Effect of Population Heterogenization on the Reproducibility of Mouse Behavior: A Multi-Laboratory Study

In animal experiments, animals, husbandry and test procedures are traditionally standardized to maximize test sensitivity and minimize animal use, assuming that this will also guarantee reproducibility. However, by reducing within-experiment variation, standardization may limit inference to the specific experimental conditions. Indeed, we have recently shown in mice that standardization may generate spurious results in behavioral tests, accounting for poor reproducibility, and that this can be avoided by population heterogenization through systematic variation of experimental conditions. Here, we examined whether a simple form of heterogenization effectively improves reproducibility of test results in a multi-laboratory situation. Each of six laboratories independently ordered 64 female mice of two inbred strains (C57BL/6NCrl, DBA/2NCrl) and examined them for strain differences in five commonly used behavioral tests under two different experimental designs. In the standardized design, experimental conditions were standardized as much as possible in each laboratory, while they were systematically varied with respect to the animals' test age and cage enrichment in the heterogenized design. Although heterogenization tended to improve reproducibility by increasing within-experiment variation relative to between-experiment variation, the effect was too weak to account for the large variation between laboratories. However, our findings confirm the potential of systematic heterogenization for improving reproducibility of animal experiments and highlight the need for effective and practicable heterogenization strategies.

Institutional Factors Affecting Agricultural Land Markets. Factor Markets Working Paper No. 16, February 2012

This paper analyses the main institutional factors affecting the rental and sales markets for agricultural land. Particular attention is paid to the effects of the common agricultural policy on land markets, and more specifically the underlying mechanism through which agricultural subsidies are capitalised into land values and farmland rents. This paper also provides a broad overview of the empirical studies that estimate the impact of agricultural support policies on land rents and land prices. Various other fundamental factors that affect agricultural land markets are discussed, such as land market institutions and regulations, transaction costs, credit market constraints and levels of profitability, the legal means of contract enforcement and land use alternatives.

Reducing suffering in experimental autoimmune encephalomyelitis (EAE)

This report is based on discussions and submissions from an expert working group consisting of veterinarians, animal care staff and scientists with expert knowledge relevant to the field. It aims to facilitate the implementation of the Three Rs (replacement, reduction and refinement) in the use of animal models or procedures involving experimental autoimmune encephalomyelitis (EAE), an experimental model used in multiple sclerosis research. The emphasis is on refinement since this has the greatest potential for immediate implementation. Specific welfare issues are identified and discussed, and practical measures are proposed to reduce animal use and suffering. Some general issues for refinement are summarised to help achieve this, with more detail provided on a range of specific measures to reduce suffering. © 2013 Elsevier Inc.

Developing accurate models of the human airways

Objectives Particle delivery to the airways is an attractive prospect for many potential therapeutics, including vaccines. Developing strategies for inhalation of particles provides a targeted, controlled and non-invasive delivery route but, as with all novel therapeutics, in vitro and in vivo testing are needed prior to clinical use. Whilst advanced vaccine testing demands the use of animal models to address safety issues, the production of robust in vitro cellular models would take account of the ethical framework known as the 3Rs (Replacement, Reduction and Refinement of animal use), by permitting initial screening of potential candidates prior to animal use. There is thus a need for relevant, realistic in vitro models of the human airways. Key findings Our laboratory has designed and characterised a multi-cellular model of human airways that takes account of the conditions in the airways and recapitulates many salient features, including the epithelial barrier and mucus secretion. Summary Our human pulmonary models recreate many of the obstacles to successful pulmonary delivery of particles and therefore represent a valid test platform for screening compounds and delivery systems.

A low mortality, high morbidity Reduced Intensity Status Epilepticus (RISE) model of epilepsy and epileptogenesis in the rat

Animal models of acquired epilepsies aim to provide researchers with tools for use in understanding the processes underlying the acquisition, development and establishment of the disorder. Typically, following a systemic or local insult, vulnerable brain regions undergo a process leading to the development, over time, of spontaneous recurrent seizures. Many such models make use of a period of intense seizure activity or status epilepticus, and this may be associated with high mortality and/or global damage to large areas of the brain. These undesirable elements have driven improvements in the design of chronic epilepsy models, for example the lithium-pilocarpine epileptogenesis model. Here, we present an optimised model of chronic epilepsy that reduces mortality to 1% whilst retaining features of high epileptogenicity and development of spontaneous seizures. Using local field potential recordings from hippocampus in vitro as a probe, we show that the model does not result in significant loss of neuronal network function in area CA3 and, instead, subtle alterations in network dynamics appear during a process of epileptogenesis, which eventually leads to a chronic seizure state. The model’s features of very low mortality and high morbidity in the absence of global neuronal damage offer the chance to explore the processes underlying epileptogenesis in detail, in a population of animals not defined by their resistance to seizures, whilst acknowledging and being driven by the 3Rs (Replacement, Refinement and Reduction of animal use in scientific procedures) principles.

