Laboratory Animal Science 48 (4)

Laboratory Animal Science 48 (4)
1998

1998 ACLAM Forum: ÒBioethics and the Use of Laboratory Animals: Ethics in Theory and Practice. Laboratory Animal Science 48 (4): 323.
Co-chair Dr A Lanny Kraus opened the forum and he stated that the aim of the forum was to bring together morally serious and credible proponents and opponents of animal based research for an open examination for the issue. Dr Beauchamp, who traced the philosophical and ethical underpinnings of the debate, examined four principles that govern the use of humans in research, including respect for autonomy, non maleficience, beneficience and justice. Dr Andrew Rowan discussed ethical principles for use of animals research.
Religious beliefs about the status of animals and their use in research were reviewed by Dr James V. Parker (Judeo-Christian) and Dr Chris Chapple (eastern religions).
Dr Peter Singer asserted that utilitarianism is the appropriated basis for evaluation of the appropriateness of animal use in research. Dr Jerry Tannenbaum discussed the legal implications of rights and challenged the notion that if people assume a duty or obligation. Dr David DeGrazia rejected the utilitarianism versus rights debate and argued for use of moral pluralism. Dr Roberts Veatch discussed the ethical controversies surrounding end-of-life decisions. Dr Bernard Rollin and Dr Jon Gordon were discussing issues associated with genetic engineering research. Dr Rowan reviewed public attitudes about the use of animals. Dr Tom Hamm implores people to open animal research facilities to tours and to volunter to help train school teachers to understand the necessity of using animals in research.
All the summaries will be published by ACLAM.
No questions

Pelvic Limb Paresis In A Safari Cat. Laboratory Animal Science 48 (4): 325.
The Safari cat is an animal model of Glucose 6 phosphate dehydrogenase (G6PD) cellular mosaicism which is used to study of hematologic neoplasms. This was a Clinical Problem Solving Exercise article which is new to LAS. It involved a 14yo intact, female,Safari cat that presented with right pelvic limb weakness which became progressively worse requiring euthanasia. Lab results: FeLV, FIV, FIP. TOXO, all negative. CBC/Chem panel unremarkable. X-rays: some ankylosing spondylitis in the thoracic and lumbar spine. Microscopic lesions in the Rt. femoral nerve revealed a homogenous lymphoid infiltrate (CD3 expressing) T cells. Lymphosarcoma is the most common hematopoietic neoplasm in the cat and is usually associated with FeLV. Lymphomatous infiltration of peripheral nerves has been described in at least two other species. Chickens with Marek's disease and humans with T cell lymphomas.
QUESTIONS:
1. The Safari cat is a model for this disease:
     A. Hypertrophic cardiomyopathy
     B. Glucose-6-phosphate dehydrogenase cellular mosaicism
     C. Spontaneous hyperthyroidism
     D. None of the above
2. Glucose-6-phosphate dehydrogenase cellular mosaicism is used to study
the clonal nature of hematologic
     neoplasms.     T of F
3. Safari cats result from crossing DSH (felis catus) and a wild Geoffroy
cat (Leopardus geoffroyi). T or F
QUESTIONS:
1.- Differential diagnosis with pelvic limb weakness
2.- T/F.It is rare that only the peripheral nerves ( peripheral
neuropathy) are involved in the lymphsarcoma
ANSWERS:    1. B        2. T       3. T
ANSWERS:
1.- Disordes of spinal cord and trauma, diabetes mellitus, hyperthyroidism
2.- F

