Q30. Hamsters: answers
1. There are 4 genera of hamsters we use in research:Name the genera – and might as well name the species too…
- Which is the smallest? Dwarf or Djungarian female hamster
- Which one does NOT hibernate and which one is a TRUE hibernator? Phodopus do not hibernate. Cricetus is the only TRUE hibernator.
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| Dwarf or Djungarian 9-45g (NB it is striped - goes white in winter) Phodopus campbelli or sangorus |
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| Mesocricetus auratus (golden or Syrian) 120g |
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| Chinese (striped) 30-45g Cricetulus griseus |
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| Armenian or gray 40-80g (2N=22...double usual) (Cricetulus migratorius) |
2. We know that hamsters have ‘special’ cheek pouches:
- What is special about them and why are they that way? They are immunologically privileged so will accept transplants of foreign tissue. No lymphatic drainage and rich in mast cells.
- What are they often used for? Tumor growth, microvascular studies, vascular smooth muscle function.
Golden (Syrian) Hamsters
3. True or false?
- They hibernate True. They do hibernate but they are not true hibernators.
- They need to fatten up before hibernation. False
- If they wake during hibernation they won’t go back into hibernation. False - they wake up periodically to eat. This is why they are not true hibernators.
4. What is different about the anatomy of the stomach in hamsters compared to other rodents? They have a non-glandular forestomach similar to ruminants. The esophagus enters between the forestomach and the glandular stomach.
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| From studyblue |
5. What characteristics of their pulmonary system makes them a good model for smoke inhalation studies? They have a limited number of glands so are good for bronchitis studies, spontaneous cancer is rare so can be used for carcinogenesis, and they are resistant to pulmonary infection and can decompose nicotine so are used for smoke inhalation studies.
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| From U of Utah |
7. What is the cause of “wet tail” in hamsters? Classic 'wet tail' or proliferative enteritis is caused by Lawsonia intracellularis - it is commoner in young hamsters and often precipitated by stress. However other bacteria can cause signs of wet tail... can you name some?
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| proliferative enteritis from ResearchGate.net |
8. Tyzzers disease in hamsters has a triad of pathologic signs: what are they? Clostridium piliforme (formerly Bacillus piliformis) causes multifocal necrotizing hepatitis, ileitis and myocardial necrosis. Can be precipitated by use of broad spectrum antibiotics. See info at DORA
9. What is unique about the hamster Helicobacter aurati (compared to mouse and rat Helicobacter species). Most rodent helicobacters are found in the large intestine and liver. H aurati causes inflammation of the stomach adn cecum. H aurati reference
10. At what ABSL must hamsters be kept if they are infected with lymphocytic choriomeningitis? Is this the same as in mice? ABSL-3 (compared with ABSL-2 for mice).
11. Hamster parvovirus is nearly identical to which mouse parvovirus? MPV-3
12. Name three differences (2 anatomic, one epidemiologic) between the 2 cestodes that hamsters are susceptible to.. (Rodentolepis nana and Hymenolepis diminuta)
(i) R nana Rostellum has hooklets ie is armed, H diminuta is not
(ii) R nana egg has polar filaments (between outer and inner membrane), H diminuta does not.
(iii) Size: R nana is less than 40mm x1mm H diminuta is 20-60mm x 4mm. R nana comes from the Latin for dwarf "nanos" hence is the "dwarf tapeworm", whereas H diminuta is just diminutive I guess..).
(iv) R nana usually has a direct life cycle (though it can also transmit via a secondary host) and can also autoinfect, whereas H diminuta requires a secondary host - usually a beetle.
Remember: R nana is often called Hymenolepis nana (eg CDC website) so do not confuse with H diminuta!
Other Hamsters
14. Which species is highly susceptible to oncogenic viruses? – can you name (3 or 4) oncogenic viruses? Djungarian hamsters. Eg Rous sarcoma virus (RSV), Simian virus 40 (SV40) , Human adenovirus-12 (Adeno-12)
15. In general, when breeding hamsters, the male should be taken away for raising the young. Which hamster species is the exception and you should leave the male to help raise the young?
Dwarf hamster Phodopus campbelli
Q29. Zebrafish: answers
1. Which characteristics of zebrafish development make them so attractive for studying embryonic development?
a. High fecundity
b. External fertilization
c. Organ visibility – transparency
d. Rapid embryo development
e. Sexual maturity at 2-4 months
2. What is the genus and species and native location
a. Brachydanio rerio
b. Ganges in India and Southern India
3. Zebrafish water quality parameters
a. pH (6.8-7.2)
b. temperature (24-28 C)
c. conductivity (3-500 microseimens)
d. hardness (Ca and Mg between 80 and 200 ppm)
4. True or false, ammonia , nitrite and nitrate are very toxic to fish
c. Organ visibility – transparency
d. Rapid embryo development
e. Sexual maturity at 2-4 months
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| From CSM.com |
2. What is the genus and species and native location
a. Brachydanio rerio
b. Ganges in India and Southern India
3. Zebrafish water quality parameters
a. pH (6.8-7.2)
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| conductivity meter from Grainger |
c. conductivity (3-500 microseimens)
d. hardness (Ca and Mg between 80 and 200 ppm)
4. True or false, ammonia , nitrite and nitrate are very toxic to fish
a. F - Although ammonia and nitrite are very toxic, nitrate is not very toxic but needs to be controlled to avoid overgrowth of algae.
