Evaluation of Cognitive Activity in Experimental Animal1

The use of animals in research and education dates back to the period when humans started to look for ways to prevent and cure ailments. Most of present day's drug discoveries were possible because of the use of animals in research. The dilemma to continue animal experiments in education and research continues with varied and confusing guidelines. However, the animal use and their handling vary in each laboratory and educational institution. It has been reported that the animals are being subjected to painful procedures in education and training unnecessarily. The extensive use of animals in toxicity studies and testing dermatological preparations has raised concerns about the ways animals are sacrificed for these “irrelevant experiments”. On the other side of the coin are scientists who advocate the relevant and judicious use of animals in research so that new discoveries can continue. In this review, we discuss the evolution of the use of animals in education and research and how these have been affected in recent times owing to concerns from animal lovers and government regulations. A number of computer simulation and other models have been recommended for use as alternatives to use of animals for pharmacology education. In this review we also discuss some of these alternatives.

Public consultation in the evaluation of animal research protocols

One response to calls for increased openness in animal research is to make protocols publicly accessible, but it is unclear what type of input the public would provide if given this opportunity. In this study we invited public responses to five different research projects, using non-technical summaries intended for lay audiences. Our aim was to assess the potential for this type of public consultation in protocol review, and a secondary aim was to better understand what types of animal research people are willing to accept and why. US participants (n = 1521) were asked (via an online survey) “Do you support the use of these (insert species) for this research”, and responded using a seven-point scale (1 = “No”, 4 = “Neutral”, and 7 = “Yes”). Participants were asked to explain the reasons for their choice; open-ended text responses were subjected to thematic analysis. Most participants (89.7%) provided clear comments, showing the potential of an online forum to elicit feedback. Four themes were prevalent in participant reasoning regarding their support for the proposed research: 1) impact on animals, 2) impact on humans, 3) scientific merit, and 4) availability of alternatives. Participant support for the proposed research varied but on average was close to neutral (mean ± SD: 4.5 ± 2.19) suggesting some ambivalence to this animal use. The protocol describing Parkinson’s research (on monkeys) was least supported (3.9 ± 2.17) and the transplant research (on pigs) was most supported (4.9 ± 2.02). These results indicate that public participants are sensitive to specifics of a protocol. We conclude that an online forum can provide meaningful public input on proposed animal research, offering research institutions the opportunity for improved transparency and the chance to reduce the risk that they engage in studies that are out of step with community values.

Skin swabbing protocol to collect DNA samples from small-bodied fish species

Fish species are commonly used as experimental models in the laboratory. DNA is routinely collected from these animals to permit identification of their genotype. The current standard procedure to sample DNA is fin clipping, which involves anaesthetising individuals and removing a portion of the caudal fin. While fin clipping reliably generates good quality DNA samples for downstream applications, there is evidence that it can alter health and welfare, leading to infection and impacting on the fish’s behaviour. This in turn can result in greater variation in the data collected. In a recent study we adapted a skin swabbing protocol to collect DNA from small-bodied fish, including sticklebacks and zebrafish, without the use of anaesthetics or sharp instruments. A rayon-tipped swab was used to collect mucus from the flank of the fish, which was then used for DNA extraction. We subsequently demonstrated that compared to fin clipping, skin swabbing triggered fewer changes in stress axis activation and behaviour. We also found that data collected from fish that had been swabbed were less variable than data from fish that had been fin clipped, potentially allowing smaller sample sizes in experimental groups after using this technique, and thereby reducing animal use. Here we provide a detailed protocol explaining how to collect DNA samples from small laboratory fish using skin swabs.

The Development of Sleep/Wake Disruption and Cataplexy as Hypocretin/Orexin Neurons Degenerate in Male vs. Female Orexin/tTA; TetO-DTA Mice

Narcolepsy Type 1 (NT1), a sleep disorder with similar prevalence in both sexes, is thought to be due to loss of the hypocretin/orexin (Hcrt) neurons. Several transgenic strains have been created to model this disorder and are increasingly being used for preclinical drug development and basic science studies, yet most studies have solely used male mice. We compared the development of narcoleptic symptomatology in male vs. female orexin-tTA; TetO-DTA mice, a model in which Hcrt neuron degeneration can be initiated by removal of doxycycline (DOX) from the diet. EEG, EMG, body temperature, gross motor activity and video recordings were conducted for 24-h at baseline and 1, 2, 4 and 6 weeks after DOX removal. Female DTA mice exhibited cataplexy, the pathognomonic symptom of NT1, by Week 1 in the DOX(-) condition but cataplexy was not consistently present in males until Week 2. By Week 2, both sexes showed an impaired ability to sustain long wake bouts during the active period, the murine equivalent of excessive daytime sleepiness in NT1. Body temperature appeared to be regulated at lower levels in both sexes as the Hcrt neurons degenerated. During degeneration, both sexes also exhibited the Delta State, characterized by sudden cessation of activity, high delta activity in the EEG, maintenance of muscle tone and posture, and the absence of phasic EMG activity. Since the phenotypes of the two sexes were indistinguishable by Week 6, we conclude that both sexes can be safely combined in future studies to reduce cost and animal use.