Shigella Infection in Macaque Colonies: Case Report of an Eradication and Control Program. Laboratory Animal Science 48 (4): 330.
This article/case report discusses the improved management and diagnostic procedures adopted by this facility to eradicate and control Shigella in their colony. 12% of their imported monkeys entering their facility tested positive for Shigella.
Improved procedures include:
     1. More frequent microbiological monitoring of incoming animals, animals with diarrhea, and following stressful procedures (weaning, post surgical, and following transport to another area).
     2. Improved sanitation practices. They altered the cages so that the monkeys no longer had access to drainage from other cages.
     3. Improved quarantine procedures where all monkeys were treated if one tested positive. An all in, all out policy of management was adopted.
     4. Treatment consisted of Amoxicillin 15 mg/kg q. 48 hours x three treatments. Trimethoprim/Sulfonamide, tetracycline and enrofloxacin have also been reported to be effective.
Fun facts about Shigella:
     Shigella is an important occupational zoonosis. Shigella infection can be eliminated by the use of antibiotics while Campylobacter spp. were difficult to eradicate. Shigella flexneri is the most common species encountered in primate colonies. Clinically, may see acute colitis with the passage of liquid stools containing blood, mucous, and necrotic colonic mucosa. Histpathologically, may see colonic mucosal herniation.
Questions
1. What portion of the GI tract does Shigella typically infect?
     a. Stomach
     b. Duodenum
     c. Jejunum
     d. Ileum
     e. Colon
     2. T/F Shigella is a significant zoonotic disease that has been frequently transmitted from nonhuman primates to humans.
Answers
1. e Colon
2. True

Enterohepatic Lesions in SCID Mice Infected with Helicobacter bilis. Laboratory Animal Science 48 (4): 334.
Helicobacter hepaticus and H. bilis have been associated with disease and are the most prevalent species in laboratory animals. H. hepaticus has been associated with rectal prolapse, chronic proliferative typhlocolitis and proctitis in immunodeficient mice. In immunocompotent animals, H. hepaticus has been found in the liver and intestine, also. C.B.-17 scid/scid was found to develop inflammatory bowel disease when inoculated with H. hepaticus. H. bilis was associated with mild hepatitis in aged mice, hepatoma in aged CBA/CA mice, and inflammatory bowel disease in defined flora ICR-scid/scid mice inoculated intraperitoneally with the organisms. In this paper, H. bilis also caused chronic active hepatitis and proliferative typhlitis in conventional flora C.B.-17 scid/scid mice by gastric inoculation of the organisms.
In this paper, some mice with histopatholgic hepatic lesions were negative by PCR of liver specimens. Mice with proliferative chronic inflammatory bowel disease of the cecum also have mild chronic mucosal epithelium inflammation in the proximal colon; however, the disease was subclinical. In this study, hepatic lesions were more prevalent and more severe in male mice. Additionally, mild hyperplastic lesions were observed at the ileocecal junction in some Helicobacter-free C.B.-17 scid/scid mice.
Questions:
1. In this paper, what sex has more severe and prevalent hepatic lesions:
(a) female; (b) male; (c) both
2. It has been documented that PCR can detect as few as ____ Helicobacter organisms.
(a) 5; (b) 10; (c) 100; (d) 20
3. What two Helicobacter species are most prevalent in laboratory animals?
(a) H. muridarum & H. rodentium (b) H. hepaticus & H. bilis (c) H. cinaedi & H. Cholecystus (d) H. trogontum & Flexispira rappini
Answers: b; a; b.

Fatal Tramatodiasis in Research Turtles. Laboratory Animal Science 48 (4): 340.
During a 5 year period, 16 freshwater turtles (Trachemys scripta elegans and Chrysemys picta) were purchased for research purposes and died spontaneously.
    Clinical Signs: Consisted of lethargy, constant swimming sideways, hemiplegia, and necrotic lesions on the carapace.
   Gross Necropsy Findings: The abdominal cavity in some turtles contained clear to red-tinged fluid; the liver had small areas of necrosis, and fibrin adhered to the capsule; and nematode parasites were observed in a majority of the gastrointestinal tract. Other lesions observed included cardiac necrosis, pneumonia, hepatic lipidosis, and pancreatic and spleenic necrosis.
    Histologic Findings: Fluke eggs were found in most organs and were associated with small blood vessels which had vilus-shaped intimal proliferation in the aorta and mesenteric arteries. There were large areas of necrosis and hemorrhage in the pancreas, spleen, liver and lung. A granulomatous response was also observed in the submucosa of the small intestine. Fluke eggs were seen in vessels of the brain.
    Eggs are deposited as the adults migrate through the tissues and deposit eggs that eventually migrate. The migration of the adults and the eggs elicit an acute inflammatory response. The pathogenicity of the fluke Spirochis parva is related to the secondary infections caused by widespread egg deposition and blockage of small vessels in the small intestine causing necrosis and a bacteremia. The diagnosis of this fluke is made from egg identification since adults are difficult to find. The eggs can be found by compressing lung tissue between a glass slide and coverslip or by fecal examination. This parasite requires a snail as an intermediate host to complete the lifecycle.
    Treatment: Praziquantel, 8 mg/kg orally is a safe anthelminthic to use in turtles. However, this drug crosses the blood brain barrier and the remaining research turtles in this case were not treated since this drug could have an effect on the tissue used in the research.
Zoonosis: Turtles carry Salmonella and Arizona and should be isolated from other laboratory animals. Handlers should take protective measures to prevent infection.
Questions:
1.      What was the genus and species of fluke that produced the trematodiasis?
2.      What was the cause of the pathogenicity?
3.      What drug could be used to treat this fluke?
Answers:
1.      Spirorchis parva
2.      Secondary infections are caused during widespread egg deposition and blockage of small vessels in the small intestine causing necrosis and a bacteremia.
3.      Praziquantel (8 mg/kg of body weight)

