Handheld digital equipment for weight composite distress pardigms: new considerations and for rapid documentation and intervention of rodent populations. Contemporary Topics 40(4), 11.
Abstract: This paper describes the use of a Palm-Pilot linked to a computer with an MS-Access data-base. The purpose of this paper was to demonstrate that using new technology it may be possible to increase the amount of individual monitoring of laboratory rodents, and either extend an endpoint or provide more timely intervention which would result in more valuable scientific intervention.
Palm-Pilots were used and a form was used to input data about individual rodents. Data included identification of rodent, numerical descriptions of attitude, gait, posture, respiratory pattern and skin color. It then had tables to input starting weight, and , daily weight and from those calculated % weight loss.
Rodents were given a summary score which put them in one of three categories, favorable status, veterinarian to be called sometime during day, or immediate need for veterinary care. These Palm Pilots were used in a laboratory animal setting as well as an exotic animal veterinary practice. On average the record collection for each individual rodent took 3 minutes, compared with 8 minutes for a hard written record.
Using the same Palm Pilot system a second form for veterinary intervention was designed. The veterinarian could check all the animals scores and determine which animals needed to be seen immediately, and which could be seen later in the day.
The author ends by stating that this system may refute some of the concerns in the industry about the high cost of observing, documenting, and reporting of rodent procedures.
Questions: 1. Using the Palm Pilot an average record was scored in ? minutes.
a. 6
b. 3
c. 8
d. 11
True or false
2.Percentage weight loss has been proposed as a standard for scoring systems to enhance humane study design?
3. The Animal Welfare Act covers rats, mice and birds.
4. Increased monitoring of animals may result in the minimum duration of distress and pain.
5. Use of Palm Pilots can complement scientific objectives rather than obstruct them,
Answers: 1.b
2.t
3.f, (this answer subject to change!)
4.t
5.t
Recovery from carotid artery catheterization performed under various anesthetics in male, Sprague-Dawley rats. Contemporary Topics 40(4), 18.
Abstract: Procedures such as indwelling catheters tomonitor cardiovascular parameters or collecting blood samples are known to alter circadian rhythms of heart rate, temperature and activity. The authors compared the time to recovery from carotid artery catheterization between three common anesthetics; 1)sodium pentobarbital, 2) halothane, or 3) ketamine, xylazine and acepromazine. Radio-telemetry transmitters were implanted in the abdominal cavities of male, Sprague-Dawley rats with the attached catheter inserted in the terminal aorta. Chronic indwelling catheters were placed aseptically in the left common carotid artery under one of the anesthetics; ketamine/zylazine/acepromazine i.p., sodium pentobarbital i.p., or halothane by face mask.
Body weights, food and water consumption, blood pressure, heart rate and the rat's activity in the home cage were used to evaluate time to recovery.
Anesthesia and caroticd artgery catheterization decreased body weights and reduced food and water consumption in rats. During the light phase of the daily photoperiod, heart rates and mean arterial blood pressures (MAP) of all rats were significantly elevated regardless of anesthetics. Activity in the home cage was not significantly affected by the anesthesia and surgery except on the third day post-surgery when the animals anesthetized with the ketamine mixture showed a significant increase in activity. Nocternal heart rates and MAP were not affected by the surgery in pentobarbital or halothane anesthetized rats; however, rats anesthetized with the ketamine mixture had a reduced heart rate and increased MAP on the night folowing surgery.
Activity during the dark phase was decreased in halothane- and ketamin-anesthetized rats.
The authors conclude that recovery depends on each anesthetic used, but in general 2-4 days should be allowed for recovery from this procedure.
Questions: 1. Which of the followings is NOT an OUTBRED?
a. Fischer 344
b. Sprague-Dawley
c. Long-Evans
d. Holtzman
2. True/False: A reduction in food intake and body weight has been shown to increase sympathetic tone and reduce parashypathetic tone.
Answers: 1. a.
2. False: A reduction in food intake and body weight has been shown to reduce sympathetic tone and increase parasympathetic tone.
