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Laboratory Animals Flashcards

1
Q

Three Regulatory Bodies Associated with Welfare of Lab Animals

A
  1. USDA Animal Welfare Act
  2. Office of Laboratory Animal Welfare
  3. Guide for Care and Use of Laboratory Animals
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2
Q

USDA Animal Welfare Act

A

o All warm-blooded species except laboratory bred mice, rats, birds
o Animal Welfare Regulations (AWRs) dictate housing requirements, care during transport, enrichment requirements

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3
Q

Office of Laboratory Animal Welfare

A

NIH funded research facilitates –> all animals involved in research, teaching

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4
Q

Guide for Care and Use of Labratory Animal

A

o Minimum welfare standard for laboratory animals for husbandry, housing, etc
o Reference document for all other regulations

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5
Q

Three Rs

A

o Reduction
o Replacement
o Refinement

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6
Q

Pathogens-Free Species

A

free of pathogens, environmental pathogens
o Biological safety cabinets, sterilized equipment
o GLOVES ALWAYS

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7
Q

Challenges Assoc with Small Patient Size

A

Difficult IV access: intraperitoneal (IP), IM, SC
 IP: high failure rate – inadvertent drug delivery into fat, SQ, gut tissue
 Need insulin/TB syringes, 25-30g

High metabolic rate – rapid drug metabolism

Prone to hypothermia

High surface area to body weight ratio –> increase heat loss
o Ventilation: need appropriately sized circuit, equipment

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8
Q

Rodents - General Features

A

Obligate nasal breathers

Nocturnally active – almost all feeding/drinking occurs during dark phase
 Post sx pain, stress during light phase can suppress activity
 Short fasting periods: hypoglycemia, dehydration

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9
Q

Rodents - central/head features

A

o Protruding eyeballs: lubricant (ointment less useful)

o Lack complete circle of Willis (anastomosis of arteries at base of brain): susceptible to cerebral ischemia upon ligation of common carotid

o Idiopathic epilepsy, hereditary diabetes reported

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10
Q

Rodents - Resp Changes

A

High chest wall compliance

Low residual capacity

Robust Bohr effect: small changes in pH have stronger effect on hemoglobin ‘s affinity for oxygen

Single left lobe, right separated into four or five lobes

Rats, hamster: cardiac striated muscle extends along pulmonary vessel walls, into lung tissue making them susceptible of spread of infection between agents

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11
Q

Blood Volume in Rodents

A

60-80mL/kg (6-8% BW)
 Blood collection = 10% BW
 Transfusion trigger: PCV <20%

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12
Q

Laboratory Mice

A

Large incisors with broad tongue  difficult intubation

Fast, agile, do not acclimate well to handling – hand restraint requires knowledge, practice
 Commercial restrainers, transfer boxes/containers

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13
Q

Laboratory Mice Drug Access

A

o IV access: lateral tail veins, submandibular venous plexus (blood draw only), orbital sinus
o SQ: flank, scruff

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14
Q

Rat Features

A

No gall bladder

Heart blood supply largely extra coronary from branches of mammary, subclavian arteries

Large incisors with broad tongue  difficult intubation

Acclimate well to handling, intelligent
 Hand restraint requires knowledge, practice
 Commercial restrainers, transfer boxes/containers
 “Tickling” acclimation

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15
Q

Rats IV Access

A

IV access: lateral tail veins, saphenous vein, jugular vein

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16
Q

Hamsters

A

o Cheek pouches with orifices near the angle of mouth: source of aspirated food material if not addressed prior to anesthesia
o Don’t usually vomit
o Hibernating: become mildly acidotic
o Anesthetic mortality = 3.7%

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17
Q

Drug Access Hamsters

A

o IV: jugular, cephalic, saphenous (medial or lateral)
o IO: tibial crest, intertrochanteric fossa of femur, ileal crest, proximal humerus

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18
Q

Induction in Rodents

A

Rapid induction, easy titration of ax
 Can be challenge to maintain sterility, control ax if one person doing everything

Mask induction: technically easy to perform, not routinely intubated

Increased exposure to personnel

Decreases variations btw subjects –> minimize # of animals used in a study
o Rapid recovery vs injectables

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19
Q

Orden et al 2021 (VAA)

A

MACsevo in rats unchanged by CBD alone, MAC reduction by morph not enhanced by CBD (CBD + opioids = synergistic effect)

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20
Q

Propofol Use in Rodents

A

o Propofol if IV (rats)
 IP admin: inconsistent depth, not recommended

Requires IVC Placement

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21
Q

Telazol in Rodents

A

High Mortality Rate

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22
Q

Tribomethanol (mice) (TBE)

A

 Common in-vitro oocyte implantations
 Peritonitis, high mortality of incorrectly stored or prepared
 When successful, 15-20’ ax