5. Why should the temperature of the tank room be maintained slightly higher than tank water?
a. To minimize condensation on colder surfaces
6. When retrofitting a fish facility into a room on an upper floor – what construction detail should not be overlooked?
5. Why should the temperature of the tank room be maintained slightly higher than tank water?
a. To minimize condensation on colder surfaces
6. When retrofitting a fish facility into a room on an upper floor – what construction detail should not be overlooked?
a. Weight bearing capacity – per the blue book 40 x10 gallon aquaria weigh nearly 2 tons.
7. What is the best method to control excess algae in the aquarium?
7. What is the best method to control excess algae in the aquarium?
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| aquarium snails |
Limiting sustenance (food and light) is the best method. Freshwater snails can, and often do, overrun the aquarium and are remarkably hard to eliminate.
a. Limit food to that which the fish can eat in 5 minutes YES
b. Freshwater snails to eat the algae NO
c. Limit light to limit growth of algae YES
8. What is the commonest cause of fish becoming ill due to infectious disease?
a. Poor water quality. Fish can thrive in the presence of many pathogens providing their water quality is excellent.
9. Which Mycobacterium species are commonest in aquarium fishes? Can they infect people?
a. M marinum, fortuitum, chelonae
b. Yes – people generally get self limiting skin disease, known as 'fish-handlers granuloma.' However those who are immune suppressed may get life-threatening systemic disease.
10. What are the commonest bacterial infections?
a. Secondary infections due to ubiquitous opportunists such as Aeromonas, Hydrophila, Flexibacter columnaris, Flavobacterium spp and Pseudomonas spp.
11. Which is the commonest nematode and what is the characteristic feature of its eggs?
a. Pseudocapillaria tomentosa. Polar plugs in eggs.
12. Which is the commonest flagellate disease and what characteristic behavior is seen with infected fish?
a. Piscinoodinium spp. AKA velvet or gold dust disease. Infect skin and gills.
b. Flashing (rubbing against objects)due to itching
13. What is the commonest fungal disease caused by and is it usually primary or secondary? What is the prognosis?
a. Saprolegnia spp.
b. Secondary to poor water quality or a primary pathogen. Poor prognosis
14. What is the commonest ciliated protozoan parasite of skin?
a. Ichthyophthiirius multifiliis (“ich” pronounced "ick")
15. Which common ciliate protozoan parasites infect gills? Is it common in the lab?
a. Trichodinids
b. No – common in pond fish
16. When treating a tank with antibiotics, what should be done with respect to external filtration? Which drug could be photoinactivated?
a. Remove the carbon filter, which could sequester or inactivate the treatment
b. Oxytetracycline
17. Which diseases can be treated by salt water immersion and raising tank temperature?
a. Piscinoodium
b. Ich
c. Trichodinosis
18. Which disease requires depopulation and intensive environmental treatment with bleach to prevent reintroduction?
a. Mycobacteriosis
19. Malachite green is a popular treatment for fish infections, which should not be used in the laboratory. Why not?
a. Teratogenic and mutagenic properties. Particularly as zebrafish are often used for mutagen studies.
20. Not to belabor the point, but what is the commonest cause for sick fish?
a. Poor water quality/environment.
References:
Common Diseases of Laboratory Zebrafish in
a. Limit food to that which the fish can eat in 5 minutes YES
b. Freshwater snails to eat the algae NO
c. Limit light to limit growth of algae YES
8. What is the commonest cause of fish becoming ill due to infectious disease?
a. Poor water quality. Fish can thrive in the presence of many pathogens providing their water quality is excellent.
9. Which Mycobacterium species are commonest in aquarium fishes? Can they infect people?
a. M marinum, fortuitum, chelonae
b. Yes – people generally get self limiting skin disease, known as 'fish-handlers granuloma.' However those who are immune suppressed may get life-threatening systemic disease.
10. What are the commonest bacterial infections?
a. Secondary infections due to ubiquitous opportunists such as Aeromonas, Hydrophila, Flexibacter columnaris, Flavobacterium spp and Pseudomonas spp.
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| From researchgate.net |
11. Which is the commonest nematode and what is the characteristic feature of its eggs?
a. Pseudocapillaria tomentosa. Polar plugs in eggs.
12. Which is the commonest flagellate disease and what characteristic behavior is seen with infected fish?
a. Piscinoodinium spp. AKA velvet or gold dust disease. Infect skin and gills. |
| from freefishadvice.com |
13. What is the commonest fungal disease caused by and is it usually primary or secondary? What is the prognosis?
a. Saprolegnia spp.
b. Secondary to poor water quality or a primary pathogen. Poor prognosis
14. What is the commonest ciliated protozoan parasite of skin?
a. Ichthyophthiirius multifiliis (“ich” pronounced "ick")
 |
| From wikipedia |
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| Ich on a neon |
15. Which common ciliate protozoan parasites infect gills? Is it common in the lab?
 |
| From researchgate.net |
a. Trichodinids
b. No – common in pond fish
16. When treating a tank with antibiotics, what should be done with respect to external filtration? Which drug could be photoinactivated?
a. Remove the carbon filter, which could sequester or inactivate the treatment
b. Oxytetracycline
17. Which diseases can be treated by salt water immersion and raising tank temperature?
a. Piscinoodium
b. Ich
c. Trichodinosis
18. Which disease requires depopulation and intensive environmental treatment with bleach to prevent reintroduction?
a. Mycobacteriosis
19. Malachite green is a popular treatment for fish infections, which should not be used in the laboratory. Why not?
a. Teratogenic and mutagenic properties. Particularly as zebrafish are often used for mutagen studies.
