DISCLAIMERS
ACLAM has not reviewed or sanctioned this presentation No information presented is known to be included in the ACLAM JAALAS 2020 Board Examinations past or present Information presented is believed to be reliable and correct though not guaranteed for accuracy Francis J. Sun, DVM, DACLAM, MBA Not every article is included in this review Laboratory Animal Workshop 2021 I used the 2nd edition Blue Book
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General study tips ACLAM certification site
Know your Role Delineation Document Laboratory Animal Medicine, 3rd Edition
Allocate your time accordingly The 3rd edition of Laboratory Animal Medicine is currently available in electronic format, with the hard cover version available soon. As it is an Trust yourself with your study habits ACLAM sanctioned textbook, it is considered fair game for the 2016 exam. When studying, please realize that there are over 3,000 questions Groups vs individual in our database. It will take the exam committee longer than a single Practice exams or note cards exam cycle to update every reference; however all questions for the 2016 exam that have the 2nd edition of the Blue Book as a reference Rest and health lifestyle are more beneficial for the last couple of will be fact checked against the 3rd edition of the Blue Book and days than cramming updated as necessary. Exam Material Cut Off Date The annual ACLAM Exam may contain questions on materials from any relevant publications up to Dec 31 of the year prior to the exam (Dec 31, 2019 for the 2020 exam).
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ACLAM certification site ACLAM certification site
ACLAM 2021 Certification Examination Reference List : This reference list provides the sources of the majority of the examination Domain 1: Management of Spontaneous and Experimentally questions for the 2021 exam. Journal articles used for the exam are typically those published within the last 5years. Induced Diseases and Conditions 25% This list is not meant to be all‐inclusive (e.g., it does not indicate the only Domain 2: Management of Pain and Distress 10% resources candidates should use to prepare for an upcoming examination). Note that the ACLAM Series Blue Books are heavily referenced and are Domain 3: Research 20% considered core references. Journals Domain 4: Animal Care 24% • Comparative Medicine, bimonthly publication of the American Domain 5: Regulatory Responsibilities 19% Association for Laboratory Animal Science, Memphis, TN. • Institute for Laboratory Animal Research (ILAR) Journal, a quarterly Domain 6: Education 2% publication of ILAR, the National Academies, Washington, DC. • Journal of the American Association for Laboratory Animal Science (JAALAS), the bimonthly publication of the American Association for Laboratory Animal Science (AALAS), Memphis, TN.
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1 The Journal Article
Abstract : Problem/issue, findings Introduction : informative about species, models, diseases, etc. Pictures! plus captions and conclusions JAALAS Volume 59, Discussion Article may not meet requirements to be testable material but could jog your memory about something you should go back and review that is clearly in the RDD No1 Poll Everywhere setup/test
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Blood Pressure Reference Intervals for Ketamine-sedated Influence of Dietary Vitamin A and Iron Deficiency on Rhesus Macaques Hematologic Parameters and Body Weight of Young Male Wistar Rats 103 male and female rhesus of various body condition and ages Why studied? Micronutrient deficiency prominent public health concern Reference Interval – usually central 95% of values Vit A and iron 2 devices at 3 anatomic sites Child development Hypothesis : vit a and iron must be taken together to ensure adequate weight Device –no difference (abstract vs discussion) gain and hematologic parameters Location BP arm 9 10 Effect of Repeated Exposure to Isoflurane on Nest Building and Burrowing in Mice Developed a refined scoring method for nest-building activity Nest building and burrowing have been recommended as noninvasive tools in the assessment of welfare in mice Hypothesis : burrowing and nest building would decrease after The scoring system was based on Answer B exposure to any of 3 stressors and scoring method would have agreement between observers A)The amount of time the mouse stayed in the square and x spots B)The height and width of the nests C)The temperature achievable and firmness D)How well hidden the mice would be Found their system to be objective. Nest building decreased after iso exposure but not to a second exposure. Burrowing did not change. Questioned applicability to welfare assessment 11 12 2 Tell-tale TINT : Does the Time to Incorporate into Nest Test Utility of Automated Feeding Data to Detect Social Instability Evaluate Postsurgical Pain or Welfare in Mice in a Captive Breeding Colony of Rhesus Macaques: a Case study of Intrafamily Aggression Use of ethologic framework of behavioral time-budgets is a Captive breeding colonies in large outdoor multimale, multifemale practical strategy for identifying pain in mice. social groups of several matrilineal families. When mice fail to perform a highly motivated behavior, this suggests Automated feeders in use with RFID micro chips implanted in NHPs that they are budgeting energy toward more biologically essential Do females or males determine the dominance hierarchy in rhesus activities (sleep, eat, drink) macaques? TINT : when mouse retrieves new nesting material and incorporates it Hypothesis : that inappetance among socially housed macaques may into an existing nest within 10 minutes be indicative of increased social stress and give evidence before