Monitoring the welfare of sheep in organic and conventional farms using an ANI 35 L derived method

The present study was undertaken to evaluate the inter-observer reliability of a welfare monitoring scheme to be applied to sheep, and compare the welfare state of the animals between 10 organic and 10 conventional sheep farms. Two trained observers performed recordings. A graded point protocol was used, that relies on five sheets mostly derived by the Animal Needs Index, which is mainly based on resource-based parameters. Therefore, in the fifth sheet animal-based parameters, deemed relevant to sheep welfare, were taken into account. In particular, the following animal-based variables were assessed: integument alterations, animal dirtiness, hoof overgrowth, lameness and lesions, which where scored on the basis of their prevalence (number of affected animals/numbers of observed animals), longevity (age in years), and mutilations, such as de-horning and caudotomy, evaluated in terms of presence/absence. No significant differences were observed between organic and conventional farms in terms of ANI scores, housing characteristics and animal-based parameters. This result was not surprising, as most of the farms, both conventional and organic, based their farming systems on an extensive use of the land by grazing animals. The monitoring protocol proved to be feasible (the mean time needed to perform the assessment of welfare was 85 min per farm) and reliable: a significant Spearman's correlation coefficient between observers was observed for total score and all sheets. As to animal-based parameters, body condition could not be assessed visually due to the presence of flee in winter; the correlation between observers was significant for integument alterations, animal dirtiness, hoof overgrowth and lameness, whereas inter-observer reliability was not significant for lesions. This result indicated that more training is needed for the assessment of lesions in order to increase the reliability of the measure. In addition, we suggest visiting farms in early summer, soon after shearing, in order to make easier the detection of lesions and the assessment of body condition.     

Techniques of embryo transfer and facility decontamination used to improve the health and welfare of transgenic mice

'Reduction' and 'Refinement' can be achieved in transgenic mouse studies by re-deriving transgenic mouse lines and subsequently maintaining them under high standards of husbandry in a unit with restricted access. This report describes the initial steps of a project to improve the health and welfare of transgenic mice at the European Molecular Biology Laboratory (EMBL), by re-deriving transgenic lines as microbiologically defined animals to be maintained in a barrier unit in a newly constructed animal facility. A pilot study showed that it was possible to transfer embryos obtained from contaminated donor mice in the old facility to specific pathogen free recipients housed in a ventilated cabinet in the new unit, without concomitant carry over of disease. The offspring born following embryo transfer were of high health status and did not show any evidence of contamination with any of the pathogens present in the mice in the old animal unit. Antibodies to various murine viruses (mouse hepatitis virus (MHV), rota virus, reo-3 virus, Theilers encephalomyelitis virus, adenovirus) and parasites were present in sentinel animals from the old animal house whereas the re-derived animals were found to be free of virus antibodies and parasites. Therefore the methods used were considered to be successful in terms of disease prevention and enhancement of welfare. The barrier unit was sterilized without the use of formaldehyde or related substances, to minimize the risks to personnel and to the environment from using potentially dangerous substances. From the results of in vitro and in vivo screening, the protocol for sterilization described here was found to be effective in achieving microbiological sterility of the barrier unit and was cost effective.     

A guide to defining and implementing protocols for the welfare assessment of laboratory animals: eleventh report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement

The refinement of husbandry and procedures to reduce animal suffering and improve welfare is an essential component of humane science. Successful refinement depends upon the ability to assess animal welfare effectively, and detect any signs of pain or distress as rapidly as possible, so that any suffering can be alleviated. This document provides practical guidance on setting up and operating effective protocols for the welfare assessment of animals used in research and testing. It sets out general principles for more objective observation of animals, recognizing and assessing indicators of pain or distress and tailoring these to individual projects. Systems for recording indicators, including score sheets, are reviewed and guidance is set out on determining practical monitoring regimes that are more likely to detect any signs of suffering. This guidance is intended for all staff required to assess or monitor animal welfare, including animal technologists and care staff, veterinarians and scientists. It will also be of use to members of ethics or animal care and use committees. A longer version of this document, with further background information and extra topics including training and information sharing, is available on the Laboratory Animals website.     