20. Not to belabor the point, but what is the commonest cause for sick fish?
a. Poor water quality/environment.
References:
Common Diseases of Laboratory Zebrafish in
Available as a free chapter download from Sciencedirect.com.
Blue Book: Biology and Management of Zebrafish in Laboratory Animal Medicine 2nd Edition. 2002. Chapter 19. pp. 862-882. Keith Astrofsky, Robert Bullis and Charles Sagerstrom.
1. What was the key development that allowed immunodeficient mice to be engrafted with components of the human immune system?
a. Immunodeficient mice bearing mutations in the IL2 receptor common gamma chain (IL2rgnull) The common gamma chain constitutes an important component of receptors for IL2, IL4, IL7, IL9, IL15 and IL21 and is indispensable for high-affinity binding and signaling of these cytokines.
2. What are the 3 immunodeficient mouse strains that together with this mutation are used as models to engraft human tissues. (common name and nomenclature please!)
NB I have underlined the superscripted parts because I can't do superscripts here...
a. NOD.Cg-PrkdcscidIl2rgtm1Wjl (NSG),
b. NODShi.Cg-PrkdcscidIl2rgtm1Sug (NOG),
c. 129S4- Rag2tm1FlvIl2rgtm1Flv (commonly referred to as BALB/c-Rag2null IL2rgnull mice or BRG)
3. Which model (common name) is engrafted by injection of peripheral blood leukocytes? What are 2 disadvantages of this model?
a. Hu-PBL-SCID rapid engraftment one week, but only CD3+ cells, GVHD within 4-8 wks
4. Which model (common name) is engrafted by IV or intrafemoral injection of human SCID-repopulating cells? What is the advantage of this model over the previous one? What is a deficiency in this model?
a. Hu-SRC-SCID. Advantage: Supports engraftment of a complete immune system.
b. Disadvantage: Granulocytes, platelets and rbc generated in BM only in v. low numbers. Also the T cells are matured in the thymus so are H2 not HLA restricted.
5. Which model (common name) uses human fetal tissues and what is the deficiency of this model?
a. Bone marrow/liver/thymus “BLT” model that is established by transplantation of human fetal liver and thymus under the kidney capsule and IV injection of autologous fetal liver HSCs.
a. Advantages: T cells are HLA restricted and mucosal immunity develops.
b. Disadvantage: Wasting GVHD-like syndrome
6. Humanized mice are frequently used to study HIV. Which 2 humanized mouse models recapitulate the disease and which one is most commonly used? Why?
a. HSC and BLT models recapitulate. BLT most commonly used because of higher engraftment and presence of mucosal immunity allows study of vaginal and rectal transmission.
7. Epstein Barr virus is studied using humanized mice. EBV affects 90% of humans worldwide. Which cells are transformed by EBV and which immune cell is normally responsible for eliminating these transformed cells? List immunological and autoimmune disorders that can result from EBV if these cells are not removed.
a. EBV infects epithelial cells. B cells required for elimination.
b. EBV causes Lymphoproliferative disorders, Burkitt’s lymphoma, Hodgkin lymphoma, nasopharyngeal carcinoma, gastric carcinoma, rheumatoid arthritis, multiple sclerosis.
8. Most humanized mouse models have used human cell lines for engraftment, however unlike in patients, cell lines and the environment they grow in is homogenous. In the next step forward, patient cancer cells are being grown in humanized mice. Which of the 3 models is required for growth of human cancer cells? What is the common name of this model?
a. Human fetal cells are required (ie BLT).
b. PDX ie patient derived xenograft
9. List 3 (or more) remaining problems to be solved in development of future humanized mouse models.
a. Reduction of remaining mouse innate immunity
b. Development of lymph node architecture
c. Identification/addition of human factors required for optimal cell differentiation that are absent in mice
d. Decrease or prevent GVHD
e. Validation of the models containing patient tumors
Reference: Annu Rev Pathol. 2017 Jan 24; 12: 187–215.
1. Which strain has long been used as a source of ES cells for targeted mutations and why is this a problem?
129. It's a problem because there are at least 16 different substrains of 129 - designated P for parental, S for steel allele on the Kitl gene which is homozygous lethal, T for the line that carries the teratoma gene and X for the line that is genetically contaminated. The original source of ES cells was the 129/ Sv (Sv for Stevens laboratory). ES cells generated from mice of different 129 strains can have significant genetic differences.
2. List 2 strains that have mutations in the dysferlin gene and what condition (s) results?
SJL/J - was a spontaneous mutation Dysf im for inflammatory myopathy. The mutation causes a lack of dysferlin protein, and the inflammatory myopathy causes progressive loss of muscle mass and strength, hindlimb clasping etc. by 8 months of age. It is a model for limb girdle muscular dystrophy 2B.
