Journal of the American Association for Laboratory Animal Science
Volume 47, Number 4, July 2008

ORIGINAL RESEARCH
Biology
Blaha and Leon.  Effects of Indomethacin and Buprenorphine Analgesia on the Postoperative Recovery of Mice, pp. 8-19

Task 1, K6
Task 2 K1 – 7
Task 3, K5

SUMMARY: The authors compared the effects of two analgesic regimens post intraperitoneal implantation of a telemetry device in 59 male C57Bl/6J mice.  Dosing twice with indomethacin at 1 mg/kg was shown to have fewer adverse effects than dosing with buprenorphine (0.3mg/kg) and once with indomethacin (1 mg/kg) post surgically.

Objective measurements for the determination of pain in rodents are limited. The telemetry device was used to monitor physiologic changes.  Indomethacin was used for this study because it has been shown not to adversely affect circadian rhythm and activity levels in mice, nor affect platelet levels, and it can be administered orally.  Buprenorphine was selected primarily due to its popularity among post surgical protocols.  

Animals were randomized for surgical time among groups, anesthetized with isoflurane, prepped, had the procedure between the hours of 0830 to 1400 and received their first dose of analgesic by injection, with a second dose given orally the following morning at 0900. Control groups (no surgery, or surgery plus vehicle administration), were similarly randomized and dosed.  Oral dosing was achieved by voluntary consumption of an indomethacin soaked pina colada flavor treat.  It should be noted that ALL mice consumed the treat readily within 1 hour of presentation. 

Parameters evaluated included body weight, food and water intake, temperature and activity counts.  The authors note that they recognized increased food intake following cage/food changes as a normal phenomenon. All surgical groups decreased in body weight for a period of 2-4 days following surgery.  The Bup/Indo group lost significantly more than the Indo-Indo group.  By day 4, all groups were identical, yet did not regain presurgical weights within 13 days following surgery.  Both analgesic groups consumed more water than control groups on day 1 post op, but no group returned to presurgical levels within 13 days.    The nonsurgical animals dosed with Bup-Indo decreased water intake on day 1 post injection, and normalized by day 3.  Food intake was decreased in all groups following injections on day 1, with the bup-indo group decreasing significantly more than other groups.  By day 2, all groups were equally decreased in food intake from intakes for a time period after cage/food change, but within range otherwise.  Water intake decreased in the Bup–Indo group on day 1.  Circadian patterns of temperature and activity were not altered by treatment.  However, during the nocturnal cycle, the indo-indo group was significantly more active than its control group, and the bup-indo group had significantly higher temp ranges during these times.    

QUESTIONS:
1.	Does indomethacin treatment significantly prolong a post surgical decrease in activity?  Yes/No
2.	How does transmitter implantation affect bodyweight, exclusive of post operative treatment?  
A. 	Decreases for 2 days  
B.  	Decreases for 1 week  
C. 	Decreased for 13 days 
D 	Increases body weight in analgesic treated groups E increases body weight in untreated groups
3.	How does analgesic administration affect body weight loss after surgery?  
A. 	Ameliorates changes completely 
B. 	Ameliorates changes only when NSAIDs are used  
C. 	Ameliorates changes only if an opiate is used  
D. 	Ameliorates changes incompletely
4.	Which analgesic group had the most significant change in recovery of water consumption on day 1 after surgery?  

ANSWERS:
1.	No
2.	C
3.	D
4.	Indo/Indo

Lamon et al.  Adverse Effects of Incorporating Ketoprofen into Established Rodent Studies, pp. 20-24

Task 4 - Treat Disease or Condition as appropriate 
Primary Species - Rat 

SUMMARY: Ketoprofen administered postoperatively to rats undergoing ovariectomy caused gastric lesions in several animals.  In the first experiment, 19/32 SD rats showed clinical signs of lethargy, anorexia, diarrhea , hunched posture, periorbital and perinasal porphyrin staining, and ruffled coats within 7 days of one postoperative injection of ketoprofen; 12 rats given saline had no clinical signs. Necropsy showed gastric ulceration and peritonitis which was confirmed with histopathology. Dosage was recalculated to assure accuracy, other avenues investigated including contamination of the ketoprofen to rule out other possible reasons for the lesions. 

