Question | Answer |
Characteristics of Research Animals? | -small
-easy to handle
-short life span
-short gestation period
-large litter size |
Types of research animals? | mice, rats, zebrafish, drosophila (flies), nematodes (worms), 80% rodents-of that 60% mice |
Types of research lab animals involved in? | inheritance, cancer, tissue transplants, understanding body & behaviour |
why use inbred mice? | results then do not vary within the species, eg. strain background, then need fewer animals and less time/cost of experiment. |
How long does it take for an inbred strain to be developed? | 5 + years |
Can you duplicate an inbred strain? | no, this means that once you lose the strain it is lost forever and if you were in the middle of an experiment-say goodbye to you research because you will have to start over. |
Housing environment-animal flora? | Can control flora to get rid of harmful ones to animal health and to get rid of ones that disrupt research |
What are transgenic breeds? | they contain the DNA of unrelated organisms that have been artificially introduced. |
Transgenic process? | 2 fertile animal produce an embryo and a transgene is added to the nucleus of the embryo. This embryo is implanted into a pseudo pregnant individual (from mating of sterile couple) and this individual produces transgenic offspring. |
knock in gene | gene sequence is added at a particular locus of a DNA sequence to create a new individual. transgenic |
knock out gene | target gene is disrupted or inactivated, loss of that gene activity which changes the phenotype (look)of the animal. |
When did lab animal research become a regulated system and what act? | In 2000, gene technology act to ensure personal and community risks are reduced. |
What are the 4 Levels of Containment? | PC1-no risk to operator or community
PC2-no risk operator, perceived risk community
PC3-risk to operator and community
PC4-high risk to operator and community |
Variable to consider in animal housing-new house? | -accepted health status (from open to SPF to gnotobiotic-germ free), consistent environment-light, temp, bedding, nesting & consistent diet |
Why use Zebrafish? | -fecund-produce lots of embryos
-embryos are free living
-fast embryonic development
-technology to manipulate their genes is rapid and cheap
-optical transparency
-genetic similarity to humans
-permeability to water borne chemicals |
Difference between zebrafish & mice? | fish-outside embryos, compared to mice- in utero, chemical uptake from water (fish)to chemicals given to mother, mice need surgery for tissue transplants-direct for fish, observation-real time for fish whereas fixed direct for mice m-model f-phenotype |
What areas are zebrafish standards lacking in? | health monitoring, water quality and nutrition |
General standards for keeping zebrafish? | 10 fish/litre
28.5 degrees Celsius-water, room-24
14/10 hrs light/dark like monsoonal
<0.02 NH3, <50 NO3, <0.1 NO2
barren tanks |
Peak Zebrafish production? | 6-12months (just after reach sex maturity)
1yr-tail is off
live for 18-24months |
Life stages of Zebrafish? | Embryo (0), alevin (4), Fry (9), Fingerling (28) and adult (60) |
Rotifers? | -feed on microalgae
-gut loadable nutrition
-common in commercial
-easy to culture
-marine so cant establish in a system
This is in comparison to a paramecium |
Paramecium? | Bacteriovore
limited nutrition
common in hobby's
easy to culture
freshwater so can be established in a system |
Alevin lifestage? | swim bladder enlarges
exogenous feeding starts |
Zebrafish health maintenance? | -daily mortality check
-weekly health inspection
-weekly scheduled terminations
-constant water quality checks
-2x daily CSL equipment checks |
Disease Control-conventional? for zebrafish | -horizontal transmission (fish-fish) by checking aquarium design, embryo bleaching, new animal isolation, tank change limits
-vertical transmission (mother-fry)
-pathology testing by early detection of disease and options for facility isolation |
import disease control for zebrafish -quarantine? | -if lifetime-for first 7 days interactions recorded on QTR's and only offspring allows to be removed from area by QAD approval
-for post release-they are tracked and monitored by whole tank |
Recirculating Aquaculture Systems-benefits and limitations? | Benefits- self contained, more control over environment, increased biosecurity, minimal water exchange
limitations-water quality-bio,chem,mechanical filtration |
Controlling water quality? | -life support condition
-source of water: municipal-town, reverse osmosis or natural
-effluent processing |
Important water quality factors? | pH-7.5, temp 28.5, CaCO3 >100 DO >6, sality 0.25-0.75, conductivity target 300-1200, hardness tolerance-Ca, Mg, HCO3, CO3 |
Municipal water benefits and limits? | Benefits-cheaper, available, minimal waste, optional culture use
Limits-government regulations on quality, limited pathogen control, limited control over culture condition, needs pre-treatment |
Reverse Osmosis water benefits and limits? | Benefits-user specified product quality, good pathogen control, good culture control over condition
Limits-equipment, higher cost, needs conditioning with sea salt (to increase CaCO3 and conductivity) & hardness generators (increase pH), increased waste |
Effluent process? | Holding tanks-mechanical filtration-biological filtration-oxygenation & CO2 removal-chemical filtration-disinfection.