A/J mice have a Dysf prmd mutation, causing progressive muscular deficits primarily in the proximal limbs, similar to the dysferlin deficient human diseases.
3. Which strain is known for has a high incidence of thymic T cell lymphoma and what is the genetic reason? Which immunodeficient strain is also has a high incidence of T cell lymphoma? Which strain has a high incidence of B cell lymphomas?
AKR mice have high incidence of T cell lymphoma before 1 year of age and die by 18m. Due to a number of endogenous retroviruses such that the mice are viremic from birth. SCID and NOD SCID mice also have high incidence of T cell lymphomas. BALB/c mice get B cell lymphomas.
4. Which strain has a high incidence of cardiac abnormalities including epicardial mineralization and cardiomyopathy and is widely used for infectious disease studies?
Arguably this could apply to several mouse strains- e.g. DBA (up to 90% with cardiac calcinosis by 1 year and susceptible to many mouse diseases) or C3H (dystrophic cardiac calcinosis, susceptible to many infectious agents). However BALB/c probably best fits the bill (age related cardiomyopathy plus calcinosis plus studied in a wide range of infectious diseases).
5. Which 2 strains with high and low mammary tumor incidence (respectively) were developed from an A albino female to a DBA male?
C3H (high mammary tumors) and CBA (low mammary tumors). Note the original cause of high mammary tumors was exogenous MMTV, which has been eliminated from most strains, but endogenous retroviruses still result in later onset mammary tumors.
6. Which strain historically had a very high incidence of mammary adenocarcinomas due to MMTV?
C3H.
7. Which strain is known for its resistance to many infectious agents but susceptibility to mouse adenovirus 1 and encephalomyelitis?
C57BL/6
8. Which induced disease model is the DBA mouse known for and what is the human condition?
Induced type II collagen-induced arthritis as a model for rheumatoid arthritis.
9. List 2 strains susceptible to seizures
FVB( lethal epileptic syndrome, spontaneous and induced by stress or noise) and DBA (audiogenic seizures between 14 and 42 days).
10. List 2 strains that exhibit acallosity
129 and BALB/c
YABA
ORF
Dengue
Marburg
Ebola
Hanta
LCM
B Virus
Hepatitis A
SIV
Measles
Newcastle Dis
Influenza
Rabies
Which disease(s) ?
1. Is/are generally lethal in new world primates yellow fever (as opposed to african monkeys where the disease is mild), also measles causes severe disease
2. Are transmittable by aerosol hantavirus, measles, Newcastle disease, influenza are all primarily transmitted by aerosol. Aerosol transmission has been demonstrated in yaba Marburg, Ebola.
3. Bats are a/the reservoir host rabies, Ebola and other filoviruses.
4. Cause tumors Yaba, a yatapoxvirus. Causes benign histiocytomas
5. Squirrels are significant reservoirs Monkeypox - specifically Funisciurus (african striped squirrel) and Heliosciurus (sun squirrel)
6. smallpox vaccine is protective monkeypox(in humans and also used in monkeys)
7. The reservoir host is probably NOT the african green monkey Marburg virus - has 100% case fatality rate in african green monkeys - reservoir hosts usually have mild or subclinical disease. Bats are suspected.
8. Transmitted by Aedes mosquitos yellow fever and Dengue
9. Rodents are the reservoir hosts hantavirus, LCM In the US, the rodents that carry Hantavirus are
Cotton Rat (Sigmodon hispidus), Deer Mouse (Peromyscus maniculatus), Rice Rat (Oryzomys palustris), White-footed mouse (Peromyscus leucopus) - although it's difficult to tell the white footed mouse and deer mouse apart.
10. Have been transmitted by corneal transplants rabies
A couple of things can be taken away from this rough and ready table summarized from the rat blue book with some updates):
- most parasites found in wild rats can also be found in multiple other species with the exception of Trypanosoma lewisi and Hepatozoon muris
- most parasites found in wild rats cause some clinicopathology which could most certainly affect research, however they are extremely unlikely to be found in modern laboratory rats
- those 'other' parasites that are found in lab rats are not thought to be pathogenic, but in fact very little is known about them
- interestingly, although Spironucleus, Entamoeba, Tritrichomonas and Giardia muris are all found in rats, a prominent vendor for rodent testing only lists Spironucleus and Entamoeba in its rat panel, although the mouse panel also lists Tritrichomonas and Giardia.
1. At what light intensity (lux) is there a probability of phototoxic retinopathy?
The exorbital and intraorbital lacrimal glands produce tears. Harderian gland secretions are both apocrine (budding off part of the cytoplasm similar to sweat glands) and holocrine (similar to sebaceous glands where the plasma membrane is disrupted and the cells contents are discharged)
RBB page 96)
1. In the US, the Animal Welfare Act (AWA) was passed in 1966. What were the 3 purposes of the Act?
The Act (Public Law 89-544) was intended to prevent pets being stolen and sold for research. Specifically "To authorize the secretary of agriculture to regulate the 1)transportation, 2)sale and 3)handling of dogs, cats and certain other animals intended for the purposes of research or experimentation or other purposes. [History of the AWA here]
(bonus -what was the name of the English Pointer whose emaciated picture in Life Magazine Illustrated ignited a storm of public opinion?) Both Sports Illustrated and Life Magazine featured articles about pets being stolen and held in deplorable conditions at dealers before being sold for research. The pointer pictured in the Life magazine article was called Lucky and was later rescued. Pepper was the dalmation who was recognized by his owner in a newspaper picture after having been stolen from the yard of his home, and was featured in the Sports Illustrated article that started the public outcry.
