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Global trends of animal ethics and scientific research

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This article deals with global trends and guidelines followed by different countries in scientific research. The various objectives of animal research, pros and con and alternative method for animal research by the use of 3Rs are disused here it is felt that however animal research is indispensible in several research but in certain alternative are alternately effective means of animal rights. This article reviews various aspects which are related to this issue.
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Journal of Medicinal Plants Studies 2017; 5(2): 96-105
ISSN (E): 2320-3862
ISSN (P): 2394-0530
NAAS Rating 2017: 3.53
JMPS 2017; 5(2): 96-105
© 2017 JMPS
Received: 01-01-2017
Accepted: 02-02-2017
Manshu Jain
School of Pharmacy, Suresh
Gyan Vihar University, Mahal
Jagatpura, Jaipur, Rajasthan,
Ritu Gilotra
School of Pharmacy, Suresh
Gyan Vihar University, Mahal
Jagatpura, Jaipur, Rajasthan,
Jitendra Mital
Ayushraj Enterprises Pvt. Ltd.,
Village Mansinghpura, Dehmi
Begus Road, Ajmer Road Jaipur,
Rajasthan, India
Jitendra Mital
Ayushraj Enterprises Pvt. Ltd.,
Village Mansinghpura, Dehmi
Begus Road, Ajmer Road Jaipur,
Rajasthan, India
Global trends of animal ethics and scientific
Manshu Jain, Ritu Gilotra and Jitendra Mital
This article deals with global trends and guidelines followed by different countries in scientific research.
The various objectives of animal research, pros and con and alternative method for animal research by
the use of 3Rs are disused here it is felt that however animal research is indispensible in several research
but in certain alternative are alternately effective means of animal rights. This article reviews various
aspects which are related to this issue.
Keywords: Animal models, Ethics, Guideline, Cons
Humans have benefited from research involving animals, in every medical achievement in the
past century. Developments in the many treatments such as diabetes, leukaemia and heart
surgery transplants have been possible through the use of animals in research. The majority of
the scientific community consider that the use of animals in research justified. The public also
now accepts the use of animals in research due to the following reasons:
They believe now there is no unnecessary suffering to the animals;
The research is for serious medical or life-saving purposes;
There is no reliable and informative alternative to their use. It is important to recognise that
work on animals in research, whose benefits to health and in treatment of disease are not
evidenced firstly. This is highly useful as it provides or gives the groundwork or basic
structure for medical advances. The moral for animal’s research should be to minimise animal
suffering and maximise the benefits to medicine, health, agriculture and fundamental
understanding of body.
Animal research only takes place within a framework of robust controls. UK legislation
requires refining the procedure to keep the minimum suffering of animals, and the use of
animals should be that much only which gives meaningful results and the use of animals with
non-animal alternatives methods is also appropriate. However, sometimes the use of non-
animal methods may not be useful for some types of research. The majority of research using
animals is about 84% in which the rodents are important subject for the experiment. Basic
research and drug development for 82% of all procedure is safely performed comparing to the
remaining 18% research [13-15].
The use of animals in research can be alternative research method evidence that research by
using animal is unnecessary. Non-animal methods, such as tissue culture, computer modelling,
research using human test subjects and population studies, are used and have utility for
scientific and medical research. All the research guidelines which are for animal welfare had
very high value for scientific and medical understanding. This is because a research study
sometime needs more than knowing how individual molecules, cells or tissues behave, but
then too it is unethical and illegal to expose human or animal new medicines knowing that it is
benefited or not, so the only alternative method is useful for study of the new entity for health
improvement and for treatment of diseased condition. The guidelines which work for animal
welfare or for ethical research on animal give the many guidelines on the different level such
as Royal Society for animals welfare they give numerous bodies, including government
departments, funding agencies, charities and discussion meetings [16, 17].
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Pros and Cons for Animals
Animals are essential for research which is use in cures and
treatments of diseases that affect human.
A better life and health is found by the experiment on the
animals, but with this good thing comes a bad one too that is
experiment on animals some time which lead to chronic
toxicity or death of the animal. There are many people and
community who
opposing the use of animals in experiments by claiming it as
non-ethical way to have an experiment on animals.
This section helps us to know the pros and cons of animal in
Pros [1]
Animal testing is useful as it contribute in life-saving by
cures and treatments in human.
Case Study: The California Biomedical Research Association
states that nearly every medical breakthrough in the last 100
years has resulted directly from research using animals.
Experiments in which dogs had their pancreases removed led
directly to the discovery of insulin, critical to saving the lives
of diabetics.
Animal research has also contributed to major advances
in understanding and treating conditions such as breast
cancer, brain injury, childhood leukaemia, cystic fibrosis,
malaria, tuberculosis, leprosy, blood cancer, insulin and
many others, and in the instrumental development of
pacemakers, cardiac valve substitutes, and anaesthetics.
Much research cannot be possible without animal studies.
For Example: "we wouldn't have a vaccine for hepatitis B
without chimpanzees”, a disease that kills more than 14,000
people every year in the United States.
Animals are always used when ethical considerations
prevent the use of human subjects, the lives of human
volunteers and patient cannot be put in danger
unnecessarily. The World Medical Association
Declaration of Helsinki states that human trials should be
preceded by tests on animals.
It will be very unethical to perform experimental
procedures on human beings before tested on animals,
such as in the genetic manipulation experiment to directly
impose on human subjects before animal testing.