the onset of overt aggression Hypothesis that negative TINT would correlate with other measures of pain Findings suggestive that stress-induced inappetance and/or increased night-feeding in key individuals may help detect emerging instability Confounded by analgesia from anesthetic (Ketamine/xylazine) Caloric decrease in deposed subfamily Confounded by lack of response to rescue analgesia (buprenorphine) Caloric decrease predicted severity of trauma 13 14 Comparing Mouse Health Monitoring Between Soiled-bedding Sentinel and Exhaust Air Dust Surveillance Programs Several pathogens are unreliably detected with soiled bedding sentinels or poorly transmitted by soiled bedding What does this image show in use? Ans D Which was not one of the organisms in the comparison? Answer A A)Hepa Filter A) EDIM B) Rodentibacter C) Mouse Norovirus D) Helicobacter B)Temperature and humidity sensor C)Air freshener Soiled bedding yielded equivalent detection as Exhaust Air dust with which organism that was compared? Answer C D)Collection media A) EDIM B) Rodentibacter C) Mouse Norovirus D) Helicobacter 15 16 Comparing Mouse Health Monitoring Between Soiled-bedding Evaluation of 3 Alcohol-based Agents for Presurgical Skin Sentinel and Exhaust Air Dust Surveillance Programs preparation in Mice Three prep agents MNV is a non enveloped RNA virus in which family? Answer C Iodine povacrylex and isopropyl alcohol – denatures proteins and damages bacterial DNA A) Adenoviridae B) Parvoviridae C) Caliciviridae D) Herpesviridae Chlorhexidine gluconate in isopropyl alcohol – denatures protein and Mainly shed in feces disrupts bacterial membranes Helicobacter – gram negative, microaerophilic organism found in Chlorhexidine gluconate solution – disrupt bacterial membranes gastroinstestinal flora and mainly transmitted fecal-oral No additional loss of core body temperature Rodentibacter – was called Pasteurella pneumotropica No evidence of dermatitis or skin irritation Transmitted poorly through dirty bedding All three prep agents were as effective as triple application of Respiratory-Sendai, F. rodentium povidine and alcohol in alternating fashion Unstable and enveloped virus 17 18 3 Effect of Cranial Flexion of Pelvic Limbs on Interlaminar Length of the Lumbosacral Space in Sternally and Laterally Recumbent Juvenile Duroc and Adult Yucatan Pigs The images to the right depict multiplanar sagittal reconstruction of Null Hypotheses the lumbar spine of a Yucatan pig in lateral recumbency Cranial hyperflexion of pelvic limbs would not increase LSS and LSA of What is the orientation of the pelvic limbs in the bottom image? the pigs in sternal or right lateral recumbency Answer A Sternal positioning would not increase LSS and LSA compared to lateral A) Cranial hyper flexion Findings B) Caudal hyper flexion Hyperflexion of pelvic limbs increases LSS and LSA in sternally recumbent adult Yucatan, laterally recumbent adult Yucatan, and juvenile Duroc C) Neutral with leg in extension Hyperflexing the pelvic limbs of juveniles in sternal recumbency did not D) Neutral with leg flexed at knee increase LSS 19 20 Institutional Animal Care and Use Committee postapproval Monitoring Programs : A Proposed Comprehensive Classification Scheme PAM – Post approval monitoring as a means to ensure researcher compliance and oversight 2008 ILAR – any effort focused on determining what happens to animals after IACUC approval JAALAS Volume 59, has been granted Animal Welfare regulations – after approval of proposed activities, IACUC facility inspections must be conducted at least once every 6 months and continuing reviews be conducted not less than annually Guide – considers in broadest sense, consisting of all types of protocol monitoring after the No2 IACUC’s initial protocol approval Includes continued protocol review, lab inspections, vet or IACUC observations of procedures, observations by animal care, external regulatory inspections IACUC, veterinarian, animal care, and compliance staff can all perform PAM Classification – suggest a formal description and specify the following distinctions Protocol review vs procedural observations Formal vs informal Announced vs unannounced Random vs targeted 21 22 Genus and species? This is a bird’s eye view of a zebrafish tank. What does this image show was being tested as a preference? Answer C A)Colors B)Water flow rate C)Enrichment D)Feed allocation 23 24 4 Enrichment Preferences of Singly Housed Zebrafish Enrichment Preferences of Singly Housed Zebrafish (Danio rerio) (Danio rerio) Showed a preference for front of the tank when caretakers entered room Strongest enrichment preference shown was for? Authors suggested the importance of simulated movement because Answer A a static image of a zebrafish scaled to size did not elicit a preference A) mirrored paper B) PVC pipe C) marbles D) tulle Zebrafish are a shoaling and scototaxic species Hypothesized that position of fish in the tank would be Shoaling - Any group of fish that stays together for social reasons is altered by various enrichment strategies compared to said to be shoaling, and if the shoal is swimming in the same direction barren tank and that the effect would be greater when together, it is schooling enrichment imitated social contact Scototaxis - A preference for, and nd Initiated 2 study based on these findings to assess value subsequent movement towards darkness of conspecifics (single or group) Zebrafish are a primary species in the RDD Found that the test subject preferred to be on the side of the tank closest to conspecifics most significantly when it Embryonic development, genetic methods, husbandry, ammonia- was just another single fish nitrate cycle, common diseases, etc. 25 26 Adoption of Exhaust Air Dust Testing in SPF Rodent