Contemporary Topics 38 (1)
1999

Effects of crowding on the social behavior of cuttlefish (Sepia officinalis). Contemporary Topics 38 (1): 49.
This study evaluated the effect of cage density on behavior. The "small tank" housed 2 males and 4 females with 0.29 in.2 per fish. The "large tank" housed 3 males and 3 females with 4.87 m2 per fish. Common stocking density in laboratory facilities is about 3 adults/m2 (i.e. similar to small tank). The cuttlefish show a variety of body patterns for crypsis and communication and these were part of the behavior repertoire evaluated.
Results: Females usually sat on bottom of the periphery. Males generally swam in the middle or sat on the bottom at the periphery. In the small tank, 2 males clearly avoided each other, the females avoided other females, and male and females overlapped. This was similar in the large tank but more spaced out. The fish spent 93% of their time without another cuttlefish within 2 body lengths. Cuttlefish in the large tank consistently ate more food per day. The subjects hovered 70% of the time. Males were more sedentary. Fish in the large tank were more sedentary. Only males showed strong zebra pattern, usually when swimming. The zebra pattern was more frequently seen in the small tank and with males more than females. All other body patterns were more frequent in the large tank. There were 3 times more displacements in the small tank. Both tanks displacing usually males displacing females. There were 3 times more Intense Zebra Displays (agonistic encounter) in the small tank and these were more directed at females.
Interpretations/Conclusions: Explanation for why fish in small tank ate less - more stressed. Believed to be benthic bottom dwellers in the natural environment. In lab culture, soft substrate is not provided to simplify care and cleaning. This lack probably influences settling behavior resulting in increased hovering and decreased amount of time resting on the bottom. The increase hovering in the small tank might be a sign of distress due to overcrowding. Displaced cuttlefish sometimes injured themselves by jetting into side of the tank. Effect of difference in aggression on health is significant. Females in both tanks were continually harassed by males. Intense Zebra Displays were a good indicator of heightened aggression by males. Separating the males from the females as soon as the male begins exhibiting Intense Zebra Displays could decrease the incidence of injury to females in the captive situation. This species is probably semi-solitary. Perhaps they are solitary most of the year and aggregate solely during the reproductive season.
Questions:
1. Genus and species of the cuttlefish?
2. What is a "benthic" fish? What is crypsis?
3. What strategy was proposed to reduce aggression within the tanks?
Answers:
1. Sepia officinalis
2. Benthic fish are those that are bottom feeders/dwellers. Crypsis is a body pattern that causes the animal to blend into the backround (camouflage).
3. Remove males as soon as they start displaying Intense Zebra Displays towards the females. Only keep together during the reproductive season.