Animals themselves get benefit from the results of animal
testing, such as if vaccines were not tested on animals,
millions of animals would have died from rabies,
distemper, feline leukaemia, infectious hepatitis virus,
tetanus, anthrax, and parvo virus.
Animal research is highly regulated, with laws and
guidelines to protect the animals in laboratory and
research by mistreatment. Such as Animal Welfare Act
(AWA) since 1966, these give minimum housing
standards for research animals and also work for regular
inspections by veterinarians.
Animal researchers treat animals humanely, both for the
animals sake and to ensure reliable test results which will
be helpful for human too, as all experiment perform
under approval of by an Institutional Animal Care and
Use Committee (IACUC)
Commercially VS Research use of animal.
Example: 1. People in the US eat more than 9 billion
chickens and 150 million cattle, pigs and sheep annually, as
compare to them only 26 million animals are used for
research, in which 95% of which are rodents, birds and fish.
As compare to them we eat more than 1,800 times the number
of pigs than the number used in research.
Similarly, the use of animal in leather preparation is more
than the animal use in research it is approximately more than
15% as compare to research.
Cons [1]
Animal testing is cruelty of the human for the animals.
As the animal is forced for feeding, forced inhalation,
food and water deprivation, prolonged periods of
physical restraint, fasting for long period of time, and
give them the infliction of burns and other wounds to
study the healing process, the infliction of pain to study
its effects and remedies, and "killing by carbon dioxide
asphyxiation, neck-breaking, decapitation, or other
Is really an inhuman behaviour by human towards the
For Example: The Draize eye test, is another method used by
cosmetics companies to evaluate irritation caused by
shampoos and other products, in this the rabbits being
incapacitated in stocks with their eyelids held open by clips,
sometimes this test take the long-time not only hours but days
too, so they cannot blink the eyes during products testing.
During LD50 the animal was killed to test the toxicity
they were sacrificed after the experiment due to severe
condition happened during experiment.
Case Study: The US Department of Agriculture (USDA)
reported in 2010 that 97,123 animals suffered pain during
experiments while being given no anaesthesia for relief,
including 1,395 primates, 5,996 rabbits, 33,652 guinea pigs,
and 48,015 hamsters, is this correct with animal?
Alternative testing methods can be used to replace the
need for animals. Drugs that pass animal tests are not
necessarily safe.
Case Study 1: The 1950s sleeping pill thalidomide, which
caused 10,000 babies to be born with severe deformities, was
tested on animals prior to its commercial release. Later tests
on pregnant mice, rats, guinea pigs, cats, and hamsters did not
result in birth defects unless the extremely high dose of drug
was administered.
Case Study 2: Animal tests on the arthritis drug Vioxx
showed that it had a protective effect on the hearts of mice,
yet the drug went on to cause more than 27,000 heart attacks
and sudden cardiac deaths before being pulled from the
95% of animals used in experiments are not protected by
the Animal Welfare Act.
Such As: Animal Welfare Act does not cover rats, mice, fish
and birds, which comprise around 95% of the animals, used in
research they, are small or not necessary as animal what the
The Animal Welfare Act has not succeeded in preventing
horrific cases of animal abuse in research laboratories.
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Case Study: In Mar. 2009, the Humane Society of the United
States (HSUS) found 338 possible violations of the Animal
Welfare Act at the federally funded New Iberia Research
Centre (NIRC) in Louisiana.
Medical animal research may still have been made
without the use of animals but, there is no enough money
and resources to use the alternative method, for this the
community working for animal welfare and common
people should take the action by participating in the
animal welfare issue.
Statements from Animal Rights Extremist Groups are as
“I don’t think you’d have to kill too many [researchers]. I
think for five lives, 10 lives, 15 human lives, we could save a
million, 2 million, 10 million non-human lives.”
–Jerry Vlasak, spokesperson for SHAC and ALF
“In a war you have to take up arms and people will get killed,
and I can support that kind of action by petrol bombing and
bombs under cars, and probably at a later stage, the shooting
of vivisectors on their doorsteps. It’s a war and there’s no
other way you can stop vivisectors.”
–Tim Daley, ALF
Why Use Animals In Research?
The experiment on animal is due to several reasons
First, in some research by studying animal behaviour, we
can gain an understanding of human behaviour.
In a similar manner by understanding the impact of
different or many environmental factors such as
crowding, cognitive stimulation, enrichment, or
deprivation on animal behaviour, by this we understand
how these environmental factors affect human behaviour.
For Example: The work of Harlow and Zimmerman (1959),
In their experiment, infant monkeys was separated from their
mothers and kept under two different surrogate conditions; in
one condition the surrogate “mother” was constructed of wire
with a wooden head, while in the other condition the wire was
covered with terry cloth to provide softness and warmth. The
experimenters were interested in seeing whether the primary
function of attachment to the mother was simply for survival
as she provides food. What the studies found was that,
regardless of which “mother” provided the food, the baby
monkeys spent more time clinging to the terry cloth “mother.”
By this experiment they concluded that attachment is not
merely for survival (food) but for contact comfort: something
warm and soft to cling. However, the studies also found that,
regardless of which type of surrogate the monkeys had, all
developed problems later on. They were either inappropriately
aggressive or timid, and the females tended to neglect or
abuse their own young. Can we conclude from this study that
attachment between human infants and their mothers also
goes beyond the simple need for food and survival? In other
words, we can say the findings of the Harlow et al. studies to
human children and hypothesize that a lack of opportunity for
contact comfort, or attachment, will also have an adverse
effect on human development? The studies conducted by
Harlow et al., for example, could not have used human
participants for obvious ethical reasons.