Lymphoproliferative Disease Induced by Murine Herpesvirus-68. Laboratory Animal Science 48 (4): 344.
A novel herpesvirus, designated Mouse Herpesvirus-68 (MHV-68), was isolated from the brains of bank voles in Slovakia. The virus is considered a gamma-2 herpesvirus due to its close homology. Following experimental intranasal infection of mice, MHV-68 replicates in the lung for about 7 days and then is rapidly cleared. A subsequent latent viral infection can be found predominantly in splenic B lymphocytes. After 18-24 months, about 10% of the animals will develop lymphomas of several tissues, but predominantly of the spleen and mesenteric lymph nodes. If treated with cyclosporin A, the number of mice developing lymphomas increases to about 60%.
As the virus induces lymphomas of B-cell origin and its incidence increases after immunosuppression, MHV-68 is proposed as a model for Epstein-Barr virus-mediated lymphoproliferative disease. Epstein-Barr virus in humans is associated with nasopharyngeal carcinoma, Burkett's lymphoma, Hodgkin's disease and post-transplantation lymphoproliferative disease.
Questions:
1. MHV-68 latent infections are found predominantly in splenic ______ (cell type).
2. After 18-24 months, lymphomas develop in 2 main tissues. Name them.
3. Name a Herpes virus that induces Burkett's Lymphoma and Malignant Lymphoma in the cotton topped marmoset.
Answers:
1. B lymphocytes
2. Spleen and mesenteric lymph nodes
3. Herpesvirus ateles, Herpesvirus saimiri

Comparison of Sensitization to Crude and Purified House Dust Mite Allergens in Inbred Mice. Laboratory Animal Science 48 (4): 346.
The purpose of this study was to develop a murine model of house dust mite allergy, a common cause of bronchial asthma and allergic rhinitis in humans.
    All strains of mice tested responded to crude mite extract, but there were different sensitivities to purified mite allergens. This may be because crude mite extract contains many antigens which cause non-allergic inflammation by activating complement or platelet activating factor.
   The availability of many strains of mice with different sensitivity to mite antigens and a detailed gene map of mice allows investigation of the genetic basis of house mite allergy.
    A/J mice were high responders; BALB/c were low responders
QUESTION:
What killed bacteria is used to induce IgE synthesis and organ sensitivity to foreign antigens?
ANSWER
Bordetella pertussis