A second experiment was performed utilizing 24 rats, 12 received ketoprofen postoperative, 12 received saline as controls. No rats showed clinical signs or developed lesions noted upon necropsy. These animals were noted to be from a different barrier than the rats treated in the original experiment. 

A third experiment was performed utilizing rats from both barriers as in the first two experiments. In this experiment, 12 rats, 6 from each barrier, were given ketoprofen, 12 rats, and 6 from each barrier received saline.  3/6 rats showed similar clinical signs as the rats in the initial experiment. Necropsy confirmed inflammatory gastroenteritis, ulcerative and necrotizing gastroenteritis with bacterial septicemia and acute hepatic necrosis. These three animals were confirmed to be from the same barrier as the rats in the initial study.

No lesions or clinical signs developed in the rats from the other barrier or from the control rats from the first barrier. 

There may be some phenotypic differences in the SD rats from each barrier that could explain the susceptibility of these animals to the adverse effects of ketoprofen. 
Dosages of ketoprofen administered were within the acceptable reference range found in the veterinary compendia. 

QUESTIONS: 
1.  	T/F The cyclooxygenase (Cox) pathway has two isoenzymes (Cox1 and Cox2) which differ in function: Cox 1 is a protein that acts as an enzyme to produce prostaglandins in the endothelium and has a protective effect on the mucosa of the GI tract, kidney and various tissues in the body, Cox 2 is induced by inflammation and acts as an enzyme to produce prostaglandins during tissue damage. Ketoprofen is an NSAID that selectively inhibits COX 2. 
2.	Chronic use of NSAIDS can cause: 
a. 	Gastric mucosal damage 
b. 	Renal necrosis 
c. 	Hepatitis 
d.  	All of the above 
3. 	T/F   There are no alternatives to pain relief aside from using the appropriate NSAID. 

ANSWERS: 
1. 	F  - Ketoprofen affects both pathways 
2. 	d - All of the above 
3.  	F - One alternative is utilizing opioids. 

Azar et al.  Effect of Housing Rats in Dim Light or Long Nights on Heart Rate, pp. 25-34

Species:  Rat (Primary)
Domain/Task: 4

Objective:  To evaluate if housing laboratory animals, rats, under lighting conditions that more closely mimic their natural environment improves their well-being.
1.	Test the hypothesis that housing rats in a dimly illuminated environment will decrease HR under normal conditions as well as after acute procedures are preformed
2. 	Confirm previous research showing that long nights reduce heart rate
3. 	Determine whether strain or sex influence the effects of the 2 different lighting schemes

Variables examined included the effects of dim light (12:12 h light: dark photocycle with 10 1xillumination) or a long-night (8:16 h light: dark photocycle with 200 1x illumination) photocycle on rat’s heart and on their heart rate in response to various acute procedures.  Sprague Dawley (SD) and spontaneously hypertensive rats (SHR) from Harlan were evaluated.  Heart rate was monitored via implanted radiotelemetry transmitters.