Many of the drug and treatment was developed through
the use of animal research are used and it also help in the
treatment of animals themselves.
Example: Diseases such as tuberculosis, scarlet fever, and
polio, not to mention high blood pressure, would continue to
claim millions of lives. In addition, According to Perkins,
90% of the animals used in research are rats, mice, and fish,
and are bred solely for research purposes.
Misguided attempts on animals can also have tragic
consequences for the animals themselves.
For Example: In a recent attack on the animal research
laboratory at the University of Minnesota, the animals were
“liberated” and set free on a field near the school. Many of the
animals were found wandering by the road, hardly a safe
environment for animals, and several of the stolen rats were
found dead.
On the basis of this type of issue, animal research does
continue, and it is governed by ethical guidelines for
research involving human participants is regulated.
Alternative Method to Avoid Use of Animal
“Alternatives” as encompassing any subjects, protocols, or
technologies that ‘(replace the use of laboratory animals
altogether, reduce the number of animals required, or refine
existing procedures or techniques so as to minimize the level
of stress endured by the animal” [4, 5].
Vitro (in glass) testing, such as studying cell cultures in a
petri dish, can produce more relevant results than animal
testing on different rodent and non-rodent species
because human cells can be used. Micro dosing can be
administered to the human who gives the appropriate
result on human volunteers, whose blood is then
Artificial human skin, such as the commercially available
products Epi Derm and Thin Cert, can be used as it is
made from sheets of human skin cells grown in test tubes
or plastic wells which can produce more useful results
than testing chemicals on animal skin as some times
result varies animal and between human.
The animals use in scientific research is act as tool in
improving the understanding of how biological systems work
both in health and disease condition. The developments of
new medicines are for both humans and animals, and for the
protection of our environment. Hence, it was properly
regulated by use of animals in improving the health and lives
of humans and animals and to the safety and sustainability of
our environment.
This research supports the UK’s world-class research base on
the environmental, agricultural, medical and other life
sciences. The alternative method improves health and
diseased condition of patients and animals in the UK by the
outcomes of research.
National Health Service is the primary source of research and
testing on animals such as by the licensing of new
medicines, and the development of new and safe vaccines and
for the detection and control of infectious diseases.
For Example
The development of monoclonal antibody had completely
transformed our ability to treat diseases including breast
and other cancers.
Journal of Medicinal Plants Studies
Rheumatoid arthritis and multiple sclerosis treatment was
also developed by the use of this technology.
It also supports a number of highly skilled jobs in research on
the base of an attractive regulatory environment.
Economically the life sciences sector contributes over £50bn a
year to the UK economy with agriculture contributing £9bn
and it underpins the UK’s £26bn sector providing us with
healthy and wholesome food.
In 2010, the Government made a commitment to reduce the
work on use on animals in scientific research. It encompasses
Replacement, Reduction and Refinement (the 3Rs) more
broadly, putting them at the science-led approach.
Implementing of the 3Rs requires in every research proposal,
which include:
Animals are replaced with non-animal alternatives
wherever possible,
The number of animals used is reduced to the minimum
needed to achieve the results sought, and
For those animals which must be used, the procedure
should be refined as much as possible to minimise the
suffering of animal.
The scientific imperative for developing new approaches to
research and development is very strong. Similarly, there are
concerns about the utility of animal studies for testing
environmental chemicals.
For Example: Animals are exposed to too much higher doses
than typical human exposures making interpretation difficult.
An international science organisation called for the
development of assays that are more predictive to human
biology. Attention has focused on non-animal technologies; it
include tissue engineering, stem cells, and mathematical
modelling [5].
On 18 July 2012, Written Ministerial Statement set our
ambition to deliver the commitment such a science-led
programme, spearheaded by relevant government departments
and agencies, working together where appropriate, and by the
National Centre for the Replacement, Refinement and
Reduction of Animals in Research (NC3Rs), an organisation
with record in promoting the 3Rs in animal research.
Plan demonstrates how taking advantage of technological
developments, including increased computer power and wider
scientific innovation, to provide the perfect environment and
catalyst for scientific change and progress which will help to
reduce the use of animals in research. The Delivery Plan set
out the Government three strategic by putting the 3Rs at the
science-led programme:
advancing the use of the 3Rs within the UK;
using international leadership to uptake and adoption of
3Rs approaches globally;
Understanding and awareness about the use of animals
where no alternatives exist.
Strategic priorities are considered in detail in Section 2 of this
Delivery Plan.
In this Strategy UK Life Sciences was published in 2013
whose aim is to make the UK the location for research and
development together for manufacturing. This Delivery Plan
shows how 3Rs makes a real contribution to realising benefits
for humans, animals, environment and sustained economic
Hence: for all these above points we use the alternative
methods which help to sustain growth of economic, animals
and humans and environment too [6].
3Rs is proposed by Charles Hume, founder of the Universities
Federation for Animal Welfare (UFAW) in 1954,
consideration alternatives was made for animal testing on
laboratory animal experiments. R.L. Burch and the zoologist
W.M.S. Russell is the microbiologist who publishes the work,
"The Principles of Humane Experimental Technique" in
London in 1959, and the book also defined the animal testing
alternatives as “The Three R's: Refinement, Reduction, and
Replacement [7, 8]
Laboratory animals are manly used in following areas:
Biomedical Researchers: use animals to understand the
body metabolism and catabolism, disease and health
condition of the body, and to develop new vaccines for
treatments of various diseases.