Facilities Use of Flavored Tablets of Gabapentin and Carprofen to Attenuate Postoperative Hypersensitivity in an Incisional Pain Model in Rats (Rattus norvegicus) Authors cite references that CAR bacillus, some helicobacters, Surgery – LCMV, Sendai, fur mites and Pasteurella species as not readily Plantar surface of the left hindpaw incised to access flexor digitorum transmitted by soiled bedding. brevis which was elevated and incised longitudinally Concern for residual DNA on racks leading to false positives though Skin incision closed with single interrupted horizontal mattress suture this group found additional washing and decontamination of the Rats received placebo tablet, placebo or Buprenorphine SR injection, racks unnecessary for their use gabapentin tablet, carprofen tablet, and combination Limited comparison of swab versus media sampling revealed no gabapentin/carprofen tablet evidence that one was more effective than the other Behavioral test Von Frey – mechanical stimuli Hargreaves – what type of stimuli? Answer A A) thermal B) chemical C) electrical D) musical 27 28 Use of Flavored Tablets of Gabapentin and Carprofen to Continuous Rate Infusion of Alfaxalone during Ketamine- Attenuate Postoperative Hypersensitivity in an Incisional Pain Xylazine Anesthesia in Rats Model in Rats (Rattus norvegicus) Showed a significant reduction in mechanical hypersensitivity can Evaluated alfaxalone as a CRI after Subcutaneous Ketamine- be achieved by providing gabapentin or carprofen PO Xylazine injection Both attenuated mechanical hypersensitivity for 3 days Atipamezole given after trial to both alfaxalone and no alfaxalone groups Thermal hypersensitivity was not attenuated with either compound Alfaxalone – neuroactive steroid Combination tablet of gabapentin and carprofen did not attenuate mechanical hypersensitivity on Day 1only Day 2 and 3 During trial tested front and hind withdrawal, surgical stimulation and HR, RR, SpO2, temperature, time to stand Ingestion of combo tablet was lower on Day 1 No rats in the alfaxalone group showed any withdrawal Rats had been receiving tablets for 3 days prior to procedure to Only 6 of 16 KX alone group lost hind limb withdrawal preload and decrease neophobia No difference in HR, RR, SpO2, temperature or time to standing differed between groups 29 30 5 Continuous Rate Infusion of Ketamine Hydrochloride and Dexmedetomidine for Continuous Rate Infusion of Alfaxalone during Ketamine- Maintenance of Anesthesia during Laryngotracheal Surgery in New Zealand White Xylazine Anesthesia in Rats Rabbits (Oryctolagus cuniculus) General – NZW are used as models of structural and functional consequences of vocal-fold vibration This study tested the anesthetic regimen for a model of hoarseness Article noted that other references showed alfaxalone to cause Answer C various levels of cardiorespiratory depression and muscle A) Vocal cord paresis B) Dysarthria C) Dysphonia D) Dystopia fasciculations Needed a regimen that provided Alfaxalone does not provide any analgesic properties Surgical depth of anesthesia To prevent pain responses Local autonomic laryngeal responses Which was not noted as a study limitation Answer C A) unidirectional anesthesia adjustment B) only male rabbits C) only 1 dosage used D) non randomized phrophylactic use of lidocaine topically 31 32 Quantitative and Qualitative Behavioral Measurements to Assess Pain in Axolotls (Ambystoma mexicanum) Evaluated which of these regimens? Answer A What is genus and species? A)Buprenorphine injection, butorphanol immersion B) Butorphanol injection, buprenorphine immersion C)Buprenorphine injection, MS222 immersion D)Butorphanol injection, Eugenol immersion Axolotls used actively in regeneration research often involving surgical injury Recent sequencing of 32Gb genome Quantitatively applied Von Frey anesthesiometers and acetic acid test as noxious stimuli 33 34 Quantitative and Qualitative Behavioral Measurements to Acepromazine and Chlorpromazine as Pharmaceutical-grade Assess Pain in Axolotls Alternatives to Chlorprothixene for Pupillary Light Reflex (Ambystoma mexicanum) Imaging in Mice Studies of isofluorane anesthetized mice in visual studies may use Previous studies primarily in frogs (Xenopus and Rana) and newts chlorprothixene to reduce isofluorane levels needed for sedation (Notophthalmus) Chlorprothixene is an antagonist at dopamine, serotonin, histamine, muscarinic acetylcholine and alpha adrenergic Von Frey receptors Curves of axolotl and mucous coating made difficult It is been discontinued as a pharmaceutical grade compound in the US, though still available in Europe Acetic Acid test Guide, pharmaceutical grade, “…should…be used when Used pipettor and small volumes for precise applications available for all animal-related procedures” Acepromazine and Chlorpromazine are phenothiazines with similar Tested at multiple time points up to 48 hours after analgesics given backbone to chlorprothixene. No significant difference in behavioral assessments (feeding, HR was significantly lower in saline group tapping, touches, novel object, inline jet) Mice on Isofluorane without sedatives had significantly slower PLR AAT had reproducible, concentration dependent pain response but Pupillary drift occurred more often in saline tx group no difference between control and analgesia groups Authors believed Acepromazine and Chlorpromazine could substitute for Chlorprothixene 35 36 6 Female Urine-induced Ultrasonic Vocalizations in male C57BL/6J Mice as a Proxy Indicator for Postoperative Pain Mice produce ultrasonic vocalizations which are > 20KHz (above what humans can hear) Ultrasonic vocalization (USV)more commonly produced in nonaggressive, same-sex and heterosexual interactions. What is being applied? Answer