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23
Q

Urethane

A

Ethyl carbamate, long-lasting, stable sx ax (6-10hr)

Few CV, resp AEs

Non-recovery procedures: CNS studies
* Depressed spinal reflexes, EEG comparable to sleep patterns, ANS reflexes preserved

Water soluble, metabolized by liver into ethanol+carbamic acid, renal excretion

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24
Q

Urethane MOA

A
  • GABAA 23%
  • a1 glycine R agonist 33%
  • NMDA R antag 10%
  • AMPA R antag 18%
  • +/- AChR?
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25
Adverse Effects of Urethane
IP: endocrine, metabolic effects = peritonitis, necrosis of abdo contents, massive leakage of plasma in pleural cavity Carcinogen: PPE, proper disposal
26
alpha-chloralose
 Numerous dose-dependent excitatory, inhibitory CNS effects  Long-lasting light ax with minimal CV effects  Prolonged induction, recovery – insufficient depth for sx
27
Analgesics in Rodents
Buprenorphine most commonly used: 35% of rodents that receive analgesia Consider SR formulas – up to 72hr in mice, rats * 40% rats skin irritation Poor PO avail 5-10%, significant first pass hepatic metabolism --> PO DOA 1-2hr  SE: pica behavior
28
SE Opioids in Rodents
o Immunosuppression with very high doses of opioids for prolonged periods, potential effects on tumor growth
29
Ventilation in Rodents
rodents normally maintain themselves at 1:1IE  1:2, 1:3 to minimize effects of PPV (prolong expiratory time) on CV system
30
Depth in Rodents
o Depth: pedal withdrawal, auricular reflexes o Minor changes in heart rate, respiratory rate equal indicators of changes in depth
31
CV Monitoring in Rodents
Pulse oximetry: pulsatile flow of RBCs past probe required  HR > 400bpm may cause difficulty in acquiring reading ETCO2: requires intubation, limitations with both mainstream and side stream BP: Constraints with BP cuffs DT small patient size High heart rate can decrease accuracy of oscillometric readings Difficult to find artery for catheterization
32
Positioning of Rodents
little horses – weight of organs on diaphragm limits appropriate depth of breath  Ensure straight head slash neck for better air flow in non intubated patients
33
Pain Assessment in Rodents
Grimace scores validated for mice, rodents Pain assessment: hunched posture, bruxism, grimace, tachycardia
34
Recovery of Rodents
--Ensure warming, avoid excessive fluid admin to minimize hypothermia = recover, eat quickly --Home cage: no particulate bedding to avoid airway obstruction --Often housed in corn cob bedding, replace with paper towel --Keep separate from others while recovering: will attack, esp males Clean off all detergents, chemicals from skin --Grooming: may consume toxic small quantities from fur Gel based hydration, electrolyte packs in cage
35
Common Peri-Anesthetic Complications with Rodents
o Hypothermia o Insufficient depth of ax, movement – esp with mask maintenance if move nose from cone o Excessive depth o Labored respirations +/- resp arrest o Inadvertent inj into spleen, GIT via IP  Delayed onset, slow recovery o Difficulty monitoring: specialized equipment, patient visualization
36
Features of the Mouse Grimace Scale
Orbital tightening: closing of eye = pain Nose bulge: bulging on nose bridge, squinting of face = pain Check bulge: see nose bulge Ear Position: rotate outward, backwards away from face - space btw ears increased Whiskers: pulled back or stand on end, lose downward curve - pointed up
37
Features of the Rat Grimace Scale
Share same features of mouse grimace scale EXCEPT have flattening of nose, cheeks not bulging with pain
38
Gent et al 2019 (VAA)
Xe less aversive than CO2 when using 30% chamber vol/min fill rate, could improve welfare of mice  Did not show jumping, freezing behaviors, decreased locomotion  EEG: sedative effects from Xe, heightened arousal from CO2
39
Rabbit Mortality Rate under GA
1.39-4.8%, ill rabbits >7% o Prey species, adept at hiding illness
40
Drug Access in Rabbits
IV access: marginal auricular, saphenous, cephalic Auricular: risk necrosis IO: trochanteric fossa of femur, greater tubercle of prox humerus, wing of ilium, tibial tuberosity
41
Chung et al 2022 (VAA)
ears that had EMLA cream applied for >60’ before catheter attempt = 10x more odds of successful catheter placement, EMLA or lido cream 10’ prior = 3x
42
Rabbits and Fasting
Fasting **not required/recommended** – inability to vomit, impt for normal GI motility  Alterations to gut flora, ileus, enterotoxemia  Functional ileus = fatal
43
Rabbits and Neophobia