Industry: uses animals to test the effectiveness and
safety of consumer products
Such As: for drugs, cosmetics, household cleaning products,
pesticides, industrial chemicals, and for many more test.
Educational: uses include dissecting earthworms or
frogs in biology class which are now rarely used, as well
as in advanced training of surgical techniques for
veterinary and medical students.
Psychological Or Social Behaviours Research: is used
for study of animals to learn more about a given species,
its biology and behaviour. They may learn from the
special skills or abilities of an animal as well such as
animal models were used.
For Example: Navy researchers have studied dolphin
echolocation--their built-in biological sonar system to
improve the human-made sonar systems used on board ships.
In the research laboratory an animal may be poisoned;
deprived of food, water or sleep; skin and eye irritants were
applied; subjected to psychological stress; deliberately
infected with disease; brain damaged; paralysed; surgically
mutilated; irradiated; burned; gassed; force fed and
electrocuted [10, 11].
Replacement of Animals in Medical Experiments (FRAME)
considered that the animal experimentation is unacceptable,
but the abolition of all animal in experiment is not possible.
New consumer products, medicines, and industrial and
agricultural chemicals can be tested in order to identify
potential of human and animal health, and to the environment.
Alternative methods fall into three broad categories. These are
called the 3 Rs
Reduction, and
Replacement is what most people think of when you say
"alternatives to animal testing"
The animals are replaced, either by methods that don’t
involve animals at all that are called as absolute
Journal of Medicinal Plants Studies
By those that use only the cells or tissues of animals that
is called as relative replacement.
Such As: Many replacement alternatives involve in vitro or in
glass techniques, where the studies are perform with cells or
tissues in culture. If the cells come from human beings, it's
absolute replacement, and if from animal it is called as
relative replacement [10].
Replacement also means replacing 'higher' animals with
'lower' animals.
Such As: Microorganisms, plants, eggs, reptiles, amphibians,
and invertebrates may be replaced by the warm-blooded
animals in many experiments.
Live animals may be replaced with non-animal models, i.e.
dummies for dissection in teaching the animal or the human
body structure, mechanical or computer models, audio-visual
aids, or in vitro modelling [12].
Replacement utilizes the pre-existing knowledge for new
system to look similarities and for using less expensive
animals or models to have research on agents for toxicity
or mutagenicity [10].
Disadvantages for replacement is that if any model are
dependent on pre-existing information, the complex
physiology and pathology of organism was not known.
Hence, research on new biological processes must utilize
a living organism; unfortunately, replacement isn't
always an option [12, 10].
Example of Replacement: of a replacement alternative
is no longer considered an alternative it has become the
norm. Not too many years ago, if a woman wanted to find
out if she was pregnant, she'd have to get a laboratory test
that involved killing a rabbit. Now, she can buy a small
kit over-the-counter that tests her urine for certain
chemicals the rabbits have been replaced [10, 11].
Reductions mean minimizing or reduce the number of animals
needed to perform an experiment or teach a concept.
In research testing some test cannot perform without
animals, at least at this research time. Researchers can
also work by reduce the number of animals used in a
given study.
Sometime the uses of far fewer animals are given valid
results. These methods include performing pilot studies
to determine the potential problems in an experiment
before numerous animals are used.
Designing a study is done by utilizing number of animals
by our own control, it include to have a maximum
amount of information from each animal, or gathering
many or much data from more than one experiment
Statistician consult should be taken for numbers of
animals required to achieve significance result during
result, or by Minimizing variable condition of animal
which affect the result such as, disease, stress, diet,
genetics, etc.
The appropriate species of animal also useful for the data
collection, and Replacement should be done whenever it
is required or possible [9-10].
Refinement means refining experimental protocols to
minimize pain or distress to the animal whenever it is possible
for the experiment.
For those animals that do undergo testing, scientists may
refine their methods to lessen or eliminate pain, distress,
or suffering and to make the animals more comfortable.
Examples of refinement which include it for the
identification pain and distress and making plans and
executed for preventing or relieving it.
During measuring the toxicity of a compound or survival
can be determined not by death just by chronic toxicity.
Such As: A tumour achieves a certain size; at the time of
course death is predictable. Subsequent experiments may be
utilized the earlier endpoint of tumour size or clinical signs of
toxicity, rather than death as the endpoint.
Refinement Can Be Done By: [8, 10]
Receiving adequate or proper training for performing a
By using proper handling technique for animals.
By ensuring that drug doses are correct and drugs should
not expired,
The performed experiment on the animal is reasonable
for that species.
Using analgesic sand anaesthetics for potentially painful
Performing surgeries procedures aseptically to prevent
Performing only a single major survival surgery on any
one animal, whenever possible.
Different Alternative Methods
Alternatives to animal use in biomedical and behavioural
research fall into four broad categories:
Continued, but modified, animal use, including a
reduction in the number of animals used, improved
experimental design and statistical analyses of results,
substitution of cold-blooded for warm-blooded
vertebrates, substitution of laboratory mammals for
domestic or companion mammals, and reduction of pain
or experimental insult;
use of living systems, including in vitro cultures (of cells,
tissues, and organs) embryos, invertebrates, micro-
organisms, and plants;
Use of non-living systems by using chemical or physical
systems; and
Computer simulation.