Testing and comparison of non-opioid analgesics in amphibians. Contemporary Topics 40(4), 23.
Abstract: Minimal research addresses effective analgesics in non-mammalian vertebrates. However, there is a substantial need for this information for practicing veterinarians and animal care personnel at research institutions. The use of non-opioid analgesics is often preferable over opioid agents since the former requires less regulations and recordkeeping.
This paper describes the relative analgesic effects of numerous non-opioid agents in Northern grass frogs (Rana pipiens). Northern grass frogs of both genders, with an average weight of 26g, were obtained from a commercial source. They were housed in groups of 48 in free-flowing tap water and fed live crickets 3-4 times per week. Animals were then randomly assigned to individual cages two days prior to the study to allow time for acclimation. Drugs tested included: antipsychotics (chlorpromazine and haloperidol), a benzodiazepine (chlordiazepoxide), partial opioid agonists (buprenorphine and butorphanol), a barbiturate (pentobarbital), a histamine antagaonist (diphenhydramine), and NSAIDs (indomethacin and ketorolac). Drugs were mixed with sterile saline (0.9%) to yield nmol/g doses of the active base and administered in a volume of 10ul/g body weight. [Dosages tested are listed in the paper] Morphine (which has previously been shown to be a potent analgesic in frogs subjected to the acetic acid test) was administered to one group of animals as a comparison. Drugs were injected subcutaneously into the dorsal lymph sac. Each animal received either a drug or a saline control injection.
Pain thresholds/nociceptive thresholds were measured with the acetic acid test that consists of applying increasingly strong concentrations of 11 serial dilutions of glacial acetic acid to the dorsum of a frog's thigh. Once the dilution is applied, the frog is observed for a wiping response of either hindlimb. If a response is not observed after 5 seconds, the acid solution is rinsed off with distilled water and then the next higher dilution is applied to the opposite limb. The nociceptive threshold (NT) is assigned based on the lowest concentration of acid that elicits a wiping response. NT were evaluated before administration of saline or drug and then at 1, 3, and 5 hours after administration.
Since the acetic acid test relies on a motor response and since some analgesics may produce sedation or motor dysfunction, the motor function of animals that showed significant analgesic effects were evaluated at 5 hours post drug/saline administration by observation of the corneal reflex, the righting reflex, and the hindlimb withdrawal reflex.
Results: None of the drugs tested produced greater than 80% analgesic effect at doses that were not lethal within 24 hours. Morphine produced the greatest analgesic effect. Significant analgesic effects were evident by the 1 hour test period and did not significantly change by the 5 hour time point. Peak analgesic effects for the highest doses of the more potent agents were morphine> chlorpromazine> chlordiazepoxide> buprenorphine>diphenhydramine> haloperidol. Specific dosages are given in the paper. Although the study was not designed to evaluate lethal dosages of these drugs, the authors did note that many agents possessed a narrow therapeutic range. In addition, the authors warn that since the acetic acid test is based on a weak noxious stimuli applied cutaneously, the dosages of drugs shown to produce analgesia in this study may not be sufficient or appropriate to treat pain from surgical procedures, visceral structures, or pain which is neuropathic in origin. Also, one would have to be very cautious extrapolating drug dosages from this study for use in other species.
Questions: 1. Name the genus and species of the Northern grass frog.
2. Describe the acetic acid test.
3. Where would you inject a frog if you wished to inject the dorsal lymph sac?
Answers: 1. Rana pipiens
2. The acetic acid test involves the application of increasingly strong concentrations of 11 serial dilutions of glacial acetic acid to the dorsum of a frog's thigh. Once the dilution is applied, the frog is observed for a wiping response of either hindlimb. If a response is not observed after 5 seconds, the acid solution is rinsed off with distilled water and then the next higher dilution is applied to the opposite limb. The nociceptive threshold (NT) is assigned based on the lowest concentration of acid that elicits a wiping response.
3. The dorsal lymph sac is most often accessed between the scapulas. Injecting the dorsal lymph sac is very similar to administering a subcuticular injection in this
Rescue of transgenic mouse line by transplantation of a frozen-thawed ovary obtained postmortem. Contemporary Topics 40(4), 28.
