Contemporary Topics 43(1)

Effects of Freund’s Complete Adjuvant on the Physiology, Histology, and Activity of New Zealand White Rabbits  pages 8-13
Purpose of study: To test the hypothesis that Freund’s Complete Adjuvant would not adversely affect the well-being of research rabbits.
Background: Freund’s complete adjuvant has been used for decades to stimulate polyclonal antibody production in rabbits. In 1997 the USDA discouraged the use of FCA due to the potential to cause pain and distress. The study presented here is based upon a pilot study investigating the effects of FCA on physiologic and behavioral parameters.
Materials and Methods: 32 NZW rabbits were divided into three groups to evaluate serum concentration of corticosterone, activity, food consumption, body weight, body temperature, WBC count, degree of induration around injection sites, antibody titers, and histopathology. Dorsum was shaved a prepared before compounds were administered subcutaneously and intradermally.
The three groups were:
Group 1: Immunized with Freund’s homogenized with keyhole limpet hemacyanin (KLH), then boostered 28 days later with Freund’s incomplete adjuvant (FIA) with KLH
Group 2: Immunized with KLH mixed with saline for initial and booster immunization with FIA.
Group 3: Control group -  no immunizations
Results: 
Body weight: all rabbits gained wt at an equivalent rate
Body temp: all rabbits had temps within normal range
Food consumption: no difference between groups
Activity: varied between individuals, but not groups
WBCs: higher for rabbits in group 1, but still within range for normal rabbits
Corticosterone: no significant difference in mean concentrations among the groups
Induration: group 1 had marked induration (but no ulcers or abscesses. Other groups had none.
Antibody response: Both immunization groups (1&2) developed antibody responses to KLH. Group 1 was 10x higher than group 2. Control rabbits did not have titers.
Histologic changes: All experimental groups had pulmonary lesions were proliferation of bronchial-associated lymphoid tissue, microgranulomas, mixed cellular accumulations, aggregations of heterophils, and interstitial hemorrhage. In group1 granulomatous inflammation was found at the injection sites of  11/12 rabbits.
Discussion: Female rabbits were chosen due to tractability, ability to mount more vigorous immune responses, and are more sensitive to lower doses of antigen. No fever or ulcers/abscesses indicate that technique was sterile. The use of Freund's in rabbits should not be discouraged. 
Questions:
1.How are immunogens prepared?
2. How does Freund’s differ from Freund’s incomplete antigen? 
3.KLH stands for? 
Answers:
1.by mixing KLH with adjuvant or saline between two glass syringes connected by a stopcock until homogenized. 
2.Freund’s has the mycobacterial cell wall components. 
3.keyhole limpet hemacyanin

Comparison of Environment and Mice in Static and Mechanically Ventilated Isolator Cages with Different Air Velocities and Ventilation Designs pages 14-20
 	The purpose of the study is to compare environmental conditions of 5 different mouse caging designs.  The designs consisted of 4 homemade ventilated caging systems and 1 static microisolator cage (all cages are shoebox style from Ancare).  The conditions that were evaluated were air velocity, temperature, relative humidity, bedding weight change, airborne dust, NH3, and CO2.   For each cage system, 5 mice were housed.
 	The 4 different ventilated cages consisted of two variations in mechanical ventilated supply air velocities (40 or 200 feet/min with air volume exchange rate of 9.3 L/min each) and two variations in air exhaust systems.  Exhaust air was either forced through a filter in the micro-isolator lid or air was forced through a single exhaust port located in narrow side of cage top.       
 	Air velocities were measured using an omni-directional probe, relative humidity with a thermohygrometer, airborne particle size with a laser particle counter, CO2 with an infrared analyzer, and NH3 with a Matheson-Kitagawa pump.  Cage locations of behaviors and bedding mounds were also recorded.
 	Relative to ventilated caging styles, the static isolator caging had higher mean levels of relative humidity, CO2 and NH3 concentrations.  These intracage parameters generally related more to air exchange rate than ventilation design or velocity.  In general, it was noted that when mean intracage NH3 levels are high (> 25 - 50 ppm), there is low air exchange, high air relative humidity, and high bedding moisture content.  Another factor that should be associated with NH3 levels in mouse cages is bedding pH.   In this study, chipped hardwood bedding was used and it is not as effective as ground corncob bedding for controlling NH3 emissions.
 	When compared with mice in mechanically ventilated cages, the water consumption and body weight gain of the mice housed in the static isolator cages were lower.  
Temperatures, particulate levels and feed consumption did not differ significantly between cage types. Locations of bedding mounds and behaviors were similar in all cage treatments.        
Questions:
1) When comparing mechanically ventilated caging to the static isolator caging, which of the following statements regarding environmental parameters is true: 
a.	Static microisolator had higher air velocities than mechanically ventilated caging 
b.	The microenvironmental temperature in the static caging was significantly higher than that of the ventilated cages 
c.	The static caging style had increased measurements in NH3, relative humidity, and CO2, relative to mechanically ventilated caging 
d.	In the mechanically ventilated cage, body weight gain and water consumption were lower as compared to the mice housed in the static cages
2) T/F  Chipped hardwood bedding is more effective than corncob bedding for controlling NH3 emissions.  
Answers:
1) Answer: C
2) Answer: False.  Corncob is more effective than hardwood bedding for controlling NH3 emissions.