Avaliação do efeito do liraglutida nos tecidos muscular e adiposo e nos parâmetros bioquímicos em camundongos Swiss submetidos à dieta de cafeteria e a atividade física

Obesity is a chronic metabolic disease characterized by adipose tissue formation excess leading to an increase in body fat mass, of multifactorial origin, produced mainly by poor eating habits combined with a sedentary lifestyle. Data consider obesity as a serious disease that affects the world's population, ranking fifth in death rates. Faced with this situation, individuals seek, increasingly, means to lose weight with less physical effort and food. In 2009 and 2010 the drug liraglutide was lauched in order to reduce weight in individuals with diabetes mellitus type 2, thus avoiding the emergence of other diseases. The aggravating factor is that obese nondiabetic individuals are making use of this substance, even if its use is not authorized by ANVISA (Brazilian Health Surveillance Agency). Thus the objective of this research is to evaluate the effect of liraglutide for muscle or fat tissues and biochemical parameters in Swiss mice submitted to cafeteria diet and physical activity. The study was approved by the Ethics Committee on Animal Use - CEUA (nº003 Protocol / 2014). For this study 74 animals (Swiss mice) were used, divided as follows: in the initial phase of this study, we carried out a pilot study (n = 10) divided into a control group (PCON) (n = 5) and cafeteria group (PCAF) (n = 5), in order to evaluate a cafeteria diet which was both attractive to the animals and that could provide an increase in adipose tissue. After the induction of the diet, animals were euthanized and as a result, the animals in the PCAF group showed an intra-abdominal adiposity 0.74 ± 0.05 g, taken as the parameter for increasing fat in animals. Subsequently the study base was conducted for this research where animals were used (n = 64) divided into 2 groups: the Cafeteria Study Base Group (EBCAF) divided as follows: cafeteria + exercise + liraglutide (CEL) (n = 8), cafeteria + exercise + saline (CES) (n = 8), cafeteria + liraglutide (CL) (n = 8) and cafeteria + saline (CS) (n = 8). The Chow Study Base group (EBR) was divided into: exercise + liraglutide (EL) (n = 8), exercise + saline + (ES) (n = 8), liraglutide (L) (n = 8) and saline solution (SS) (n = 8). All animals went through the submission process to the cafeteria diet, followed by exercise protocol through swimming and treatment with the test substance intraperitoneally (200 mg / mL / kg). After the treatments, the animals were euthanized and had the following parameters evaluated: the muscle tissue mass, adipose tissue mass and biochemical parameters. It was observed that the processing done with the exercise-associated liraglutide reduced adipose tissue mass significantly (0.32 ± 0.05 g) compared to the saline group (0.53 ± 0.07 g). There were no changes in the muscle tissue of the group which was treated and exercised (1.39 ± 0.03 g) compared to the saline group (1.33 ± 0.03 g). Regarding biochemical parameters it was evident that there were changes in these parameters. Interesting to note that, although blood glucose values have been changed, the animals did not become diabetic. Thus, it appears that physical activity together with liraglutide is eficcient to the loss of intraabdominal adipose tissue and the maintenance of lean body mass thereby generating a satisfactory result in the pursuit of quality of life and disease prevention.

Avaliação do efeito do liraglutida nos tecidos muscular e adiposo e nos parâmetros bioquímicos em camundongos Swiss submetidos à dieta de cafeteria e a atividade física