Model of Emotional Stress in Rats. Laboratory Animal Science 48 (4): 353.
The release of arginine vasopressin (AVP) is increased by physiologic stimuli but is suppressed by emotional stress. AVP is thought to induce the release of adrenocorticotropin hormone (ACTH). Low frequency foot shock has historically been used to suppress AVP release but this technique makes blood collection difficult during the procedure. The authors devised a model where rats could observe other rats fighting with each other and blood samples from the spectator rats were collected via a swivel jugular catheter. The purpose of the study was to determine if observation of intraspecies aggression (emotional stress) can serve as a model for suppression of release of AVP. The authors conclude that the noxious stimuli of nearby fighting was followed by suppression of AVP release as a result of the emotional stress.
     In order to induce aggressive behavior in the fighting rats, they were given 6-hydroxydopamine (6-OHDA) in the lateral ventricles of the brain and a pressure "clip" was applied to the tail. In addition to hematological parameters, spectator rats were observed for excessive grooming, face washing, scratching, and "freezing up" as signs of emotional stress.
     Reviewers note-there was a lengthy discussion on Compmed about this article and the use of aggression between the rats to induce emotional stress.
Questions
1. What procedures were done to increase intraspecies aggression?
2. Release of arginine vasopressin (AVP) is thought to _____________emotional stimuli and _____________during physiologic stimuli.
        a. increase, decrease
        b. increase, increase
        c. decrease, increase
        d. decrease, decrease
3. Name another method to induce emotional stress.
Answers
1. Inject 6-OHDA in the lateral ventricles and apply a tail clip will induce aggression.
2. c
3. Low frequency foot shock.

Hemodynamic Response to Anesthesia in Pregnant and Nonpregnant ICR Mice. Laboratory Animal Science 48 (4): 357.
In order to study murine cardiovascular development and hemodynamics in the mouse embryo, the authors determined the relationships between minute ventilation and arterial blood gas content, changes in mean arterial blood pressure (BP) and heart rate (HR) in pregnant and nonpregnant mice to determine "adequate ventilation" during short-term anesthesia. Anesthetic protocols compared the hemodynamic effects of a single dose of pentobarbital (80 mg/kg, i.p.) or repeated doses of ketamine and xylazine ((150:12.5 mg/kg; 90:7.5 mg/kg). Righting reflex and recumbent time were evaluated.
Normalized minute ventilation for mechanicel ventilation/tracheotomy was 3.5 ml/min-1/g-1 for nonpregnant and 3.0 for pregnant mice. Mean BP and HR was higher in pregnant than in nonpregnant for all protocol. Pentobarbital induced higher tachycardia and hypotension than the other protocols. Ketamine/Xylazine induced small changes in HR and BP but provide adequate anesthesia for superficial surgical procedures in pregnant and nonpregnant.
Questions
1) In order to preserve uteroplacental flow in pregnant mice, what is the best anesthesia protocol?
a) Pentobarbital
b) Combination of Ketamine/Xylazine
c) Both a & b
d) None of the above
2) T/F. The decrease in BP and HR after injection of Ket/Xylazine is likely because of its central alpha-adrenergic receptor action.
Answers
1) b
2) True

Glucocorticoid-Resistant B-Lymphoblast Cell Line Dervied from the Bolivian Squirrel Monkey (Saimiri boliviensis boliviensis). Laboratory Animal Science 48 (4): 364.
The goal of the study was to develop a continuous cell line from squirrel monkeys that expresses the species specific phenotype of impaired sensitivity to glucocorticoids. The cell line would then serve as a source of unlimited supply of donor DNA, mRNA, and proteins so that actual squirrel monkeys won't have to be used. Only 1 other cell line is available from squirrel monkeys, lung fibroblast DPSO 114/74 cells and they have a limited life span. In humans, transformation of B-lymphocytes into continuous lines of B-lymphoblasts is possible with Epstein-Barr Virus (EBV). In this study, the transformation of B-lymphocytes from the Bolivian squirrel monkey (Saimiri boliviensis boliviensis) is described. 30 ml of blood from an adult male S. boliviensis was obtained and centrifuged for separation. The buffy coat was further separated to isolate out lymphocytes. The lymphocytes were placed in a culture flask with EBV from Cotton-top tamarin B95-8 cells (as a source of EBV) with the addition of 10 ug cyclosporin A. This was incubated for 6 wks. until colonies of cells developed. Clusters of cells were expanded and cloned twice by limiting dilution method. A clone 4D8 was isolated. The karyotype of the 4D8 clone was 44 which is the squirrel monkey karyotype with an XY sex chromosome composition. By scanning EM, the cloned cells were round, similar in size and had numerous short villous processes on their surface. By TEM, cells had a nucleus that was round to oval with abundant euchromatin and a thin peripheral rim of heterochromatin, large nucleoli, and some cells had EBV viral particles in the cytoplasm. 4D8 cells reacted with antibody to CD20, a B-cell-specific marker. Most cells stained for LMP1, a latent infection associated gene product; and some had nuclear reactivity with anti-ZEBRA (BZLF1) antibody. Glucocorticoids were shown to not affect the growth of 4D8 cells and blunt the expression of FKBP51 mRNA when compared to HL cells (human EBV-transformed B-lymphoblast cell line). In conclusion, they were able to develop a continuous cell line from squirrel monkeys by transformation of B-lymphocytes with EBV with a decreased sensitivity to glucocorticoids. It can be used to study the molecular basis of glucocorticoid resistance in the squirrel monkey without use of additional animals.
Questions:
1) How was the continuous cell line established?
2) How was it confirmed that this cell line was from a squirrel monkey?
Answers:
1) B-lymphocytes were isolated from a squirrel monkey and cultured with
EBV to transform them into B-lymphoblasts using cotton-top tamarin B95-8
cells as a source for EBV.
2) The new clone 4D8 was tested by karyotyping (2N=44), SEM and TEM
(cells consistent with B-lymphocytes), immunophenotypic and
immunocytochemical characterization (antibodies react with B-cell
markers), glucocorticoid responsiveness (4D8 cell growth was not
affected by glucocorticoids or glucocorticoids blunted FKBP51 mRNA
expression).

