Journal of the American Association for Laboratory Animal Science
Volume 46, Number 3, May 2007

ORIGINAL RESEARCH
Biology
Ruben et al.  Effects of Bedding Substrates on Microsomal Enzymes in Rabbit Liver, pp. 8-12

Task 3 - Provide Research Support, Information and Services (Biomethodology)
Primary Species: Rabbits

SUMMARY: Bedding substrates such as pine has been found to induce microsomal hepatic enzymes such as cytochrome P450 in rodents.  The purpose of this study is to determine whether cytochromes P450 2B and 3A are induced in rabbits housed on softwood bedding. There were three groups: negative control (no bedding), paper exposure group, pine exposure group, and two positive control groups which were housed without bedding (one group had rifampin and another had pentobarbital).  The Erythromycin N-demethylase assay was used on liver samples to determine the presence of CYP450 3A and the 7-Pentoxyresorufin-O-deethylation assay for CYP450 2B.  The enzymes levels did not reach statistical significance as compared to the positive control groups. 

QUESTIONS:
1.	The erythromycin N-demethylase assay is used to detect the presence of __________?
2.	The 7 Pentoxyresorufin-O-deethylation assay is used to detect the presence of __________?

ANSWERS:
1.	Cytochrome P450 3A
2.	Cytochrome P450 2B

Ferland et al.  Effect of Buprenorphine on Intracerebral Collagenase-induced Hematoma in Sprague-Dawley Rats, pp. 13-18

SUMMARY: The objective of this study was to evaluate the effects of buprenorphine on a collagenase-induced intracerebral hemorrhage in rats by evaluating blood biochemistry, motor behaviors, brain edema and histopathology. This hematoma model was developed by Rosenberg and colleagues in which collagenase is injected sterotaxically into the caudoputamen nucleus. This model reproduces many characteristics of cerebral hemorrhagic processes of traumatic brain injury seen in mammals and humans.  Serum biochemistry parameters showed no change after surgery in controls and buprenorphine-treated animals. At 48 h after collagenase injections, the performance of treated rats on the rotarod treadmill test was not significantly different from that of untreated rats, but the neurological exams of treated rats showed significantly improved performance. Although the volume of the hematoma was reduced with buprenorphine, the number of necrotic neurons in the penumbra was significantly increased. The data indicates administration of buprenorphine led to neurological and histopathological differences in a rat model of intracerebral hemorrhage, and data from such studies should be interpreted carefully if an opioid analgesic is used to minimize pain.

QUESTIONS
1.  	The intracerebral collagenase injection is a model for what?
2.  	In the current study, the use of buprenorphine post-op statistically affected which of the following compared to controls?
a.	Blood biochemistry parameters
b.	Brain water content
c.	Rotorod score
d.	Volume of hematoma

ANSWERS:
1.  	Cerebral hemorrhagic process of traumatic brain injury 
2.  	d

Cloud-Hansen et al.  Thirteen-lined Ground Squirrels (Spermophilus tridecemlineatus) Harbor Multiantibiotic-resistant Bacteria, pp. 17-20

Task 4: Develop and manage animal husbandry programs
Species: Tertiary

SUMMARY: Facilities that use wild caught species should be aware of potential health concerns and educate their staff accordingly.   This article cites an example of one facility which uses wild caught thirteen - line ground squirrels for hibernation studies and found tetracycline and chloramphenicol resistant Morganella morganii along with multi resistant Stentotrophomonas maltophilia as normal flora of the cecum.  Both of these bacteria have zoonotic potential with poorly understood modes of transfer.  

QUESTIONS:
1.	What research is Spermophilus tridecemlineatus used for? 
a. 	Sleep studies  
b. 	Behavioral studies 
c. 	Hibernation studies  
d. 	None
2. What is not true regarding Spermophilus tridecemlineatus? 
a. 	The genome has not been sequenced  
b. 	It is used for hibernation studies
c. 	Handling may pose a health risk  
d. 	The species is generally wild caught

ANSWERS: 
1.	c    
2.	a

Perumal et al.  Effects of Ketamine and Thiopentone Anesthesia on Serum Lipid Parameters in Adult Bonnet Monkeys (Macaca radiata), pp. 21-23