D USV that is NOT reproducible and heavily described in literature are? Answer B A) Pup isolation A)Compound to test copper toxicity B) Away from nest C) Intruder B)Dye for identification method D) Males in presence of female urine C)Preparation for photodynamic therapy Vasectomized mice that did not receive analgesia produced fewer FiUSV compared to baseline Production and quality of FiUSV depends on strain, social status, previous D)Healing clay for UD heterosexual experience Authors suggest FiUSV can be used as proxy indicator of pain in well-controlled experiment but not practical as general cage-side assessment Found Buprenorphine SR at 0.6mg/kg provided pain relieve after vasectomy as evidenced by unaltered FiUSV when mice compared to baseline 37 38 Elevated Arsenic and Lead Concentrations in Natural Healing Elevated Arsenic and Lead Concentrations in Natural Healing Clay Applied Topically as a Treatment for Ulcerative Dermatitis Clay Applied Topically as a Treatment for Ulcerative Dermatitis in Mice in Mice Ulcerative dermatitis is a common laboratory mouse skin condition Arsenic Many different treatments have been reported including natural Not taken up by tissues clays Filtered from blood and excreted rapidly in urine Clays are composed mainly of minerals for the illite group Lead Authors found high concentrations of arsenic and lead in the clays Does accumulate in tissues Mice often ingested through grooming Hepatic and renal concentrations of lead were elevated in clay Authors believe with increasing trace element exposure from clay treated mice compared to controls would increase in organs in following order Authors discouraged use of clay treatment in studies of lead toxicity, Kidney>liver>heart>brain heme biosynthesis or alpha2macroglobulin function 39 40 Comparison of Rectal and Infrared Thermometry Temperatures in Anesthetized Swine (Sus scrofa) Chose lateral canthus of the eye and base of the ear Infrared thermometry (IRTM) non contact method to measure temperature Proportional bias exists between IRTM and rectal temps Compared rectal temp to IRTM in swine IRTM underestimated at lower values and overestimated at higher Based on CT scan showing vasculature, what two locations were Limitations chosen to be sampled by IRTM? 2-3mm depth of penetration Study was heavily controlled sedated, precise distances and locations IRTM could potentially be used as a quick, non-invasive screen 41 42 7 Volume 3 Starts off with several review articles JAALAS Volume 59, Rodent Euthanasia Methods Intraperitoneal Injection of Sodium Pentobarbital as a Method of Euthanasia in Laboratory Rodents No 3 No pictures but points to other areas of study AVMA Euthanasia Guidelines CO 2 Updates Neonates Large groups Physical separation Assessing efficacy 43 44 Evaluation of Effective and Practical Euthanasia Methods for Hematologic Values of Jamaican Fruit Bats (Artibeus Larval African Clawed Frogs (Xenopus laevis) jamaicensis) and the Effects of Isoflurane Anesthesia Used to model Middle east respiratory syndrome, dengue virus, Zika, Tacaribe Existing guidance offer recommendations for egg and adults stages Dengue is a flavivirus but not larval stage Vector for dengue? Answer B Compared MS222, eugenol, and rapid chilling exposure for 15 minutes A) Amblyomma tick B) Aedes mosquito Tadpoles at NF (Nieukwop and Faber) stages 46, 47, and 49 C) Glossina fly 6g/L MS222 100% effective D) Tunga flea Eugenol at 800uL/L variable by age group and batches of stock solutions Jamaican Fruit Bat (JFB) belong to order Chiroptera Chilling was completely ineffective Zoonoses from JFB only directly observed from rabies, Hendra and Nipah (henipaviruses) 45 46 Using Vaginal Impedance Measurement to Identify Proestrus in Rats Given Luteinizing Hormone Releasing Hormone (LHRH) Blood collection from the propetagial (cephalic) vein Agonist Compared parameters in sedated and awake collection WBC, monocytes and lymphocytes were lower in unsedated bats Vaginal impedance is measurement of electrical RBC, platelet parameters not different between groups resistance of the vaginal mucosa What value in kiloohms is considered the threshold to indicate proestrus? Answer B A) 1 B) 3 C)30 D) 300 Value > 3 kiloohms indicated proestrus. The 3 other estrus stages have values lower than 3kΏ (PAUSE) Found serial measurement resulted in much lower false positives and negative breeding rates 47 48 8 What rat estrus cycle stage is pictured in B? Pharmacokinetic Profiles of Gabapentin after Oral and Answer A Subcutaneous Administration in Black-tailed Prairie Dogs A) Estrus B) Proestrus C) Diestrus D) Metestrus (Cynomys ludovicianus) Diurnal, terrestrial rodent with high degree of social organization Associated with human monkeypox outbreak in 2003 Used in studies monkeypox, Yersinia, Francisella, hepatobiliary disease, clostridial diarrhea, oxygen consumption, hibernation Evaluated oral and subcutaneous administration of gabapentin Peak plasma within 5h of administration Oral administration had longer half life than SQ though not statistically significant different Found likely to maintain plasma concentration at half maximum effective concentration for 12 hours 49 50 Investigation of Various Intramuscular Volumes Delivered to the Semimembranosus Muscle of Cavia porcellus Assessing dispersion leakage of IM injection Assess pain and pathology associated with IM injection of saline Results suggest volumes of 150 and 300 ul remain within the target Used iohexol and CT imaging for the leakage assessment muscle Injection into semi-membranosus muscle Pain and pathology did not appear to increase as IM volume Of volumes tested 300ul showed the least expansion/leakage and increased most rapid absorption Used iohexol injections and CT imaging Larger