Biomedical and behavioural had focus attention on drug
development by using alternatives rather than animal
methods. Alternative methods in research show advantages
that can be based on the prospect as follow i.e. on scientific,
economic, and humane which include including:
The number of animals used can be reduced;
Animal pain, suffering, and experimental insult can be
Reduction in investigator-induced and art factual
physiological phenomena;
Using the process which provide more quickly result;
To perform ability for replicative protocols on a routine
Journal of Medicinal Plants Studies
cost of research reduction;
alter conditions for flexibility with experimental protocol;
At the same time, these methods are fraught with inherent
disadvantages, including:
Ability of organismal growth process reduces;
Ability to study cells, tissues, and organ systems reduces
by acting in concert;
Biochemical and metabolic pathways cannot studies;
Behaviour study on cell tissue or organ cannot possible;
Study of recovery of damaged tissue cannot possible;
Interaction between the organism and its environment
ability reduces;
Decrease ability to study of idiosyncratic or species
specific responses;
Drug effect on male and female-specific phenomena
cannot be possible; and Different alternative methods are
as follow:
Organ Culture
In history of research, investigators have attempted success in
every major and minor organ of mammalian body in variety
of research purposes. In last few years, techniques were
improved, such as artificial blood media system which helps
in, successful organ culture. The applications of organ culture
include the study of protein synthesis in lactating mammary
tissue [21] and by using human placentas in toxicology studies
[18]. Whole organs are not use in long-term in vitro culture.
The complexity of whole organs makes it impossible for
nourishment and for normal function of growth without any
external support. Cryostat sections of organ are studies into
organ-specific test for metastatic tumour cells. These in vivo
events reflect the use of whole animals in a very active
research area [20]. Whole mammalian embryos are used basic
development in biological research and other experiment,
which have been cultured in vitro for other purposes.
Protocols included the effects of hormones and teratogens [22].
Tissue Culture
Tissues from humans and animal species can be studied in
tissue culture. The progress have been achieved in 1907, by
R.G. Harrison who studied first frog tissue outside the body
for weeks, which had changed the tissue culture from an art of
science. Bacteria or virus are alive for more than few hours to
gain a better result in the antibiotic study. The tissue culture
was extended by controlling bacterial contamination. In tissue
culture, isolated tissue of a living organism is maintained by
various cells which are arranged as they were in the original
organism. Tissue cultures are both “better” and “worse” than
a single cell type. They act as better in the manipulation in
different natural environment and different cell types which
show interaction similar to in vivo, as they are worse in more
difficult condition to maintain. Tissue-culture experiment
requires animal sacrifice, as they are alternative to numerous
sections of animal adjoining tissue to use. By this way, two or
more research can be performed in the single animal tissue,
rather than a number of individual animals. Tissue culture is
successfully used in biomedical research. In neurology,
embryonic rat tissues are used to examine the destruction of
dopamine neurons [25], which is prior to development, as
monkey act as best model for the study of degenerative and
the other study to degenerative effect observed in humans.
Cell Culture
It is not a new technique; development and application of cell
culture are seen in many past year in different fields. Cell
culture every ethic of biomedical research, as well as in
clinical practice.
The approaches of biomedical research by cell culture method
are as seen:
Eggs and sperm of many species are used in the study of
fertilization and early development [28, 19].
Mutants are used to understand membrane associated
proteins which are essential for knowing animal cell
function [29].
In oncology, bacterial recombinant DNA induces in the
transformation of white blood cells which is used to
observed infection, cancer, and rheumatoid arthritis [30].
Steroid metabolism is studied by using rat epididymis
cells [31].
A kidney cell of monkey was use to demonstrate the
metabolic effects of many drug [26].
Surgical research is use to cultured human epitheliums
for permanent coverage of large burn wounds has moved
from the lab- oratory into clinical trials [32].
In immunology, studies on antibody synthesis and
response have been bolstered by the Nobel-prize-winning
elucidation of monoclonal antibodies.
Biologists have developed techniques for the controlled
disruption of cells that can leave many organelles intact or
allow the harvesting of selected intracellular membranes.
These fractions have proved to be invaluable in the search for
information at the molecular level.
For Example
Microsomal membrane fractions from rat and human
liver have been used in comparative anaesthesia research
The most unique uses of subcellular fractions involves
the bringing together of mixed species systems in
biochemical studies of protein transport across
intercellular membranes.
Researchers studying intracellular protein translocation
used dog pancreatic microsomes, bovine pituitary and
rabbit reticulocyte messenger RNA, bacterial nuclease,
and a wheat germ cell free system to elucidate the
structure of the signal recognition particle [33].
Ethical Issue [12, 34, 35]
Many ethical issue works in animal research which are
explained in the different issue as quarry with solution:
‘Are humans or animal status are different?’
People in large number believe that humans have the high
moral status, then animal they believe animals are inferior
then humans. However, others disagree and say that this
discrimination is having no justification in it. Status of
humans and animals can be discussed in following ways:
One characteristic i.e. self-awareness or rationality.
Capacity of suffering by animal, which characterised as is
an animal’s sentience, or to understand its pain, suffering
and pleasure.
Social relationships of animal should also considered,
while many animals differ in these characteristics from
humans, and also different between animal to animal.
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‘How can we know how much animals suffer?’
A recent data show that the amount of pain suffered by
animals is more as people think; it is very difficult to
understand human concepts and emotions about pain, distress,
fear, happiness or affection to animal. Even if we can say that
animals have similar brain activity as humans, which means
pain suffering in the animal is in the same way.
‘Can we justify making animals suffer?’