Effects of space allocation and housing density on measures of wellbeing in laboratory mice: a review

In the majority of countries where there are legislative requirements pertaining to the use of animals in research, figures are quoted for minimum cage sizes or space allocation to be provided per animal. These figures are generally based on professional judgement and are in common usage. However, there is a growing trend and expectation that welfare science should inform regulatory decision-making. Given the importance of the potential welfare influences of cage size on the animals themselves, this paper presents the latest scientific knowledge on this topic in one of the most commonly used animals in research, the mouse. A comprehensive review of studies in laboratory mice was undertaken, examining the effects of space allocation per animal and animal density on established welfare indicators. To date, animal density studies have predominated, and the effects of space allocation per se are still relatively unclear. This information will guide those involved in facility management or legislative review, and provide a more solid foundation for further studies into the effects of routine husbandry practices on animal welfare.     

“What’s wrong with my monkey?” Ethical perspectives on germline transgenesis in marmosets

The birth of the first transgenic primate to have inherited a transgene from its parents opens the possibility to set up transgenic marmoset colonies, as these monkeys are small and relatively easy to keep and breed in research facilities. The prospect of transgenic marmoset models of human disease, readily available in the way that transgenic laboratory mice are currently, prompts excitement in the scientific community; but the idea of monkeys being bred to carry diseases is also contentious. We structure an ethical analysis of the transgenic marmoset case around three questions: whether it is acceptable to use animals as models of human disease; whether it is acceptable to genetically modify animals; and whether these animals’ being monkeys makes a difference. The analysis considers the prospect of transgenic marmoset studies coming to replace transgenic mouse studies and lesion studies in marmosets in some areas of research. The mainstream, broadly utilitarian view of animal research suggests that such a transition will not give rise to greater ethical problems than those presently faced. It can be argued that using marmosets rather than mice will not result in more animal suffering, and that the benefits of research will improve with a move to a species more similar in phylogenetic terms to humans. The biological and social proximity of monkeys and humans may also benefit the animals by making it easier for scientists and caretakers to recognize signs of suffering and increasing the human motivation to limit it. The animal welfare and research impacts of the transition to marmoset use will depend very much on the extent to which researchers take these issues seriously and seek to minimize animal harm and optimize human benefit.     

The Concept of Animal Welfare at the Interface between Producers and Scientists: The Example of Organic Pig Farming

In organic farming animal welfare is one important aspect included in the internationally agreed organic principles of health, ecology, fairness and care (IFOAM 2006), reflecting expectation of consumers and farmers. The definition of organic animal welfare includes—besides traditional terms of animal welfare—‘regeneration’ and ‘naturalness’. Organic animal welfare assessment needs to reflect this and use complex parameters, include natural behaviour and a systemic view. Furthermore, various parties with seemingly conflicting interests are involved, causing ethical dilemmas, such as the use of nose rings for outdoor sows (impaired animal welfare vs. destruction of humus). Solutions can only be found when foundational concepts are translated and applied to practical situations. On-farm animal welfare assessment and implementation of improvement strategies are increasingly relevant scientific areas. They combine on-farm welfare assessment, identification of key problem areas and connected risk factors. Constant communication between all parties is crucial for success. Animal health and welfare planning is one application of this approach, which was carried out on Austrian organic pig farms as well as organic dairy farms in seven European countries. The projects included welfare assessment, feedback and benchmarking as a tool for communication between farmers, advisors and scientists. Finally goals were set by the farmer and improvement strategies applicable to organic farming were implemented. This included prevention of disease by management strategies instead of routine treatment with pharmaceutical products. It appeared that next to problem structuring, multidisciplinary problem solving demands good communications skills to relate animal welfare science to value reflections.     

Transgenic mice: beyond the knockout

Transgenic mice have had a tremendous impact on biomedical research. Most researchers are familiar with transgenic mice that carry Cre recombinase (Cre) and how they are used to create conditional knockouts. However, some researchers are less familiar with many of the other types of transgenic mice and their applications. For example, transgenic mice can be used to study biochemical and molecular pathways in primary cultures and cell suspensions derived from transgenic mice, cell-cell interactions using multiple fluorescent proteins in the same mouse, and the cell cycle in real time and in the whole animal, and they can be used to perform deep tissue imaging in the whole animal, follow cell lineage during development and disease, and isolate large quantities of a pure cell type directly from organs. These novel transgenic mice and their applications provide the means for studying of molecular and biochemical events in the whole animal that was previously limited to cell cultures. In conclusion, transgenic mice are not just for generating knockouts.     