Results:
1.  HR Under Undisturbed Conditions:
1	HR of undisturbed SHR male rats housed in dim or long nights were significantly lower vs. the control group
2	HR of undisturbed SD female rates was not significantly affected
2.  HR responses to acute husbandry procedures:
1	Gentle handling→SD males and SHR females in dim light had lower HR vs. same rats in long nights had no change in HR; SD females and SHR males had no change under both conditions
2	Cage change→SHR males and females had lower HR compared to SD males and females for both lighting conditions (similar to handling variable)
3	Stressful procedures →Overall, variable results. 
4	Intruder Rat:  SHR male HR increased/were higher in both lighting conditions whereas SD male were not affected;  SHR females had no change compared to SD females had reduced HR in the long night group.
5	Removal of Established cage mate:  SHR male rats were unaffected vs. SD males had lower HR in long night groups; Both SHR and SD female rats were significantly reduced
3.  HR responses to acute experimental procedures:
1	Neither dim light nor long nights affected the HR responses to hand restraint and SQ injections; SHR males had significantly smaller responses than did SD males, but this effect not seen in either female strains.
2	Dim light enhanced HR responses of SHR male rats but no significant effect on SD males whereas SD female rats had enhanced HR response but no effect on SHR females.
3	Long nights significantly reduced HR response of SD male rats and enhanced SD females, whereas there was no effect seen in SHR male or female rats.
4	Hand restraint and tail vein injection:  HR responses were not significantly affected by altered lighting and only long nights significantly reduced HR response of SHR male rats to IP injection; *In general, SHR male and female rats had lower responses to both procedures vs. SD males and females
4.    HR responses to acute stressful procedures:
1	Urine/Fecal odor:  HR of male SD and SHR were not altered by dim light or long nights.  In contrast, HR of SD were lower in both conditions (SHR no alteration).
2	Female Urine/Fecal odor:  Male SD and SHR had similar HR responses but those of SHR females were significantly greater vs. SHR males
3	Dried rat blood:  Lighting conditions did not alter HR response of SD or SHR male rats.  Dim light reduced the HR response of SD female rates (SHR no alteration).
4	Physical restraint:  Dim light reduced the HR response of SD males (SHR no alterations).  Long night enhanced HR response of SHR males and also SHR females.  The HR response of SD females was not affected by either lighting condition.

Discussion
1	Results show housing rats under dim light or long night conditions significantly reduced HR of SD male and SHR male and female rats when undisturbed
2	HR decreased to only some of the acute husbandry, experimental, or stressful procedures when housed in dim light and long night cycles.
3	Both HR during undisturbed conditions and HR responses after acute procedures varied with strain and sex:  Male rats had lower HR than females and SHR rats had lower HR than SD when undisturbed
4	The decreasing effect of altered lighting on HR in undisturbed animals is consistent with the possibility that rats housed in 12:12 h light: dark at 200x are stressed to some degree
5	The comparison of light treatments and strain during undisturbed periods indicate that HR of SHR rats were reduced more by altered light than SD rats
6	Overall, lighting effects appeared to be related to type of procedure, strain, and sex of rats; Females were affected more by altered light than were males
7	The dim light treatment may mimic the lighting condition of a rat burrow whereas the long night treatment may mimic a bright day in the middle of winter but without cold temperatures
    
QUESTIONS:
1. 	Which of the following represent stress effects:
a.  	Enhanced secretion of hormones of the HPA hypothalamic-pituitary axis
b. 	Increased/suppressed hormones of HPA like oxytocin, prolactin and growth  hormone
c.  	Enhanced activity of autonomic nervous system (sympathetic)
d.  	Alterations in GI, cardiovascular, and thermoregulatory systems  
2.  	True or false:   Female rats generally have greater stress responses versus male rats
3.  	Results of this study show housing rats under dim light or long night conditions significantly reduced HR of SD male and SHR male and female rats when undisturbed

ANSWERS:
1.  	a,b,c,d
2.  	True
3.  	True

Harvey et al.  Hematology and Plasma Chemistry Reference Intervals for Mature Laboratory Pine Voles (Microtus pinetorum) as Determined by Using the Nonparametric Rank Percentile Method, pp. 35-40

Domain 1:  Management of Spontaneous and Experimentally Induced Diseases 
and Conditions

Task 3:  Diagnose disease or condition as appropriate

Tertiary species: other rodents

SUMMARY: The objective of this study was to establish clinically relevant normal hematological and plasma biochemical values in the nonreproductive adult pine vole.  Pine voles, in contrast with meadow and motane voles, are monogamous and exhibit cooperative breeding, pair bond formation and biparental care.  Vole species have historically served as a model for studies in behavioral neuroscience and endocrinology, and pine voles have been more recently evaluated as a model for infection with /Rickettsia rickettsii/, the causative agent of Rocky Mountain Spotted Fever.

 Most reference intervals for laboratory species have been determined using the mean and standard deviation of a reference group.  This method is not recommended by clinical laboratory medicine organizations because many biological analytes are not normally distributed around a mean and calculated reference intervals can result in nonphysiologic values.  The current preferred method is the rank percentile method, which was used to determine 95% and 50% interfractile reference intervals in this study.  This method is valid regardless of the data distribution, but it can provide misleading results if the number of subjects is less than 50.