Research Progression of the Genus Merremia: A Comprehensive Review on the Nutritional Value, Ethnomedicinal Uses, Phytochemistry, Pharmacology, and Toxicity

The genus Merremia Dennst. ex Endl. (Convolvulaceae) is a rich source of structurally diverse phytochemicals with therapeutic relevance. This review presents the first comprehensive, up-to-date information and research progression on the nutritional value, ethnomedicinal uses, phytochemistry, pharmacological activities, and toxicity of the genus Merremia. Using the key search term “Merremia”, relevant documents and information were retrieved from electronic databases. Relevant documents were uploaded in RStudio with installed bibliometric software packages and used for data retrieval, tabulation, and network visualization. Bibliometric analysis revealed that ca. 55% of the studies related to Merremia were published in the last decade, which can be grouped into four thematic areas: (i) drug formulation, (ii) taxonomy, (iii) chemical analysis, and (iv) treatment of diseases. Ethnomedicinal uses, phytochemistry, and biological activities studies showed that species in the genus are promising medicinal plants with various pharmaceutical potentials. However, clinical studies to validate the efficacy of the reported bioactivities and the mechanisms underlying the various activities are lacking and should constitute a future research focus. Additionally, reports on the nutritional and antinutritional constituents of Merremia species revealed that the species meet high nutritional quality criteria for animals and are therefore suitable for inclusion in livestock diets. The few available investigations on toxicity indicated that most Merremia species are safe for human and animal use but not with prolonged chronic administration.

Issues of Animal Welfare Regulations on Pharmacy Education and Research in India

Biomedical research is essential to the health and well-being of our society. Animal use for biomedical research has a long history and is routinely performed in new drug discovery and development processes. Animal experiments are an integral part of the curriculum for students in the life sciences, including pharmacy, to learn how to conduct animal experiments. These experiments may cause pain and distress to the animals. Laws and regulations have been enacted to make it illegal to cause undue pain or suffering to animals. These guidelines provide that due and full consideration should be given to alternative technologies not involving animal testing. Despite the movement to minimize animal use in research, pieces of evidence show that there has been a continuous increase in the worldwide use of laboratory animals over 10 years, from 115.2 animals to 192.1 million. The lack of suitable animal-alternative technologies and unavailability of required infrastructures are some of the reasons for animal use. As per directives of the University Grant Commission, the Pharmacy Council of India has decided to prohibit animal experimentation in pharmacy education. This adversely affected teaching and research activities in pharmacy institutions. As a result, the number of seats available for the postgraduate course (Master of Pharmacy) in Pharmacology is decreasing every year. In 2021, the highest number of seats are available for Pharmaceutics (9510, 35%) followed by that for the Pharmacology (4620, 17%). This article mainly focuses on the background of Indian legislation for animal experimentation and the impact of these regulations on animal experiments for pharmacy education and research in India.

Making room for the 3Rs principles of animal use in ecology: potential issues identified through a survey

Research on animals is one of the most controversial ethical issues in our society. It is imperative that animal welfare is being considered and the harm and distress to animals used in research is minimized. This could be achieved through implementation of the so-called 3Rs principles for animal research, which are now implemented in many legislations worldwide. These principles serve as a basis for research without the use of animals (Replacement), with as few animals as possible (Reduction), and in which the animal’s welfare is as good as possible (Refinement). While there has been a lot of focus on implementation of these principles, only a few studies have documented the knowledge and adoption of the 3Rs among researchers. One field that has been particularly neglected is ecological research, which can involve many practices that affect animal welfare. Moreover, the knowledge, experience, and attitudes about animal use in ecological research and education has never been examined before. In order to close this gap, I conducted a survey among European ecologists. Responses from 107 respondents from 23 countries revealed that lethal and invasive research methods are prevalent, and that more than half of the respondents have never heard of the 3Rs principles for animal research. Major concerns are also the lack of calculation of the minimum sample size and widespread of dissection classes as part of education. Additionally, most respondents experienced ethical doubts about their research, and did not receive any training in animal welfare or ethics. These findings revealed that it is necessary to implement rigorous standards for ecological research and enforce the implementation of the 3Rs principles. Furthermore, the evaluation of current educational practices in ecology is urgently needed.

A review on: Alternatives to animal experimental models

An English solicitor, Jeremy Bentham, disdained the unregulated use of livestock, in the literature of 1780, which cast doubt on the inability to pay morale to animals. Animal experiment was fundamental to scientific science during his life, but it has also been a question of heated popular and intellectual discourse from decades. Since then, general understanding and attitudes have strengthened against the unethical or cruel application of animals for research purposes. Russell & Burch provided in 1959 the principles set out during UFAW of the clinical lab method. The alternative measures, replacements, or non-animal steps shall be used as alternatives for live animal methods, or methods of measurement without live animal use. The term 'alternative' refers to approaches or strategies that substitute the overall use of laboratory animals, minimize the number of animals used, or enhance a current system or technology to reduce animal stress. In recent years, the creation of alternatives to animals has been accelerated by designing projects with the intention of developing and introducing alternatives. This report summarizes the key potential approaches to animal science. An integrated use of these techniques will offer an insight into the minimal use of animals for experimental research.

Establishment of a Newborn Lamb Gut-Loop Model to Evaluate New Methods of Enteric Disease Control and Reduce Experimental Animal Use

Enteric infectious diseases are not all well controlled, which leads to animal suffering and sometimes death in the most severe cases, in addition to economic losses for farmers. Typical symptoms of enteric infections include watery diarrhea, stomach cramps or pain, dehydration, nausea, vomiting, fever and weight loss. Evaluation of new control methods against enteric infections requires the use of many animals. We aimed to develop a new method for an initial in vivo screen of promising compounds against neonatal diseases such as cryptosporidiosis while limiting experimental animal use. We therefore adapted an in vivo method of multiple consecutive but independent intestinal loops to newborn lambs delivered by cesarean section, in which endotoxin responsiveness is retained. This new method allowed for the screening of natural yeast fractions for their ability to stimulate immune responses and to limit early Cryptosporidium parvum development. This model may also be used to investigate host–pathogen interactions and immune responses in a neonatal controlled environment.