Oral Anticoagulant Therapy and International Normalized Ratios in Swine. Laboratory Animal Science 48 (4): 371.
The prothrombin time (PT) test is commonly used to monitor the effectiveness of oral anticoagulant therapy. A major variable affecting PT results is the specific thromboplastin reagent used. To account for the varying sensitivities of different products, each preparation of thromboplastin is assigned an international sensitivity index (ISI). This index can be used to compare any given preparation with an international reference standard, (a WHO reference plasma). To further standardize reported values between laboratories, it has been recommended that PTs be reported as international normalized ratios (INR). The INR =((patients PT)*(ISI))/mean PT of normal range. This paper reports the INR results, using 2 different analyzers and 2 different thromboplastin reagents, from a group of Yucatan miniature swine at 7 and 14 days of oral anticoagulant therapy (coumadin). In brief, there was a large difference in values obtained by the two methods. (Note that the authors did not try to determine how much of the variance was due to differences in the analyzers and how much was due to the different thromboplastin reagents.) Still, I believe the carry home message here is that caution must be use when comparing PTs obtained from different labs even when reported as INRs. For those interested, reference 4 is a good readable source of info on dealing with the problems inherent in using INRs and for the real obsessive/compulsive, reference 13 (page 115, not page 134 as listed) describes in agonizing statistical detail how to predict confidence intervals for INR measurements in your own lab.
Questions:
1. How is the PT determined?
2. What clotting factors are Vitamin K sensitive?
Answer:
1. The prothrombin time is the time required for a fibrin clot to form when
plasma is added to a thromboplastin-calcium mixture.
2. factors II, VII, IX and X

Myocardial injury in the mouse induced by transthoracic cauterization. Laboratory Animal Science 48 (4): 374.
The authors wished to induce myocardial injury in mice, so that eventually genetically modified mice could be studied after induction of infarction. Thoracotomy and ligation of the left anterior descending coronary artery was fraught with complications, largely due to the small size of the animals. Therefore, they attempted to induce myocardial injury by application of electrocautery, a technique first described in dogs in 1978. Ten SPF male Swiss-Webster mice were anesthetized with ketamine, xylazine and acepromazine for a brief aseptic procedure. An incision was made in the left fourth intercostal region through skin and underlying musculature down to the level of the intercostal muscles. A standard 20-guage needle was blunted, the distal 5mm bent at a 90 degree angle, and the shaft insulated with a length of PE tubing. The bent end of the needle was introduced through the thoracic wall until it rested on the beating heart. Cutting electrocautery (40W, 1 sec) was used to injure the myocardium. Seven to 10 days later the hearts were examined histologically. Nine out of ten surviving mice had evidence of left ventricular tissue damage.
The authors surmise that damage occured in two ways: by direct electrical or thermal injury, and/or by causing vascular occlusion resulting in infarction distal to the site.
Questions
1. Describe a way to induce myocardial damage in a heart the size of a mouse's.
2. Name two ways in which application of electrocautery to the surface of the heart might induce damage to the myocardium, creating a model of infarction.
Answers
1. Instead of doing a thoracotomy and trying to isolate and ligate the left anterior descending coronary artery, do the procedure blindly by inserting a sheathed needle through the left 4th intercostal space and apply electrocautery.
2. Electrocautery may damage the heart muscle by either thermal/electrical injury, or by causing vascular thrombosis and infarction.