2. The Public Health Service Act pre-dated the AWA. When was it first passed? 1944. What was its purpose? Preventing the introduction, transmission and spread of communicable diseases from foreign countries into the United States. nb.Why did I include this? - because there can be confusion between the names - it is The Health Research Extension act of 1985 (PL 99-158) that establishes guidelines for the proper care and treatment of research animals and is the legislative mandate for the Public Health Service Policy, and hence the Guide for the Care and Use of Laboratory Animals. PL 99-158 is administered through the Office of Laboratory Animal Welfare (OLAW)
3. Which government agency is responsible for enforcement of the AWA? Animal and Plant Health Inspection Service (Animal Care)
4. Which species are specifically excluded from the AWA? "farm animals used for food or fiber (fur, hide, etc.); coldblooded species (amphibians and reptiles); horses not used for research purposes; fish; invertebrates (crustaceans, insects, etc.); or birds, rats of the genus Rattus, and mice of the genus Mus that are bred for use in research. Birds (other than those bred for research) are covered under the AWA but the regulatory standards have not yet been established." From the APHIS website accessed 11/2017
5. True or false: The AWA regulates...
6. True or false: The AWA is enforcible through criminal penalties False, the penalties are civil penalties. What is the difference? Civil penalties are fines, whereas criminal penalties can be fines, imprisonment or even execution. However the civil fines are not insignificant: they can be $10,000 per violation per animal per day!
8. What is the purpose of Policy 12? It requires investigators to consider alternatives to procedures that cause more than momentary or slight pain or distress and requires a written narrative describing methods used to to determine the availability of alternatives (as defined by the 3 R's)
9. What is the Act that allows enforcement of the PHS Policy? Health Research Extension Act (PL 99-158) of 1985 - this should come immediately to mind as I already gave you this answer!
10. How do the requirements for IACUC composition differ between the AWA and the PHS policy:
PHS : 5 members including one veterinarian, one practicing scientist, one non-scientist, one not affiliated with the institution
AWA: 3 members including a chairman, a veterinarian and a non-affiliate, and if there are more than 3 members, no more than 3 can come from the same administrative unit.
11. Give examples of research animals that are NOT covered by either the AWA or the PHS policy. AWA regulates all warm blooded animals with specific exceptions (see above). PHS policy regulates all vertebrate animals that are supported by the PHS. Therefore animals excluded under the AWA such as rats mice or birds bred for research, or cold blooded animals, or farm animals used in food or fiber research) that are not supported on PHS or NIH grants are NOT covered.
1. Depending on how you classify them, nonhuman primates can be divided into suborders based on retention of primitive characteristics (prosimians meaning "before primates" and anthropoids or "resembling humans") or based on shared derived characteristics (Strepsirrhines - from the Greek meaning "twisted nose" and Haplorrhines - meaning "simple nose"). Strepsirrhines have wet noses and haplorrhines have dry noses.
a. May have prehensile tail platyrrhines
b. cheek pouches catarrhines
c. ischial callosities catarrhines
d. "toilet claw" prosimians (bonus - do you know which digit?)
e. largely nocturnal prosimians (bonus - do you know the exceptions? - and which platyrrhine is nocturnal?)
f. Fissured nose prosimians
g. fused mandible platyrrhines and catarrhines
h. Hindlimbs longer than forelimbs prosimians
i. Can be bipedal catarrhines (specifically the great apes)
j. All arboreal platyrrhines (catarrhines are terrestrial or arboreal or both)
k. All carnivorous tarsiers
l. Stereoscopic color vision all anthropoids
m. Flat nose with flared nares platyrrhines
n. Require vitamin D3 platyrrines
3. Platyrrhines can be divided into families Callitrichidae and Cebidae.
a. Which family has many CITES listed genera? Callitrichidae and Cebidae both have a fair number of CITES listed spp. in both appendix I and appendix II)
b. What does CITES stand for? the Convention on International Trade in Endangered Species of Wild Fauna and Flora What specifically does it place controls on international trade in listed species
c. What is the unique feature of the placenta in Callitrichidae? - they usually have twins, the hemochorial placentas have vascular anastomoses and undergo continuous hematopoiesis so there is stem cell transfer between the 2 fetuses which are thus chimeras
d. Which genus is often used for malaria research? Aotus Why? susceptible to human and other NHP malaria spp. Saimiri are also used for e.g. vaccine development.
e. Which genus has naturally occurring atherosclerosis? Saimiri
f. Which genus was sent into space? Saimiri
f. Squirrel monkeys are often classified according to whether the hair pattern on their face resembles a gothic arch or a roman arch. Which genera fall into each, and which of these is used in malaria research? oops - meant to say which species - of those commonly used, S sciureus and S oerstedii and S ustus have gothic (or pointed) arches and S boliviensis has a round or roman arch. S boliviensis is used in malaria research.