cycle |
What is Mechanic filtration? | -removes solids
-allows healthy biofilter growth
-increases disinfection efficiency
-considerations-whether it isolates or removes the waste (welfare consideration), consumables, technical skill for operators, automation-cost |
What is Biological filtration? | -dedicated media
-media allows bacteria growth by providing a substrate
-these bacteria breakdown waste products |
2 types of bacteria in biological filtration | hetertrophic-use dissolved carbonaceous material
chemosynthetic-use ammonia and nitrite and are 5 times slower |
Ammonia as a waste in fish? | excrete waste as ammonia NH3 through gills and it flows down its concentration gradient to outside environment, if outside is greater than >0.05ppm then it is toxic to the fish due to this gradient |
What is Chemical filtration? | Dissolved wastes are harder to remove, so use activated carbon or foam fractionation/protein skimming. |
What is activated carbon-chemical filtration? | -uses process of adsorption to trap pollutants in the carbon substrate pore structure. Removes Cl, Cu (copper), dissolved organic compounds and colloidal solids |
What is protein skimming-chemical filtration? | uses process of adsorption, hydrophobic pollutants bind to microbubbles-air or ozone and rise in a column to the top where form foam that is dislodged into a foam stream. |
Disinfection-water for zebrafish? | -reduces micro-organism populations
-uses UV or Ozone
-UV only stops reproductivity, has a short effectiveness time
-ozone is more effective than UV, but higher risk which can be reduced if it is used in a skimmer |
What is UQBR? | -lab animal housing and care facility
-animal importation services
-transgenic services
-tissue collections
-embryo rederivations
-colony management |
Role of Vet Tech in UQBR? | -assist researcher
-know basic technical procedures for animal life
-know the lab and animal health technical procedures
-maintain/manage the colony
-husbandry, colony management, facility management & technical procedures |
Husbandry? | -sexing
-identifying pregnancy
-basics: food water etc
-necropsies
-restraint
-provide enrichment
-health checks |
Common Health Problems? | -ringtail
-barbering (missing bits of hair)
-malocclusion (very long teeth)
-ulcerative dermatitis
-prolapse (red and inflamed vagina)
-fight wounds
-abnormal enlarged bladder
-tumor
-dwarfism
-hydrocephaly (enlarged brain due to fluid) |
Colony Management? | -monitor breeder performance
-replace breeders if needed
-keep reserve breeders
-follow a breeding protocol
-produce required animal numbers
-reduce wastage-protocol (replace, reduce, refine)
-maintain colony: genetic control
-raise concerns |
3 R's in Colony Management? | -replace
-reduce
-refine |
Facility Operations? | -dirty cages need to be knocked out
-cages washed, then autoclaved
-stores topped
-lots of water bottles filled
-clean cages need bedding |
Technical procedures? | -injections (IP,IM,SC,IV)
-blood collection (saphenous vein-leg, dorsal pedal vein-paw, retro-orbital bleed, cardiac puncture, sub mandibular
-euthanasia (CO2, cervical dislocation, lethabarb)
-tissue collection- ear notch and toeing-identification |
Why do we need record keeping? | -legislative reason
-research requirement
-welfare reason |
UQBR Supervisor role? | -assist in overall facility management
-staff/student supervision
-specialized animal husbandry & surgical procedures
-ensure biosecurity
-general husbandry
-assist handling courses
-provide reports on operational activities
-monitor AEEC no. etc |
Why do we need lab rodent anaesthesia? | -immobilization
-muscle relaxation
-unconsciousness |
When do we use anaesthesia and what are the considerations? | -husbandry and experiments
-considerations-animal welfare AEC approval
- length
- depth
- research outcome requirements
- recovery or terminal |
Types of anaesthesia? | -general (gas or injection)
-local (topical like emia or injection) |
Advantages and disadvantages of general gas anaesthesia? | Advantages: oxygen administration during anaesthesia, accurate control over depth, rapid induction and recovery
disadvantages: specialized equipment, good ventilation and scavenging system, requires training. |
Advantages and disadvantages of general injection anaesthesia? | advantages: minimal equipment, rapid induction, titratable dosing, some reversible
disadvantages: predetermined dose, need good restraint, individual variation, slow absorption in non IV routes |
Anaesthetic Monitoring? | -limited equipment
-anaesthetic depth
-reflexes
-muscle tone
-body temp
-hydration
-respiration
-mm colour |
What are the two types of analgesia used? | -NSAIDs
-Opioids |
Common disease continued? | -bacteria or viruses
-extoparasites: shown by bite marks with lines on stomach area where inhabit
-endoparasites: worms |
What law are lab animal specifically ruled under? | Australian code of practice for the care and use of animals for scientific purposes |
What is humane killing and euthanasia? | Humane killing-ending a life with no top pain and stress
Euthanasia-humane killing that is in the welfare interests of the individual and to alleviate pain and stress |
When is humane killing used? | -at the end of the study when need to obtain tissue samples
-when the animal can no longer be used for breeding purposes
-when stock are not suitable or not required |
When is euthanasia used? | -when pain or stress levels of animal expected to exceed normal levels and cant prevent
-health and wellbeing concern |
Recommended euthanasia? | -CO2
-sodium pentobarbitone (lethabarb) |
Accepted euthanasia but with reservations? | -isofluorine
-cervical dislocation
-decapitation
-stunning |
Not accepted euthanasia? | -CO
-Nitrogen
-chloroform
-microwave irradiation
-ether
-hydrogen cyanide
-decompression
-rapid freezing |
Euthanasia Considerations? | -avoid distress
-produce rapid unconsciousness
-reliable, reproducible, irreversible
-appropriate for age and species
-minimum restraint
-code of practice legislation requirements
-compatible to study
-simple- administer
-safe- operator |
Main Codes governing use of animals in lab?- in QLD | -QLD animal care and protection act 2001
-Australian code care and use of animals science research
-code of practice for the care and housing of lab mice, rats, guinea pigs and rabbits
-NHMRC CODES |
what do the 3 Rs mean-replace, reduce, refine? | Replace- try and obtain results and purpose of experiment without the use of animals
Reduce-reduce no of animals use, but so still have comparable results and same amt of info
Refine: minimize pain and enhance wellbeing |
Animal Ethics? | -actions are considered right or wrong
-determines what can and cant do in research experiments (as unethical)
-need to apply for approval
-animal cruelty and protection act 2001
AEC chairman, vet, researcher, welfare person and no associate |