The role of the veterinary staff in mouse breeding colony management

The rapid increase in the production and use of transgenic mice has been a boon for biomedical research and a challenge for the animal care and use programs responsible for providing housing and medical care to these animals. The authors suggest ways in which the veterinary staff can successfully organize and manage transgenic mouse breeding programs to reduce uncontrolled breeding and the problems associated with it.     

SHORT COMMUNICATION: Double pronuclei injection of DNA into zygotes increases yields of transgenic mouse lines

Transgenic mice are increasingly used for gene function and regulation studies of mammalian genes. A major limitation is the necessity to produce a large number of founder animals to obtain one line with the desired expression pattern. We developed a method, the 'double pronuclei injection', that doubles the yield of transgenic mouse lines obtained from each injection session, thereby reducing the time, effort and costs of generating transgenic mice. Three transgenic vectors were microinjected into the male and female pronuclei of zygotes. Approximately half of the resulting born mice were transgenic. This represented a 60% increase in the yield of founders per injected zygote, and a 100% increase in the yield of transgenic mice per born animal, when compared to yields obtained using single pronucleus injection. This method should prove useful for generating large numbers of transgenic mice for gene regulation studies and for conditional gene ablation     

The effect of daily disturbance on the breeding performance of mice

The United Kingdom Home Office Code of Practice for the housing and care of breeding animals requires that, 'the general well-being of all animals must be checked at least once daily'. However, excessive daily disturbance of rodent breeding colonies could be counter-productive to animal welfare if it increases pre-weaning mortality. An experiment involving 100 breeding cages of BALB/c mice compared daily inspection of the mouse cages, but without disturbing the mice within the nest, with daily inspection in which every individual was studied even if this involved removing the cage lid and disturbing the nest. No statistically significant differences were found between the two groups in breeding performance or pre-weaning mortality, though the disturbed group produced marginally fewer offspring and had slightly higher mortality. Average weaning weight did not differ between the groups, but sexual dimorphism at weaning was significantly increased in the disturbed group. It is concluded that there are unlikely to be any welfare benefits in an inspection regimen that involves disturbance of breeding mice, provided the cage is inspected daily.     

筑巢材料在实验大、小鼠环境丰荣中的应用

相对于标准鼠盒,实验大、小鼠更喜好有筑巢材料的“环境丰荣”鼠盒.向标准鼠盒中放入筑巢材料,可以为大、小鼠提供筑巢、躲避、攀爬、休息的环境.因此,在实验大、小鼠饲养过程中,提供筑巢材料,是一个简单有效的丰富实验动物环境,提高实验动物福利的方法.本文就筑巢材料应用背景、近年来在啮齿类实验动物环境丰荣中的应用、对实验动物福利的影响三方面做以综述,并对今后筑巢材料在改善实验动物福利的应用前景作以展望.     

Time to Integrate to Nest Test Evaluation in a Mouse DSS-Colitis Model

Severity assessment in laboratory animals is an important issue regarding the implementation of the 3R concept into biomedical research and pivotal in current EU regulations. In mouse models of inflammatory bowel disease severity assessment is usually undertaken by clinical scoring, especially by monitoring reduction of body weight. This requires daily observance and handling of each mouse, which is time consuming, stressful for the animal and necessitates an experienced observer. The time to integrate to nest test (TINT) is an easily applicable test detecting disturbed welfare by measuring the time interval mice need to integrate nesting material to an existing nest. Here, TINT was utilized to assess severity in a mouse DSS-colitis model. TINT results depended on the group size of mice maintained per cage with most consistent time intervals measured when co-housing 4 to 5 mice. Colitis was induced with 1% or 1.5% DSS in group-housed WT and Cd14-deficient mice. Higher clinical scores and loss of body weight were detected in 1.5% compared to 1% DSS treated mice. TINT time intervals showed no dose dependent differences. However, increased clinical scores, body weight reductions, and increased TINT time intervals were detected in Cd14 ^-/- compared to WT mice revealing mouse strain related differences. Therefore, TINT is an easily applicable method for severity assessment in a mouse colitis model detecting CD14 related differences, but not dose dependent differences. As TINT revealed most consistent results in group-housed mice, we recommend utilization as an additional method substituting clinical monitoring of the individual mouse.     