Comparison of Fertility Between Intracytoplasmic Sperm Injection and In Vitro Fertilization with a Partial Zona Pellucida Incision by Using a Piezo-Micromanipulator in Cryopreserved Inbred Mouse Spermatozoa pages 21-25
Summary: 
Embryo cryopreservation is one solution for preserving mouse strains.  Unfortunately, 200 early-pregnancy mice are needed to generate the requisite 500 embryos.  Thus, a colony must be expanded before it can be reduced.  Another solution is cryopreservation of spermatozoa.  There are considerable strain differences in fertilization rates using frozen-thawed spermatozoa.  Cryopreserved BALB/c and C57BL/6 spermatozoa have poor fertilization rates.  The authors recently reported the assisted reproductive technique (ART) of intracytoplasmic sperm injection (ICSI).  In this manuscript, they compare ICSI and conventional in vitro fertilization (IVF) with a different technique:  in vitro fertilization with a partial incision of the zona pellucida by a piezo-micromanipulator (ZIP/IVF).  For BALB/cA, C57BL/6, and B6C3F1 cryopreserved spermatozoa, the fertilization rates of manipulated oocytes by ZIP/IVF were higher than for ICSI.  Embryonic development to term for oocytes fertilized by cryopreserved spermatozoa was not significantly different between ZIP/IVF and ICSI for C57BL/6 and B6C3F1.  The overall efficiency of mouse production in ZIP/IVF was higher than for ICSI and conventional IVF in C57BL/6 and B6C3F1.  ZIP/IVF requires 3.3 times less manipulation than ICSI.  The authors propose that ZIP/IVF is a useful assisted reproductive technique for IVF of ova by cryopreserved spermatozoa.
Questions: 
1.      BALB/c and C57BL/6 cryopreserved spermatozoa have ________ fertility. 
a.      high 
b.      low 
c.      average compared to other inbred strains 
d.      no 
2.      True or False:  In the mouse, in vitro fertilization with a partial incision of the zona pellucida by a piezo-micromanipulator (ZIP/IVF) requires less manipulation than intracytoplasmic sperm injection (ICSI).
Answers: 
1.      B 
2.      True 