Abstract: The authors had previously shown that it was possible to transplant ovaries within 2 hours after death of a transgenic dam into nontransgenic ovariectomized littermates and to obtain viable transgenic offspring. Other investigators had shown that frozen ovarian tissue was able to restore fertility to ovariectomized females after transplantation. In this study, the authors showed that it is possible to cryopreserve ovaries from dead mice within 2 hrs after death and to obtain fertile mice after thawing and transplanting that ovarian tissue.
Two cryopreservation techniques were examined: one method used a controlled-rate freezer while the other method did not. The former method is called the Szteins method, while the latter is called vitrification method or the Rall and Fahy method. Both methods appear to work. However, the number of litters obtained from the Rall and Fahy method was probably due to the lack of penetration of cryoprotectant into ovarian tissue.
The cause of death is important. In these studies mice were euthanized by cervical dislocation. In transgenic colonies, death may be 'natural' and factors such as poor blood flow, declining body temperature, etc., may affect results. In addition, when mice with lymphoma were grafted, then a similar neoplasia developed in recipients.
Rescue of valuable mutant strains can be accomplished by cryopreservation of ovaries that are retrieved within 2 hours of death.
Questions: 1. True/False: Spermatozoa recovered as long as 24 hours after death from dead mice and stored at 22 degrees Centigrade can be thawed and can fertilize oocytes.
2. True/False: Oocytes can be obtained from dead mice as long as 2 hours after death, cryopreserved, thawed, and transplanted into similar mice to obtain fertility.
3. Name two methods of cryopreservation and identify what differentiates them.
Answers: 1. True
2. True
3. 1). Szteins method: uses rate controlled freezing; time consuming
2. Rall and Fahy: does not use rate controlled freezing.
Exclusion of mouse hepatitis virus from a filtered, plastic rodent shipping container during an in transit field challenge. Contemporary Topics 40(4), 32.
Abstract: This study was developed to test and verify the practice of using filtered shipping boxes to protect animals from contamination during transport. Although this practice is currently a standard accepted practice in lab animal science, no information has been available in the literature to demonstrate that any of these boxes actually accomplish this task.
Twelve (12) plastic shipping boxes with filters and tight-fitting lids were used as test shipping containers and six (6) cardboard shipping boxes without filters were used as control shipping containers. Each container was stocked with 10 x 10-12 week old female, adventitious disease-free Swiss Webster mice from Taconic Labs. All 18 shipping boxes and their contents were sterilized prior to placement of the animals. The boxes were then transported to a facility that housed a breeding colony of conventional mice enzootically infected with four murine viruses - mouse hepatitis virus (MHV), mouse parvovirus (MPV), mouse encephalomyeltis virus (GDVH) and epizootic diarrhea of infant mice (EDIM). The shipping boxes were placed on the floor on the conventional mouse room overnight (15 h) then transported to a commercial testing laboratory, at which the animals were aseptically removed and were housed in microisolation cages. After 28 days, the animal's sera was tested for antibody to all four murine viruses using ELISA as a primary test and IFA retesting for any samples that yielded high background noise to the control sample.
All serum samples from mice held in the 6 non-filtered cardboard control boxes were positive for MHV, although negative for MPV, GDVH and EDIM.. Sera from all mice held in the plastic filtered boxes were negative for antibody to any of the four viruses.
This study demonstrates that at least one type of filtered shipping container protects mice from a field challenge of MHV. To the best of our knowledge, this is the first documentation of any microbial efficacy testing conducted on filtered shipping containers for laboratory animals.
Questions: 1. Why did the control mice sero-convert only to MHV?
2. By what mechanism does one virus interfere with subsequent infection by a second virus?
3. Can one assume all filtered shipping containers are protective against contamination during shipping procedures?
4. Second to the integrity of the shipping container, what is one other very important step in animal transport that provides protection
from contamination exposure?