Many people believes that it is wrong to make animal suffer
without reason; but some people think that there is some
condition more than the interests of people outweigh the
interests of an animal, this justify animal suffers more than
In research using animals, the costs, in the form of suffering,
are largely experienced by the animals while any benefits
arising from the research are largely directed to human
Guidelines for Ethical Research [36-39]
The UK has regulation on research involving animals
worldwide. Law of the Animals are (Scientific Procedures)
Act 1986. This law was act to protect animal by cause “pain,
suffering, distress or lasting harm” must be licensed.
‘Protected’ animals include all vertebrates (mammals, birds,
reptiles, amphibians and fish, and octopus). All procedures
must carry three separate licenses for the premise; the
researcher and the project.
Licences for research involving animals
Premise Licence: The place where research is conducted
must meet standards of animal housing and care and is
regularly subject to inspection by the Animals (Scientific
Procedures) Inspectorate. In 1999, there were 296 designated
premises in the UK.
Personal Licence: Any person conducting animal procedures
must be competent to do so and must have completed an
accredited training course in order to be awarded a licence.
This licence specifies which procedures can be conducted and
on which species. In 1999, there were around 13,700 active
personal licences in the UK.
Project Licence: This defines the types of animals which
may be used, the estimated number to be used, and the
procedures that may be carried out. The licence application
includes details of the purpose of the project, an initial
welfare assessment and a limit of severity for each procedure.
In general, project licences should only be granted if:
The research cannot be done without animals;
The likely costs of the research are weighted against the
potential benefits in a cost benefit assessment, and the
benefits are deemed to be greater;
The minimum number of animals will be used;
Researchers and technicians are trained and experienced
in animal testing;
Research premises have facilities for animal care.
Inspectors visit laboratories to assess licence regulations.
There are currently 25 inspectors in the UK, who make more
than 2,100 in every year. The independent Animal Procedures
Committee (APC) was established to advise the Secretary of
State on the operation of the 1986 Act.
Testing Guidelines
Testing guidelines are developed for a variety of reasons:
To allow results of various test substances or species to
be easily compared,
To encourage the use of certain protocols so that testing
need not be repeated, and
To facilitate the work of those who design and carry out
Many organizations differently developed their own testing
FDA Guidelines Involving Whole Animal Testing
OECD Guidelines Involving Whole Animal Testing
Pesticide Assessment Guidelines Involving Whole-
Animal Testing
Ethical guidelines of animal testing are:
CPCSEA Committee for the Purpose of Control and
Supervision on Experiments on Animals
CARE Community on Animal Research and Ethics
GCUASR Guidelines For Care And Use Of Animals In
Scientific Research
NACLAR National Advisory Community for Laboratory
Animal Research
APA American Psychological-al Association
Table 1: Different Community Work for the Ethical Guidelines for Animal
Full Form
Committee for the Purpose
of Control and Supervision
on Experiments on
Community on Animal
Research and Ethics
Guidelines For
Care And Use Of
Animals In
National Advisory
Community for
Laboratory Animal
American Psychological-
al Association
India Washington, DC
Indian National
Australia, Canada,
New Zealand, US, US
Guidelines is to promote
the humane care of animals
used in biomedical and
behavioural research and
testing with the basic
objective of providing
specifications that will
enhance animal well-being,
Important parts of these
endeavours are teaching
and research on the
behaviour of nonhuman
animals, which
contribute to the
understanding of basic
principles underlying
The interest of
society and the
welfare of
Guidelines set out
the responsibilities
of all the parties
involved in the care
and use of animals
for scientific
purposes, in
accordance with
The acquisition, care,
housing, use, and
disposition of nonhuman
animals in research must
be in compliance with
applicable federal, state,
and local, laws and
regulations, institutional
Journal of Medicinal Plants Studies
quality in the pursuit of
advancement of biological
knowledge tl1at is relevant
to humans and animals.
behaviour and to
advancing the welfare of
both human and
nonhuman animals.
widely accepted
scientific, ethical
and legal
policies, and with
international conventions
to which the United
States is a party.
Table 2: Points Covered In Different Guidelines
Veterinary care Justification of
the research
Sources of
experimental animals
General principles for the care and use
of animals for scientific purposes
Justification of the
Animal procurement Personnel
Laboratory animal
husbandry and
Animal housing and management Personnel
Quarantine, stabilization
and separation
Care and housing
of animals
Transport of
laboratory animals Procurement and transport of animals Care and Housing of
Laboratory Animals
Animal care and
technical personnel
Acquisition of
Anaesthesia and
euthanasia Staff at housing and research facilities Acquisition of
Laboratory Animals
Functional areas Experimental
Disposal of animal
carcasses Veterinary care Experimental
Animal husbandry Field research Laboratory animal
Responsibilities of institutions and the
institutional animal care and use
committees (IACUC)
Field Research
Record keeping Educational use
of animals Legal provision Responsibilities of investigators Educational use of
nonhuman animals
Laboratory animal ethics - - Responsibilities of teachers -
Table 3: Brief Explanation of All the Guidelines
Daily observation of
Care should be done
according to
"American Veterinary
Medical Association
(AVMA) Panel on
The post-operative
holding rooms, cages
and the care should be
under the supervision
of a qualified
Investigators must
have ready access to
veterinary care for
the animals at all
Care should be done
according to
"American Veterinary
Medical Association
(AVMA) Panel on
of the
No animal should be used
for experimentation for
more than 3 years unless
adequate justification is
Research should be
undertaken with a
clear scientific
purpose. There should
be a reasonable
expectation for
Research should be
undertaken with a
clear scientific
All animals found in
a moribund state
must be euthanized
unless there is
specific justification
to do otherwise.
