relate the various risk classes of these organisms to the categories of physical containment for recombinant DNA (rDNA) organisms according to the OECD report [OECD Report (1986) Recombinant DNA Safety Considerations, Paris].In view of the differences in the numbering systems of the EFB for risk classes (1–4) and the OECD system of Good Industrial Large Scale Practice (GILSP) and containment categories 1–3, the former have been given alternative names. Relationships of the EFB-Classification of Microorganisms according to risk and OECD safety precautions have been defined (Table 1).Report prepared by the Working Party on Safety in Biotechnology of the European Federation of Biotechnology (EFB)     

Functional priorities in LCA and design for environment

Aim, Scope and Background  The interest in environmental questions has increased enormously during the last decade. Environmental protection has become an issue of strategic importance within the manufacturing industry and many companies are now working in the field of Design for Environment (DFE). The main purpose of DFE is to create products and services for achieving a sustainable society. Designers are widely believed to have a key role in adapting products to a sustainable society and one of the major instruments in the context of Design for Environment is Life Cycle Assessment (LCA). However, product development creates particular challenges for incorporating environmental issues that combine functional and environmental assessment. A natural and important part of product design is to define and analyse the functions of the product. Consequently, the functional unit in LCA is a core issue in DFE. Most recent research in DFE has focused on how to reduce the environmental impact of products throughout their life-cycle by addressing environmental aspects, while little attention has been given to the functionality of the product. Additionally, early product development phases, so called re-think phases, are considered to have the influence on major changes in products in general. These phases have thus the highest potential for changing products and product systems towards a sustainable development. Main Features  This paper discusses an extended functional representation in design for environment methods to evaluate sustainable design solutions, especially in early (re-think) phases of product design. Based on engineering-design science and several case studies, a concept has been developed describing how functional preferences can be visualised in design for environment and product development. In addition, the functional unit in LCA is discussed. The concept is called Functional Profile (FP) and is additionally exemplified in a case study on radio equipment. Discussion  The new functional characterisation concept helps identify functional priorities in design for environment. The Functional Profile is a structured, systematic and creative concept for identifying the necessary functions of a new product. The FP is envisioned to complement existing design for environment methods, not to replace them. Instead of being a product-development tool or method, the concept is an approach that increases understanding of inter-reactions between functional characteristics of products and their environmental characteristics, which furthermore facilitates trade-off decisions. One of the objectives behind the concept is to highlight the importance of balancing functional requirements and environmental impacts, presenting both the advantages and disadvantages of the product. Outlook  A second paper will be produced to complement the functional-environmental characterisation concept in early product development phase, presenting the environmental characterisation part and illustrating correlations between the functional and environmental sides.     

Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNAs from Vibrio cholerae O1, O139, and non-O1 strains.

Enterobacterial repetitive intergenic consensus (ERIC) sequence polymorphism was studied in Vibrio Cholerae strains isolated before and after the cholera epidemic in Brazil (in 1991), along with epidemic strains from Peru, Mexico, and India, by PCR. A total of 17 fingerprint patterns (FPs) were detected in the V. cholerae strains examined; 96.7% of the toxigenic V. cholerae O1 strains and 100% of the O139 serogroup strains were found to belong to the same FP group comprising four fragments (FP1). The nontoxigenic V. cholerae O1 also yielded four fragments but constituted a different FP group (FP2). A total of 15 different patterns were observed among the V. cholerae non-O1 strains. Two patterns were observed most frequently for V. cholerae non-01 strains, 25% of which have FP3, with five fragments, and 16.7% of which have FP4, with two fragments. Three fragments, 1.75, 0.79, and 0.5 kb, were found to be common to both toxigenic and nontoxigenic V. cholerae O1 strains as well as to group FP3, containing V. cholerae non-O1 strains. Two fragments of group FP3, 1.3 and 1.0 kb, were present in FP1 and FP2 respectively. The 0.5-kb fragment was common to all strains and serogroups of V. cholerae analyzed. It is concluded from the results of this study, based on DNA FPs of environmental isolates, that it is possible to detect an emerging virulent strain in a cholera-endemic region. ERIC-PCR constitutes a powerful tool for determination of the virulence potential of V. cholerae O1 strains isolated in surveillance programs and for molecular epidemiological investigations.     

Carbon dioxide for euthanasia: concerns regarding pain and distress, with special reference to mice and rats

Carbon dioxide (CO2) is the most commonly used agent for euthanasia of laboratory rodents, used on an estimated tens of millions of laboratory rodents per year worldwide, yet there is a growing body of evidence indicating that exposure to CO2 causes more than momentary pain and distress in these and other animals. We reviewed the available literature on the use of CO2 for euthanasia (as well as anaesthesia) and also informally canvassed laboratory animal personnel for their opinions regarding this topic. Our review addresses key issues such as CO2 flow rate and final concentration, presence of oxygen, and prefilled chambers (the animal is added to the chamber once a predetermined concentration and flow rate have been reached) versus gradual induction (the animal is put into an empty chamber and the gas agent(s) is gradually introduced at a fixed rate). Internationally, animal research standards specify that any procedure that would cause pain or distress in humans should be assumed to do so in non-human animals as well (Public Health Service 1986, US Department of Agriculture 1997, Organization for Economic Cooperation and Development 2000). European Union guidelines, however, specify a certain threshold of pain or distress, such as 'skilled insertion of a hypodermic needle', as the starting point at which regulation of the use of animals in experimental or other scientific procedures begins (Biotechnology Regulatory Atlas n.d.). There is clear evidence in the human literature that CO2 exposure is painful and distressful, while the non-human literature is equivocal. However, the fact that a number of studies do conclude that CO2 causes pain and distress in animals indicates a need for careful reconsideration of its use. Finally, this review offers recommendations for alternatives to the use of CO2 as a euthanasia agent.