Answers: 1. Several reasons can be factors: #1 MHV is one of the most infectious murine viruses; #2 MHV can interfere with infection by other murine viruses (at least one); #3 possibility of inadequate exposure to the other three viruses in the environment used in the experiment.
2. Interferon
3. No, since protection would be dependent upon the filter medium used, the number of animals per shipping container, and how tightly the container could be sealed.
4. Removal of the animals from the container requires appropriate decontamination procedures for the exterior of the shipping container and scrupulous aseptic technique during housing.
The effects of overnight fasting, feeding or sucrose supplementation prior to necropsy in rats. Contemporary Topics 40(4), 36.
Abstract: Male or female rats were either given no food, regular chow, or sugar cubes 17 hours prior to CO2 euthanasia. Necropsy was preformed and histology was prepared and a number of Clin Path Parameters were compared between the three groups. The most remarkable finding in sucrose fed animals was increased glycogen storage in the liver. Other affected parameters included a higher neutrophil count in sucrose fed animals, Blood glucose ranged 3.0-6.1mM/L highest in sucrose fed animals, lowest in fastened animals. BUN ranged from 4.6 - 20.2 mg/dL highest in fed animals lowest in sucrose fed animals, Alkaline Phospahatase ranged from 91-236 U/L, reference range 34-353 U/L was lowest in fastened animals and highest in Chow fed animals, Alanine Aminotransferase (ALT) ranged from 38-89 U/L, reference range 13-74 U/L highest in sucrose fed animals lowest in fastened animals.
Questions: 1. What was the most significant histological effect of 17 hours of sucrose feeding in rats prior to necropsy?
2. 17h fasting puts BUN, ALP and ALT outside reference values. T or F
3. Blood glucose values were ______ in sucrose fed animals
Answers: 1) Massive deposition of glycogen
2) False
3) highest
Postnatal development of penile NADPH diaphorase in male rats (Rattus norvegicus): an indicator of erectile function. Contemporary Topics 40(4), 41.
Abstract: NO is considered to be one of the neuromodulators of penile erection by causing cavernous smooth muscle relxation. Nicotinamide adenine dinucleotide phosphate diaphorase (NADPH) is one enzyme involved in the synthesis of NO. Staining of biopsy specimens for NADPH has been used as a diagnostic tool for neurogenic impotence. In this study, immunohistochemistry was performed on penile tissues of rats ranging in age from 1-65 days of age. Staining was selective for NADPH and the concentration of NADPH was quantified by counting the number of positive granules per section. The concentration of NADPH increased from 0-200 in the corpus cavernosum and from 0->800 in the dorsal nerve. The greatest increase was seen in rats between 30-50 days of age which correlates with the age rats develop erections. NADPH along with other known neuromediators (vasointestinal peptide [VIP], calcitonin gene-related peptide [CGRP] and neuropeptide Y [NPY]) modulate penile erection in rats and other species. NADPH, VIP and CGRP cause tumescence and NPY causes detumescence.
Questions: 1. What is the name of the enzyme that produces NO?
2. At what age do rats develop the ability to produce erections?
Answers: 1. Nicotinamide adenine dinucleotide phosphate diaphorase
2. 30-40 days of age
Chronic myelocytic leukemia in a juvenile Rhesus macaque (Macaca mulatta). Contemporary Topics 40(4), 44.