Cryopreservation Methodologies:  Freezing Location and Protocol Alter Development of Mouse Embryos,  page 26-31
The authors evaluated three different freezing protocols for preservation of two cell mouse embryos, and how the position of the embryos in the cooling chamber affected their successful thawing and development.  The freezing chamber had a round configuration with liquid nitrogen entering at the rear of the chamber near the 12 o’clock position.  Generally one would assume uniform temperature within the chamber, but the authors found that there was slight variation within the chamber and that the cells kept at the 3 o’clock position developed best.   They also made the observation that one must work very quickly once the straws containing embryos were cooled when moving to a storage chamber as even a brief period of partial thawing could influence later survival and development.
Cryopreservation procedure:
1.	Superovulate mice, mate, and collect embryos
2.	Treat embryos with cryoprotectant media 
a.	PBS and human serum from day 10 of a woman’s cycle (human serum)- 15-20 sec. At 37 C.
b.	First loution plus ppropanediol – 15 minutes at room temperature.
c.	Solution B plus sucrose – 5 minutes at room temperature
3.	Load embryos into 0.5 ml plastic straws containing the third freezing solution, heat seal ends, and allow 10 min. resting time at room temp.
4.	Insert straws, in a horizontal position into the freezing chamber of a Planer freezer (model 10.10, T.S. Scientific).
5.	The freezer is programmed to cool the embryos.  Three protocols were used (note that each stage is called a Ramp):
a.	Ramp 1:  23 C to –7 C at –2 C/min.; Ramp 2:  hold at –7 for 5 min. for seeding (seeded 2 min into ramp with 3 min post-seeding soak); Ramp 3:  -7 to –30 at –0.3/min; Ramp 4:  -30 to –180 at –50/min.  Ramp 5:  hold at –180 for 10 min, then remove embryos and plunge into liquid Nitrogen for storage.
b.	Differs from “a” at Ramp 2:  hold at –7 for 20 min. (seeded 5 min. into ramp with 15 min. soak) and ramp 4:  hold at –30 for 10 min before embryos plunged for storage.
c.	Differs from “a” except used Ramp 2 of “b”
d.	For programs b and c the position in chamber were significant but not for a.  
6.  Thawing – embryos removed from storage, thawed at room temp for 40 sec., moved through four decreasing osmolality solutions (4 thawing solutions of decreasing osmolality), then placed in a 37 C incubator with 5% CO2 and 95% air at 100% relative humidity, left for 72 hours, then number of embryos at blastocyst stage determined.
Questions:
1.	What is the primary reason for the multi-stage cooling and subsequent thawing of embryos in the cryopreservation process? 
A.	To prevent dehydration of cells
B.	To prevent rupture of cell structures due to ice crystal formation within cells
2.	Which of the following variables were evaluated with this study, select all that apply?
A.	Order of loading straws into the freezing chamber
B.	Freezing protocol, time and rate of cooling
C.	Position of straws in the freezing chamber
D.	Methods of thawing
3.	Why is cryopreservation of embryos important in laboratory animal medicine, select all that apply?
A.	It employs more technicians
B.	Allows unique genetic strains/stocks to be maintained for future use.
C.	Saves space, money, and manpower over maintaining breeding colonies.
D.	Is a method for development of knockout or  transgenic mice.
Answers:
1.	B
2.	B and C
3.	B and C

A Comparison of Xylazine and Medetomidine in an Anesthetic Combination in New Zealand White Rabbits  pages 32-34

SUMMARY: 
New Zealand White rabbits are a common species used in cardiothoracic research. A common anesthetic protocol for such procedures is that of intramuscular (IM) xylazine, ketamine and acepromazine followed by intubation and isoflurane.  However this regimen often demonstrates hypotension and may not be preferred as there are no preemptive analgesics.  This article discusses a comparison of two other anesthetic protocols used in rabbits: Xylazine-ketamine-buprenorphine and medetomidine-ketamine-buprenorphine.  
	It was found that the medetomidine protocol resulted in higher systolic, diastolic and mean blood pressures.  The return of the palpebral reflex took longer using the medetomidine protocol (23 +/-5 mins) compared with the xylazine (6 +/-3 mins).  The return of the pedal reflex took longer using the medetomidine protocol (46 +/-10 mins) compared with the xylazine (11 +/-3 mins).  The return of the righting reflex took longer using the medetomidine protocol (166 +/-19 mins) compared with the xylazine (127 +/-8 mins).  There was no significant difference between the two protocols in the return of spontaneous breathing, extubation time and heart or respiratory rates.
	Overall, it was found the medetomidine-ketamine-buprenorphine Anesthetic protocol used for rabbits provided safe and effective anesthesia, provided a better cardiovascular profile, maintained stable blood pressures, induced sufficient anesthetic depth and duration and provided preemptive analgesia.
QUESTIONS:
1. Genus and species of New Zealand White rabbits?
2. What class drugs are medetomidine and xylazine?
3. Which of the following statements is/are true
A. Anesthetized horses showed no difference in blood pressure between medetomidine and xylazine treatments.
B. Anesthetized swine showed no difference in blood pressure between medetomidine and xylazine treatments.
C. Anesthetized dogs showed no difference in blood pressure between medetomidine and xylazine treatments.
D. Anesthetized rabbits showed no difference in blood pressure between medetomidine and xylazine treatments.
ANSWERS:
1. Oryctolagus cuniculus
2. Alpha 2 agonist.  NOTE that medetomidine has a 10-fold greater affinity for alpha-2 binding sites.
3. A and B are true, C is false as dogs have been reported to have increased blood pressures with medetomidine.  D is false as rabbits have been reported to have increased blood pressures with medetomidine as indicated in this article.