(reluctant to eat, try new foods): offer palatable high energy, high water content supplements
44
Rabbits and Corprohagia
(eat stool): avoid inadvertent prevention by Elizabethan collar
45
Anticholinergics in Rabbits
Atropinease: present in 1/3-1/2 rabbits, faster metabolism of atropine o Glyco recommended
46
Which opioid has been shown to have no effect on GI motility in rabbits?
Buprenorphine
47
Reabel et al 2022 (VAA
Blind vs endoscopic techniques both techniques assoc with laryngeal damage, blind orotracheal intubation assoc with higher number of attempts, endoscopic groups required more dexmed/alfax
48
Anatomical Challenges with Rabbit Intubation n
 Long, narrow oral cavity with fleshy tongues  Long incisors  Difficult visualization  Very sensitive to laryngeal stimulation – vasovagal reflex  Obligate nasal breathers  Epiglottis entrapped on dorsal surface of soft palate (beneficial with nasotracheal intubation)
49
other techniques to intubate rabbits
Supraglottic airway device (VGEL), mask, nasotracheal intubation
50
Inhalants in Rabbits
**More sensitive to vasodilatory effects** of inhalant anesthetics than other species: **hypotension occurs at levels of 1 MAC** **Dopamine, phenylephrine tend to be ineffective – NE** Fentanyl, lidocaine, ketamine CRIs
51
Blood Pressure Measurement in Rabbits
Oscillometric moderately accurate FL normal-low pressures, less sensitive at detecting hypotension
52
Desprez et al 2022 (VAA)
vs cd aortic ABP in sevo ax rabbits, cuff with 41% ratio (2cm) on thoracic limb = best estimation of MAP vs cuff with 50% ratio (2.5cm), measurements on hind limb
53
Assessment of Depth in Rabbits
sx palpebral absent, corneal reflex present, deep/reg resp pattern, toe pinch absent (reliability: HL > FL)
54
Blood Glucose in Rabbits
prognostic indicator: stressed rabbits increased BG vs non stressed High BG at presentation with obstructive ileus vs non
55
Edis et al 2022 (VAA)
rabbits undergoing OVH in theatre temp of 28*C = significantly higher temp vs 23*C, did not affect return to feeding/defecating
56
Location for IBP in Rabbits
central auricular artery
57
Features of the Rabbit Grimace Scale
Orbital Tightening Cheek Flattening Nostril Shape Whisker Shape, Position Ear Shape, Positionin
58
Features of the Bristol Rabbit Pain Scale
1. Demeanor - looking around vs non responsive 2. Locomotion - active vs not 3. Posture - comfortable vs tense, stiff, hunched 4. Ears - freely move toward sound vs no moment 5. Eyes - open vs closed/tight 6. Grooming - grooms vs no
59
Guinea Pigs and Chinchillas: Mortality Rate
3.8% for GPs, 3.3% for chinchillas
60
Respiratory Features of GPs, ChC
 Lung fields: small due to combination of comparatively large hearts, profuse GI volume  High chest wall compliance  Low FRC  More alveoli with thinner diameters to maximize oxygen exchange
61
Vomiting in GPs, ChCs
Don’t usually vomit: lack coordinated brainstem response to complete emetic episode Also reduced muscularity of diaphragm, long abdominal esophagus length, small abdominal esophageal circumference, medial position of esophagus in stomach
62
GP, ChC: fasting
Fasting not required/recommended – inability to vomit, impt for normal GI motility  Alterations to gut flora, ileus, enterotoxemia  Normally retain food in oropharynx Can consider fasting zero to 8 hours for food, zero to two hours for water due to reduction of food material in oropharynx, reduce incidence of passive regurgitation, potentially reduced GI volume (probably negated by cecum) Neophobic, coprophagic
63
Premedication in GP, ChC
o Prior to PMs: rinse oral cavity, clear mouth of food material using cotton tip applicators 30 to 60 minutes prior to anesthesia
64
Anatomic Challenges of GP, ChC Intubation
 Long, narrow oral cavity  Difficult to visualize  **Palatal ostium** – central opening btw caudal tongue, soft palate (2-3mm)  Deviation of tube = damage to velopharyngeal folds
65
How to Intubate GP, ChC
Endoscopic assisted – rigid endoscope, otoscope, vaginal speculum Guidewire advanced into airway Airway mucosa = fragile  Potential for blood to obscure view, aspiration of blood, laryngeal trauma, tracheal tears, airway obstruction DT swelling
66
Alvarez et al 2022 (JAVMA)
SQ alfax 5mg/kg – reliable sedation for non-painful procedures in GPs, midaz 0.5mg/kg + alfax 5mgkg = longer, deeper sedation, more significant in adults
67
Mask Induction for GPs, ChCs
particularly irritating to GPs if used for mask induction o Iso: ocular, nasal discharge > sevo: less lacrimation o Least irritating = halothane
68
Monitoring for GPs, ChCs
Same as Rodents
69
NHP