Research should be
undertaken with a
clear scientific
purpose. There should
be a reasonable
expectation for
Sources of
Institution Biosafety
committee and
Institutional Animal
Ethics Committee.
The USDA and local
ordinances should be
consulted for source
of animal.
The only authentic
source of getting right
type of animals for
research should be
from recognized
scientific animal
A licensed or
otherwise legally
permitted source.
The USDA and local
ordinances should be
consulted for source
of animal.
Care and
housing of
Housing different species
in separate rooms;
however, cubicles,
laminar-flow units, cages
that have filtered air or
separate ventilation, and
isolators shall be suitable
The facilities housing
animals should meet
or exceed current
regulations and
guidelines (USDA,
1990, 1991), and are
required to be
inspected twice a year
(USDA, 1989).
It is important that
they are housed in an
isolated building
located as far away
from human
habitations as possible
and not exposed to
dust, smoke, noise,
wild rodents, insects
and birds.
Animals should be
housed in a manner
that facilitates the
expression of
behaviour and
minimises stress–
induced behaviours.
The facilities housing
animals should meet
or exceed current
regulations and
guidelines (USDA,
1990, 1991), and are
required to be
inspected twice a year
(USDA, 1989).
The Animal House
should maintain records
of chemicals, Breeding,
stock, purchase, staff,
sales records
Record should be kept
for the breeding, stock,
purchase, staff, sales
records, death and
animal use in
Record should be
available to the
personnel providing
post-operative care.
The size and nature of a
facility will determine
whether areas for
separate service functions
are possible or necessary.
Research conducted
in populated areas
should be done with
respect for the
property and privacy
Of the inhabitants of
the area.
The area should be
according to the
Research conducted
in populated areas
should be done with
respect for the
property and privacy
Of the inhabitants of
the area.
Journal of Medicinal Plants Studies
The global trend for the use of animals in research is
indispensable in several field hence the use of animal in
research should be justify as the animal use in research maybe
ethical in some way where alternative methods are not known
or very expensive. The different methods which are
alternatively used are sometimes beneficial sometimes not as
seen in animal use too. Hence if experiment were performing
on animal should be ethical as given in the guidelines for
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Response to the House of Lords Report on Animals in
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to exclude GM animals that are used for breeding
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presentation of the annual statistics to eliminate any
scope for misunderstanding”.
Full-text available
The role of oxygen in skin care is well known for assisting in skin rejuvenation and repair. This work is an unprecedented scientific and clinical effort to understand changes in percent skin oxygen saturation level (StO2) with topical application of a range of oxygen therapeutic skin care products, and in a course of skin breakout, using a newly developed three-wavelength system in a trial involving a total of eight volunteers. This technology is able to provide measurable positive effects on facial StO2 of the recruited volunteers one week after the topical application of oxygen creams with mean (standard deviation, SD) relative change in StO2 of 5.8 (4.6) %. Meanwhile a case study using this system on an individual with untoward skin condition revealed a drastic change in the mapping of StO2 values from the formation of papule to pustule, and to its encrustation. This optical technology could be a significant breakthrough in the dermatology research by redefining pharma-cosmeceutical research design and process in the development of skin care products by evaluating the efficacy of these products in improving skin oxygen level. In the future, this system may have major applications in the translational research of human physiology and pharmacology.
Full-text available
The natural history of the mannose 6-phosphate receptor was examined by radiolabeling cells in monolayers or in suspension; the receptor was isolated by immuno- or affinity precipitation followed by polyacrylamide gel electrophoresis. The receptor was found to contain asparagine-linked oligosaccharide chains and phosphorylated serine residues. Newly made receptor was sensitive to endo-beta-N-acetylglucosaminidase H (endo-H) and was slowly converted to a mature endo-H resistant form; phosphate was found on the mature receptor only. The receptor had an apparent molecular weight of 215,000 at all times, as determined under reducing and denaturing conditions; unreduced receptor had a greater electrophoretic mobility, suggesting the presence of intrachain disulfide linkages. The synthesis of immunoreactive receptor occurred with a lag of 50 min and of functional receptor with a lag of 70 min, indicating a requirement for some post-translational event(s) for acquisition of immunoreactivity and binding activity. Maturation of asparagine-linked oligosaccharides was not the requisite modification, since endo-H sensitive or deglycosylated receptor bound to both antibody and to insoluble phosphomannan; however, much less immunoreactive and functional receptor was detected in the presence of tunicamycin. Immunoprecipitable [3H]leucine-labeled receptor was degraded with a t1/2 of 16 h and 6 h for cells in monolayers and suspension, respectively, whereas 32P was lost with a corresponding t1/2 of 2.3 and 4 h. A pool of cell surface mannose 6-phosphate receptor was identified by separation on Percoll gradients as well as by iodination of cells with 125I; receptor in this pool was resistant to endo-H and had a t1/2 similar to that of the total [3H]leucine-labeled receptor, even in the presence of a saturating concentration of ligand. During endocytosis, ligand (beta-galactosidase) and 125I-receptor separated, the ligand accumulating within lysosomes. These results are consistent with current concepts of recycling of the mannose 6-phosphate receptor.