Method to Quantify Tail Vein Injection Technique in Small Animals pages 35-38

Summary:
	The authors discuss the present lack of standard method to quantify technician performance in the injection of animals or to validate and verify the administration of test compounds in the in vivo systems as injection errors, which are often not readily recognized, can greatly impact the outcome of pre-clinical research studies. As a result, unrecognized misadministration of test compounds can render a high cost to the biomedical community.
	In this study, the authors propose six criteria for a reagent designed to asses tail vein injection technique in small animals and suggest a reagent, colloidal gold labeled with the stable isotope 197Au, that satisfies these criteria, thereby describing and validating for the first time a method to quantify technical compliance in tail vein injection. In an application of this reagent, the authors show the degree of variation experienced by technologists performing tail vein injection procedures in mice.
	In this study, mice were manually restrained and received tail vein injection. One hour later mice were euthanized, various organs including the injection site (Tail) were collected, and their gold content was quantified by neutron activation.
	Three experienced technologist were involved in this study. Total of 30 mice were used to test their tail vein injection proficiency. Misinjection is defined as when 10% of the intended volume remain in the tail.
	Two molecular weight markers were prepared and mixed to allow Simultaneous injection into the tail vein of mice. Ethylene diamine tetraacetic acid-biotin conjugate (EB1; 30ug) was charged with 99mTc(4) and diluted with saline to a final volume of 1 ml.
	Based on the above criteria, 12 of 30 injection were misadministered. Two with Tech 1, three with Tech 2 and seven with tech 3. Although the supervisor was able to correctly rank the injection skills of the three technologists used in this experiment, she greatly underestimated the extent and degree of  injection failure for the procedure.
	Finally, the result of this study illustrate the potential problem associated with the technical compliance with this common laboratory procedure and suggest that there is a need to validate injection methods and a need to monitor technical competence. Application of reagents similar to colloidal gold and the methods presented will facilitate the development of improved methods of teaching injection technique and monitoring technical quality in the laboratory setting.
No questions.

In Vivo Micro Computed Tomography of Subchondral Bone in the Rat
After Intra-articular Administration of Monosodium Iodoacetate.  Pages 39-43