 Results for biochemical and hematologic analysis of cardiac blood taken at euthanasia for 112 pine voles are summarized in Tables 1 and 2 on page 38.  There were no clinically significant differences between the sexes.  Younger animals had a lower hematocrit, higher mean corpuscular volume (MCV), and lower mean corpuscular hemoglobin concentration (MCHC) than older animals, which was similar to other mammals whose neonates and young juveniles have a mild transient macrocytic hypochromic anemia.  Two hematologic analyzers were used in the study, but only platelet counts differed when comparing values from the different analyzers.  Pine voles have a lower MCV, higher RBC, higher BUN, much higher ALT, and lower glucose and phosphorus concentrations relative to rats and mice.

QUESTIONS:
1. 	T/F:  In contrast to other vole species, pine voles are polygamous
      and solitary.
2. 	Microtine rodents have historically served as a model for which
      type of studies?
 a. 	Salmonella infection
 b. 	Behavioral neuroscience
c. 	Behavioral endocrinology
d. 	b and c
3. 	Pine voles have recently been evaluated as a model for which
      rickettsial disease?
a. Ehrlichiosis
b. Babesia
c. Lyme disease
d. Rocky Mountain Spotted Fever
4. 	According to this study, what is the current preferred method of
      calculating reference intervals for laboratory species?
5. 	Compared to rats and mice, pine voles exhibit higher levels of
      which analytes?

ANSWERS:
1. 	False, pine voles are monogamous and exhibit cooperative breeding
      and pup care
2. 	d
3. 	d
4. 	The rank percentile method
5. 	RBC, BUN and ALT

Wimsatt et al.  Long-term and Per Rectum Disposition of Clarithromycin in the Desert Tortoise (Gopherus agassizii), pp. 41-45

SUMMARY: In this article, the authors summarize three different PK studies done using clarithromycin (CLARI) in the desert tortoise. The desert tortoise is susceptible to chronic upper respiratory infections associated with Mycoplasma agassizii. Affected tortoises show symptoms of oculonasal discharge and swelling of the eyelids initially and can, after a prolonged infection, suffer from weight loss and opportunistic infections. Mycoplasma sp. are susceptible to macrolide antibiotics such as CLARI. The latter was chosen as it is more stable in response to stomach acids and is better absorbed in the presence of food than erythromycin.

All of the tortoises used in the study were wild-caught in the area of St. George UT and confirmed positive for Mycoplasma infection by enzyme immunoassay. The turtles are listed as "threatened" species and because of their infection status, they could not be released and thus were maintained as a teaching/research colony. The tortoises were outfitted with cardiac access ports for blood sampling. Tortoises have increased activity with increasing daylight (Jan/Feb) with courtship behavior occurring in the spring (increased eating patterns and gastrointestinal transport time are seen during this period). A second smaller surge of courtship behavior is seen between Aug- Sep followed by a slower eating/GI transit period for the winter. 

In the first study the authors looked at the PK profile of CLARI following administration of a single oral dose of CLARI. A total of 6 tortoises were used and the study was done in mid-February. The tortoises were dosed once at 15 mg/kg and blood collected at 4, 8, 12 hours and then daily until day 14. This study resulted in consistent plasma levels in 4 of the 6 tortoises with a Cmax of 1.69 ug/mL and a Tmax of 6 hours. Quantifiable levels were seen for 48 h in 2 tortoises. One tortoise showed very low values at 12 h which rebounded at 24 h. Clarithromycin levels were below the threshold of detection by day 14.

The second study evaluated repeat dosing of CLARI in the same 6 tortoises and started immediately after the first study. The tortoises were dosed at 15 mg/kg orally every 3.5 days for 6 months. 