Task 3 - Provide research support, information and services 
SUMMARY: Serum cholesterol, triglycerides, very low-density lipoprotein (VLDL) and high-density lipoprotein (HDL) were measured by authoanalyzer in ten female bonnet monkeys aged 12-14 yrs while under two different anesthesia regimens after a baseline (pre-anesthesia) collection.. The first day, samples were collected while animals were sedated with  ketamine hydrochloride at 15 mg/kg IM. The next day, thiopentone sodium at 25 mg/kg was administered IV for sample collection. Purpose of the analysis was to establish baseline lipid parameters in bonnet macaques for future experiments assessing pathologic and toxicological changes in serum lipid parameters in adult bonnet monkeys.
Results of the sampling showed 7/10 animals with significantly decreased cholesterol and 8/10 had decreased HDL under ketamine. Under thiopentone, decreased serum triglycerides  and VLDL in 9/10 animals and increased HDL significantly in 9/10 animals.
QUESTIONS:
1.	Name the genus/species of the bonnet macaque. 
2.	T/F The two anesthetic regimens had a similar effect on HDL in the bonnet macaque. 
3.	T/F Bonnet macaques are readily available for research purposes but are not used because baseline serum chemistry and lipid parameters are not available.

ANSWERS:
1.	Macaca radiata 
2.	False, ketamine decreased HDL, thiopentone increased HDL. 
3. 	False - they have limited availability world-wide and published reports on hematological and biochemical values are limited.

Allison et al.  Assessment of Buprenorphine, Carprofen, and Their Combination for Postoperative Analgesia in Olive Baboons (Papio Anubis), pp. 24-31

Introduction: Humane and ethical treatment of laboratory animals requires that we recognize the signs of pain and adequately treat with appropriate analgesia.  There is a lack of published information outlining the assessment of post-operative pain in non-human primates.  This study outlined a method to assess signs of post-operative pain in Olive baboons.  Primarily, this study evaluated the efficacy of both a commonly used analgesic and one not typically used in NHP’s, as well as the combination of the two.  Buprenorphine is an opioid that is commonly used postoperatively in primates due to its long duration of action.  Carprofen is an NSAID that is a COX2 selective inhibitor which spares the homeostatic effects of COX1 prostaglandins.  Carprofen has a fairly long duration of action as well.  The combination of Buprenorphine and Carprofen can provide multimodal analgesia by controlling pain via 2 different pathways. 

Materials and Methods:  12 female olive baboons (Papio anubis) ranging in age from 8 to 18 years and weighing 14-20 kg were used in the study.  Housing and enrichment followed the recommendations of the Guide and the facility was approved by AAALAC.  Each animal served as its own control.  All baboons underwent surgery outlined in other IACUC approved protocols; no surgeries were conducted solely for the purposes of the analgesia study.  The following data points were collected during baseline and/or postoperative studies: radiotelemetry data (heart rate and body temperature), body weight, food consumption, serum cortisol levels (postoperatively only), urine cortisol levels (postoperatively only), behavioral observations (31 min time intervals noting locomotion, foraging, posture, and checking).

Data Collection:  Baseline analgesic studies were performed either before or 2 months after surgery. Baseline studies started on the first day of analgesic treatment (day 1) through day 6 of the study.  Animals received analgesic treatment days 1-3 and did not receive analgesic treatment days 4-6.  Animals were then placed in the following treatment groups: IM injection of buprenorphine 0.01 mg/kg (n=4), IM injection of carprofen 2.2 mg/kg (n=4), or a combined IM injection of 0.01 mg/kg buprenorphine and 2.2 mg/kg carprofen (n=4).  All analgesics were administered preoperatively, at 1600 postoperatively, and twice daily for 3 additional days after surgery.  Animals were also videotaped for 31 minute intervals at 3-4 hours after the morning analgesic treatment and at 1-2 hours prior to the afternoon analgesic treatment.  These time periods were used to ascertain whether any behavioral differences existed between the time of maximal and minimal analgesic benefit. 