Phenotype Characterization and Welfare Assessment of Transgenic Rodents (Mice)

Methods of transgenesis in vertebrate animals in the laboratory involve the stable addition or selective substitution of defined genes into the germline. Although there is a continuous and remarkable development in transgenic technology-the quality of transgenes, gene-targeting vectors, and experimental approaches-the consequences for the phenotype induced by this kind of experimental mutation cannot be completely predicted. This is especially true for the majority of transgenic animals who are generated by pronuclear injection. Randomly integrated foreign DNA may increase the risk of disturbing normal physiological processes, resulting in discomfort possibly crucial to the animals' welfare. Only a careful and comprehensive phenotype and welfare assessment can determine (a) whether apparent and relevant changes of phenotype (genetic burdens) occur and (b) how to deal with them in terms of the 3Rs (replace, reduce, refine). Two structured forms, data record and characterization, have been developed and are proposed for a routine characterization and assessment of newly generated transgenic rodents. Data record form and characterization operate, respectively, at individual (health monitoring) and strain levels (representative sample of individual life histories).     

Constitutive expression of IL-2Rbeta chain and its effects on IL-2-induced vascular leak syndrome

IL-2-induced vascular leak syndrome (VLS) is an important mechanism explaining the toxic effects of this cytokine and limiting its therapeutic use. We previously characterized a mouse model of IL-2-induced pulmonary VLS used to demonstrate that NK lymphocytes are involved in early/acute phase VLS (after one IL-2 injection). We also showed that NK cells and polymorphonuclear neutrophils (PMN) are involved in the late/chronic phase of the syndrome (after four daily IL-2 injections). In this study we use our mouse model to evaluate the role played by the IL-2 receptor (IL-2R) in VLS induction. Mouse and human IL-2R are different since the mouse IL-2Rbeta chain does not recognize IL-2. Here, we compare the acute and late VLS responses in human IL-2Rbeta transgenic and C57BL/6 wild type mice. Parameters linked to early phase VLS (bronchoconstriction and PMN mobilization) are enhanced in human IL-2Rbeta transgenic mice. By contrast, parameters used to measure late events (protein leakage and edema) are similar in human IL-2Rbeta transgenic mice and C57BL/6 wild type animals. However, after four IL-2 injections, the cellular content of the bronchoalveolar lavage fluids was different between the two types of animals. This study also characterizes a humanized animal model that could be further used to study human IL-2 activity and side effects in vivo.     

Constitutive expression of IL-2Rbeta chain and its effects on IL-2-induced vascular leak syndrome

IL-2-induced vascular leak syndrome (VLS) is an important mechanism explaining the toxic effects of this cytokine and limiting its therapeutic use. We previously characterized a mouse model of IL-2-induced pulmonary VLS used to demonstrate that NK lymphocytes are involved in early/acute phase VLS (after one IL-2 injection). We also showed that NK cells and polymorphonuclear neutrophils (PMN) are involved in the late/chronic phase of the syndrome (after four daily IL-2 injections). In this study we use our mouse model to evaluate the role played by the IL-2 receptor (IL-2R) in VLS induction. Mouse and human IL-2R are different since the mouse IL-2Rbeta chain does not recognize IL-2. Here, we compare the acute and late VLS responses in human IL-2Rbeta transgenic and C57BL/6 wild type mice. Parameters linked to early phase VLS (bronchoconstriction and PMN mobilization) are enhanced in human IL-2Rbeta transgenic mice. By contrast, parameters used to measure late events (protein leakage and edema) are similar in human IL-2Rbeta transgenic mice and C57BL/6 wild type animals. However, after four IL-2 injections, the cellular content of the bronchoalveolar lavage fluids was different between the two types of animals. This study also characterizes a humanized animal model that could be further used to study human IL-2 activity and side effects in vivo.     