Blood samples were collected at 0.5, 2.5 and 3.5 months during the continuous dosing period. Results showed median CLARI plasma levels of 3.32, 3.93, 4.79 ug/mL at 0.5, 2.5 and 3.5 months, respectively. The active metabolite (14-OH) seen in humans was never detected. Levels of CLARI remained at 2 ug/mL or higher for all time points consistent with therapeutic values in other species. No adverse effects were observed during the treatment period and the animals showed complete remission of their clinical signs for up to 9 months after the treatment period.  The accumulation of CLARI in the plasma was not expected based on previous study results.

Because oral dosing of chelonians can be difficult and stressful for the animals, the researchers wanted to explore the possibility of per-rectum dosing as an alternative. The third study evaluated per-rectum dosing of CLARI in a different group of 7 tortoises. The dosing was done at 0,12,24,36 h and then once daily thereafter for 8 days. Blood was collected at dosing and 4, 8, 12, 24,36, 48,60, 72 h and at 14 and 15 days post-dosing. While per-rectum dosing resulted in CLARI absorption, plasma levels were low with only 1 animal exhibiting levels of 1 ug/mL. None of the tortoises had plasma levels in the target range of 2 ug/mL or higher and 2 of the tortoises had only one detectable CLARI value each.

Based on previous studies, the GI transit time of tortoises is 12 days but may be longer during the early portion of the year. Small intestinal transit typically takes around 3 days and the absorption profiles seen in this study are consistent with small intestine as the primary site of uptake. The authors theorize that either protracted large intestinal absorption or cystoenteric reflux contributed to values seen beyond the
3 day time point. The authors note that GI transit time is also impacted by diet and thus the plasma values could vary depending on the diet fed.

The authors conclude that long term dosing of CLARI is safe and effective in this species. Per-rectum administration would require higher doses that may not be feasible in a clinical setting. The accumulation of the CLARI over time was not predicted by the single dose study that was initially done and the authors caution readers to keep this in mind as much of the dosing done in exotic species is based on extrapolation from single dose studies.

QUESTIONS:
1. 	What is the genus and species of the desert tortoise?
2. 	Which of the following accurately describes the breeding cycle of desert tortoises?
a. 	One breeding cycle triggered by shortened light cycle (breeding in late fall/early winter) 
b. 	One breeding cycle triggered by lengthening light cycle (breeding in Jan/Feb)
c. 	Two breeding cycles, the first triggered by lengthening light cycle (spring) and the second at the end of summer (Aug-Sep.) 
d. 	Two breeding cycles, the first triggered by lengthening light cycle (spring) and the second in mid winter (Feb)
3. 	True or False: Gastrointestinal transit time for the desert tortoise varies with season 

ANSWERS:
1. 	Gopherus agassizii
2. 	c.
3. 	True; diet content can also impact the transit time

Experimental Techniques
Johnson et al.  A New Method for Respiratory Gating During Microcomputed Tomography of Lung in Mice, pp. 46-56

Task 3- Provide Research Support, Information, and Services; K1 

SUMMARY: The primary aim of this study was to determine whether the use of a controlled multiple-breath hold respiratory gating technique enhanced the quality of uCT images of the murine lung compared with those from an alternative, more conventional, respiratory gating method and a free-breathing method.  The secondary aim was to explore the biologic effects of anesthesia and handling associated with these imaging procedures.

4 scan techniques in 2 genetically altered mice were performed.  Mice with a 129/SV background were used, and the strain was developed by targeted insertion of a vector carrying k-ras alleles.  This allele causes them to typically present with microscopic lung tumors at 2 to 3wks of age.  CD-1 mice were in addition to the 129/SV mice were used in the secondary experiment to evaluate stress applied to mice during an imaging session.

It was found the small (diameter, less than 1 mm) lung adenomas were visualized most easily in the 2 breath-hold mode scans, somewhat blurred in delay-triggered scans, and difficult to detect in free-breathing scans.  A right lung lobe fissure that was present was best displayed in images obtained with cyclic breath-hold gating.  Normal lung tissue was most clear in the 720-view breath-hold gated scan, and less clear in other views.  The use of multiple-breath-holding gating technique enhanced the quality of the uCT lung images, particularly because of its abilities to increase the contrast between lung and soft tissues and to arrest respiratory motion during each X-ray exposure.  The delay-triggered scans, with pressure ventilation of 16 cm H2O, offered better lung contrast and better overall image quality than did free-breathing scans.  It was also determined that breath-hold gated scans best arrested respiratory motion during imaging and therefore best depicted organ boundary, followed by the free-breathing and delay-triggered scan techniques, respectively.  