Results:    Baseline and postoperative heart rates, body temperatures, and overall activity were increased during daylight hours. These are expected findings in diurnal animals.  It appeared that the heart rate increased during days 1-3 postoperatively for the buprenorphine treatment group.  1 animal in the Buprenorphine treatment group had an increased heart rate during the postoperative study.  This increased heart rate was likely indicative of pain in this animal and influenced the results of this treatment group.  The other 3 animals in this group had postoperative heart rates that were not significantly different from their baseline heart rates.  Postoperative temperature increased in all groups during days 1-3, most likely due to inflammation and healing of the surgical site.  Activity was decreased overall in the postoperative period.  Particularly, postoperative buprenorphine activity was decreased as compared with baseline buprenorphine activity. This is expected due to the lingering effects of anesthesia and surgery. Food consumption was decreased in all groups on day 1 compared to days 2-7.  Animals in all treatment groups maintained normal body condition. Urine cortisol demonstrated a significant increase on postoperative day 1. Serum cortisol data were not statistically significant.  Behavioral observations showed that standing was decreased in the postoperative period compared with the baseline observations. A decrease in standing on day-4 for the buprenorphine treatment group as compared with days 2 and 3 was seen.  The same animal that had an elevated heart rate in this group also had a significant decrease in standing behavior which may have skewed the standing data for the buprenorphine treatment group.  The data suggests that buprenorphine did not provide adequate analgesia in this particular animal. Decreased checking was seen in the postoperative period compared with baseline due to animals being less alert after anesthesia and surgery.  Findings suggested that 3 days of treatment were sufficient to control postoperative pain.

Conclusions: A BID dosing for 3-days with carprofen or buprenorphine + carprofen provided adequate and reliable analgesia for all baboons.  However, buprenorphine alone only provided adequate analgesia in 3 of 4 animals.  Therefore, these authors suggest that carprofen alone or buprenorphine + carprofen as more effective alternatives to buprenorphine alone for postoperative analgesia.

QUESTIONS:
1.	Which physiological and/or behavioral variables can be monitored when assessing pain in the Olive Baboon? 
A. 	Heart rate
B. 	Standing
C. 	Yawning
D. 	Temperature
E.  	A, B, and D  
2.	True/False.  Carprofen alone or in combination with buprenorphine is more effective for postoperative analgesia for baboons than buprenorphine alone? 
3.	Buprenorphine is a commonly used opioid analgesic in NHP’s because: 
A.	It is inexpensive 
B.	It has anti-inflammatory properties 
C.	It has a long duration of action (6-12 hr) compared to other opioids 
D.	A and B 

ANSWERS:
1. 	E
2. 	T
3. 	C

Morgan and Glowaski.  Teaching a New Method of Rabbit Intubation, pp. 32-36

SUMMARY: This paper compares training veterinary technology students on two different methods of intubating rabbits.   Two groups of students were trained to intubate rabbits with each technique.  These two groups of students were taught the same techniques in a different order.

Students were trained to intubate rabbits by the "blind method."  With this method, a rabbit was held by its head in sternal recombency.  An endotracheal tube was passed partially through the rabbit's oral cavity.  When breath sounds were heard and the rabbit inhaled, the endotracheal tube was passed to the lower respiratory tract, timing movement of the tube with the opening of the laryngeal inlet.

The second technique involved placing a rabbit in lateral recumbency with its feet towards the student and the nose pointed towards the dominant hand of the student.  The rabbit's head had to be maximally dorsoflexed (see Fig 1 article).  The student's subordinate hand lightly held the head in dorsoflexion on the table, while the student's dominant hand introduced the endotracheal tube behind the incisors.  The student was coached to look for condensation within the lumen of the endotracheal tube, indicating exhalation (see Fig 2 article).  The student slowly advanced the tube, looking for the tube to clear of condensate on inhalation (see Fig 3 article), then fog on exhalation.  The tube entered the trachea at the point of maximal inspiration.  Coughing was an indication of successful intubation.
 
Students were taught to check endotracheal tube placement regardless of which technique was used.  These findings indicate successful placement:  unassisted respiration, condensation upon a metallic object, and proper movement of the thorax.

Authors found regardless of which technique was trained first, mastering the second technique took less time than did the first technique.  All students preferred the novel lateral recombency technique.  This decision was based on ease of intubation in noisy environments and comfort of the rabbit.

QUESTIONS:
1.  	Which is not a significant complication of rabbit intubation?
a.	Laryngeal trauma
b.	Tracheal rupture
c.	Vagal stimulation
d.	Breath holding
e.	Death
2.	Which are clinical signs of stage 3 anesthesia?
a.	Lack of muscle and jaw tone
b.	Loss of gag and pedal reflexes
c.	Decreased respiratory and heart rate
d.	Presence of palpebral reflexes
e.	a, b, and c above
f.	a, b, c, and d above
3.	Which are evidence of proper endotracheal tube positioning?
a.	Coughing on placement of tube
b.	Unassisted respiration
c.	Condensation on a metallic object
d.	Proper movement of thorax
e.	 All of the above
4. 	According to the article, which intubation technique was preferred by students?