The purpose of this study was to determine if in vivo three-dimensional microCT was of use for detecting progressive changes over time to the subchondral bone (femorotibial joint) of Wistar rats treated with MIA.  Rat femorotibial joints were imaged at day 0, 1, 7, 14, 28, and 56 days post-MIA.   Images at 14 days post-MIA showed mild sclerosis of the tibial plateau.  Mild surface contour alterations, in addition to mild sclerosis was noted at day 28.  Images at day 56 post-MIA showed marked sclerosis of the tibial plateau with marked alteration of the tibial plateau surface contour and osteophyte formation.  It was demonstrated that temporal progression and severity of the changes noted by microCT correlated to the changes seen using histologic analysis.  Correlative quantitative measures of subchondral trabecular bone mineral density were also demonstrated.  It was determined that microCT is a viable alternative to the use of histologic analysis to track the progression of osteoarthritis in an animal model.  With the advantage of high-resolution and high-through-put capabilities, in vivo microCT has the potential to be a valuable tool in the analysis and screening of animal models of osteoarthritis used to identify disease therapeutics.
Questions
	1.	True or False.  Joint pain does not correlate with the severity of joint disease.
	2.	Subchondral bone changes were evident by 
			a.	7 days post-MIA
			b.	14 days post-MIA
			c.	21 days post-MIA
			d.	56 days post-MIA
	3.	What is MIA and what is its mechanism of action?
	4.	Why is the subchondral bone of interest in osteoarthritis therapeutic research?
Answers
	1.	True
	2.	B
	3.	Monosodium iodoacetate.  MIA (monosodium iodoacetate, iodoacetic acid) reacts with cysteine groups to form a carboxymethylated protein that inhibits glyceraldehydes-3-phosphate activity in articular chondrocytes, resulting in disruption of glycolysis and eventual cell death.
	4.	Although cartilage degradation is fundamental to osteoarthritis, it is not the site of origin for the pain, due to the lack of innervation within the tissue.  In contrast, the periosteum and subchondral bone are heavily innervated.  

Ocular Melanona in a Long Evans Rats pages 44-46

Summary:  During a long-term study, a rat exposed to half-amplitude band fluorescent blue light (434 to 475 nm) and receiving a calcium channel blocker developed an ocular mass.  By visual examination, a presumptive diagnosis of ocular melanoma was made.  Subsequent surgical removal of the eye, followed by histopathology, confirmed the lesion as a highly pigment epitheliod melanoma.
Questions: 
1.	True or False.  The majority of ocular melanomas observed in rats have been amelanotic.
2.	True or False.  Calcium channel blockers promote apoptosis.
3.	Intraocular melanomas TYPICALLY exhibit (benign, malignant) behavior
Answers
1.	True
2.	False
3.	benign
 
Spontaneous Uterine Leiomyosarcoma in a Rhesus Macaque  pages 47-49

This paper describes the detection and work up of a uterine leiomyosarcoma in a 12 year old female rhesus macaque.  These types of tumors are mainly reported in humans.  Eker rats are used as a model for benign and malignant smooth muscle tumors found in the uterus and cervix.
Authors used percutaneous needle biopsy, radiography, ultrasonography, and finally exploratory celiotomy to investigate this tumor.  Due to a poor prognosis (numerous adhesions and tumor size), the macaque was euthanized during surgery.  The mass was removed and sent for histopathology processing.  The tumor had histological features similar to that observed in other leiomyosarcomas.
Uterine leiomyosarcomas are very cellular, composed of smooth muscle cells found in intersecting bundles.  These tumors tend to be solitary, large, and poorly circumscribed with a soft fleshy consistency and some necrosis.  Clinical signs can be absent or signs of bleeding, pain, and/or urinary incontinence can be detected.  These tumors can be distinguished from uterine leiomyomas (fibroids) in that fibroids are firm, solitary or multiple, and well-circumscribed.
Questions:
1.  What type of rodent(s) is/are used in models of smooth muscle tumors seen in the uterus and cervix?
     a.  SD rats                    c.  BN rats                      e.  none of the above
     b.  Eker rats                 d.  two of the above
2.  Which of the following are clinical signs of leiomyosarcomas?
     a.  no clinical signs          c.  bleeding                    e.  all of the above
     b.  pain                        d.  urinary incontinence
3.  Which criteria are assessed by biopsy samples to determine malignancy of uterine tumors?
     
     a.  mitotic index             c.  degree of cytological atypia      e.  all of the above
     b.  presence of necrosis  d.  a and b above
4.  Which is the most common site for metastasis of leiomyosarcomas?
     a.  liver                          c.  bones
     b.  lungs                        d.  none of the above
5.  What is the rule out (differential diagnosis) for this type of uterine leiomyosarcoma, and what is the main difference between the two types of tumors?
Answers:
1.  b            2.  e            3.  e             4.  b      
5.  uterine leiomyoma (fibroids)   Fibroids are benign, firm, and well-circumscribed.