Neither the initial temperatures nor heart rates after initial induction with isoflurane varied significantly from those during the free breathing or intubation session.  Immediately after injection of medetomidine, heart rate dropped significantly compared with that after initial induction with isoflurane.  It was also found that final heart rate was significantly lower in the breath–hold gated group than the free-breathing group.

So in summary, intubation and regulated ventilation of mice can dramatically increase the quality of CT imaging of the lung; in particular, visualization of lesions smaller than 1mm in diameter can be improved substantially.  

QUESTIONS:
1.	What is the leading cause of death in anesthetized mice?
2.	Why is Microcomputed tomography (uCT) particularly well suited to imaging the lung?
3.	What are the 2 distinct advantages offered by using respiratory gating when imaging mouse lungs? 

ANSWERS:
1.	Hypothermia, and it generally becomes a concern once the internal temperature drops below 25 degrees Celsius.    
2.	The X-ray attenuation of lung tissue is very different from that of typical soft tissue, this characteristic provides inherent contrast in uCT images.
3.	1) Inflating the lungs to a degree not normally achieved in free-breathing anesthetized mice enhances the contrast between the lungs and soft tissue on CT.   
	2) Controlling the breathing rates and synchronizing them with the individual exposures that make up the scan data minimizes the effects of increased motion from the larger inspiration volumes.

CASE REPORTS
Sorrell et al.  Secondary Acute Anterior Uveitis with Hyphema in a Purpose-bred Kitten, pp. 57-60

SUMMARY: A 7 wk old domestic shorthair kitten developed an acute case of anterior uveitis and hyphema. This case report describes the prescribed treatments and diagnostics used to determine the etiology.  The vendor health summaries indicated that the cat colonies were seronegative for FIV, FIP, FeLV, Calicivirus, Herpesvirus, Coronavirus, panleukopenia virus, syncytial virus, Toxoplasma gondii, and Chlamydia psittici.

The following diagnostic tests were performed: flurorescein dye test, complete blood count, chemistry, blood pressure, tonometry exam, serologic titers for Bartonella henselae, feline leukemia virus antigen, and feline immunodeficiency virus antibody. In addition an antibody test was done for feline immunodeficiency virus. All tests were within normal limits.

The kitten was treated with 1% prednisolone acetate ophthalmic suspension every 6h, 1% atropine sulfate every 12 hours, and meloxicam 0.25mg daily.  The atropine, meloxicam, and prednisolone was tapered off once the abnormalities were nearly resolved.

Causes of feline anterior uveitis can be divided into 3 main categories: primary (ocular), systemic (infectious and noninfectious), and idiopathic.  Causes of hyphema include congenital malformations, blunt or penetrating trauma, coagulopathies, systemic hypertension, neoplasia, uveal or retinal neovascularization, and iatrogenic causes.  After the diagnostic workup it was determined that this cat’s condition most likely was due to an impact o the head that resulted in a tear of the ciliary body with subsequent hyphema and miosis.

QUESTIONS:
1.	T/F  Anterior uveitis associated with systemic disease can be unilateral.
2.	T/F  Topical atropine sulfate produces cyclopegia and mydriasis.
3.	Clinical presentation of anterior uveitis is nonspecific and can include all of the following except:
a.	Blepharospasm
b.	Photophobia
c.	Third eyelid protrusion
d.	Epiphora
e.	Miosis
f.	Mydriasis

ANSWERS:
1.	T
2.	T
3.	f

Snider and Reichard.  Preputial Obstruction and Urine-induced Cellulitis Due to Nonnutritive Suckling in a Male White-tailed Deer (Odocoileus virginianus) Fawn, pp. 61-63

Tasks 1, 2 & 6
Species: tertiary

SUMMARY: White tailed-deer, Odocoileus virginianus,  not being a commonly seen laboratory animal species, are found in increasing fashion at different research institutions, usually for study of infectious disease.  In the past they have been used on studies looking at tick-borne disease, chronic wasting disease, and mycobacterial disease.  There is little literature available as to the husbandry requirements for white-tailed deer, especially in confined housing necessary for many research protocols.  