volumes were not associated with greater pathology or pain as majority of damage was attributed to the needle 51 52 Reference Intervals for and the Effect of Sample Handling and Reference Intervals for and the Effect of Sample Handling and Sex on Rotational Thromboelastometry in Health Adult Pigs Sex on Rotational Thromboelastometry in Health Adult Pigs The elastic properties of a clot are measured from formation until ROTEM – Rotational thromboelastometry lysis by immersing a pin into a blood sample The elastic properties of a clot are measured from formation until Pig had high serum fibrinogen concentration compared to humans lysis Evaluated the effects of sex, sampling order, and agitation on Allows evaluation of the entire coagulation cascade including ROTEM results platelet function and fibrinolysis No effects by sex or agitation Sampling order had an effect on clot formation time (CFT) and Extrinsic Assay panel 53 54 9 Vertebral heart Score in Rhesus Macaques (Macaca mulatta): Generating Normal Reference Intervals and Assessing its Validity for Identifying Cardiac Disease JAALAS Volume 59, Vertebral heart scoring is a semiquantitative method to assess the presence and severity of cardiomegaly on thoracic radiographs Measurements along short and long axes compared to the number No 4 of thoracic vertebrae present Hypothesized that NHP with cardiac disease on echocardiography would also have higher VHS 55 56 Behavioral Effects of Cage size and Environmental Enrichment in New Zealand White Rabbits Housed rabbits in 3 different size cages Ethogram constructed to quantify behaviors and compare activity between cage sizes Then explored if enrichment could promote the Did they find that VHS could be used as an isolated screening tool expression of active behaviors in standard size to identify rhesus macaques needing further cardiovascular cages examination? B- NO Also analyzed the fecal glucocorticoids after A) Yes B) No shipping or surgery to see if enrichment The sensitivities and specificities of the VHS thresholds are not mitigated effect of stressors sufficiently robust to avoid a high-probability of false positives and negatives Doesn’t help distinguish between types of cardiac disease Not significantly different between control and hypertrophic cardiomyopathy animals 57 58 Room Decontamination Using Ionized Hydrogen Peroxide Fog and Mist Reduces Hatching Rates of Syphacia obvelata Ova Study found housing in larger cages spent more time performing active exploratory behavior but less what? Answer A Syphacia Obvelata A)Grooming B) Resting Life cycle direct and completed in 12-15 C) Digging days D) Rearing What sign is indicative that an ova has E) Frenetic Random Activity Period hatched in picture D? Answer C Enrichment devices produced active exploratory behavior in standard housing similar to that in large housing A)Striations Rearing up in extreme vertical position called ‘telescoping’ which was only seen in B)Color large housing C)Opercula Frenetic random activity periods is thought to be a behavioral expression of excitement D)Banana shape FRAP did not increase with enrichment devices No increase in fecal glucocorticoids after shipment or surgery General - > size > species specific behavior; enrichment in standard cage had positive behavioral change too 59 60 10 Room Decontamination Using Ionized Hydrogen Peroxide Fog Prewarming Followed by Active Warming is Superior to Passive and Mist Reduces Hatching Rates of Syphacia obvelata Ova Warming in Preventing Hypothermia for Short Procedures in Adult Rats (Rattus norvegicus) Under Isoflurane Anesthesia According to the article, the most important promoter of Collections from perianal tape test of mice purchased from a pet hypothermia during anesthesia is? store A)Cold liquids used to prep surgical sites Ova exposed to varying whole room fogging for different lengths of time or different number of applications of surface misting B)Anesthetic effect on hypothalamic threshold (handheld applicator) C)Temperature of the surgical suite Although complete inhibition was not achieved, their results showed D)Vagal effect on frontal lobe a statistically significant reduction of ova hatching across all 3 iHP exposed groups Anesthesia disrupts hypothalamic threshold which normally triggers a thermoregulatory autonomic response to small drops in temperature This allows warm core blood to distribute to periphery and lose heat to the environment 61 62 Prewarming Followed by Active Warming is Superior to Passive The Stability and Efficacy of Tricaine Methanesulfonate Warming in Preventing Hypothermia for Short Procedures in (MS222) Solution After Long-Term Storage Adult Rats (Rattus norvegicus) Under Isoflurane Anesthesia 3 groups Ester-type local anesthetic agent absorbed through gills or skin Acts on muscle and peripheral and central nervous systems Control Highly lipid soluble and readily crosses the cell membrane to block Pre-warm and passive heat (blanket) sodium channels which then limits membrane excitability and action Pre -warm and Active heat (temp controlled heat pad) potential transmission Primarily excreted unchanged through the gills but can be metabolized Pre-warming defined as 1% increase over baseline core temp through acetylation in liver, kidney, blood and muscle depending on species Found pre-warming and passive heat was minimally effective in maintaining normothermia No histologic lesions in MS222-related organs Hypothesized and found that MS222 is chemically stable and remains Active heat more effective an effective anesthetic agent when stored for up to 6 mo at 4C or less (100mg/ml stock) Powder may cause skin, eye, or respiratory irritation/damage but not labeled a carcinogen 63 64 Effect of Nearby Construction Activity on