Welfare issues of genetically modified animals

Genetically engineered animals have opened new frontiers in the study of physiology and disease processes. Mutant animals offer more accurate disease models and increased precision for pathogenesis and treatment studies. Their use offers hope for improved therapy to patients with conditions that currently have poor or ineffective treatments. These advantages have fostered an increase in studies using mice in recent years, a development viewed with alarm by those who oppose the use of animals in research. Scientists point out that the mice are replacing more sentient species, such as nonhuman primates, and are increasing the quality of research being conducted. They assert that study of genetically engineered animals will eventually permit decreases in numbers of animals used in research. Nevertheless, the increase in use of genetically altered animals presents many challenges in reviewing protocols and providing care. Identification and resolution of any welfare problems is a responsibility that is shared by institutional animal care and use committee, veterinary, animal care, and research staffs. To identify potential welfare concerns, a database such as TBASE () can be searched to learn what has been reported for established mutant lines. In addition, newly created lines should be monitored by a surveillance system and have phenotype assessment to identify the effects of altering the genome. Methods of ensuring welfare can include treatment of conditions produced, restriction of gene expression to tissues of interest or to certain time periods, and establishment of endpoints for removing animals from a study before problems appear.     

Phenotyping of genetically engineered mice: humane, ethical, environmental, and husbandry issues

The growing use of genetically engineered (GE) mice in scientific research has raised many concerns about the animal welfare of such mice. The types of welfare concerns may differ within the three stages that comprise the establishment of GE animal models: development, production, and research use. The role and impact of the members of the research team on these concerns may also vary with each stage. To make both scientific and animal welfare decisions at each stage, it is necessary to have a thorough knowledge of the animal model-in this case, the phenotypic expression of the GE animal. Phenotype screening is the analysis of visible or measurable characteristics of an animal that result from the genotype and its interaction with the environment. Phenotypes expressed that are relevant to the research program are usually carefully investigated; however, those that may have an impact on the animal's welfare but have little or no impact on the disease process under study are often less carefully studied. Thorough analysis and documentation of the animal welfare aspects of phenotype provide the research team with the information they need to control the environment to minimize negative animal welfare effects. Such information is also essential to allow members of the institutional animal care and use committee to perform necessary cost:benefit ethical review of proposed GE animal studies. Investigators who submit information about models for publication should document all aspects of a phenotype, including the area of scientific interest as well as those areas that affect animal welfare, for clarity and for subsequent research with the respective models.     

Using conjoint analysis to weight broiler welfare variables based on slaughterhouse data

The Council of the European Union adopted the welfare directive for broiler chickens in 2007 (Commission of the European Communities, 2007). The directive defines minimum demands and objectives for broiler production ranging from stocking density to welfare. This study found that the level of nonhuman animal welfare can be determined, for example, with welfare indexes that have different weight values for welfare variables. A total of 20 poultry experts from slaughterhouses, the feed industry, administration, and research received simulated data on 24 flocks. This included data for 5 welfare variables based on postmortem meat inspection information on commercial flocks and based on literature: mortality during rearing, foot-pad score, and the incidence of ascites, dermatitis, and cachexia in routine meat inspection. The experts ranked these flocks from the worst to the best according to animal welfare. Conjoint analysis weighted the relative importance of the welfare variables for broiler welfare. The study based relative importance for broiler welfare on 20 expert opinions: 24.6% for mortality, 35.9% for foot-pad score, 19.7% for the incidence of ascites, 15.7% for the incidence of dermatitis, and 4.1% for the incidence of cachexia. In the future, an index for broiler welfare evaluation could be developed based on the results of this study.     

Using Conjoint Analysis to Weight Broiler Welfare Variables Based on Slaughterhouse Data

The Council of the European Union adopted the welfare directive for broiler chickens in 2007 (Commission of the European Communities, 2007). The directive defines minimum demands and objectives for broiler production ranging from stocking density to welfare. This study found that the level of nonhuman animal welfare can be determined, for example, with welfare indexes that have different weight values for welfare variables. A total of 20 poultry experts from slaughterhouses, the feed industry, administration, and research received simulated data on 24 flocks. This included data for 5 welfare variables based on postmortem meat inspection information on commercial flocks and based on literature: mortality during rearing, foot-pad score, and the incidence of ascites, dermatitis, and cachexia in routine meat inspection. The experts ranked these flocks from the worst to the best according to animal welfare. Conjoint analysis weighted the relative importance of the welfare variables for broiler welfare. The study based relative importance for broiler welfare on 20 expert opinions: 24.6% for mortality, 35.9% for foot-pad score, 19.7% for the incidence of ascites, 15.7% for the incidence of dermatitis, and 4.1% for the incidence of cachexia. In the future, an index for broiler welfare evaluation could be developed based on the results of this study.