This article was a report of a case of distal preputial swelling resulting in obstruction a subsequent urethral rupture in a young male white-tailed deer fawn.  The fawn was in-house for a research study on Anaplasma phagocytophilum along with 8 other fawns.  The fawns arrived to the facility at 2-3 days of age and were housed indoors in pens, some individual and some in pairs.  The development of the featured fawn was normal up until 73 days of age.  He presented with severe depression and anorexia accompanied by intermittent bruxism and low-grade fever.  As the fawn was pair-housed, he was then isolated and managed conservatively with hand feeding a nursing care.  A little more than 24 hours later, diffuse swelling of ventral abdominal skin was noticed, and urine output was absent for a period of time as well.  Blood work revealed an elevated BUN, leukocytosis with marked neutrophilia and monocytosis, and elevated fibrinogen.  Interpretation of blood work in cervids can be challenging because of variations due to sex, age, time of year, geographic location, nutritional status, and method of restraint used during the collection.  Because of this challenge, control samples are often drawn for comparison from other deer in the same group.  The original tentative diagnosis was urethral rupture secondary to urolithiasis, as this has been seen in a documented case in the past although in this case, no signs or uroliths were found on urinalysis.

Due to this animal not being a viable candidate for study, euthanasia was elected.  On necropsy, the preputial obstruction and urine-induced cellulitis was discovered.  It was theorized that this was due to nonnutritive suckling by this animal’s pen mate, causing the trauma.  Nonnutritive suckling is commonly seen in captive fawns, and well documented in calves.  This occurs in the face of provision of adequate nutrition, and is thought to be an instinctive behavioral need and promotes physiologic well-being.  The nonnutritive suckling is normally not associated with injury or illness.  Due to the outcome of this report, this behavior may have to be actively managed through individual housing or creative control to avoid future issues with captive fawns.  

QUESTIONS: 
1.	On what types of laboratory animal studies are white-tailed deer most commonly found?
2. 	What is the best way to interpret cervid hematology?  What factors make it so variable?
3. 	What are two rule outs for a young fawn with anuria, preputial swelling, depression, and fever?
                                            
ANSWERS:
1. 	Infectious disease studies such as tick borne disease, chronic wasting disease, and mycobacterial disease
2. 	Compare to controls in the same group.  Variable factors include sex, age, time of year, geography, nutrition, and sample collection restraint.
3.  	Urolithiasis obstruction  and trauma due to nonnutritive suckling by a pen mate

Gozalo et al.  Visceral and Neural Larva Migrans in Rhesus Macaques, pp. 64-67

Task 1 - Prevent, Diagnose, Control, and Treat Disease; K3 Parasitology
Species:  Primary, Rhesus (Macaca mulatta) 

SUMMARY: This is a case report describing an outbreak of Baylisascariasis in otherwise clinically normal rhesus macaques.  21 captive-born, 3-5 year old rhesus were imported into a laboratory setting.  These were animals bred and raised in an outdoor breeding facility on an island in the southeastern US.  These animals underwent a 90 day quarantine period that included routine serologic, bacteriologic, and parasitic testing.  Special fecal testing was performed to rule out GIT parasites.  Campylobacter and Shigella spp. were cultured, but all other tests were normal.  Complete blood counts and clinical chemistries were within normal limits.  There was no evidence of clinical disease, especially parasitic disease.  These animals were used in an encephalitic viral study and were euthanatized routinely as per study design.  On necropsy, there were no gross lesions present.  However, on histological analysis, 13 of the 21 (62%) animals had large ascarid larvae within eosinophilic granulomas in mesenteric and pancreatic lymph nodes.  Of the 13 affected animals, 7 also had brain granulomas, with 4 of those having the same larvae as the lymph nodes.  The 3 animals with brain granulomas and no larvae seen are suspected to have been due to the angle or section of the tissue submitted.  Again, there were no clinical signs in these animals.