Obesity is a chronic metabolic disease characterized by adipose tissue formation excess leading to an increase in body fat mass, of multifactorial origin, produced mainly by poor eating habits combined with a sedentary lifestyle. Data consider obesity as a serious disease that affects the world's population, ranking fifth in death rates. Faced with this situation, individuals seek, increasingly, means to lose weight with less physical effort and food. In 2009 and 2010 the drug liraglutide was lauched in order to reduce weight in individuals with diabetes mellitus type 2, thus avoiding the emergence of other diseases. The aggravating factor is that obese nondiabetic individuals are making use of this substance, even if its use is not authorized by ANVISA (Brazilian Health Surveillance Agency). Thus the objective of this research is to evaluate the effect of liraglutide for muscle or fat tissues and biochemical parameters in Swiss mice submitted to cafeteria diet and physical activity. The study was approved by the Ethics Committee on Animal Use - CEUA (nº003 Protocol / 2014). For this study 74 animals (Swiss mice) were used, divided as follows: in the initial phase of this study, we carried out a pilot study (n = 10) divided into a control group (PCON) (n = 5) and cafeteria group (PCAF) (n = 5), in order to evaluate a cafeteria diet which was both attractive to the animals and that could provide an increase in adipose tissue. After the induction of the diet, animals were euthanized and as a result, the animals in the PCAF group showed an intra-abdominal adiposity 0.74 ± 0.05 g, taken as the parameter for increasing fat in animals. Subsequently the study base was conducted for this research where animals were used (n = 64) divided into 2 groups: the Cafeteria Study Base Group (EBCAF) divided as follows: cafeteria + exercise + liraglutide (CEL) (n = 8), cafeteria + exercise + saline (CES) (n = 8), cafeteria + liraglutide (CL) (n = 8) and cafeteria + saline (CS) (n = 8). The Chow Study Base group (EBR) was divided into: exercise + liraglutide (EL) (n = 8), exercise + saline + (ES) (n = 8), liraglutide (L) (n = 8) and saline solution (SS) (n = 8). All animals went through the submission process to the cafeteria diet, followed by exercise protocol through swimming and treatment with the test substance intraperitoneally (200 mg / mL / kg). After the treatments, the animals were euthanized and had the following parameters evaluated: the muscle tissue mass, adipose tissue mass and biochemical parameters. It was observed that the processing done with the exercise-associated liraglutide reduced adipose tissue mass significantly (0.32 ± 0.05 g) compared to the saline group (0.53 ± 0.07 g). There were no changes in the muscle tissue of the group which was treated and exercised (1.39 ± 0.03 g) compared to the saline group (1.33 ± 0.03 g). Regarding biochemical parameters it was evident that there were changes in these parameters. Interesting to note that, although blood glucose values have been changed, the animals did not become diabetic. Thus, it appears that physical activity together with liraglutide is eficcient to the loss of intraabdominal adipose tissue and the maintenance of lean body mass thereby generating a satisfactory result in the pursuit of quality of life and disease prevention.

The use of non-animal alternatives in the safety evaluations of cosmetics ingredients by the Scientific Committee on Consumer Safety (SCCS)

In Europe, the safety evaluation of cosmetics is based on the safety evaluation of each individual ingredient. Article 3 of the Cosmetics Regulation specifies that a cosmetic product made available on the market is to be safe for human health when used normally or under reasonably foreseeable conditions. For substances that cause some concern with respect to human health (e.g. colorants, preservatives, UV-filters), safety is evaluated at the Commission level by a scientific committee, presently called the Scientific Committee on Consumer Safety (SCCS). According to the Cosmetics Regulations, in the EU, the marketing of cosmetics products and their ingredients that have been tested on animals for most of their human health effects, including acute toxicity, is prohibited. Nevertheless, any study dating from before this prohibition took effect is accepted for the safety assessment of cosmetics ingredients. The in vitro methods reported in the dossiers summited to the SCCS are here evaluated from the published reports issued by the scientific committee of the Directorate General of Health and Consumers (DG SANCO); responsible for the safety of cosmetics ingredients. The number of studies submitted to the SCCS that do not involve animals is still low and in general the safety of cosmetics ingredients is based on in vivo studies performed before the prohibition.

Glycerol as a by-product of biodiesel production in Brazil: Alternatives for the use of unrefined glycerol

On January 1 2008, Brazil included yet another element into its energy matrix: biodiesel. The predominant biodiesel production process involves a phase of transesterification that yields glycerol as a by-product. The use of this glycerol is limited since it is considered an unrefined raw material that must be refined for its various types of use. Several studies have addressed identification of possible uses for unrefined glycerol. Given the diversity of uses, an overview is necessary. The purpose of this work is to present alternatives currently being considered for the use of unrefined glycerol as a by-product of biodiesel production, aiming to contribute to the sustainable consolidation of the biofuel market. Exploratory research was carried out to identify these viable alternatives for the use of this by-product. The possibilities include the production of chemical products, fuel additives, production of hydrogen, development of fuel cells, ethanol or methanol production, animal feed, co-digestion and co-gasification, and waste treatment among others. The present research reveals that there are promising possibilities for the use of unrefined glycerol, which may help consolidate the sustainability of the biofuel market. (C) 2012 Elsevier Ltd. All rights reserved.