ANSWERS:
1. 	d
2.	e
3.	e
4.	Novel lateral recumbency technique

Barbaric et al.  A New Coat Color Mouse Line for Testing Germline Transmission of Embryonic Stem Cells while Retaining an Inbred Genetic Background, pp. 37-40

SUMMARY: Most gene-targeted mice are produced on a mixed genetic background of C57BL/6 and substrains of 129/Sv. Mating chimeric mice containing 129/Sv-derived embryonic stem cells that are wild type at the agouti locus (A) in a nonagouti (a) donor genetic background with inbred C57BL/6 mice that are homozygous for the nonagouti allele allows the use of coat color to detect germline transmission. Agouti pups from such a cross indicate germline transmission of embryonic stem cell-derived genetic material. However, 129/Sv substrains and C57BL/6 are genetically and phenotypically quite different and, consequently, differing genetic contributions of the 2 backgrounds may influence the phenotype under investigation. To avoid this problem yet maintain the usefulness of the coat color system in detecting germline transmission, authors generated a new strain of mouse by selectively introducing the nonagouti locus into a 129/Sv inbred background. This mouse strain contains 129/Svderived genetic material almost entirely except for a small region surrounding the nonagouti allele. Germline transmission can be detected in the usual manner, but the agouti offspring will be almost identical to 129/Sv inbred mice. Thus, the system allows the generation of gene-targeted mutations on a 129/Sv genetic background.

QUESTIONS:
1.	What is the main reason to produce gene target mice on a mixed genetic background 
a)	To detect germline transmission by coat color
b)	To introduce heterosis
c)	To avoid dead of litters
d)	To increase reproductive ability
2.	What does type of crossing is recommended for introduction of autosomal genetic material 
a)	Speed congenic
b)	Standard congenic backcross
c)	Standard breeding brother x sister
d)	Outbreeding techniques

ANSWERS:
1. 	a)
2. 	b)

Experimental Technique
Aoto et al.  Production of Progeny Mice by Intracytoplasmic Sperm Injection of Repeatedly Frozen and Thawed Spermatozoa Experimental Technique, pp.41-46

Task 3 - Provide Research Support, Information, and Services
Primary Species: Mus musculus

SUMMARY: This paper was on the experimental technique of intracytoplasmic sperm injection of repeatedly frozen and thawed spermatozoa to successfully produce viable progeny with laboratory mice.  Sperm cryopreservation and assisted-reproduction techniques, such as intracytoplasmic sperm injection (ICSI) have been shown to be successful in other studies, however the number of possible progeny is limited due to the finite number of cryopreserved sperm preparations.  Frozen spermatozoa is more economical and efficient than using frozen embryos for IVF or ICSI due to the fact that more spermatozoa are produced than embryos or oocytes.  The ability to refreeze and reuse sperm preparations would extend the utility of each cryopreserved preparation.  The study used mouse sperm subjected to repeat freezing and thawing in TYH medium already commonly used for in-vitro fertilization techniques.  ICSI is more successful than IVF when using repeatedly frozen and thawed sperm because it can solve infertility issues due to loss of mobility of defective sperm.  Three inbred strains of laboratory mice (C57BL/6J, BALB/cA, and C3H/HeN) were reproduced by ICSI after reuse from spermatozoa frozen and thawed 10 times.  The reuse of cryopreserved spermatozoa can extend the opportunities for reproduction of progeny from cryopreserved sperm and can increase the utility of cryopreserved preparations as bioresources.

QUESTIONS:
1.  	What does ICSI stand for?
2.	Why is ICSI more successful than IVF for repeatedly frozen and thawed spermatozoa?
3.  	Why is using frozen spermatozoa more efficient than frozen embryos or oocytes?

ANSWERS:
1.  	Intracytoplasmic Sperm Injection
2.  	Because it can solve the infertility issues created by repeat freeze/thaw cycles on the mobility of sperm.
3.  	Because more sperm are generated per mouse and can be successfully refroze and thawed to extend the preserved sperm samples for many more breedings.

