Maintenance of laboratory animals as per CPCSEA guidelines

BP408P Pharmacology I Practical / S Y B Pharmacy Notes

Good Laboratory Practices (GLP) for animal facilities is intended to assure quality maintenance and welfare of animals used in laboratory studies while conducting biomedical and behavioural research and testing of products.

1. GOAL

The goal of these 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, quality in the pursuit of advancement of biological knowledge that is relevant to humans and animals.

2. VETERINARY CARE

Adequate veterinary care must be provided and is the responsibility of a veterinarian or a person who has training or experience in laboratory animal sciences and medicine.
- Daily observation of animals can be accomplished by someone other than a veterinarian; however, a mechanism of direct and frequent communication should be adopted so that timely and accurate information on problems in animal health, behaviour, and well being is conveyed to the attending veterinarian.
- The veterinarian can also help the establishment in designing appropriate policies and procedures for ancillary aspects of veterinary care, such as use of appropriate methods to prevent and control diseases (e.g. vaccination and other prophylaxis, disease monitoring and surveillance, quarantine and isolation), operative and post-operative care, diagnosis and treatment of diseases as well as injuries. reviewing protocols and proposals, animal husbandry and animal welfare; monitoring occupational health hazards containment, and zoonosis control programs; and supervising animal nutrition and sanitation. Institutional requirements will determine the need for full-time or part-time or consultative veterinary services.

3. ANIMAL PROCUREMENT

All animals (like cattle, buffalo, sheep, goat, pigs, equine etc.) must be acquired lawfully as per the CPCSEA guidelines. Small animals and dogs can be procured from registered breeders. Large animals can be procured from farm, farmers or as per guidance of wild life department, as is done in case of macaques. Cats can be bred for their use. Rodents can be imported from abroad after necessary licence from Director General of Foreign trade (DGFT) is obtained for import.

A health surveillance program for screening incoming animals should be carried out before purchase to assess animal quality. Methods of transportation should also be taken into account
- Each consignment of animals should be inspected for compliance with procurement specifications, and the animals should be quarantined and stabilized according to procedures appropriate for the species and circumstances.

4. QUARANTINE, STABILIZATION AND SEPARATION

Quarantine is the separation of newly received animals from those already in the facility until the health and possibly the microbial status of the newly received animals have been determined. An effective quarantine minimizes the chance for introduction of pathogens into an established colony. The duration at quarantine in small lab animals is from one week to one month and large animals allowed up to 6 weeks (cat, dog, monkey, etc). However, duration of quarantine can be increased depending on type of infection / suspected infection noticed in the animals.
Effective quarantine procedures should be used for non-human primates to help limit exposure of humans to zoonotic infections. The period varies from 2 to 3 months depending on the reaction of TB testing. Any macaque found positive for TB for at least two times and shows signs of weight loss or ill health should be euthanized as is practiced internationally to prevent spreading of TB to workers and other macaques.
Regardless of the duration of quarantine, newly received animals should be given a period for physiologic, psychologic and nutritional stabilization before their use. The length of time stabilization will depend on the type and duration of animal transportation, the species involved and the intended use of the animals.
Physical separation of animals by species is recommended to prevent interspecies disease transmission and to eliminate anxiety and possible physiological and behavioural changes due to interspecies conflict.
Such separation is usually accomplished by housing different species in separate rooms; however, cubicles, laminar-flow units, cages that have filtered air or separate ventilation, and isolators can be used as suitable alternatives.
In some instances, it shall be acceptable to house different species in the same room, for example, if two species have a similar pathogenic status and are behaviourally compatible.
Separate set of personnel should be identified for taking care of these infected (sick) animals and other workers should be restricted from entering in to the facilities unless otherwise required and after handling these animals they should not be handling any other animals in the facilities