Endothelial Function, Sensitivity to Nitric Oxide, and Potassium Channel Activity in the Middle Cerebral Arteries of Rats Assessed the effect of nearby construction activity on Reponses of rat MCA to acetylcholine and sodium nitroprusside Acetycholine – vasodilator Sodium nitroprusside – Nitric oxide donor Two monitoring systems Other papers of deleterious effects on nearby construction Comercially available unit Reduced food intake Constructed Raspberry-Pi Plasma ACTH, corticosterone, aldosterone doubled During construction This study found animals evaluated after construction completed Vasodilator responses to acetycholine and SNP were abolished had intact MCA responses to acethycholine and SNP MaxiK single channel current opening frequency and open state probability dramatically decreased 65 66 11 Effects of Giardia lamblia colonization and Fenbendazole Treatment on Canine Fecal Microbiota Authors found that Fenbendazole treatment of Giardia did impact Giardia lamblia (duodenalis) the gut microbiota of canines? Answer B-False, there was minimal 2 life stages impact Trophozoite - “Found in diarrhea of infected dogs and cats, but do not A) True B) False survive for a prolonged period outside the host” Cyst – Resistant in environment. Can survive several months outside of host Giardia-infected mice have been found with altered GM Suggested that giardiasis may alter resident microbiota Fenbendazole often used to treat giardia Recently fenbendazole used to treat mouse pinworms showed minimal GM change Study to assess GM based on giardia status and fenbendazole treatment 67 68 Study found no consistent difference in B-diversity, alpha-diversity JAALAS Volume 59, and richness of GM based on Giardia status Study did find significant differences of GM between? Answer D A) Gender No 5 B)Age C)Breed/background D) Housing facility FBZ had minimal effect on the canine GM composition, richness or diversity 69 70 A Nonsurgical Embryo Transfer Technique for Fresh and Cultured Blastocysts in Rats Describes a non surgical device (rNSET) used to deliver fresh or cultured blastocyst stage embryos to the uterine horn of pseudopregnant female rats Estrus cycle synchronization of pseudopregnant female recipients with IP injection of PCR and RT-PCR overview luteinizing hormone-releasing hormone antagonist 4 days prior to mating Review of published Anesthesia Protocols used to Create Ischemia Oxytocin used to dilate the cervix by IP injection of 2IU 3.5 days post coitum. Most had cervix dilation by 2hr post injection Reperfusion Myocardial Infarcts in Swine Other uses of device AI in mice Deposit bacteria into uterine tissue What difference was noted between cervix of mice and rats when using the device? Answer A A)Cervix of rats not dilated at optimal transfer time without oxytocin injection B)Cervix more easily damaged in rats C)Cervix much further away proportionally in rats D)Dual cervix in rats 71 72 12 Compressed Paper as an Alternative to Corn Cob Bedding in Mouse (Mus musculus) Cages Corn cob bedding which can Be consumed, altering feed conversion Shown that rats on corn cob spend less time in slow-wave sleep Based on the article, which statement is false? Answer B Contain phytoestrogens A) rNSET did not induce a change in fecal corticosterone levels Unprocessed agricultural byproduct B) rNSEt group showed a short pause in weight gain compared to all the other groups Other research confounding reports C) There was no significant difference in birth rates seen between SET Compressed paper and rNSET when Blastocysts transferred Contains significantly lower initial endotoxin and coliform counts D) Birth rates by rNSET of cultured blastocysts was significantly less than Has greater absorption capacity SET transfer 73 74 Compressed Paper as an Alternative to Corn Cob Bedding in Cage Position and Response to Humans in Singly-housed Mouse (Mus musculus) Cages rhesus Macaques (Macaca mulatta) CP has higher total and immediate absorbent capacities Common traditional laboratory housing is 2-tiered design CP decreased the number of early cage changes required Many NHPs arboreal and use verticality to escape predators Had lower overall intracage ammonia levels and lower rates of Light levels also lower in bottom cages ammonia production No effects of tier on behaviors Study examines the response of rhesus macaques to attempted positive social interaction Tests hypothesis that responses to humans are more negative in bottom tier inhabitants 75 76 Cage Position and Response to Humans in Singly-housed Construction of an Affordable Open-Design Recirculating rhesus Macaques (Macaca mulatta) Zebrafish Housing System Study provides no support for a relationship between cage row and Zebrafish (Danio rerio) response to humans (hand feeding) Family Cyprinidae Subjects had been housed on different rows at different times Found naturally in Pakistan, India, Bangladesh, Nepal,Myanmar No relationship in behaviors observed to proximity to door Adult females may spawn every 2-3 day Did find significant sex differences in reactions to humans The white, circular pieces shown in the picture are for? Answer C Which of these best describe female behaviors compared to males in the study? Answer B A) Environmental enrichment A) more likely to take treat and more likely to lip smack B)Growing artemia to feed the zebrafish B) less likely to take treat and more likely to show fear C)Growing denitrifying bacteria C) more likely to show aggression and less likely to take treat D)Mechanical filtration Increased age and tenure were associated with greater treat taking 77 78 13 A Comparison of Ketamine or Etomidate Combined with Xylazine for Intraperitoneal Anesthesia in Four Mouse Strains Mice on which anesthetic protocol experienced more