Method for Continuous Infusion into the Portal Vein of Mice. Laboratory Animal Science 48 (4): 379.
The authors developed a methodology for continuous infusion into the portal vein in order to facilitate delivery of gene therapy to hepatocytes. They found that 6 ml/24 hours could be tolerated for two weeks with no morbidity. They tested saline. However, retroviral-mediated gene transfer requires actively dividing cells. Hepatocytes can be induced to divide by 70% hepatectomy, hepatic expression of toxic gene products, or infusion of growth factors. The method they developed involved permanent placement of a portal vein catheter consisting of a 5 foot lenth of sterile tubing. The catheter exited the mouse at the neck, where it was attached to a button attached subcutaneously. The catheter was attached to a low-torque swivel mounted on a ring stand above an open mouse cage, so the mice could be unrestrained.
Q: What are some methods for inducing hepatocyte division?
A: hepatectomy of 70% of the liver, expression of toxic gene products, or
infusion of growth factors.

Vesicular Epidermal Rash, Mucosal Ulcerations, and Hepatic Necrosis in a Cynologus Monkey. Laboratory Animal Science 48 (4): 384.
A male Macaca fascicularis(88111) with a full body rash was reported to the clinical veterinarian at the Washington Regional Primate Research Centre. The monkey had been housed in a large room with others macaca and he had grooming contact with his female partner. The male 88111 and seven animals housed in his cage bank were moved on May 1, 1977 to a smaller room relatively isolated from the rest of the facility. Physical exam: On initial observation monkey 88111was found to be bright, alert and ative, despite his full body rash. The exam revealed a lean male with epistaxis from the left nostril and hemorrhagic, maculopapular to vesicular rash. The multifocal lesions over most of the body were present in the axillary inguinal, facial and gingival areas. None of the primates housed with monkey 88111 had similar clinical signs. Blood work, serum chemistry, antibody titers and biopsies were taken. The morning of May 9 ,1977 monkeys oral lesions had progressed to large necrotic plaques and suffered cardiopulmonary arrest.
Pathological Findings
Gross lesions: Cutaneus and mucosal lesions and foci scattered though the heart, adrenals and testis and red foci in liver.
Histological: Skin, tongue and gingiva showed multifocal necrosis, epithelial syncytia, and vesicle formation, with numerous epithelial cells containing eosinophilic intranuclear inclusions. The liver contained coalescing necrotic foci with intranuclear inclusion present in hepatocytes, biliary epithelium and vascular endothelium. Necrosis and inclusions were also seen in the spleen, adrenal cortex, gastrointestinal apithelium and urinary bladder
Blood: neutrophilic leukocytosis, trombocytopenia and marked elevation in liver enzymes. Antibody tests to Cercopithecine herpesvirus-1, simian retrovirus, herpes simplex-1, and 2, simian varicella, monkey pox, and human varicella-zoster were all negative.
Diagnosis: Simian Varicella Virus (SVV) and alphaherpesvirus. This diagnosis was supported when a virus isolated from a biosy of a vesicular lesion was reported. On may 8, 1997 three of the seven animals housed with 88111 were positive for SVV but not other primate of WaRPRC showed sings of SV.
This ocurrence of SW is unusual because there was only one clinical case. However, the three animals had positive antibody screens without a history of clinical disease and the grooming parther seroconverted in absence of clinical signs. Viral latency in the dorsal root ganglia has been demostrated in primates without clinical varicella
Questions
1.- What is the classic presentation of SVV?
2.- Why is this ocurrence of SVV unusual?
Answers:
1.- Vesicular Epidermal Rash, Mucosal Ulcerations, and Hepatic Necrosis
2.- This ocurrence of SW is unusual because there was only one clinical case