Stereotactic surgery and long-term maintenance of cranial implants in research animals. Contemporary Topics 38 (1): 56.
This article reviews commonly used neurosurgical techniques and offers suggestions for managing or preventing problems with these cranial implants. Principles of Stereotactic Surgery including implantation and fixation of experimental devices or probes are presented. Techniques for Long-Term Maintenance of Recording Chambers.
QUESTIONS:
1. Name several components of a permanent neurosurgical implant.
2. The two components of the stereotactic apparatus which anchor the skull and control the angle and the position of the skull are the _____ and the ______. The _____ are inserted first.
3. (True or False) Long narrow tipped ear bars are recommended for survival and long-term procedures whereas blunt-tipped ear bars should be used in anesthetized animals that will not be allowed to recover.
4. List two aspects of anesthetic management for stereotactic surgery.
5. Name and define 3 landmarks for stereotactic surgery. In rats, which landmark is morre accurate for locating forebrain structures and which for brainstem structures?
6. (True or False ) Brain coordinates for specific target locations vary considerably depending on sex, age, weight, and strain of animal.
7. After several weeks of stable attachment movement of the headpiece suggested bone degeneration (but osteoporosis could have caused earlier detachment). What should be evaluated to prevent reoccurence of this problem?
8. When performing a Pinealectomy where should a rectangular section of the skull be removed?
a. Between the sagittal and the coronal sutures
b. Between the sagittal and the frontal sutures
c. Between the sagittal and the lambdoid sutures
d. Between the sagittal and the ethicon sutures
e. Between the sagittal and the bobbit sutures
9. (True or False) Acidulated Fluoride applied to the skull does not strengthen the bone and decreases the retention of headpieces in adult rats.
10. Acrylic Cement and Methyl Methacrylate Solvents may cause also
EXCEPT
a. reactive fibrous layer between the cement and the bone
b. blood between the cement and the bone
c. hypotension
d. formation of emboli
e. anaphylactic reactions
11. (True or False) To reduce the likelihood that the electrode will introduce microorganisms into the brain, 70% ethanol solution with a sterile saline flush or a chlorine dioxide 10 minute immersion.
12. Removal of granulation tissue from the surface of the dura to provide long-term maintenance of recording chambers is known as ________ ________ .
ANSWERS:
1. Name several components of a permanent neurosurgical implant.
Mounting Screws
Dental Acrylic Cement
Threaded Posts
Electrical Connectors
Micromanipulators
Trephine Hole
Recording Chamber
Electrode
Cannula
2. The two components of the stereotactic apparatus which anchor the skull and control the angle and the position of the skull are the ear bars and the incisor bar . The ear bars are inserted first.
3. ( False) Blunt- tipped ear bars are recommended for survival and long-term procedures whereas long narrow ear bars should be used in anesthetized animals that will not be allowed to recover.
4. List two aspects of anesthetic management for stereotactic surgery.
Reducing the level of general anesthetic when removing the ear bars from the animal at the end of the surgery.
Use of local anesthetic ointments or solutions at the skin margins and skull surface during surgery can reduce the depth of general anesthesia required intra-operatively.
5. Name and define 4 landmarks for stereotactic surgery.
Interaural Line - an imaginery line that passes through the brain at the midpoint of the external auditory meatus.
Stereotactic Zero - the intersection of the Interaural Line and the Midsagittal Plane.
Bregma - the midline intersection of the sutures lines between the frontal and parietal bones (intersection of the coronal and sagittal skull sutures from the Blue Book).
Lambda - the midpoint of the best fitting curve passing long the lambdoid suture which is located between the parietal and the occipital bones (intersection of the lambdoidal and sagittal sutures from the Blue Book).
In Rats -
Bregma - Forebrain
Lambda or Interaural Line - Brainstem
6. (True ) Brain coordinates for specific target locations vary considerably depending on sex, age, weight, and strain of animal.
7. After several weeks of stable attachment movement of the headpiece suggested bone degeneration (but osteoporosis could have caused earlier detachment). What should be evaluated to prevent reoccurence of this problem? Sterile technique of surgeon.
8. When performing a Pinealectomy where should a rectangular section of the skull be removed?
c. Between the sagittal and the lambdoid sutures
9. (False) Acidulated Fluoride applied to the skull does not strengthen the bone and decreases the retention of headpieces in adult rats. Acidulated Fluoride applied to the skull strengthens the bone and improves the retention of headpieces in adult rats.
10. Acrylic Cement and Methyl Methacrylate Solvents may cause also
EXCEPT
b. blood between the cement and the bone
11. (True) To reduce the likelihood that the electrode will introduce microorganisms into the brain, 70% ethanol solution with a sterile saline flush or a chlorine dioxide 10 minute immersion.
12. Removal of granulation tissue from the surface of the dura to provide long-term maintenance of recording chambers is known as dural scraping.

High fungal spore load in corncob bedding associated with fungal induced rhinitis in two rats. Contemporary Topics 38 (1): 64.
Two immunocompetent F344 rats housed on nonautoclaved corncob contact bedding in standard polycarbonate cages were diagnosed with fungal-induced rhinitis after being on a toxicology study. The animals presented with slight dehydration, inspiratory stridor, wheezes, or death with no premonitory signs. Gross necropsy revealed purple right cranial lung lobe, and mild serosanguinous nasal discharge. Histopathology revealed multifocal areas of congestion and atelectasis and prominent aggregates of neutrophils and fungal hyphae consistent with an infection with Aspergillus in the nasal turbinates. The authors looked at 2 common contact rodent bedding as the possible source of the fungal infections, corncobs and Aspen chips. Bedding from one Aspen chip manufacturer and 2 corncob bedding manufacturers was analyzed from both autoclaved and nonautoclaved samples. Bedding was mixed with sterile saline and aliquots were cultured on Rose Bengal agar plates and incubated for 2-4 days. 99.4% of the colonies isolated were from the nonautoclaved corncob bedding with the remaining 0.6% from the autoclaved corncob and the Aspen chip both autoclaved and nonautoclaved bedding. The fungi isolated included *Cladosporidium, Acremonium, Penicillium, Aspergillus, Fusarium, and Scolobasidium. The study showed that the bedding could definitely have been the source of fungi and that animals housed on nonautoclaved corncob bedding may be exposed to large numbers of fungal spores. Quality control data on spore counts in corncob bedding range from 800-80,000 spores/g. This study demonstrated a 700-5440 spores/g count. Non autoclaved corncob bedding is contraindicated for psittacine husbandry because of the potential for respiratory aspergillosis. There are already a few reports in the literature of aspergillosis in rats. They suggest autoclaving or gamma-irradiating bedding prior to use the significantly decrease the fungal spore load although it will not eliminate the presence of mycotoxins.
Questions:
1. What was the suggested media for performing the fungal cultures?
2. Which species of fungus is reported in the literature as causing supperative rhinitis in rats?
Answers:
1. Rose Bengal agar plates
2. Aspergillus