Stender et al.   Establishment of Blood Analyte Intervals for Laboratory Mice and Rats by Use of a Portable Clinical Analyzer, pp. 47-52

Task : 1- Prevent, Diagnose, Control and Treat Disease (Diagnostic tests) and Task 3- Provide Research Support, Information and Services (Biomethodology)
Species: Primary- mice and rats

SUMMARY: Portable clinical analyzers are used in human medicine and veterinary medicine. These instruments are popular due to their automatic calibration and quality control, need for low or no maintenance, convenience and versatility. Small sample volumes of approximately 3 drops of whole blood can yield multiple test results in 2 minutes. Typically the blood is collected in lithium heparin anticoagulant. Many blood parameters may be measured, including electrolytes, glucose, clotting times, acid base status and blood gases.

This paper hypothesized that portable clinical analyzers would be a useful tool for antemortem clinical and phenotypic assessments of individual laboratory rodents. They established normal ranges for 7 chemistry, hematological and electrolyte markers in clinically normal lab mice and rats, and compared the portable analyzer's values with published range intervals available for these species. After establishing these ranges, they used the analyzer to confirm alterations in specific disease markers of experimental rodent models.

Strains of mice sampled for the study included BALB/c (44 mice), C3H (49 mice), and 129 mice (38 mice), of varying ages and both sexes. Blood samples were harvested via submandibular vein prior to euthanasia by CO2 inhalation. Male mice on mixed genetic background enrolled in a type I diabetes mellitus study were evaluated for elevated blood glucose levels. Diabetes had been induced with streptozotocin.

Rats sampled were Sprague-Dawley (50 rats) and NIH (26 rats), of varying ages and both sexes. F344-Tg(DTR)C354 rats transgenic for human diphtheria-toxin receptor, were inoculated with diphtheria toxin, to induce glomerulosclerosis in adult rats. Blood was evaluated for increased BUN. Blood was collected from anaesthetized rats were blood through the lateral saphenous vein, and from the glomerulosclerosis rats under anesthesia. Anticoagulated blood was drawn into sterile disposable pipettes and expressed into wells of the cartridges in accordance with manufacturer instructions.

The portable clinical analyzer used in mice or rats gave comparable values to those previously published using other analytical methods. Comparison of values that were at least 20% above or below the computed bounds from all mice or all rats revealed no significant differences among mice and rats of varying genetic backgrounds. However, differences may represent possible phenotypic differences due to experimental or genetic background variables. This study showed a broader range in all parameters measured in mice, but also had a greater number of animals sampled. The most discordant result within their study was the greater range of normal glucose in mice. This could have been related to the intracardiac method of blood collection after CO2 inhalation.

The authors had minimal problems getting test results from the portable clinical analyzer and conclude that these devices have multiple applications in laboratory animal medicine.

QUESTIONS: 
1. 	What is a possible reason that this study had wider “reference ranges” than other studies using portable clinical analyzers?
2. 	Which clinical chemistry could be altered by the method in which blood is collected?
3. 	What are 2 possible reasons behind variations in clinical chemistry and electrolyte levels in control animals’ values?

ANSWERS: 
1. 	Greater sample size/ greater number of animals sampled in this study
2. 	Glucose, from possible stress responses
3. 	Stress from handling, underlying subclinical abnormalities

Hopper and Cray.  Evaluation of a Portable Clinical Analyzer in Cynomolgus Macaques (Macaca fasicularis), pp. 53-57

SUMMARY: The authors compared values obtained from a portable clinical analyzer (i-STAT, Abbott Laboratories, Abbott Park, IL.) to their in-house institutional laboratory and published values for commonly performed tests on the blood of healthy Cynomolgus macaques. The tests chosen for comparison were glucose, urea (BUN), sodium, potassium, chloride, total carbon dioxide (TCO2), hematocrit (HCT) and hemoglobin (HGB). As has been reported in similar studies for other animals and in people, there is potential for significant disagreement between the portable analyzer and in-house institutional laboratory. Most of the differences can be explained by physiology of blood, serum and cells or variation caused by elapsed time, sample collection and handling. The portable analyzer can be a useful and valuable tool for immediate monitoring, but the clinician must be aware of the potential for differences in the values reported between the two methods. Furthermore, normal ranges should be developed for each new species of interest.
 
QUESTIONS:
1.	What is the genus and species for the Cynomolgus macaque? 
2.	Why would you expect glucose values obtained from a portable analyzer to be higher when compared to values obtained from an institutional laboratory? 
3.	Why would you expect electrolyte values obtained from a portable analyzer to be lower when compared to values obtained from an institutional laboratory?
 