The parasite was identified as a species of Baylisascaris based on size and morphology.  Due to the island’s wildlife populations (and lack of skunks) in the area that these animals were raised, the parasite was assumed to be from the raccoon and considered B. procyonis.  This parasite has been known to infect humans and cause severe neurologic abnormalities.  In fact, more than 100 species of birds and mammals have developed fatal or severe CNS disease due to B. procyonis.  The parasite has a thick shell that allows it remain infective in the soil for years.

Unfortunately, by the time of diagnosis, neurologic damage has already occurred and treatment can be unrewarding.  The drug of choice in humans and animals for treatment is albendazole and corticosteroids.  Daily administration of pyrantel salts has been used as an effective preventative in endemic areas.  The low-level of exposure in the colony is consistent with low-levels of exposure in humans.  Most human cases have been in the Midwest, Northeast, and on the West Coast where the parasite ahs its highest prevalence in raccoons.

QUESTIONS:
1.  	T or F:  Baylisascariasis is not a common problem because the parasite is very host-specific.  
2.  	T or F:  Treatment of affected animals and humans is very successful with most patients having a full recovery.
3.  	T or F:  Multiple organs systems and areas of the body can be affected in the same affected host.
4.  	T or F.  Humans can acquire Baylisascaris infection.

ANSWERS:
1.  	F.  Baylisascaris affects many different species of birds and mammals, including humans.
2.  	F. Treatment is often unrewarding because the parasite has usually caused its damage prior to diagnosis.
3.  	T.  	Larval migrans can be present in the skin, muscle, retinas, and CNS.  It can be present in multiple locations in the same host.
4.  	T. 	Human infections are well documented in many different countries.

Gozalo et al.  Dyscoria Associated with Herpesvirus Infection in Owl Monkeys (Aotus nancymae), pp. 68-71

Domain 1: Management of Spontaneous and Experimentally Induced Diseases and Conditions 
T2: Diagnose disease or condition as appropriate 
Tertiary (other nonhuman primates): Owl monkeys 

SUMMARY: Dyscoria was noted in a female owl monkey and 2 of her offspring.  The third offspring was found dead with necrohemorrhagic encephalitis.  Two male monkeys paired with the female died, 1 of which showed oral ulcers at necropsy.  Histological examination of the oral ulcers revealed syncytia and eosinophilic intranuclear inclusion bodies in epithelial cells.  Ocular examination revealed posterior synechia associated with the dyscoria in all 3 animals.  Serum samples from the female and her offspring were positive for Herpesvirus simplex (HSV) antibodies by ELISA.  Because the clinical signs and lesion of owl monkeys with HSV are indistinguishable from those caused by Herpesvirus saimiri 1 infection and because ELISAs do not discriminate between Alpha herpesviruses, the cases described in the report might have been due to infection with H saimiri 1. 

QUESTIONS:
1.  	The gestation period of Aotus nancymae is approximately 
a. 	133 days 
b. 	150 days 
c. 	164 days 
d. 	164-186 days 
2.  	In the wild, Aotus spp live as 
a. 	Large multimale-multifemale troops numbering from 50 animals up to 300 
b. 	Male-dominated, multimale-multifemale groups ranging in size from 10 to 50 members
c. 	Multimale-multifemale groups with a less stringent hierarchical dominance ranking involving males 
d. 	Monogamous pairs within an extended family that may include 2 or 3 offspring 
3.  	Define iris coloboma 
4.  	All of the following are Alpha herpesviruseses EXCEPT 
a. 	Human herpesvirus 1 
b. 	Cercopithecine herpesvirus 1
c. 	Saimirine herpesvirus 2
d. 	Saimirine herpesvirus 1 
5.  	List two methods of discriminating between Alpha herpesviruses 

ANSWERS:
1. 	a 
2. 	d 
3. 	A full thickness defect in the iris 
4. 	c 
5. 	Immunohistochemistry or RCR tests on DNA extracted from formalin-fixed or paraffin-embedded tissues using primers specific to the polymerase gene region