Abstract: A juvenile rhesus macaque was part of a study of the effects of subclinical lead exposure. The animal was administered
daily doses of lead acetate from 8 days post partum until 1.5 years of age to achieve a target blood lead concentration of about
35ug/dl. Biweekly routine blood draws were performed. At 25 months old, an abnormal hemogram was noted. The animal had a
leukocytosis with an increase in immature white blood cells (band cells, metamyelocytes, promyelocytes, blast cells). 1 week later,
serum was tested for antibodies to simian retrovirus (SRV), simian immunodeficiency virus (SIV), and simian T-cell leukemia virus
(STLV-I) of which all were negative. Bone marrow biopsy confirmed an increased number of neturophil precursors. Since the
original finding, weekly blood draws showed a continued rise in WBCs including blasts and a decline in RBCs. Chemotherapy
(25mg/kg hydroxyurea) was initiated 2 months after the original abnormal finding which improved the situation for a very short time (a
month). However, the rebound was severe and the animal developed facial swelling and petechial hemorrhages of the skin and sclera
and was euthanized. On the final blood draw, the animal had 95,000 WBC/ul with 67% blast cells and was anemic. Also at
euthanasia, plasma and WBCs were examined for SRV, SIV and STLV by coculture and PCR (all negative). The animal was
seropositive for Epstein-Barr virus (EBV)- like agents. Tumor cells from the liver were anyalyzed by PCR for bcr-ABL gene (to detect
the chromosomal abnormality associated with chronic myeloid leukemia in human patients) which was absent, but the result was
inconclusive since the primers were for a human sequence. The blood lead conc. was compared to age-matched lead-treated females
and was not statistically different.
Discussion: Chronic myeloid leukemia (CML) is defined (in part) in humans by the appearance in the peripheral blood of immature cells
of myelocytic origin and is typically treated by chemotherapy and/or allogeneic bone marrow transplantation. The production and
release of abnormal RBCs is also a common feature. In adults, the ABL gene normally located on Chr 9 is translocated to Chr 22 and
is dysregulated, becoming an integral part of the pathogenesis of most forms of CML.
In monkeys, SIV and STLV-1 have been associated with hematologic neoplasias. EBV-like agents have been associated with
lymphomas and could not be completely ruled out, but it was stated that epidemiological factors make it an unlikely etiology in this
case. However, the authors strongly suggest that the exposure to lead may have played a role in the development of myeloid neoplasia
which is previously unreported in macaques.
Questions: 1. Name three retroviruses that have been associated with immunodeficiency or neoplasia in macaques.
2. Which two retroviruses have been associated with lymphoid neoplasia in non-human primates?
3. True or False:
a. EBV is a herpesvirus
b. Infection with some form of EBV-like virus is ubiquitous in many species of primates, including humans
c. All viral variants of EBV appear to cause disease in the host.
4. Lead poisoning is known to cause chronic myeloid leukemia in humans exposed as neonates
Answers: 1. Simian Immundeficiency virus (SIV), Simian T-Lymphotropic virus (STLV) and Simian Retrovirus (SRV)
2. SIV and STLV
3.
a. True
b. True. EBV is a ubiquitous herpesvirus, limited primarily to B lymphocytes and nasopharyngeal cells of humans and NHPs
c. False
4. False
Unexpected deaths in young New Zealand White rabbits (Oryctolagus cuniculus). Contemporary Topics 40(4), 49.
Abstract: Case report-young 12 week old rabbits (1.5-2.0 kg) were received from a reputable vendor into a conventional rabbit facility. Five weeks later 4 out of 6 rabbits died within a 48 hr period. The surviving 2 rabbits were euthanized and necropsied. All 6 rabbits had green hepatic miliary abscesses and bile duct proliferation (hyperplasia). Ellipsoidal oocysts were present in the bile duct lumen and walls. The parasitic oocysts were identified as Eimeria steidae which have an ellipsoidal shape and a typical size of 37 X 21 um. The research facility was determined to be the source, since it had previously received infected rabbits and had never effectively sanitized their housing area. Routine disinfection was not adequate. Key points of paper: 1. Fecal examination- wet mount or fecal flotation will detect the oocysts. 2. Coccidiosis is one of the most common infections in non-laboratory grade, but commercial, pet and wild colonies. Many species of coccidia parasitize the GIT of rabbits. Only E. steidae parasitized the liver. 3. Life cycle is as follows: * Fecal unsporulated oocyst are shed into the environment---> they sporulate and are ingested--->they penetrate the GIT mucosa then migrate to the liver---->colonize the bile ducts (schizogony commences and then gametogony produces the unsporulated oocysts) which become hyperplastic--->infected host cells rupture and release the unsporolated oocysts into the bile ducts----> feces. * The prepatent period is 15-18 days.
Questions: No questions