Medetomidine-ketamine anesthesia in red-eared slider in turtles (Trachemys scripta elegans). Contemporary Topics 40(3), 08.
Abstract: The objective of the study was to evaluate whether an IM administered anesthetic combination could produce an adequate level of anesthesia for the examination of and/or performance of minor procedures in Red-Eared Slider turtles. Such an approach would be useful to avoid some of the difficulties inherent in anesthetizing aquatic turtles. Such difficulties include their unique physiology (can hold their breath for long periods of time making inhalant induction difficult and time consuming), slow metabolic rate (results in prolonged recovery times, from hours to days) and ability to withdraw into shell. Thirty adult Red-eared Slider turtles were divided into two groups. One group received a low dose (LD) combination of medetomidine (0.1 mg/kg) + ketamine (5 mg/kg). The other group received a high dose (HD) combination of medetomidine (0.2 mg/kg) + ketamine (10 mg/kg). Both were delivered IM to a forelimb. Turtles were monitored for heart rate and rhythm (via ultrasonic Doppler), palpebral reflex and skeletal muscle relaxation at 15 min. intervals until turtles were responsive and resisted manipulation. Cloacal temperatures were monitored throughout. At the 45 minute point each turtle was evaluated for responsiveness to an IM injection (normal saline) and endotracheal intubation. The high dose group also underwent evaluation to a small skin incision and suture placement. Anesthesia was reversed with Atipamezole (0.5 mg/kg for LD animals and 1.0 mg/kg for HD animals) at 60 minutes after MK injection. There was no significant difference between either group for heart rate or skeletal muscle relaxation score until 75 minutes after MK injection. Cloacal temperatures remained the same throughout. Palpebral scoring and hindlimb relaxation differed between groups except at one time point (T45). LD turtles recovered significantly faster (were able to return to the water 30 min. after atipamezole) than HD turtles (60 min. after atipamezole). LD turtles did not undergo skin incision and suturing due to resistance to IM injection and endotracheal intubation at the time of evaluation. It was determined that even the high dose of medetomidine / ketamine would be unsuitable for invasive surgical procedures. In preliminary trials, neither low doses of medetomidine or ketamine alone produced satisfactory anesthesia. The combination of low doses of medetomidine and ketamine was shown to produce reliable anesthesia with rapid recovery following reversal with atipamezole and presents a suitable alternative to high dose ketamine anesthesia in turtles. This combination is an easy and suitable induction regime to inhalant anesthesia for more invasive procedures. This represents a significant advantage to use of high dose (20-80 mg/kg IM) ketamine for restraint and minor procedures.
Questions: 1. What class of drug is medetomidine?
2. What difficulties are inherent in anesthetizing chelonians?
Answers: 1. Medetomidine is an alpha2-adrenergic agonist sedative with some analgesic properties. It is 10 times more specific for alpha2 recepetors than ketamine, making it more selective and causing less stimulation of alpha1 and non-alpha2 adrenoreceptor sites.
2. In addition to difficulty in handling (they withdraw head and limbs into their shells), turtles are capable of holding their breath for very long periods of time. This renders impractical almost any form of induction with an inhalant anesthetic. Also, they have very slow metabolic rates and injectable anesthetics take a very long time to be metabolized (reported recovery times following high doses of ketamine range from 24 to 96 hrs.) This makes it very difficult to recover and return the animal back to its normal aquatic environment following even minor procedures.