ANSWERS:
1.	Macaca fasicularis 
2.	Continued metabolization of glucose by blood cells in the elapsed time between sample collection and processing. 
3.	Leakage of electrolytes from cells during elapsed time and clotting centrifugation prior to analysis in an institutional laboratory.

Gamble et al.  Use of a Low-concentration Heparin Solution to Extend the Life of Central Venous Catheters in African Green Monkeys (Chlorocebus aethiops), pp. 58-60

Task 3- Provide Research Support, Information and Services
Primary Species- primate/African Green Monkey

SUMMARY: The purpose of this article is to evaluate the use of a low concentration heparin solution to increase the mean patency time of a dual-lumen central venous catheter, used in combination with a jacket and tether, for repeated blood sampling and intravenous fluid administration without the use of anesthesia and without inducing capture-related stress.  Compared to saline flush, low concentration heparin solution doubled the patency period of the catheter.

QUESTIONS:
1.	Hematology may be affected by 
a.	Ketamine anesthesia 
b.	Stress 
c.	Repeated sampling 
d.	All of the above 
2.	Ketamine and/or capture/physical restraint may alter
a.	Body temperature 
b.	Activity 
c.	Appetite 
d.	All of the above 
3.	Bupivicain hydrochloride may be used to
a.	Give perioperative /preemptive analgesia along an incision site 
b.	Minimize vasoconstriction associated with vein manipulation 
c.	a and b 
4.	Additional modification that may extend patency period include
a.	Flush catheter daily rather than every other day 
b.	Increase the concentration of the heparin solution 
c.	Increase the infusion flow rate 
d.	All of the above 
5.	Heparin may be inappropriate if measuring
a.	Hemogram 
b.	Coagulation parameters 
c.	Both 
d.	Neither 

ANSWERS:
1.	d
2.	d
3.	c
4.	d
5.	b

Sikoski et al.  Comparison of Heating Devices for Maintaining Body Temperature n Anesthetized Laboratory Rabbits (Oryctolagus cuniculus), pp. 61-63

Task 2: Prevent, Alleviate, and Minimize Pain and Distress
Species Designation: Primary

SUMMARY: Monitoring and maintaining a patient’s normal body temperature throughout an anesthetic procedure is widely accepted as standard veterinary practice.  External devices are effective in preventing surgical hypothermia.  The more common and generally accepted methods of maintaining body temperature during anesthesia are circulating warm-water blankets and forced-air warming devices.  Previous studies have been performed regarding these methods in dogs and cats, but this is the first to analyze these methods in rabbits.

15 male NZW rabbits were housed routinely and divided in to 3 groups (water blanket, forced-air warmer, and no supplemental warming).  All 15 rabbits were anesthetized with ketamine and xylazine and then received a tibial nerve resection.  The rabbit’s temperature was recorded in 5 minute intervals for 45 minutes via an esophageal probe.

The authors found that in the rabbit, the warm-water blanket was the most effective heating device, causing a slight elevation in mean body temperature by the end of the recording period.  The forced-air warming device also was effective in maintaining a normothermic condition, with only a non-significant -0.2°C drop from the baseline body temperature.  Lack of a heating device resulted in a significant -1.1°C decrease in body temperature across the 45 minute recording period.  The authors conclude that both warm-water blanket and forced-air warmer effectively maintained normal body temperature during anesthesia, whereas no heating device significantly contributed to anesthetic hypothermia.

QUESTIONS:
1.	T/F Body temperature during and after anesthesia is an important parameter to monitor in rabbits.
2.	T/F Hot water blankets are the only accepted method of external temperature control for laboratory animals.
3.	T/F If a body cavity is not entered during anesthesia, body temperature will remain within normal limits.
4.	T/F Electric blankets are the preferred method for maintaining proper temperature and have very minimal risks associated with their use.

ANSWERS:
1.	True 
2.	False 
3.	False 
4.	False 

CASE REPORTS

Green et al.  Ovarian Hyperstimulation Syndrome in Gonadotrophin-treated Laboratory South African Clawed Frogs (Xenopus laevis), pp. 64-67

Task – 1: Prevent, diagnose, control and treat disease
Species: Xenopus frogs

SUMMARY: Ovarian hyperstimulation syndrome (OHS) is a rare but possible side effect of Xenopus frogs treated with gonadotropin to maximize oocyte harvest. Signs of this syndrome in frogs include subcutaneous accumulation of fluid, coelomic distension, edema and inability to dive. On necropsy, degenerative oocytes were found free-floating within the coelomic cavity of both frogs. Three cases of OHS have been observed at this institution over the past six years. OHS is rare in human women and is associated with young age, good ovarian reserve, low body weight and polycystic ovarian syndrome. Clinical signs are similar to those seen in these frogs.