5. SURVEILLANCE, DIAGNOSIS, TREATMENT AND CONTROL OF DISEASE

All animals should be observed for signs of illness, injury, or abnormal behaviour by animal house staff. As a rule, this should occur daily, but more-frequent observations might be warranted, such as during postoperative recovery or when animals are ill or have a physical deficit. It is imperative that appropriate methods be in place for disease surveillance and diagnosis
Post-mortem examination and signs of illness, distress, or other deviations from normal health condition in animals should be reported promptly to ensure appropriate and timely delivery of veterinary medical care. Animals that show signs of a contagious disease should be isolated from healthy animals in the colony. If an entire room of animals is known or believed to be exposed to an infectious agent (e.g. Mycobacterium tuberculosis in non-human primates), the group should be kept intact and isolated during the process of diagnosis, treatment, and control. Animals suffering from contagious diseases like Tuberculosis etc. must be euthanized as is practiced internationally to prevent its spread to other animals and often animal handlers.
The isolation, quarantine and stabilization programs for newly arrived animals are necessary to provide time to assess their health status, allow them to recover from the stress of shipment and an opportunity to adapt to their new environment. The extent of these programs depends on several factors, including species and source of the animals as well as their intended use. For some animals, such as rodents obtained from reliable sources for which health status is known, visual inspection on arrival may suffice. For species such as nonhuman primates, farm animals, wild animals, dogs, cats and non- specific pathogen free rabbits and rodents, appropriate quarantine and isolation procedures must be employed.
Preventive medicine programs such as vaccinations, ecto- and endoparasite treatments and other disease control measures should be initiated according to currently acceptable veterinary medical practices appropriate to the particular species and source. Only animals of defined health status should be used in research and testing unless a specific, naturally occurring or induced disease state is being studied. Systems should be established to protect animals within the institution from exposure to diseases.

Transgenic and mutant animals may be particularly susceptible to diseases and may require special protection to ensure their health. Systems to prevent spread of disease may include facility design features, containment/isolation equipment, and use of standard operating procedures. Training of animal care and research staff is essential to prevent spread of animal diseases.
Disease surveillance is a major responsibility of the veterinarian and should include routine monitoring of colony animals for the presence of parasitic and microbiological agents that may cause overt or unapparent disease. Additionally, cells, tissues, fluids, and transplantable tumors that are to be used in animals should be monitored for infectious or parasitic agents that may cause disease in animals. The type and intensity of monitoring necessary will depend upon professional veterinary judgment and the species, source, use and number of animals housed and used in the facility.
Diagnostic laboratory services must be available and used as appropriate. Laboratory services should include necropsy, histopathology, microbiology, clinical pathology, serology, and parasitology as well as other routine or specialized laboratory procedures, as needed. It is not necessary that all of these services be available within the animal facility (Facilities from other laboratories with appropriate capabilities may be used).
Animals with infectious / contagious disease must be isolated from others by placin them in isolation units or separate rooms appropriate for the containment of the agents of concern. In certain circumstances, when an entire group of animals is known or suspected to be exposed or infected, it may be appropriate to keep the group intact during the time necessary for diagnosis and treatment, for taking other control measures, or for completion of a project.
The veterinarian must have authority to use appropriate treatment or control measures, including euthanasia in consultation with at least one more additional veterinarian if required, following diagnosis of an animal disease or injury. If possible, the veterinarian should discuss the situation with the principal investigator to determine a course of action consistent with experimental goals. However, if the principal investigator is not available, or if agreement cannot be reached, the veterinarian must have authority to act to protect the health and well- being of the institutional animal colony and workers.

6. ANIMAL CARE AND TECHNICAL PERSONNEL

(a) Animal care programs require technical and husbandry support. Institutions should employ people trained in laboratory animal science or provide for both formal and on-the- job training to ensure effective implementation of the program (Annexure-7).

7. PERSONAL HYGIENE

It is essential that the animal care staff maintain a high standard of personal cleanliness. Facilities and supplies for meeting this obligation should be provided with appropriate Personnel Protective Equipment (PPE) e.g. showers, change of uniforms, footwear etc.
Clothing suitable for use in the animal facility should be supplied and laundered by the institution. A commercial laundering service is acceptable in many situations; however, institutional facilities should be used to decontaminate clothing exposed to potentially hazardous microbial agents or toxic substances. It is acceptable to use disposable gloves, masks, head covers, coats, coveralls and shoe covers. Personnel should change clothing as often as is necessary to maintain personal hygiene. Outer garments worn in the animal rooms should not be worn outside the animal facility.
Washing and showering facilities appropriate to the program should be available. Personnel should not be permitted to eat, drink, smoke or apply cosmetics and perfumes in animal rooms. They should finish the work with animals as early as possible and sit somewhere else, outside and not in the animal rooms / areas. A separate area or room should be made available for these purposes.