Evaluation of a transdermal fentanyl system in Yucatan miniature pigs. Contemporary Topics 40(3), 12.
Abstract: Fentanyl (N-phenyl-M-(1-2-phenylethyl-4-piperidyl) propanamide) patches (Duragesic TTS) were applied to miniature swine and blood samples obtained to determine the serum concentration 0- 96 hours following application. Fentanyl interacts with m (mu) receptors in the brain, spinal cord and other tissues and is 75-100 times more potent than morphine. Patches have several advantages over injectable analgesics; continuous controlled delivery of drug for up to 72 h, minimal peaks and troughs of serum concentration, elimination of discomfort and inconvenience of repeated oral or parenteral administration, reduction of first pass hepatic clearance, decreased labor costs, and decreased stress associated with repeated handling. Disadvantages include; self removal which can result in ingestion and potentially overexposure (although fentanyl is not absorbed well in the stomach because of the acid concentration, it is absorbed in the intestines), and the absorption rate can be affected by body temperature (>40C can increase absorption by 1/3). As with injectable fentanyl, the patches can result in hypoventilation and respiratory depression and constipation. Many in the medical community including one of the manufacturers (Janssen Pharmaceutica) strongly discourage its use for postoperative analgesia in humans. The authors of this manuscript believe that the complications can be addressed with appropriate nursing care. The amount of fentanyl released is proportional to the surface area contacted by the patch, 25ug/h per 10 cm2. The effective serum concentration is similar in dogs, cats and humans (0.2-3 ng/ml plasma) and the swine dose is believed to be in the same range. As much as 44-84% of the original fentanyl can remain in the patch after its 3 day application. Therefore, attention should be directed at proper disposal. The article stated that the proper method of disposal was to fold the patch in half and flush down the toilet.
Questions: 1. Which of the following are advantages for the use of fentanyl patches for postoperative analgesia?
a. Continuous administration for 72 hours with minimal peaks and troughs in serum concentration.
b. Decreased stress associated with repeated handling
c. Decreased labor costs for daily treatment
d. All of the above.
2. What are the complications associated with the used of fentanyl patches for postoperative analgesia?
3. What is the proper method for patch disposal?
Answers: 1. D
2. respiratory depression and hypoventilation
3. fold in half and flush down the toilet.

Antinociceptive activity of clinical experience with buprenorphine in swine. Contemporary Topics 40(3), 17.
Abstract: The authors examine the effectiveness of buprenorphine as an analgesic for swine by performing an objective study using antinociceptive testing and a retrospective review of clinical records. In the nociception study, baseline skin-twitch latency to a focused light source was determined for each pig. The pigs were then divided into 3 treatment groups receiving the following doses of buprenorphine IV: 0.005, 0.01 and 0.02 mg/kg. The skin-twitch latency to a focused light source was then measured at the following time points: 15, 30, 60, 120, 180, 240, 300, 360, 420, 480, 540 and 600 minutes. In an initial portion of the study, the authors used two higher doses of buprenorphine (0.08 and 0.16 mg/kg). However, with the higher doses, none of the latency measurements were below the 40 second cut-off time so lower doses were used for the rest of the study. The authors found that there was no significant increase in skin-twitch latency with the lowest (0.005 mg/kg) dose showing that this dose is too low to provide adequate anesthesia for swine. The next lowest dose (0.01 mg/kg) resulted in increased latency up through the 300 minute time point indicating that this dose provides close to 6 hours of analgesia for swine. This is a shorter duration than the authors' clinical dosing of 0.01 mg/kg every 8 to 12 hours. The pigs given the 0.02 mg/kg dose had increased latency at all time points except at 480 minutes indicating that this dose could be used to provide approximately 10 hours of analgesia. The authors also retrospectively examined a database containing 32,500 medical records, of which 1,757 represented 264 swine with 936 individual problems requiring the use of buprenorphine. The authors examined the records for evidence of failure of buprenorphine to provide adequate analgesia. They specifically looked for clinical observations indicating signs of pain in the animal or situations where the analgesia regimen was changed presumably because the buprenorphine did not work. The authors ultimately found just 18 cases of treatment failure out of 416 where swine were treated bid or tid with 0.01 mg/kg iv or im. This is a failure rate of 4.3% which leads the authors to conclude that the clinical use of 0.01 mg/kg iv or im, bid or tid provides adequate analgesia in the clinical setting. The authors do note some exceptions to this including cases that involve inflammation, systemic disease or organ failure.
Questions: 1. Buprenorphine is classified as what type of drug:
a) opiate
b) alpha-2 antagonist
c) partial opiate agonist
d) nonsteroidal antiinflammatory
e) partial opiate antagonist
2. Buprenorphine exhibits its primary activity at which opiate receptor:
a) kappa
b) epsilon
c) sigma
d) mu
e) delta
3. The dental formula for a pig is:
a) 2(I 0/3, C 0/1, P 3/3, M 3/3)
b) 2(I 3/3, C 1/1, P 4/4, M 2/3)
c) 2(I 3/3, C 1/1, P 4/4, M 3/3)
d) 2(I 3/3, C 1/1, P 3/3, M 3/3)
Answers: Answers: 1. c 2. d 3. c