reactions of hyperexcitement? EX in BALB/C, NU Assess etomidate/xylazine as an alternative to ketamine/xylazine Which was the only protocol/strain combination found in this study to produce Also evaluated sex and strain differences reliable surgical anesthesia? Answer B Ketamine reported to have several anti-inflammatory effects A) EX in Nude mice Reduced function of cytokines, lymphocytes, neutrophils and NK cells B) EX in C57BL/6 C) KX in Nude mice Increased tumor metastasis D) KX in C57BL/6 Etomidate All groups exhibited marked hypothermia with NU most pronounced GABA receptor agonist Authors believe EX could be used in tested strains (BALB/C, C57BL/6, NU, CD1) for Hypnotic agent short, non-painful procedures Minimal effects on HR, RR, BP Sex or strain differences Not DEA controlled Decreased sedation times for CD1 Reports of pain or myoclonus during induction after IV administration Achieving surgical anesthesia in females more likely than males 79 80 Intraperitoneal Alfaxalone and Alfaxalone-Dexmedetomidine Anesthetic Effects of Intramuscular Alfaxalone-Ketamine in Anesthesia in Sprague-Dawley rats (Rattus norvegicus) Naked Mole Rats (Heterocephalus glaber) 3 Doses of alfaxalone alone and 4 doses of alfaxalone- dexmedetomidine combination Naked mole rats Found that alfaxalone alone only achieved a sedative level of Family Bathyergidae native to subSaharn Africa anesthesia Eusocial? Answer C All A-D combinations reached surgical level (based only on pedal A)Monogamous breeding pairs, shared work duties reflex) B)Polygamous breeding pairs, shared work duties Sex differences C)One ‘queen’ as the sole breeder the rest are mainly workers Females experienced longer duration of sedation than males with same D)One ‘king’ with harem and the rest are mainly workers dose Low basal metabolic rate, high hematocrit and high hemoglobin Another study suggested 3X dose needed for males Alternative routes (SQ) not considered for this study Longevity compared to other rodents 81 82 Anesthetic Effects of Intramuscular Alfaxalone-Ketamine in A Comparison of Buprenorphine, Sustained-release Naked Mole Rats (Heterocephalus glaber) Buprenorphine, and High-concentration Buprenorphine in Male New Zealand White Rabbits Evaluated tissue reaction, food consumption, fecal production Induction time was significantly longer and anesthetic duration Buprenorphine, Buprenorphine SR, Buprenorphine HC significantly shorter in male mole rats Buprenorphine μ agonist; δ,Κ antagonist Induction defined as time from injection to loss of righting reflex Reported adverse effect of long-acting Bup are sterile abscesses at injection site AK did not provide consistent surgical plane as indicated by variable responses to toe pinch 2 of the 7 in the HC group experienced nystagmus, ataxia, head tilt and circling In rodents hindlimb withdrawal may be lost at a lighter plane of Most treated rabbits lost weight anesthesia than forelimb Which group(s) had significant but transient decreases in food Males showed no loss of hindlimb and minimal loss of forelimb consumption compared to controls? Answer D Loss of palpebral reflex also used to indicate a deeper anesthetic A)Bup SR plane but difficult in rodents and subterranean species may be B)Bup HC further confusing C)Bup and HC Sex associated differences shown in alfaxalone use in other reports D)Bup SR and HC 83 84 14 A Comparison of the Efficacy and A Comparison of the Efficacy and Cardiopulmonary Effects of Cardiopulmonary Effects of 3 Different 3 Different Sedation Protocol in Otolemur garnettii Sedation Protocol in Otolemur garnettii Northern greater galago AKA small eared galago, Garnett’s greater galago, or bush baby Common Name? Nocturnal, arboreal prosimian Compared Alfaxalone Ketamine Ketamine/ Dexdomitor 85 86 Ketamine Increase HR, CO, BP Respiratory depression called apneustic breathing JAALAS Volume 59, Slow recovery, muscle damage, and excessive salivation All 3 groups efficacious in ability to immobilize Immobilization time longest in which group? Answer A No 6 A) Aflaxalone B) Ketamine C) Ketamine/Dexdomitor* Muscle tension greatest in which group? Answer B A) Aflaxalone B) Ketamine C) Ketamine/Dexdomitor* Ketamine/Dex group statistically significant lower average CK and highest glucose 87 88 Comparative Morphology and Morphometry of Blood Cells in Zebrafish (Danio rerio), Common Carp (Cyprinus carpio carpio), and Tilapia (Oreochromis niloticus) Morphometric studies defined as quantitative descriptions of a cell’s geometric structures in all dimensions. Which of the following classifications is correct? Answer C A) Zebrafish (Cyprinidae), Tilapia (Cyprinidae) What cell type is shown above from the zebrafish, and tilapia respectively? Answer D B) Zebrafish (Cichlidae), Tilapia (Cichlidae) A) vacuolated monocyte C) Zebrafish (Cyprinidae), Tilapia (Cichlidae) B) neutrophil D) Zebrafish (Ictaluridae), Tilapia (Oreodae) C) thrombocyte D) eosinophil 89 90 15 Basophils were only found readily in which of the sampled species? What procedure is shown here? Answer D Answer B A) Embryo transfer A) Zebrafish B) Carp C) Tilapia B) Viral transfection C) BrdU staining Younger erythrocytes were more? Answer A D) ICSI A) circular B) elliptical C)oblong D)angular Neutrophils were largest from which species? Answer B A) Zebrafish B) Carp C) Tilapia 91 92 Cryopreservation of Mauritian Cynomolgus Macaque (Maca fasicicularis) Sperm in Chemically Defined Medium Goal is to optimize cryopreservation of sperm from Mauritian cynomolgus Mauritian are a subspecies Small founder population Limited MHC diversity (7 haplotypes) Good for AIDS, vaccine and stem cell research Cryopreservation often results in decrease in sperm quality, motility survival and ability to fertilize Compared sperm viability