8. ANIMAL EXPERIMENTATION INVOLVING HAZARDOUS AGENTS

Institutions should have policies governing experimentation with hazardous agents. Institutional Bio-safety Committee whose members are knowledgeable about hazardous agents are in place in most of the higher-level education, research institutes and in many pharmaceutical industries for taking care of safety issues. This committee shall also examine the proposal on animal experiments involving hazardous agents in addition to its existing functions
Since the use of animals in such studies requires special considerations, the procedures and the facilities to be used must be reviewed by both the Institutional Bio-safety committee and Institutional Animal Ethics Committee (IAEC). Disposing of tissues and fluids from such used animals must also be appropriately governed as per the laid in practices of the institution / bio-safety regulation.

9. MULTIPLE SURGICAL PROCEDURES ON SINGLE ANIMAL

Multiple surgical procedures on a single animal for any testing or experiment are not to be practiced unless specified in a protocol only approved by the IAEC.
Individual animals should not be used in more than one experiment, either in the same or different projects, without the express approval of the IAEC. However, it is noted that Appropriate re-use of animals may reduce the total number of animals used in a project, result in better design of experiments, and reduce stress or avoid pain to additional animals. Animals that are used in more than one experiment should be permitted to recover fully from the first experiment before the subsequent experiment is performed. Certification of attending veterinarian is however, required before subjecting animal to the second experiment.

10. DURATIONS OF EXPERIMENTS

No animal should be used for experimentation for more than 3 years unless adequate justification is provided.

11. PHYSICAL RESTRAINT

Brief physical restraint of animals for examination, collection of samples, and a variety of other clinical and experimental manipulations can be accomplished manually or with devices be suitable in size and design for the animal being held and operated properly to minimize stress and avoid injury to the animal.
- Prolonged restraint of any animal, including the chairing of non-human primates, should be avoided unless essential to the research objectives. Less restrictive systems, such as the tether system or the pole and collar system should be used when compatible with research objectives.

Following points should be considered during handling and restraining animals:-

Animals should be handled by competent individuals trained in methods that cause minimaldistress and injury (for example, a person with a publication to his/her credit and post/experience in relevant techniques for handling animals is preferable).
The use of restraint devices is sometimes essential for the welfare of the animal and safety of the handler. Restraint devices should be used to the minimum
extent, for the minimum period required to accomplish the purpose of the experiment and be appropriate for the animal.
Tranquilisers or anaesthetics may initially be used to aid restraint but they may prolong recovery from the procedure. When these agents have been used, recovery of the animals should be closely monitored.

The following are important guidelines for the use of restraint equipments:
Restraint devices cannot be used simply as a convenience in handling or managing animals. The period of restraint should be the minimum required to accomplish the research objectives. Animals to be placed in restraint devices should be given training to adapt to the equipment, prior to initiation of the experimentation.
Provision should be made for observation of the animal at appropriate intervals. Veterinary care should be provided if symptoms or illness associated with restraint are observed. The presence of illness, or severe behavioural change should be dealt with by temporary or permanent removal of the animal from restraint related protocol.

12. LOCATION OF ANIMAL FACILITIES TO LABORATORIES

Good animal husbandry and human comfort and health protection require physical separation of animal facilities from personnel areas such as offices, break room, training and education room.
Laboratory animals are very sensitive to their living conditions. It is important that they shall be 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. The building, cages and environment of animal rooms are the major factors, which affect the quality of animals.
This separation can be accomplished by having the animal quarters in a separate building, wing, floor, or room. Careful planning should make it possible to place animal housing areas adjacent to or near laboratories, but separated from them by barriers such as entry locks, corridors, or floors.
While planning an animal facility the space should be well divided for various activities. The animal rooms should occupy about 50-60% of the total constructed area and the remaining area should be utilized for services such as stores, washing, office and staff, machine rooms, quarantine and corridors. The environment of animal room (Macro- Environment) and animal cage (Microenvironment) are factors on which the production and experimental efficiency of the animal depends. Since animals are very sensitive to environmental changes, sharp fluctuations in temperature, humidity, light, sound and ventilation should be avoided. The recommended space requirements for animal rooms, for different species

13. FUNCTIONAL AREAS

The size and nature of a facility will determine whether areas for separate service functions are possible or necessary. Sufficient animal area required to:

Ensure separation of species or isolation of individual projects when necessary; Receive, quarantine, and isolate animals; and

Provide for animal housing.