g. which genus has a semi-prehensile tail? Cebus Which genera have true prehensile tails? Alouatta (howler monkeys), Ateles (spider monkey) Brachyteles (woolly spider monkey), Lagothrix (woolly monkey), What is the difference between prehensile and pseudo-prehensile tails anyway? True prehensile tails have a tactile pad on the underside for better grip
h. What is the regulatory impact of brachiation? Brachiating species require larger cage sizes
i. Which genus is used to study gout? Lagothrix
j. What is special about the hallux in callitrichidae? It is opposable whereas the thumb is not And what is a hallux? big toe - bonus - what is scientific name of the thumb?
k. Which genus gets Vitamin E responsive hemolytic anemia? Aotus
1. Mouse pups: at what age does...
Q28: On humanized mice: answers
 |
| from: Translational Microbiome Research Forum |
1. What was the key development that allowed immunodeficient mice to be engrafted with components of the human immune system?
a. Immunodeficient mice bearing mutations in the IL2 receptor common gamma chain (IL2rgnull) The common gamma chain constitutes an important component of receptors for IL2, IL4, IL7, IL9, IL15 and IL21 and is indispensable for high-affinity binding and signaling of these cytokines.
2. What are the 3 immunodeficient mouse strains that together with this mutation are used as models to engraft human tissues. (common name and nomenclature please!)
NB I have underlined the superscripted parts because I can't do superscripts here...
a. NOD.Cg-PrkdcscidIl2rgtm1Wjl (NSG),
b. NODShi.Cg-PrkdcscidIl2rgtm1Sug (NOG),
c. 129S4- Rag2tm1FlvIl2rgtm1Flv (commonly referred to as BALB/c-Rag2null IL2rgnull mice or BRG)
3. Which model (common name) is engrafted by injection of peripheral blood leukocytes? What are 2 disadvantages of this model?
a. Hu-PBL-SCID rapid engraftment one week, but only CD3+ cells, GVHD within 4-8 wks
4. Which model (common name) is engrafted by IV or intrafemoral injection of human SCID-repopulating cells? What is the advantage of this model over the previous one? What is a deficiency in this model?
a. Hu-SRC-SCID. Advantage: Supports engraftment of a complete immune system.
b. Disadvantage: Granulocytes, platelets and rbc generated in BM only in v. low numbers. Also the T cells are matured in the thymus so are H2 not HLA restricted.
5. Which model (common name) uses human fetal tissues and what is the deficiency of this model?
a. Bone marrow/liver/thymus “BLT” model that is established by transplantation of human fetal liver and thymus under the kidney capsule and IV injection of autologous fetal liver HSCs.
a. Advantages: T cells are HLA restricted and mucosal immunity develops.
b. Disadvantage: Wasting GVHD-like syndrome
6. Humanized mice are frequently used to study HIV. Which 2 humanized mouse models recapitulate the disease and which one is most commonly used? Why?
a. HSC and BLT models recapitulate. BLT most commonly used because of higher engraftment and presence of mucosal immunity allows study of vaginal and rectal transmission.
7. Epstein Barr virus is studied using humanized mice. EBV affects 90% of humans worldwide. Which cells are transformed by EBV and which immune cell is normally responsible for eliminating these transformed cells? List immunological and autoimmune disorders that can result from EBV if these cells are not removed.
a. EBV infects epithelial cells. B cells required for elimination.
b. EBV causes Lymphoproliferative disorders, Burkitt’s lymphoma, Hodgkin lymphoma, nasopharyngeal carcinoma, gastric carcinoma, rheumatoid arthritis, multiple sclerosis.
8. Most humanized mouse models have used human cell lines for engraftment, however unlike in patients, cell lines and the environment they grow in is homogenous. In the next step forward, patient cancer cells are being grown in humanized mice. Which of the 3 models is required for growth of human cancer cells? What is the common name of this model?
a. Human fetal cells are required (ie BLT).
b. PDX ie patient derived xenograft
9. List 3 (or more) remaining problems to be solved in development of future humanized mouse models.
a. Reduction of remaining mouse innate immunity
b. Development of lymph node architecture
c. Identification/addition of human factors required for optimal cell differentiation that are absent in mice
d. Decrease or prevent GVHD
e. Validation of the models containing patient tumors
Reference: Annu Rev Pathol. 2017 Jan 24; 12: 187–215.
Q27: Mouse Strains & their Spontaneous Diseases
1. Which strain has long been used as a source of ES cells for targeted mutations and why is this a problem?
129. It's a problem because there are at least 16 different substrains of 129 - designated P for parental, S for steel allele on the Kitl gene which is homozygous lethal, T for the line that carries the teratoma gene and X for the line that is genetically contaminated. The original source of ES cells was the 129/ Sv (Sv for Stevens laboratory). ES cells generated from mice of different 129 strains can have significant genetic differences.
2. List 2 strains that have mutations in the dysferlin gene and what condition (s) results?
SJL/J - was a spontaneous mutation Dysf im for inflammatory myopathy. The mutation causes a lack of dysferlin protein, and the inflammatory myopathy causes progressive loss of muscle mass and strength, hindlimb clasping etc. by 8 months of age. It is a model for limb girdle muscular dystrophy 2B.