Increasing Supervisory Effectiveness in a Laboratory Animal Facility  pages 50-53
 
SUMMARY:  Management personnel expressed concerns that the allocation of work was not properly aligned to permit maximum efficiency in the supervisor job family.  Employees at various levels in the supervisor job family also felt that there was a need to be greater definition regarding which tasks needed to be done by which level in the organization.  Using data collection and interviews, a team of staff employees and consultants successfully evaluated the facility operation and supervision.  The information was analyzed and compared with the existing supervisory job description.  An action plan was developed and presented to management personnel to increase efficiency and to s! tratify the responsibility of the existing supervisory levels.  Supervisory reassignment was proposed. 
	Taking into consideration the relationship among the Laboratory Animal Resource Supervisors, production, and other organization functions and the environment, it was clear that the supervisory resource was being expanded outside the technical core.  Managerial skills required for successful operations should be focused on the technical core activities and directed away from the boundary systems (including non-critical duties).  This distribution would allow supervisory resource to focus on the key production functions, which for animal facility are to (1), maintain and ensure quality! animal care. (2) provide expertise and services for research needs, (3) comply with governmental regulations, and (4) achieve these functions with the minimum cost to the organization.  Vilfredo Pareto, the famous Italian engineer-economist-sociologists, was applied to the relocation of supervisory effort.  The work group’s basic assumption was that supervisor activities with the highest value (technical core) should receive maximum attention and investment, whereas those with lower value receive proportionately less.  The work group using the universal Pareto principle (80-20 rule) determined the vital few supervisory activities (20%) that focused on the key productive functions and was believed to provide the greatest (at least 80%) operational return.  By using this same rule, 75% to 85% of supervisory ef! fort was reassigned to these technical core tasks (supervision, scheduling and professional/technical resulting in more appropriate use and investment of supervisory personnel skill/experience.  In General Action Plan, the team presented to eliminate all supervisory time being used for Support Service/Clerical (9%) and to reallocate 14% Inventory time to Professional/Technical Services.  The team recommended that supervisor effort be increased from 57% to 75% to 85% in light of operational needs and the supervisory job descriptions.   The team reassigns all support service/clerical duties.  Eliminate all supervisory time being used for Support Service/Clerical.  Reallocate 14% of inventory time to Professional/Technical Services.&! nbsp; Out of 51% of 83% of top six tasks of supervisory tasks, the value is consumed in Scheduling, Inventory, Meeting, and Support Service/Clerical.  Only 32% is being allocated to Supervision and Professional/Technical Services.  The team recommended that the percentage of supervisory effort be increased to “value-added” tasks.  The team recommended specific action plan as follows: 1) The effort of supervision (95 h/week) needs to be increased by eliminating unnecessary tasks (e.g., scheduling employees for health services and safety training, completing questionnaires, surveys, etc.) or reassigning them to others.  Associate Supervisor level personnel should spend the majority of their time on the floor.  Supervisory level personnel would be their primary back-up.  2)  Scheduling (74 h/week), where possible the process for tracking and management (work order system) must be evaluated and automated.  3)  Professional /Technical Services (37 h/week) is too low, more investigator contact us needed.  This function should be by the Senior Supervisor, which back-up from the Supervisor level.  4)  Meeting (42 h/week), this effort must be minimized by establishing effective meeting schedules and processes with agenda and aims.  5)  Time Cards (12 h/week) Supervisory time should be limited to authorization only by the Associate Supervisor; the r! est should be reassigned to administrative support.  6)  Support Service/Clerical (37 h/week), this figure should be removed from supervisory time and reassigned to administrative support.  7)  Inventory (58 h/week), this process should be reevaluated, centralized approach is beneficial, entry/clerical task should be simplified and data moved to administrative support.  8)  Lock-up (8h/week), this task should be the function of the Associate Supervisory level only.  9)  Maintenance (25 h/week), this task should be the primary responsibility of the Senior Supervisor level only.  10)  Phone/E-mail (16 h/week), this figure is an acceptable amount for communication with investigators, etc.  11)  Miscellaneous (8h/week), this value appropriately account for unplanned events, emergencies, and “duties as assigned.”
	The team developed an action plan and presented to management to increase efficiency and stratify the responsibility of the existing supervisory levels.  In addition, the during overtime periods, an associate supervisor may be expected to handle the responsibilities of higher level positions.  The supervision of the animal care staff would be the main duty of the Associate Supervisor level, with support from the Supervisor level.  While interaction with the researchers and physical plant staff, representing the area in meetings, vendor interactions, etc., would be the main duties of the Supervisor and Senior Supervis! or level.  Senior Supervisors should meet separately with management for operational planning and research-critical support issues.  Human Resources have rewritten job descriptions to focus work responsibilities and define skills set for promotion as well as to provide new candidate expectations.  As the result of this study, the department’s overall clerical tasks/responsibilities were also reevaluated and more defined structure instituted, which is supervised by an Administrative Support Supervisor.  Clerical support now tracks animal orders, confirming delivery dates with the vendors, attendances tracking – electronically entering payroll for non-salaried and hourly personnel and logging absenteeism.  With the improvement of electronic system reduced clerica! l-typed duties, the purchasing of supplies and products have been greatly enhanced.  This work process plan will assist the department in coping with changing science and research growth while maintaining the highest level of quality and efficiency.  
Questions:
1)      It is not important to reassigned Supervisory effort to technical core tasks (supervision, scheduling and professional/technical resulting) and not to eliminate all supervisory time being used for Support Service/Clerical.  True or False
2)      The effort of supervision (95 h/week) needs to be increased because it is important to eliminate unnecessary tasks such as scheduling employees for health services and safety training.  True or False
3)      Associate Supervisor level personnel should spend the majority of their time on the ……….
	a. Meeting
	b. Floor
	c. Clerical duties
	d. Inventory
	e. Supervision
4)    The supervision of the animal care staff would be the main duty of ……., with support from the Supervisor level.  
5)      By using Vilfredo Pareto’s rule, the team recommended …% to  ...% of supervisory effort was reassigned to these technical core tasks (supervision, scheduling and professional/technical); resulting in more appropriate use and investment of supervisory personnel skill/experience.
6)      Interaction with the researchers and physical plant staff, representing the area in meetings, vendor interactions, etc., would be the main duties of  the …….and……. level.
7)      As the result of this study, Clerical support now tracks animal orders, confirming delivery dates with the vendors, attendances tracking –electronically entering payroll for non-salaried and hourly personnel and logging absenteeism.  True or False
Answers:
1)      False
2)      True
3)      b  Floor
4)      Associate Supervisor
5)      75% to 85%
6)	Supervisor and Senior Supervisor
7)   True