Environmental conditions in a chamber with dry ice. Contemporary Topics 38 (1): 67.
Dry ice is used as a source CO2 for euthanasia of rodents. This study was performed to evaluate conditions created in a standard 9-liter bell jar. Results indicated that 500 g ice was req uired to maintain filling rate and concentration at levels recommended by the AVMA Panel on Euthanasia; after removing a animal, the lid should be replaced for one minute before another animal is placed in. Temperature in the chamber never fell below 17C but platform temp decreased to 0C during the 80 hr time period. If chamber must be used for more than an hour, alternate between two chambers allowing a chamber to warm up to room temp before reuse.
Question:
What are the AVMA recommendations concerning CO2 euthanasia?
Answer:
The AVMA recommends a filling rate of 20% per minute with animal in chamber and for a prefilled chamber 70% CO2 is needed to produce rapid unconciousness.

Reference blood chemistry values throughout the lifespan of M. spretus. Contemporary Topics 38 (1): 70.
In this study blood chemistry values for Mus spretus are reported and analyzed with respect to age and gender. Mus spretus have been used for genetic analysis and are a promising model for gerontology research. M. spretus are one of the most genetically divergent species that still interbreeds with Mus musculus to produce at least one sex that is fertile. Of the serum chemistry parameters evaluated, only cholesterol levels varied with respect to age (cholesterol levels increased with age). Direct bilirubin levels differed significantly between female and male mice, with females having increased levels. Cholesterol is a principal building block of all plasma membranes and a key compound for the synthesis of steroid hormones and bile salts. When red blood cells are destroyed by the reticuloendothelial cells, the globin portion of the hemoglobin is split off and the heme is converted to biliverdin, which can then be converted to bilirubin. If the age- and gender-associated differences in cholesterol and bilirubin values that we observed are representative of the species, M. spretus may be an animal model for conditions such as atherosclerosis.
Questions:
1. True or False: Cross breeding of Mus spretus with Mus musculus produces at least one sex of fertile offspring.
2. What is the Telomere Hypothesis and why is Mus spretus a good species to test this?
3. Is the student's t test unpaired or paired?
4. The correct order of biochemical reactions from RBC breakdown is:
a. Hemoglobin - heme - conjugated bilirubin - free bilirubin - biliverdin
b. Hemoglogin - heme - free bilirubin - conjugated bilirubin - biliverdin
c. Hemoglobin - heme - biliverdin - free bilirubin - conjugated bilirubin
Answers:
1. True
2. The Telomere hypothesis states that in some species, telomeres, or the ends of chromosomes, shorten with age. Because M. spretus chromosomes are shorter than M. musculus, they can be used. The telomere length and telomerase activity of various M. spretus tissues differ with age.
3. Unpaired.
4. C