Stress-Related Hormonal and Metabolic Responses to Restraint, with and without Butorphanol Administration, in Pre-Conditioned Goats. Laboratory Animal Science 48 (4): 387.
This study was performed to evaluate the stress related hormones and glucose changes in animals that experienced non-pain related stress. These stressors may include fear, anxiety, and change in environment. The goats were pre-conditioned for 3 months prior to the start of the study. For the course of the study they were placed in a stachion previously exposed to. All samples were collected from this pen. Just prior to the start of the study a catheter was place for blood collection. The administration of butorphanol lowered glucose level and increased heart rate, respiratory rate and rectal temperature. Baseline free fatty acid levels were higher than at any other time. This was due to the effects of norepinepherine release as a result of the catheter placement which stimulates lipolysis. One drawback to the interpretation of the data from this study is that the criteria are specific for each stressor.
Cortisol has been used routinely for the measurement of stress response in goats. This study confirmed the importance of conditioning animals prior to studies in which physiologic, metabolic, and stress-related hormonal responses are evaluated.
Question
1. What effect did butorphanol on the plasma glucose level on the goats in this study?
Answer
1. Plasma glucose levels were lowered after botorphanol administration.

Systemic Yeast Infection in Owl Monkeys ( Aotus vociferans): Ante-Mortem Screening and Diagnosis by Examination of Bone Marrow Aspirates. Laboratory Animal Science 48 (4): 391.
Three adult non-geriatric wild-caught owl monkeys (from Peru) from a large colony had an unclassified yeast organism found after euthanasia or death. Multiple yeast cultures on Sabouraud Dextrose Agar, SABHI Agar with blood, and SABHI Agar with Chloramphenicol and Cycloheximine were negative.   However, paraffin-embedded formalin-fixed stained tissues (spleen, liver, kidneys, lymph nodes, heart, lungs, adrenal glands, and bone marrow) demonstrated myriad spherical to oval, thick-walled, uninucleate yeast cells 8-10 microns in diameter.   A few cells had narrow-based budding of single daughter cells. Chains of organisms were not formed. Yeast cells were common both free in the tissue and inside macrophages, but inflammatory infiltrates were absent. The yeast cell morphology was consistent with Histoplasma capsulatum var duboisii. The yeast cells tested from one animal were positive by FA testing to H. capsulatum and Blastomyces dermatitidis using a bivalent conjugate; however, the serum tested from this animal was negative by immunodiffusion and complement fixation for antibodies to both yeast species.
   Bone-marrow aspirates were performed on 15 apparently health monkeys from this colony in an effort to screen for subclinical systemic yeast infection. Smears were fixed and stained with Dif-Quik and examined at x400 mag. Fixed bone marrow clots were paraffin-embedded, sectioned at 5 microns, and stained with H&E and GFS. One of these monkey had yeast cells detected in his bone marrow. The description of the yeast was similar to that of the 1st three monkeys.   On the H&E stain the 1-micron thick capsule was clear. The GFS stain imparted a magenta color to the cell wall against the yellow counter stain, enhancing visibility of the yeast cells.   The monkey remains asymptomatic 8 months later, with no treatment. The yeast remains unclassified. Although its morphology was most consistent with Histoplasma capsulatum var duboisii, the absence of cellular reaction, inability to grow the organism on artificial media and the fact that the monkeys were from South America are inconsistnet with that organism. Other yeast species were considered, but none completely fit the profile of the investigated organism. Much remains to be learned about this yeast. Bone-marrow aspirates should be considered for screening healthy wild-caught owl monkeys or working up debilitated or anemic animals. Routine histological examination of surgically removed visceral organs is also suggested.
Questions:
1. True-False: Infection with Histoplasma capsulatum var duboisii is
commonly diagnosed in humans and nonhuman primates from South America.
2. True-False: It is recommended to give newly obtained wild-caught
owl monkeys a prophylactic course of Amphotericin B to eliminate
subclinical systemic yeast infections.
3. Which of the following known yeasts cannot be grown on artificial
media?
a) Histoplasma capsulatum var duboisii
b) Blastomyces dermatitidis
c) Loboa loboi
Answers:
1. False. It is generally found in humans or animals in or originating
in Africa, and has not been reported from South America.
2. False. Broad-spectrum antifungal agents such as Amphotericin B are
excreted principally by the kidneys and have the potential to cause
renal damage. Since renal disease is common in owl monkeys, this course
of treatment could be problematic.
3.    ( c ) Loboa loboi. Its host/geographic range is restricted to
humans in South and Central America, with the exception of dophins off
the Florida coast. It is primarily an infection of the skin and
subcutis, and does not disseminate viscerally.