Floor space needs for laboratory mice: BALB/cJ males or females in solid-bottom cages with bedding. Contemporary Topics 40(3), 21.
Abstract: This study compares the effect of 3 different cage space allowances for male and female BALB/cJ. Three mice of each sex were given 32.2, 96.9 and 129 cm2/mouse respectively. Animals housed in 1/3 of the space that is recommended by the Guide showed no adverse effects of the reduced space. The mice may have and in fact benefited from the "comfort" of less space. Female mice housed in the smallest amount of space were heavier than in the other 2 groups. No mortalities were observed in the 32.2 cm2 group while 1 or 2 mice died in each of the other groups. No abnormal amount of aggression was observed in any group. Females in the 32.2 cm2 group exhibited more sitting and grooming behaviors. The only immune response affected by the grouping was an increase in lymphocyte proliferation under 20mg/mL PHA (phytohemagglutinin). The authors conclude that floor space as limited as 32.2cm2 does not cause behavior, health, immune or performance problems for BALB/c J mice.
Questions: 1. According to the Guide, what is the recommended floor space/mouse for adult mice?
2. What is the effect of giving adult BALB/cJ one third of the floor space recommended by the Guide?
Answers: Not provided

Mealworm feeders as environmental enrichment for common marmosets. Contemporary Topics 40(3), 26.
Abstract: Insects make up a considerable portion of the diet of free-ranging common marmosets. This study examined the impact of a foraging
enrichment device, the "mealworm feeder," on the behavior of the common marmoset. Each marmoset was observed with a mealworm feeder on separate days in consecutive weeks. The mealworm feeder was mounted near the upper perch in the cage. Behavior was observed immediately after placement and again after 3 hours. Behaviors scored consisted of six categories: 1) feeder use, 2) use of other enrichment devices (ladder, swing,didhew toy), 3) sitting on the bottom of the cage, 4) sitting still, 5) stereotyped pacing and 6)play. Females demonstrated greater overall use during hour 0 than did males. Greater frequency of use was shown by subadults in family groups at 0 hour and continued at an intermediate rate at 3 hours at which time adult females and males approached zero use. Significantly decreased rates of pacing and time spent sitting when observed in association with the placement of the mealworm feeder. There was an increased use of other enrichment devices which was comprised primarily of cage furniture when the mealworm feeder was present.. The mealworm feeders did not increase the use of vertical space in the cage. Singularly housed female marmosets used the mealworm feeders more often than males; however, subadults used the feeders significantly more often than dominant females or males in family groups. The results of this study suggest that the mealworm feeder is an effective form of environmental enrichment for the common marmoset, but interest decreases after a 3 hour period.
Questions: 1. The mealworm feeders were used by which group with the greatest frequency during the entire 3 hour period?
2. What two types of stereotyped behavior were decreased when the mealworm feeder was placed in the cage?
3. The mealworm feeder did not increase ______________ use of the cage when the mealworm feeder available as an enrichment device.
4. Other enrichment device use was increased when the mealworm feeders were in the cage. True or False
Answers: 1. Subadults
2. pacing and sitting
3. Vertical space
4. True

Monkeys are frequently used as animal models of various human conditions; thus, the investigators found it of value to measure concentrations of ascorbic acid in a population of rhesus macaques to assess variations due to sex, age, and physiologic state versus humans. This short paper was aimed at publishing these values and comparing them to the same parameters in humans. Their findings may be summarized as follows: 1. Among rhesus macaques, no differences in ascorbic acid were noted between sexes. This contrasts with humans, where ascorbic acid concentrations in women of reproductive years are greater than men. 2. Pregnant and nonpregnant rhesus had similar ascorbic acid values. Values for lactating primates were slightly higher. This compares similarly to humans. 3. Rhesus exhibited an age-related decrease in ascorbic acid; juveniles had concentrations greater than "old" macaques (7-14 yrs), which were greater than "very old" macaques (16-20 yrs). This finding is similar to that of humans. Plasma ascorbic acid concentrations in a population of rhesus monkeys (Macaca mulatta). Contemporary Topics 40(3), 30.
Abstract: Monkeys are frequently used as animal models of various human conditions; thus, the investigators found it of value to measure concentrations of ascorbic acid in a population of rhesus macaques to assess variations due to sex, age, and physiologic state versus humans. This short paper was aimed at publishing these values and comparing them to the same parameters in humans. Their findings may be summarized as follows: 1. Among rhesus macaques, no differences in ascorbic acid were noted between sexes. This contrasts with humans, where ascorbic acid concentrations in women of reproductive years are greater than men. 2. Pregnant and nonpregnant rhesus had similar ascorbic acid values. Values for lactating primates were slightly higher. This compares similarly to humans. 3. Rhesus exhibited an age-related decrease in ascorbic acid; juveniles had concentrations greater than "old" macaques (7-14 yrs), which were greater than "very old" macaques (16-20 yrs). This finding is similar to that of humans.
Questions: 1. Which of the following is true regarding a recent study of ascorbic acid concentrations in rhesus macaques?
a. Among rhesus macaques, there was no difference in ascorbic acid concentrations between sexes.
b. Among rhesus macaques, ascorbic acid concentration was greater in females than males.
c. Rhesus and humans exhibit similar trends in ascorbic acid concentration related to sex, with males greater than females.
d. None of the above.
2. T/F. A recent study concluded that rhesus exhibit an age-related increase in plasma ascorbic acid concentration.
3. T/F. A recent study concluded that ascorbic acid concentration is slightly higher in lactating rhesus versus pregnant or nonpregnant rhesus.
Answers: 1. a 2. F 3. T