QUESTIONS:
1.	Which of the following is not a primary differential for OHS in Xenopus frogs?
a.	Renal failure
b.	Primary bacterial sepsis
c.	Secondary or opportunistic bacterial sepsis
d.	Heteropolaria infection
2.	Which of the following is used to treat OHS in human women?
e.	IV fluids
f.	Albumin
g.	Electrolytes
h.	All of the above

ANSWERS 
1.	D. Heteropolaria is usually a superficial infection of the skin and doesn’t usually present with systemic signs.
2.	D. Treatment in humans is supportive only

Phaneuf et al.  Porcine Dermatitis and Nephropathy Syndrome Associated with Porcine Circovirus 2 Infection in a Yorkshire Pig, pp. 68-72

ACLAM Task 1:  Prevent, Diagnose, Control, & Treat Disease
Primary Species - Pig

This was a case report of a 14 week old, 60 kg, female Yorkshire pig.  It arrived at the facility clinical ill with multifocal to coalescing raised, red, non-puritic, papular to crusted skin lesions as large as 5 cm in diameter on the rump, vulva, perineum, things, and lower handles.  Its temperature as 38.1C and it was depressed and reluctant to move.  Differential diagnoses included erysipelas, Actinobacillus suis pneumonia and septicemia, exudative dermatitis (Staphylococcus hyicus), swinepox, salmonellosis, hog cholera (Classical Swine Fever), and African Swine Fever.

The animal was euthanized.  Clinical chemistry results indicated that it was azotemic with markedly increased blood urea nitrogen (BUN) and creatinine levels.  Phosphorus was elevated and potassium was decreased.  The urinalysis results showed a moderate hematuria and proteinuria, urate crystals, and occasional leukocytes.  Hematology results demonstrated a mildly decreased RBC count, decreased hemoglobin and hematocrit, mildly decreased MCV and MCH, and a mild lymphopenia.

On gross necropsy, the kidneys were bilaterally pale and enlarged.  The lungs were mildly consolidated with multifocal, patchy, red areas.  Histopathology of the skin revealed a severe, multifocal, nonsuppurative, necrotizing, and ulcerative dermatitis and panniculitis.  There was a severe multifocal, nonsuppurative, necrotizing glomerulonephritis and interstitial nephritis.  There was also a severe, multifocal to coalescing, nonsuppurative interstitial pneumonia as well as a moderate, multifocal, interlobular hepatitis.  There was a strong + reaction to PCV2 on immunohistochemistry.

Porcine Dermatitis and Nephropathy Syndrome (PDNS) is an important emerging syndrome in North American affecting 11 to 14 week old pigs.  While the example mechanism of action is unknown, it is thought that the clinical signs are a result of a probably immune-mediated vasculitis.  Animals with this syndrome are usually euthanized.  The virus is very hardy in the environment and can be spread by fomites, clothes, and equipment.  Chlorine-releasing disinfectants, quaternary ammonia compounds, and iodophors are thought to be efficacious at killing the virus.

QUESTIONS:
1.	Porcine Dermatitis and Nephropathy Syndrome (PDNS) typically affects pigs in the following age range:
a.	2 to 6 weeks
b.	8 to 10 weeks
c.	11 to 14 weeks
d.	8 to 12 months
2.	What is the virus associated with PDNS?
a.	A picornovirus
b.	Porcine Circovirus 2
c.	Parvovirus
d.	Swine pox
3.	Hog cholera is also known as:
a.	African Swine Fever
b.	Classical Swine Fever
c.	Atrophic Rhinitis
d.	PDNS
4.	What bacterium has sometimes been associated with PDNS?
a.	Pasteurella multocida
b.	Bordetella bronchiseptica
c.	Staphylococcus hyicus
d.	Actinobacillus suis
5.	What is the causative agent of exudative dermatitis in swine?
a.	Pasteurella multocida
b.	Bordetella bronchiseptica
c.	Staphylococcus hyicus
d.	Actinobacillus suis

ANSWERS:
1.	C
2.	B
3.	B
4.	A
5.	C