and motility From osmotic change, cold shock and ice crystals when cryopreserved in chemically The image above is from a CASA IVOS II capture field. What do the sperm with defined freezing media of variable red squares represent? Answer A osmolarity and with either ethylene A)Static sperm glycol or glycerol B) Largest 25% of sperm sample C) 1st fraction sperm D) Most viable sperm 93 94 Effects on Mouse Food Consumption After Exposure to Bedding from Sick Mice or Healthy Mice Some other reports of freezing with home-made egg-yolk and TES based media with glycerol Believe yolk complicates ICSI Housing of mice in the same room as study mice from pancreatic ductal adenocarcinoma study showed decreased food intake Agglutination coincident with the cachexia experienced by the PDAC mice Clog pipette Cachexia – multisystemic syndrome involving lean mass catabolism, Obscure sperm visibility metabolic disturbances and behavioral changes such as fatigue Author’s extender based on PIPEs and TES and anorexia Stronger buffering capacity PDAC models are highly associated with cachexia Included 3 amino acids – proline, glycine, glutamine Interestingly, controls from the study experience decreased food intake Protein-stabilizing osmolyte – trehalose Found 310mOsm, 4.6% ethylene glycol and 5% glycerol best sperm Theory – empathy event and hypothesize that normal mice exposed viability to soiled bedding of cachexic mice will also consume less food 95 96 16 Metaphylactic Antibiotic Treatment to Prevent the Transmission of Corynebacterium bovis to Immunocompromised Mouse Offspring C bovis is a common bacterial pathogen among immunocompromised mouse colonies The authors constructed this device to do what? Answer C C. bovis is a facultative anaerobic, gram + bacterium A)Allow air passage of pheromones In athymic nude mice can reach a persistent carrier state and dermal B)Allow for off-gassing from sick mice hyperkeratosis, dehydration, lethargy and decreased body condition C) Sift orts from bedding Haired immunocompromised mice like NSG, can have rough hair coat, D) Provide a raised platform for eating decreased body condition, scaly skin, alopecia, conjunctivitis and erythematous pinnae Mentions depopulation, embryo transfer and cesarean rederivation as Found increase and decrease in eating at different time points eradication Did not have enough numbers to attain statistical power Testing amoxicillin-clavulanic acid in drinking water as metaphylactic treatment 97 98 Injection-site Reactions to Sustained-release Meloxicam in Sprague-Dawley Rats 6 control Sprague-Dawley received saline injection SQ Metaphylactic treatment is best defined as…? Answer D 16 received sustained-release meloxicam SQ A) treating with the goal of curing an unknown infection No control rats developed masses at site whereas all treated rats did B) treating with the goal of curing a known infectious agent C) treating with the goal of preventing possible infections D) treating with the goal of controlling an infection that is already present This study was to control C. bovis infected NSG breeders with amoxicillin-clavulanic acid to prevent post-partuition transmission AB given to qPCR positive adult breeding pairs C. bovis offspring moved to another room and confirmed negative at 10 wks and 31 wks 57% of breeders PCR positive and 34% culture positive at 10 wks DC Abx therapy alone does not eliminate established C. bovis infections 99 100 Injection-site Reactions to Sustained-release Meloxicam in Sprague-Dawley Rats Sustained release meloxicam widely available and use but adverse skin reactions are reported infrequently Prolonged time between injection and emergence of lesion 2-3d palpable The black arrow is described by the authors as…? Answer A 5-6d defined borders A)Ulceration of the epidermis 7d for erythema B)Necrotizing panniculitis Not visually apparent, remained furred C)Foamy macrophages Attributed to other etiologies D)Organizing granulation tissue 101 102 17 A comparison of Buffered Tricaine Methanesulfonate (MS-222) Cardiovascular and Metabolic Responses to Carbon Dioxide and Isoeugenol Anesthesia for Caudal Fin Clipping in Zebrafish Euthanasia in Conscious and Anesthetized Rats (Danio rerio) Compares safety and efficacy of buffered MS-222 and isoeugenol for Compared physiologic responses to CO2 euthanasia alone and with anesthesia and caudal fin clipping isoflurane preanesthesia MS-222 characterized as a local anesthetic Implanted Sprague-Dawley rats with telemetry devices Must be buffered MAP Retinotoxic and irritating to mucous membranes for personnel handling HR Isoeugenol is a constituent of clove oil Blood glucose Mechanism unknown but may inhibit sodium, potassium and calcium AVAMA Guidelines on Euthanasia 2020 increase recommended CO2 channels, inhibit NMDA receptors and potentiate GABA displacement rate to 30-70% Induction faster Stress ≠ distress Recovery longer Stress is a perturbation to an organisms physiologic homeostasis or psychologic Transfers rapidly from plasma to tissues but then slowly eliminated wellbeing Found safety and efficacy similar for anesthesia and caudal fin clipping Distress when adaptive mechanisms fail to restore homeostasis 103 104 Cardiovascular and Metabolic Responses to Carbon Dioxide Euthanasia in Conscious and Anesthetized Rats THE END MAP was significantly higher than baseline in both groups when cage lid placed over the home cage Good Luck! HR increased when lid was placed and remained stable through LOC under isoflurane but decreased quickly and drastically with just Thank you! CO2 likely to quick asphyxiation Glucose levels remained stable thru procedure Both groups affected similarly Pre-anesthesia with isoflurane did not offer any benefit to the euthanasia process 105 106 107 18