Maternal Transfer of Hantavirus Antibodies in Rats. Laboratory Animal Science 48 (4): 395.
Hantavirus affects rodents and humans. There are 20 species of Hantavirus. Old world species include Hantann, Seoul, Puumala, Dobrova. New world species include HPS (Hantavirus pulmonary syndrome), found in North America and South America. HFRS (Hemorrhagic Fever and Renal Syndrome) is a clinically similar disease to HPS, found in Eurasia and surrounding areas.
    HPS - capillary leakage is localized to the lungs. Death is due to shock and cardiac complications.
    This article confirms that there is prenatal transfer of protective antibodies to Hantavirus in rats. IgG and IgA are transferred in utero and via the milk, equally efficiently. The IgA transferred in utero lasts a shorter duration than that transferred via milk.
QUESTIONS
1.      a)      What is the family and genus species of the viral agent causing HPS?
        b)      Is this virus DS(double-stranded)DNA, SS(single-stranded) DNA, DS RNA or SS RNA?
2.      Bunyaviruses are arthropod borne. True or False
3.      Vero cells are derived from?
4.      a)      The natural reservoir of Hantavirus is?
        b)      The end stage host is?
ANSWERS
1.      a)      Family Bunyaviridae
                Genus   Hantavirus
        b)      SS RNA
2.      False, all are arthropod borne except Hantavirus.
3.      African green monkeys.
4.      a)      rodents
        b)      humans (incidental end stage host)

Arterial Blood Gas Changes in New Zealand White Rabbits During Carbon Dioxide-Induced Pneumoperitoneum. Laboratory Animal Science 48 (4): 398.
      Laparoscopic surgery with carbon dioxide insufflation to visualize the internal abdomen is common in humans. The following physiological alterations have been reported in association with carbon dioxide-induced pneumoperitoneum (CDP): 1. arterial and pulmonary hypertension, 2. reduced lung compliance, 3. increase in cardiac output, venous oxygen pressure, and venous saturation, 4. acidemia, 5. hyperkalemia, and 6. hypercapnea. The aim of this study was use the rabbit as a model for CDP in infants and to assess the physiologic changes.
      Twenty New Zealand White rabbits (both sexes) were assigned to two groups of 10. All rabbits were catheterized in the marginal ear vein and given atropine (0.29-0.40 mg/kg IV) and pentobarbital (25.2 mg/kg IV). Additional pentobarbital was administered to maintain ventilator dependence. An endotracheal tube was placed in each rabbit and connected to a ventilator (FiO2 of 40%, inspiratory pressure of 20 cmH20, 30 breaths/min). A baseline arterial gas tension (blood from auricular artery) was taken after 20 minutes.
      Ten rabbits received 100% CO2 intraperitoneally via a Veress needle inserted 1.5 cm caudal to the umbilical scar to maintain an abdomenal pressure of 8mmHg. The second group of 10 were maintained on anesthesia but without abdomenal intervention. The rabbits were anesthetized for 105 minutes. 0.3 ml samples of arterial blood were taken every 15 minutes.
       Results: "Rabbits weighing <3.5kg with CDP at 8mmHg have a statistically and clinically significant increase in PaCO2, with a consequent reduction in arterial pH, and...arterial oxygen saturation variation did not correlate with pH or PaCO2 alterations."
Questions:
1. What is the most common gas used for insufflation in laparoscopic
    procedures?
2. The amount of CO2 absorbed through the peritoneum in a constant
    intra-abdomenal infusions state is dependent on the amount of gas
    infused. T or F?
3. What can be done to counteract the increase in PaCO2 during CDP?
4. What is the genus/species of the pig?
Answers:
1. CO2
2. F Dependent on the pressure generated
3. Insrease the ventilatory rate
4. Sus scrofa