A/J mice have a Dysf prmd mutation, causing progressive muscular deficits primarily in the proximal limbs, similar to the dysferlin deficient human diseases.
3. Which strain is known for has a high incidence of thymic T cell lymphoma and what is the genetic reason? Which immunodeficient strain is also has a high incidence of T cell lymphoma? Which strain has a high incidence of B cell lymphomas?
AKR mice have high incidence of T cell lymphoma before 1 year of age and die by 18m. Due to a number of endogenous retroviruses such that the mice are viremic from birth. SCID and NOD SCID mice also have high incidence of T cell lymphomas. BALB/c mice get B cell lymphomas.
4. Which strain has a high incidence of cardiac abnormalities including epicardial mineralization and cardiomyopathy and is widely used for infectious disease studies?
Arguably this could apply to several mouse strains- e.g. DBA (up to 90% with cardiac calcinosis by 1 year and susceptible to many mouse diseases) or C3H (dystrophic cardiac calcinosis, susceptible to many infectious agents). However BALB/c probably best fits the bill (age related cardiomyopathy plus calcinosis plus studied in a wide range of infectious diseases).
5. Which 2 strains with high and low mammary tumor incidence (respectively) were developed from an A albino female to a DBA male?
C3H (high mammary tumors) and CBA (low mammary tumors). Note the original cause of high mammary tumors was exogenous MMTV, which has been eliminated from most strains, but endogenous retroviruses still result in later onset mammary tumors.
6. Which strain historically had a very high incidence of mammary adenocarcinomas due to MMTV?
C3H.
7. Which strain is known for its resistance to many infectious agents but susceptibility to mouse adenovirus 1 and encephalomyelitis?
C57BL/6
8. Which induced disease model is the DBA mouse known for and what is the human condition?
Induced type II collagen-induced arthritis as a model for rheumatoid arthritis.
9. List 2 strains susceptible to seizures
FVB( lethal epileptic syndrome, spontaneous and induced by stress or noise) and DBA (audiogenic seizures between 14 and 42 days).
10. List 2 strains that exhibit acallosity
129 and BALB/c
Q26: Viral Zoonoses
YABAORF
Dengue
Marburg
Ebola
Hanta
LCM
B Virus
Hepatitis A
SIV
Measles
Newcastle Dis
Influenza
Rabies
Which disease(s) ?
1. Is/are generally lethal in new world primates yellow fever (as opposed to african monkeys where the disease is mild), also measles causes severe disease
2. Are transmittable by aerosol hantavirus, measles, Newcastle disease, influenza are all primarily transmitted by aerosol. Aerosol transmission has been demonstrated in yaba Marburg, Ebola.
3. Bats are a/the reservoir host rabies, Ebola and other filoviruses.
4. Cause tumors Yaba, a yatapoxvirus. Causes benign histiocytomas
 |
| Sun squirrel |
 |
| Yaba |
5. Squirrels are significant reservoirs Monkeypox - specifically Funisciurus (african striped squirrel) and Heliosciurus (sun squirrel)
6. smallpox vaccine is protective monkeypox(in humans and also used in monkeys)
 |
| Cercopithecus |
7. The reservoir host is probably NOT the african green monkey Marburg virus - has 100% case fatality rate in african green monkeys - reservoir hosts usually have mild or subclinical disease. Bats are suspected.
 |
| Aedes aegypti |
8. Transmitted by Aedes mosquitos yellow fever and Dengue
 |
| cotton rat |
 |
| rice rat |
 |
| deer mouse |
 |
| white footed mouse |
Cotton Rat (Sigmodon hispidus), Deer Mouse (Peromyscus maniculatus), Rice Rat (Oryzomys palustris), White-footed mouse (Peromyscus leucopus) - although it's difficult to tell the white footed mouse and deer mouse apart.
10. Have been transmitted by corneal transplants rabies
Q25: Rat "other" parasites
 |
| Calodium hepaticum |
- most parasites found in wild rats can also be found in multiple other species with the exception of Trypanosoma lewisi and Hepatozoon muris
- most parasites found in wild rats cause some clinicopathology which could most certainly affect research, however they are extremely unlikely to be found in modern laboratory rats
- those 'other' parasites that are found in lab rats are not thought to be pathogenic, but in fact very little is known about them
- interestingly, although Spironucleus, Entamoeba, Tritrichomonas and Giardia muris are all found in rats, a prominent vendor for rodent testing only lists Spironucleus and Entamoeba in its rat panel, although the mouse panel also lists Tritrichomonas and Giardia.
Q24: Random Rat Facts Answers
1. At what light intensity (lux) is there a probability of phototoxic retinopathy?
130 -270 lux above the light level at which it was raised (rat blue book(RBB) page 315, Guide p.48)
2. Which 2 glands are responsible for tear production?
The exorbital and intraorbital lacrimal glands produce tears. Harderian gland secretions are both apocrine (budding off part of the cytoplasm similar to sweat glands) and holocrine (similar to sebaceous glands where the plasma membrane is disrupted and the cells contents are discharged)RBB page 96)
3. What is the normal hearing range of rats? How does that compare to humans?
250 - 80,000 Hz. Human hearing is 20-20,000 Hz (decreasing with age). nb. the rat blue book incorrectly states that human hearing is 16,000-20,000 Hz! Ultrasonic refers to frequencies that cannot be heard by humans ie >20,000 Hz.