The recognition that biological cells exploit the water-oil interfacial activity of certain lipids to define anatomical boundaries has, in recent years, encouraged many workers in this laboratory* and others to develop and study protein-free model membrane systems prepared with such compounds. A considerable technical advance was made more than 10 years ago, when Mueller et al. (1962a,b) first reported a method for preparing usable preparations of bimolecular membrane from membrane molecules, the so-called black lipid membranes or BLMs. The merits of this powerful technique were quickly realized, and, as with Aladdin’s lamp, a property requested was a property acquired! Indeed, it might be said that the skeptics themselves were the holders of the lamp, because it was they who, by dismissing the model as unrealistic for a succession of reasons, indicated explicitly the characteristic implicit (passive) properties of a biological membrane: before very long, the earlier convictions of the Langmuir-Hardy-Rideal schools of surface chemistry, relating the role of amphiphilic molecules to biological membrane structure, were vindicated.
This study presents evidence that estradiol, mediated through oviduct fluid, may adversely affect the development of early embryos. Two-cell mouse embryos were cultured in Whitten-Biggers medium with or without 0.2 to 20 microgram/ml of estradiol or progesterone or in mouse oviduct fluid from donors treated with estradiol or estradiol and progesterone. Embryos cultured in fluid from estrogen-dominated donors were significantly less able to develop to morula (65%) or blastocyst (14%) than were those cultured in either Whitten-Biggers medium (91% and 41%) or progesterone-dominated fluid (87% and 36%). Transfer of cultured morulae and blastocysts to uteri of pseudopregnant recipients resulted in 2/3, 4/6, and 0/7 recipients' becoming pregnant for control, progesterone-dominated, and estrogen-dominated, respectively. When estradiol or progesterone was added directly to the control medium, no inhibitory effects were observed at levels higher than those found physiologically in plasma. Protein concentration and osmolalities for representative samples of oviduct fluid were 5.2 mg/ml and 329 mOsM for estrogen-dominated fluid and 1.6 mg/ml and 339 mOsM for progesterone-dominated fluid.
The developing use of in vitro fertilization (IVF) as a human clinical procedure has prompted the exploitation of nonhuman primates to assess the chromosomal and biochemical normality of embryos produced by IVF. Of 1995 oocytes recovered from squirrel monkeys, 628 (31.5%) matured and 339 (54.0%) fertilized. Fertility can be significantly enhanced by the addition of 1 or 10 microM dbcAMP to the culture medium. Chromosome analysis of oocytes and embryos used in these studies revealed an incidence of abnormality between 7 and 25%, comparable with that found for both in vivo and in vitro fertilized embryos from other laboratory species. There is no evidence that the IVF technique increases chromosomal abnormality. There was a decrease in protein synthesis of oocytes at maturation and during the early embryonic development stages, but an increase in the rate of RNA synthesis as development progressed. There was steroid uptake in early preimplantation embryos. The temporal relationships of early embryonic developmental events in the squirrel monkey have been determined.
Binding of tumor cells to cryostat sections of host organs was studied. B16-F10 melanoma cells and reticulum cell sarcoma cells demonstrated an organ specificity in their binding in vitro that reflected the organ specificity of their metastatic distribution 25 days after intravenous injection. These results provide evidence for specific binding of tumor cells to the tissues that they selectively colonize in vivo.
Monocyte or macrophage polykaryons (MP) are seen in different tissues in various inflammatory states and in normal bone (osteoclasts). The factors controlling the formation and the function of MP are not completely understood. This study was designed to evaluate the effects of the lymphokine gamma-interferon (IFN-gamma) on human monocyte function in vitro. Purified recombinant IFN-gamma [20-200 units/ml (0.1-1.0 nM)] caused the appearance of MP in cultures of normal human monocytes cultured in 10% unheated autologous serum. The MP were noted by as early as 36 hr of culture with fusion indices of 40%-60% and up to 160 nuclei per cell. The effect was seen with both recombinant IFN-gamma and natural IFN-gamma produced by Staphylococcal enterotoxin A-stimulated lymphocytes, but IFN-alpha (leukocyte-derived and recombinant) and IFN-beta did not induce MP formation. The activity of the IFN-gamma was destroyed by heating at 56 degrees C for 4 hr, incubating at pH 2 for 3 hr, or incubating with antibody against IFN-gamma. Populations of monocytes incubated 3 days with 100 units of IFN-gamma per ml (0.5 nM) had enhanced capacity to produce H2O2 in response to phorbol 12-myristate 13-acetate and increased content of acid phosphatase and plasminogen activator. As determined by autoradiography, the MP did not incorporate [3H]dThd into their nuclei. Thus, the IFN-gamma appears to induce MP formation by a process of monocyte fusion, and to "activate" monocytes, as judged by various parameters.
Isoflurane is metabolized by both rat and human hepatic microsomal cytochrome P-450 in vitro to fluoride ion and organofluorine metabolites. The forms of rat liver microsomal cytochrome P-450 induced by phenobarbital and pregnenolone-16 alpha-carbonitrile appear to be involved in the metabolism of isoflurane, while the forms induced by beta-naphthoflavone do not. Different pathways are favored for the metabolism of isoflurane by rat and by human liver microsomes: trifluoroacetaldehyde appears to be produced from isoflurane by rat liver microsomal cytochrome P-450, while trifluoroacetate or other nonvolatile fluorinated metabolites were not. The trifluoroacetaldehyde so produced binds tightly to microsomal constituents. Human liver microsomes converted isoflurane extensively to nonvolatile fluorinated products, one of which appears to be trifluoroacetate. The proposed pathways for the metabolism of isoflurane are considered in view of the above results.