In facilities that are small, maintain few animals or maintain animals under special conditions (e.g., facilities exclusively used for housing germfree colonies or animals in runs and pens)some functional areas listed below could be unnecessary or included in a multipurpose area. Professional judgement must be exercised when developing a practical system for animal care.

Specialized laboratories or

Individual areas contiguous with or near animal housing areas for such activities as surgery,intensive care, necropsy, radiography, preparation of special diets, experimental manipulation,treatment, and diagnostic laboratory procedures containment facilities or
Equipment, if hazardous biological, physical, or chemical agents are to be used Receiving and storage areas for food, bedding
Pharmaceuticals and biologics, and supplies
Space for administration, supervision, and direction of the facility Showers, sinks, lockers and toilets for personnel
An area for washing and sterilization equipment and supplies, An autoclave for equipment
Food, and bedding; and separate areas For holding soiled and cleaned equipment
An area for repairing cages and equipment
An area to store wastes prior to incineration or removal

14. PHYSICAL FACILITIES

The physical condition and design of animal facility determine, to a great extent, the efficiency and economy of this operation. The design and size of an animal facility depend on the scope of institutional research activities, animals to be housed, physical relationship to the rest of the institution, and geographic location. A well planned, properly maintained facility is an important element in good animal care.

Housing facility should be compatible with the needs of the species to be housed.

Housing Facilities should be designed and operated to facilitate control of environmental factors to exclude vermin and limit contamination associated with the housing of animals, delivery of food, water, bedding, and the entry of people and other animals.

Housing Facilities should be maintained in good repair. Walls and floors should be constructedof durable materials with surfaces that can be cleaned and disinfected readily.
Housing Facilities should be kept clean and tidy and operated to achieve maximum possiblehygiene.
There should be a pest control programme to monitor and control vermin.
There should be adequate and appropriate storage areas for food, bedding and equipment.
Deodorants designed to mask animal odours should not be used in Housing Facilities as they may expose animals to volatile compounds which can alter metabolic processes. In addition, deodorants must not be used as a substitute for good cage and equipment cleaning practices and good ventilation.
Cleaning practices should be monitored on a regular basis to ensure effective hygiene and sanitation. This may include visual inspection, monitoring water temperatures and microbiological testing of surfaces after cleaning.
There should be proper water supply and drainage.
There should be adequate contingency plans to cover such emergencies as flooding and fire, or the breakdown of lighting, heating, cooling or ventilation.
In the interest of disease prevention and general animal welfare, access to the Housing Facilities by unauthorised persons should be restricted

Pharmacology I Practical

Second Year B Pharm Notes, Syllabus, Books, PDF Subjectwise/Topicwise

S Y B Pharm Sem III	S Y B Pharm Sem IV
BP301T Pharmaceutical Organic Chemistry II Theory	BP401T Pharmaceutical Organic Chemistry III Theory
BP302T Physical Pharmaceutics I Theory	BP402T Medicinal Chemistry I Theory
BP303T Pharmaceutical Microbiology Theo	BP403T Physical Pharmaceutics II Theory
BP304T Pharmaceutical Engineering Theory	BP404T Pharmacology I Theory
BP305P Pharmaceutical Organic Chemistry II Practical	BP405T Pharmacognosy I Theory
BP306P Physical Pharmaceutics I Practical	BP406P Medicinal Chemistry I Practical
BP307P Pharmaceutical Microbiology Practical	BP407P Physical Pharmaceutics II Practical
BP308P Pharmaceutical Engineering Practical	BP408P Pharmacology I Practical
	BP409P Pharmacognosy I Practical