On the use of correlated beta random variables with animal population modelling

We give reasons why demographic parameters such as survival and reproduction rates are often modelled well in stochastic population simulation using beta distributions. In practice, it is frequently expected that these parameters will be correlated, for example with survival rates for all age classes tending to be high or low in the same year. We therefore discuss a method for producing correlated beta random variables by transforming correlated normal random variables, and show how it can be applied in practice by means of a simple example. We also note how the same approach can be used to produce correlated uniform triangular, and exponential random variables. (C) 2008 Elsevier B.V. All rights reserved.

Foundation and Perspectives of the Use of Plant Extracts as Performance Enhancers in Broilers

Feed is responsible for about 70% of broilers production costs, leading to an increasing number of studies on alternative dietary products that benefit bird performance and lower production costs. Since the 1950s, antimicrobial additives are the most frequently used performance enhancers in animal production and their positive results are observed even in high-challenge conditions. Since the 1990s, due to the ban of the use of some antibiotics as growth promoters and the growing trend of the public to consume natural products, plant extracts have been researched as alternatives to antibiotic growth promoters. The first study that evaluated the antibacterial activities of plant extracts was carried out in 1881; however, they started to be used as flavor enhancers only during the next decades. With the emergence of antibiotics in the 1950s, the use of plant extracts as antimicrobial agents almost disappeared. There are several studies in literature assessing the use of plant extracts, individually or in combination, as antimicrobials, antioxidants, or digestibility enhancers in animal feeds. Research results on the factors affecting their action, such as plant variety, harvest time, processing, extraction, as well as the technology employed to manufacture the commercial product and dietary inclusion levels show controversial results, warranting the need of further research and standardization for the effective use of plant extracts as performance enhancers, when added to animal feeds. This article aims at presenting plant extracts as alternatives to antibiotics, explaining their main modes of action as performance enhancers in broiler production.

Exploring the use of Viagra in place of animal and plant potency products in traditional Chinese medicine

Recently, conservationists have debated whether consumers of animal and plant potency products used to treat erectile dysfunction (ED) in traditional Chinese medicine (TCM) might be switching to Viagra, consequently consuming fewer of these animals and plants. To address this question, a survey examined the medical decisions of male consumers of TCM in Hong Kong who were over the age of 50. As predicted, these consumers reported selectively switching to Western medicines to treat ED, but not to treat other health ailments. These findings provide support for the possibility that Viagra may have conservation benefits for certain species.

Neoskin development in the fetus with the use of a three-layer graft: an animal model for in utero closure of large skin defects

Objective: To assess the ability of a three-layer graft in the closuse of large fetal skin defects. Methods: Ovine fetuses underwent a large (4 x 3 cm) full-thickness skin defect over the lumbar region at 105 days` gestation (term = 140 days). A bilaminar artificial skin was placed over a cellulose interface to cover the defect (3-layer graft). The skin was partially reapproximated with a continuous nylon suture. Pregnancy was allowed to continue and the surgical site was submitted to histopathological analysis at different post-operative intervals. Results: Seven fetuses underwent surgery. One maternal/fetal death occurred, and the remaining 6 fetuses were analyzed. Artificial skin adherence to the wound edges was observed in cases that remained in utero for at least 15 days. Neoskin was present beneath the silicone layer of the bilaminar artificial skin. Conclusions: Our study shows that neoskin can develop in the fetus using a 3-layer graft, including epidermal growth beneath the silicone layer of the bilaminar skin graft. These findings suggest that the fetus is able to reepithelialise even large skin defects. Further experience is necessary to assess the quality of this repair.

Estudo da produção e aplicação de cola animal

Com as crescentes exigências do mercado e fortes restrições a nível ambiental, as indústrias modernas tendem a progredir no sentido da eficiência, sustentabilidade e rentabilidade dos seus processos produtivos. Sendo a indústria dos curtumes uma forte geradora de resíduos sólidos, é indispensável criar alternativas à valorização desses mesmos resíduos de forma a cumprir todos os requisitos ambientais. Este trabalho remete à reutilização dos resíduos, provenientes da divisão da pele em tripa após operação de descarna, com o intuito de se desenvolver um novo bio-produto apto para ser utilizado noutros sectores industriais. Os resíduos em causa consistem maioritariamente em colagéneo, que apresenta um enorme potencial para se produzir cola animal. Desta forma, este trabalho tem como objectivo principal avaliar a utilização deste tipo de resíduo na produção de um novo bio-produto, cola animal, e testar a sua aplicabilidade técnica e funcional. Com a realização deste trabalho, mostrou-se ser possível produzir um produto, a partir de retalhos da indústria de curtumes, com um perfil de carácter positivo no que diz respeito à viabilidade técnica e funcional da utilização da cola animal como eventual alternativa a um ligante, para formulações de colas de base aquosa para a indústria gráfica e do papel.