Specific gravity of whole blood in cynomolgus monkeys (Macaca fascicularis), squirrel monkeys (Saimiri sciureus), and tamarins (Saguinus labiatus) and total blood volume in cynomolgus monkeys. Contemporary Topics 40(3), 33.
Abstract: Due to animal welfare concerns, the specific gravity of whole blood (SGB) and total blood volume (TBV) are two essential hematologic parameters when establishing quidelines for frequency and maximum volume of blood to be withdrawn during animal experiments. Previously, recommendations regarding blood sample frequency and maximum volume have been established for nonhuman primates (NHPs) by using a radiolabelling technique. The purpose of this study is to clarify the normal limits of SGB and TBV in various nonhuman primate species by using simple methods. SGB measurements were determined by dropping samples of whole blood into a graded series of copper sulfate solutions of known specific gravities and noting whether drops rose or fell in each solution. TBV was measured by the Evans blue dye dilution method. The TBV was obtained after correcting for the hematocrit and the dilution factor of the Evans Blue solution. The values for the mean SGB +/- 1 standard deviation are: cynomolgus monkeys, 1.0526 +/- 0.0037; squirrel monkeys, 1.0555 +/- 0.0037; and tamarins, 1.0582 +/- 0.0020. Formulae were established to estimate TBV by using body weight (BW). The estimated TBV was approximately 6% of the BW and is within a range reported for mammals. An exception was noted in the correlation between BW and TBV in the case of obese animals due to fewer blood vessels in fat. The data presented is important when establishing NHP blood withdrawal guidelines in which the maximum volume is based on the TBV. Both hematocrit and SGB should be monitored when establishing the frequency of blood withdrawal.
Questions: 1) What are two important hematologic parameters when determining guidelines for frequency and maximum volume of blood to be withdrawn during animal experiments?
2) The estimated total blood volume was ________ percent of the NHP body weight.
3) Obesity will influence the correlation between BW and TBV - True or False.
Answers: 1) TBV and SGB
2) 6
3) True

A practical dog bed for environmental enrichment for geriatric beagles, with applications for puppies and other small dogs. Contemporary Topics 40(3), 36.
Abstract: The author describes dog bed fashioned from high-density polyethylene barrels. The barrels were originally filled with liquid cleaners and were slated for recycling after cleaning. A jigsaw was used to cut off the top of the barrel, leaving the bottom for a bed.
The primary reason these beds were constructed was due to the fragile nature of a geriatric beagle colony. Although perches were provided for these dogs, many were unable to safely use them. Thus, this bed provided a safer option. Bed liners of fleece were chosen after evaluating rubber matting, carpeting in addition to the fleece.
The author has also used these beds for weaning puppies and other small convalescent animals. Anecdotal information is provided regarding the beagles' acceptance of these beds. The author suggests that a formal study should be performed to actually quantitiate bed use and its' place in environmental enrichment.
Questions: 1. What is the space requirement of the dog according to:
a. The Guide
b. AWA
2. What is the height requirement of the dog according to:
a. The Guide
b. AWA
3. True/False: The bench in a cage is never included in computing floor space.
Answers: 1a. < 15 kg 8 sq. ft
up to 30 kg 12 sq. ft
> 30 kg > 24 sq. ft
1b. "minimum square feet of floor space be equal to the mathematical square of the sum of the length of the dog in inches (measured from
the tip of its nose to the base of its tail) plus 6 inches; then divide the product by 144.
2a. No specific height stated. The dog should be able to assume normal behavior and postural adjustments.
2b. "The height should be sufficient to allow occupant to stand in comfortable position".
3. False - The Guide: pg. 26
"Low resting surfaces that do not allow the space under them to be comfortably occupied by the animal should be counted as part of the
floor space."