Evaluation of tiletamine-zolazepam as an anesthetic in quail (Coturnix coturnix japonica). Contemporary Topics 38 (1): 73.
Tiletamine-Zolazepam was administered alone or in combination with atropine, xylazine, and levomepromazine to quail. From doses of 10-100 mg/kg of Tiletamine-Zolazepam either alone or in combination with xylazine or levomepromazine produced only hypnosis. Discrete acute focal myositis was produced initially and scarring occurred after 21 days. Even though the study showed that the anesthetics at the ranges studied did not produce anesthesia, the authors recommended its use for non-invasive and minimally painful procedures requiring chemical restraint and recumbency at 30 mg/kg.
Questions:
1. Name three compounds that are classified as arylcycloalkylamines.
2. The anesthetic state induced by dissociative drugs differs from that produced by conventional anesthetics by electroencephalographic evidence of dissociation between the :
a. Cerebral Cortex and the Peripheral Nervous System
b. Hypothalamus and the Ascending Reticular Activating System
c. Corpus Callosum and the Cuneocerebellar Tract
d. Thalamus and the Limbic System
e. All of the Above
3. True or False: Zolazepam hydrochloride is a nonphenothiazine diazepinone.
True or False: There is no such thing as a phenothiazine diazepinone.
True or False: The tranquilizers may be classified as Phenothiazines, Butyrophenone, Benzodiazepines, and Alpha-2-Agonists.
4. Cornick and Jensen evaluated tiletamine-zolazepam as an induction agent in ratites.
Define ratite. For extra credit give examples of ratites and their continent of origin.
5. True or False: With Tiletamine-Zolazepam anesthesia corneal, palpebral, pharyngeal, and laryngeal reflexes are reduced but not eliminated.
6. In Brazil, quail are considered as a reference species for the appraisal of what?
7. How was response to noxious stimuli assessed in the quail?
a. Pinching of skin for 1 sec with a hemostat
b. Pinching abdominal musculature for 1 sec with a hemostat
c. Pinching interdigital space for 1 sec with a hemostat
d. Pinching the wattles for 1 sec with a hemostat
e. a., b., and c.
f. all of the above
8. The Fisher test was used to analyze (parametric or non-parametric data) (select one).
The ANOVA followed by the Tukey-Kramer multiple comparisons test was used to analyze (parametric or non-parametric data) (select one).
9. Give the Genus and Species of the Following:
Quail
Green Heron
10. Describe the injection site lesions associated with Tiletamine-Zolazepam injection in Quail.
Answers:
1. Name three compounds that are classified as arylcycloalkylamines.
Tiletamine
Ketamine
Phencyclidine
2. The anesthetic state induced by dissociative drugs differs from that produced by conventional anesthetics by electroencephalographic evidence of dissociation between the :
d. Thalamus and the Limbic System
3. True (As stated in the article): Zolazepam hydrochloride is a nonphenothiazine\diazepinone.
True (By classification of tranquilizers): There is no such thing as a phenothiazine diazepinone.
True (By classification of tranquilizers): The tranquilizers may be classified as Phenothiazines, Butyrophenone, Benzodiazepines, and Alpha-2-Agonists.
4. Cornick and Jensen evaluated tiletamine-zolazepam as an induction agent in ratites.
Define ratite.
Ratite - a bird with a flat breastbone, esp: any of various mostly flightless birds
(as an ostrich, rhea, emu, kiwi, cassowary).
Ostrich - Africa
Rhea - South America
Emu, Kiwi, Cassowary - Australia
5. False: With Tiletamine-Zolazepam anesthesia corneal, palpebral, pharyngeal, and laryngeal reflexes are maintained.
6. In Brazil, quail are considered as a reference species for the appraisal of ecotoxicity of chemical agents in domestic and wild animals.
7. How was response to noxious stimuli assessed in the quail?
g e. a., b., and c.
a. Pinching of skin for 1 sec with a hemostat
b. Pinching abdominal musculature for 1 sec with a hemostat
c. Pinching interdigital space for 1 sec with a hemostat
8. The Fisher test was used to analyze non-parametric data.
The ANOVA followed by the Tukey-Kramer multiple comparisons test was used to analyze parametric.
9. Give the Genus and Species of the Following:
Quail - Coturnix coturnix japonica
Green Heron - Butorides virescens
10. Describe the injection site lesions associated with Tiletamine-Zolazepam injection in Quail. Acute focal myositis, hemorrhage, polymorphonuclear infiltrate, discrete fibrosis between muscle fibers (normal scarring).

Vaginal cytology of the lesser bandicoot rat (Bandicota savilei). Contemporary Topics 38 (1): 76.
The lesser bandicoot rat is a wild field rat commonly found in Thailand and is known to serve as a reservoir of several important parasites that cause human disease. The purpose of this study was to determine
estrous cycle induced changes in the vulvar appearance and vaginal cytology of colony -born B. savilei and to compare them with those of the common Laboratory rat ( Rattus norvegicus).
Five colony born animals were used. Samples were taken twice daily for 28 days . All samples were stained with a Giemsa stain. The external appearance of the vulva and the appearance of the tip of the
vaginal swabs were also recorded. The vaginal cells were quantified by percentage into three groups ( nucleated epithelial cells, keratinized epithelial cells , and leukocytes).
There were no abnormal vaginal changes or discharges. Most of the swab tips were clear. Some contained yellow or white material that seemed to correspond to diestrus and estrus. The vaginal cytology seemed to
be similar to the stages of the estrous cycle in the common lab rat.
Questions:
1. What mite larva is carried in the ear canal of the bandicoot rat .
2. What disease is this mite the vector for?
Answers:
1. trombiculid mite larvae Leptotrombidium sp.
2. scrub typhus