Technology Update - Acoustic Startle page 54-55

Startle reflex is noted in many species including humans.
	Startle is a pattern of facial and skeletal muscle contractions, 
accompanied by an elevation of heart rate, mediated by neurons in the brainstem.  It is evoked by a sudden intense stimulus which may be tactile, auditory, visual or olfactory.
	The startle response can be made stronger or attenuated by the use of cues associated with the startling event.  This provides a measure of sensorimotor learning ability.
	Acoustic startle is most commonly used in rodents.  >80 dB is usually necessary to illicit the reflex.  white noise (background) can be used at ~65dB.  Also used to startle:  Air puff, light flash, shock grid.  The startle is measured with at piezoelectric strain gauge on the floor of the chamber.  (see photo of the acoustic startle apparatus and sound attenuating chamber/foam)
	Max sound intensity should be 120 dB.  above that you can have hearing damage and the vibration interferes with testing.
	the Acoustic startle threshold is the minimal sound intensity that produces a startle response. (test of hearing)
	Prepulse inhibition:  the animal is warned of the upcoming startle stimulus, this decreases the magnitude of the startle response.
	Fear-potentiated startle:  the animal is warned of a painful stimulus, thus increasing the magnitude of the startle response.
Questions:
1) T/F  Hearing impairment is the most common sensory deficit in human populations
2) What is the noise level referenced in the "Guide" as having effects in rodents?
a) 65 dB
b) 75 dB
c) 85 dB
d) 95 dB
Answer:
1) true.
2) c.  p. 36  - eosinopenia, increased adrenal weights, reduced fertility