A simple transport system for radiation treatment of specific pathogen free mice in lifetime studies. Contemporary Topics 40(3), 39.
Abstract: Wanted to take animals to a potentially contaminated irradiator and return them to a clean facility. Also, did not want to handle the
animals while they spent 2 hours in the irradiator.
Came up with a polycarbonate jar with the following specs:
Look at the picture on page 40. The transport vessel is a modified round, straight-sided, wide-mouth gray polycarbonate jar with standard 70-mm Mason jar threads To correctly position the mouse in the gamma field, a polyethylene plug was placed in the bottom of the jar. No bedding was placed in the vessel: urine runs off the polyethylene plug and accumulates at the bottom of the jar, so the animal remains dry; feces remain on the plug. Air exchange was made possible by cutting a circular opening in the top of the plastic cap. A disc of the same filter-top material (Reemay) used on the cages was placed inside the cap; the filter-top material provides a barrier preventing particulate dust from entering the vessel. The filter was held in place with a poly-vinyl chloride (PVC) disc perforated with 6.5-mm diameter holes spaced 3 mm apart (about 46 holes per disc). The PVC disc protects the filter-top material from being damaged by mouse chewing, keeping the mouse safely contained while allowing air exchange. A small card with strain and treatment notations can be placed over the filter disc before it is screwed into place. All vessel components can be washed, disinfected, and reused. The polycarbonate jars are soaked in quaternary ammonium disinfectant, rinsed, and dried in a clean, SPF environment or are sanitized in a cage washer. Lid components are disinfected in 70% ethanol and dried in a clean fume hood.
Questions: No questions

Immunohistochemistry. Contemporary Topics 40(3), 43.
Abstract: Note- this is a short one page article and the figures are the meat of it. Take a look at it.
Immunohistochemistry (IHC)
Antigen- antibody technique used to identify antigens in histologic sections of tissues.
Uses- identifying cell surface markers to identify cell type, the cellular location of proteins (originating from host or pathogen)
Step 1:
Fixed or frozen sections are cut 4-6 microns thick and placed on a charged glass slide
Step 2:
Direct method
Antigen<--Primary Antibody=Reporter Molecule
Detect signal by color reaction of reporter molecule at antigen site
Indirect
Antigen<--Primary Antibody<--Secondary antibody=Reporter Molecule
Primary binds to antigen and the primary antibody is detected by the secondary antibody which is conjugated to an enzyme, colloid gold particle, or radiolabeled with and isotope
Indirect using avidin-biotin to increase sensitivity by amplifying the detection signal
Antigen<--Primary Antibody<--Secondary
antibody=Biotin<--Avidin<--(x3) Bitotin=Reporter Molecule
So 3 biotin w/ reporter molecules are bound for each primary antibody.
Signal can also be amplified by conjugating secondary antibody and the reporter molecule to a polymer backbone. This can incurs the sensitivity by up to 100 fold (depending on how many reporter molecules are on the backbone)
Step 3: Evaluate immuno stained slides by light microscopy for the location of the antigen of interest
Advantages- cellular localization of the antigen
IHC can be sensitive and specific
Cost efficient and rapid
Disadvantages-Fixation method (formalin) can interfere w/ antigen and prevent antibody binding
Antigen may be disrupted during fixation and require retrieval; steps
Endogenous molecules present in tissues may interfere with enzymatic steps and results in false negatives
Non specific background staining can cause false positives
Optimization can be labor intensive
Alternate methods- in situ nucleic acid detection (PCR, RT-PCR, fluorescent in situ hybridization)
Questions: 1.The use of an avidin-biotin complex in IHC increases the ___________ of the procedure.
2. When collecting tissues for IHC formalin is the fixative of choice (T/F).
3. IHC can be used to detect exogenous proteins only (T/F)
Answers: 1.The use of an avidin-biotin complex in IHC increases the sensitivity of the procedure.
2. When collecting tissues for IHC formalin is the fixative of choice (False).
3. IHC can be used to detect exogenous proteins only (False)