Diagnostic exercise: Fluffy, white, cotton candy-like growth on the fills, fins, and skin of salamanders (Ambystoma tigrinum). Contemporary Topics 38 (1): 81.
Two weeks following arrival, a shipment of 30 salamanders was exposed to increased water temperatures of 25-30oF for a few days. Five (5) of the 30 salamanders developed a milky appearance to their skin, which progressed during the subsequent 4-5 days to the appearance of fluffy, white, cotton candy-like material on the gills, fins and skin (particularly around the head). Gross lesions were not evident in other tissues or organs. Histologically, there were severe multifocal erosions and ulcerations of the skin and hyphae localized to the skin surface, dermis and subjacent muscles. Mild focal subcutaneous edema, hemorrhage, and muscular necrosis were also present. Microbiological culture of skin on cornmeal agar isolated aseptate hyphae.
Diagnosis: Saprolegniasis
Pathogenesis from Saprolegnia spp. is usually associated with environmental or social stresses to fish, amphibians, and other aquatic animals. Saprolegnia spp. are ubiquitous and spread raidly in aquatic environments. The fungus can also infect the eggs of amphibians. The preferred treatment is elimination/depopulation of affected animals, improvement of the water environment, and reduction of shipping, handling, and social stresses.
Questions:
1. What treatments for Saprolegniasis are described in the Blue Book?
2. Name 3 other fungal infections of amphibians.
3. What are the 2 most common isolates of Saprolegnia listed in the Blue Book?
Answers:
1. malachite green dips, copper sulfate dips, chloramine bath, mercurochrome bath
2. Chromomycosis, Phycomycosis (Mucormycosis), Ichthyosporidiosis
3. S. parasitica and S. ferax

Improved survival rates in mice that received prophylactic fluids after carcinogenic treatment. Contemporary Topics 38 (1): 84.
A high mortality rate attributed to dehydration was observed in a mouse model of mammary adenocarcinoma induced by 7,12-dimethylbenz[a]anthracene. In this study, the survival rates of mice given therapeutic hydration following the onset of clinical dehydration versus prophylactic administration of fluid immediately following administration of the carcinogen were compared. For both groups, saline was administered subcutaneously. Mortality in the prophylactically treated mice was 5% versus 47% in mice treated after the onset of clinical signs.
No questions

Acute paralysis caused by fibrocartilaginous embolism in a pigtail macaque. Contemporary Topics 38 (1): 87.
A female pigtail macaque (Macaca nemestrima) was evaluated for acute flaccid rear limb paralysis with reduced patellar and anal reflexes three days after being received. The animal was treated with antibiotics and corticosteriods with no improvement. The patient was necropsied and a soft tissue swelling was observed within L1 to L3. Microscopic examination revealed severe diffuse ischemic necrosis especially in the gray matter. H and E staining indicated small vessels in a nerve root contained a blue staining material indicating fibrocartilage. Fibrocartilaginous embolism is an uncommon cause of acute infarction and ischemic necrosis of the spinal cord in man and animals. It is postulated that degenerating discs may be invaded by small vessels. If pressure within disc exceeds arterial pressure, extruding disc material may enter these vessels and travel into the spinal cord. This is the first reported case in a nonhuman primate and only 50 cases have been seen in man.
No questions

Spontaneous renal cell carcinoma in a New Zealand White rabbit. Contemporary Topics 38 (1): 89.
A case report of a renal cell carcinoma in a two year old New Zealand White rabbit. Grossly, this tumor was 10 cm x 12 cm and had a 1-2 cm rim of viable tissue surrounding a hollow core which was filled with brown fluid. Tumor cells were arranged in sheets, nests, and acini and the cells were often associated with an eosinophilic hyaline matrix. Acicular spaces suggestive of cholesterol crystals were also present. No areas of metastasis were identified.
Primary renal neoplasia in animals is quite rare. This is the second reported case of a renal cell carcinoma in the laboratory rabbit.
Questions:
1. Name the four most common naturally occurring tumor types in the rabbit (in ranked order).
2. Name the Genus and species of wild (cottontail) rabbits.
3. Embryonal Nephroma can also commonly be called a ____________.
4. Name a stain that can be used to detect mineralization.
Answers:
1. (Blue book answer p. 231)
Uterine adenocarcinoma
Lymphosarcoma
Embryonal nephroma
Bile duct adenoma of the liver
This article suggests and references a paper that says embryonal nephroma's are the second most common rabbit neoplasm rather than lymphosarcoma.
2. Sylvilagus floridanus
3. Wilms Tumor
4. von Kossa, Alizarin red