New World (NW NHP) – trees – marmosets, tamarains, capuchins, tropical monkeys (owl, spider, howler, squirrel) Old World (OW NHP) – scary – Baboons, macaques, Great Apes
70
Safety Considerations with NHPs
Sensitive nature for using NHPs o Very intelligent, always thinking Escape risks Prehensile hands, fingers – may remove/reflect darts, pole syringes Strength: ability to pull, scratch, bite o Sharp teeth, long powerful limbs, +/- prehensile tail o Handling to conscious NHP avoided DT stress to animal, risk of physical injuries to operator, risk of traumatic injury to patient
71
Disease Transmission Assoc with NHP
Cercopithecine Herpesvirus B endemic in macaques – fatal to humans, PPE Tuberculosis – humans --> NHP, NHP--> humans Measles – humans --> NHP Other herpesviruses = reverse zoonotic danger to great apes, btw species
72
fasting guidelines in NHP
Monogastric: Fasting 6-8hr, 2-4hr in marmosets/tamarins  Allow access to water, electrolyte solution (Gatorade, clear Ensure)  Smaller sap-eating species prone to hypoglycemia
73
Other Considerations of NHP
Social animals: group housing  Segregate but should remain in sight/presence of conspecifics Will usually be suspicious, know something up PE, minimum database often performed under sedation, IV access via superficial veins  Gorillas: middle tibial vein (??)  NW NHP: caudal tail vein
74
Ketamine in NHP
Assoc with **long-term cognitive impairment** in NHPs if admin during sensitive period for brain development eg **in utero, during first week of life** **Tolerance**: 3 consecutive days = increased time to recumbency by 35%
75
Airway Management with NHPs
Fasted: can regurgitate, vomit  intubation recommended Visualization = challenging * Wide range of vocalization DT caudal descent of larynx (s resonance capacity), large part of tongue to be in hypopharynx – restricts visualization Rich vascularization of larynx – prone to injury
76
Other Features that Make Intubation Difficult with NHPs
Positioning: consider dorsal recumbency on tilt table to maximize alignment of PO cavity with pharynx Diameter of larynx, trachea smaller than species of equal weight Copious secretions = thicker with anticholinergic administration **Damage to vocal chords VERY significant in these species!**
77
Depth of ETT in NHPs
Proximal bifurcation of trachea, avoid passing tube past point of scapula (macaque)  Marmosets: length of trachea predicted = 0.42(craniosacrolength)
78
Analgesia in NHP
o LJ: no current scale that definitely assess pain in NHP o Tend to hide behavioral signs of mild or moderate pain from unfamiliar observers, best monitored by familiar handler or remotely o All commonly available analgesics can be used, long acting formulations preferred  Meloxicam – particularly palatable
79
Ferrets
Anesthetic mortality = 0.33% Opioids: emetic effect, consider maropitant or ondansetron  Presumed to be MAC-sparing in ferrets Best indicator of depth = jaw or masseter muscle tone Prone to both hyper and hypothermia BG monitoring: insulinomas common
80
Hedgehogs
 Mantle (keratinaceous spines), roll into ball – difficult to examine  Narrow oral pharyngeal cavity  Caudally located pharynx  Prominent tongue base and large tonsils  Epiglottis = broad with a prominent apex, normally dorsal to soft palate and must be displaced to visualize the glottis
81
Which pocket pets are obligate nasal breathers?
Rodents Rabbits GPs, ChCs NOT ferrets, SGs, HHs,
82
IV Access in HH, SG
 HH: cephalic, saphenous veins  SG: virtually impossible
83
Premedication/Sedation in HH
HH: midaz + torb, +ket to deepen/lengthen  IN midaz effective  Alfaxalone also effective SC, IM  Telazol: no real effect of flumazenil
84
Premedication/Sedation in SGs
midaz can go IN, Telazol controversial – neurologic signs, death
85
Induction with SGs, Has
Chamber induction most common, monitor posture changes SG: recumbent * May vomit HH: no longer assume defensive posture * Rarely vomit
86
Intubation for HH, SGs
o Intubation is challenging o Consider endoscopy using rigid tube or smaller, semi rigid fiber optic endoscopes and the use of stylets o Helpful to have small Miller blades o Usually need 1-2mm ETT for HH, 1-1.5mm ETT for SG o Often maintain via face mask due to difficulties intubation o Can use small rabbit V gel in larger HH (360-700g)
87
Recovery of His
brief period of tachypnea often occurs before arousal, attempted self removal of airway device
88
Recovery of SGs
self-mutilation following surgery very common, close monitoring during recovery essential

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