4. Also which kinds of rat communication calls happen in the ultrasonic range?
22KHz alarm calls
50 KHz "appetitive" or friendly calls
ref: Brudzynski SM, Behav Genetics 2005 35(1) 85-92 Principles of Rat Communication...
5. Which teeth do rats NOT have (ie of incisors, canines, premolars, molars)?
canines and premolars ( 1/1 0/0 0/0 3/3) = 16 total teeth
6. Which gland in the head is a "mixed" gland?
The sublingual gland.
The parotid and submandibular glands are serous (note the submandibular gl is mixed in humans)
The sublingual gland.
The parotid and submandibular glands are serous (note the submandibular gl is mixed in humans)
ref: Osamu Amano et al. Acta Histochem Cytochem 2012 45(5) 241-250.
7. We all know rats have no gall bladder, but where does the common bile duct enter the GI tract?
The duodenum. (RBB p.106)
8. How many chromosomes does a rat have?
42. Note humans have 46, but # of chromosomes does not bear a direct relationship to the phylogenetic scale. Eg a goat and a gypsy moth both have 60, a rhesus and wheat have 42, a badger and yeast have 32...
ref: wikipedia
9. Rat pups can absorb maternal antibodies until a) 7d, b) 15d c) 20-21d, d) birth
c. Unlike in many other species, rats and mice gain most of their maternal antibodies after birth, however absorption drops off rapidly after 20 days. The FcRn receptor is responsible.
ref: Pentsuk N, et al. An interspecies comparison... Birth Defects Res B Dev Reprod Toxicol.2009 86(4) 328-344
10. Rats do not begin breeding until a) 9-13 weeks b) 4-6 weeks c)7-10 weeks d) 11-15 weeks.
Rats usually reach puberty by 6 weeks. However females are physically not mature enough at 6wk, so 7-10 weeks (or even later) is a better age to start breeding.
11. List rat models for:
- Obesity: the outbred Zucker fatty rat is the most widely used. It is hyperphagic will become lighter if food restricted but still retains an obese phenotype. It is thought that the brain is insensitive to insulin. It is hyperlipidemic, (with high cholesterol) hyperinsulinemic and hyper triglyceridemic. It is not diabetic or hypertensive. In contrast the SHROB rat is obese, hypertensive and has a hyperinsulinemia. It is a good model for human metabolic syndrome X because it also gets renal disease (focal segmental glomerulosclerosis) typical of that disease. other diet-induce obesity models include the Osborne-Mendel rat and the DIO (dietary induced obesity prone) rat.
- Spontaneous Type I diabetes mellitus. The inbred Biomedical Research Models (BBDP) rat (aka biobreeding rat). It is a spontaneous autoimmune disease with polygenic inheritance. It is hypoinsulinemic - hyperglycemia develops between 60 and 120 days and it requires insulin to survive.
- Type II diabetes mellitus. The inbred Zucker diabetic fatty rat (ZDF rat) is the primary model. An interesting feature of this model is that the females (but not the males) require a high fat low protein diet to exhibit the phenotype. Other models include the Cohen diabetic rat, the Goto Kakazaki (GK) rat, the Otsuka Long-Evans Tokushima fatty (OLETF) rat and the Wistar Fatty rat.
- Hypertension. The inbred Spontaneously Hypertensive Rat (SHR) is the main model. It develops a systolic BP of 150mm Hg by 10 weeks and also has insulin resistance. Although the SHR model was developed from the outbred Wistar Kyoto stock, the inbred WKY rat is genetically dissimilar and not thought to be a good normotensive control. Other models include the genetically hypertensive (GH) rat, the Fawn hooded hypertensive (FHH) rat, the Lyon hypertensive (LH) rat, the Sabra hypertension-prone (SBH) and resistant rats, and the Milan hypertensive (MHS) and normotensive rats. The Dahl Hypertension salt-sensitive (SS/Jr) rat develops fulminating hypertension if given an 8% salt diet for 3-4 weeks and dies within 8 wks of being placed on the high salt diet. It develops low renin renal disease arteriolosclerosis and glomerulosclerosis identical to that of human hypertensive renal disease. - its control is resistant to salt (SR/Jr). The SS develop a survival hypertension on a low salt diet.
- Prostate cancer. The Lobund Wistar rat develops metastasizing sex-hormone influenced prostate adenocarcinoma. However it doesn't develop until a mean age of 26 months: a chemically induced version develops after 10.5 months of treatment. Also the Noble rat.
- Diabetes insipidus. The Brattleboro rat has a lack of vasopressin and produces large quantities of dilute urine. It also has high plasma renin, high angiotensin II and hypertension.
- Stroke. The Stroke prone rat was developed from the A1 and A3 substrains of the SHR rat and is called the SHRSP rat. A special Japanese style diet low in potassium and protein together with a 1% salt water accelerates development of stroke starting at 110 days.
Ref: RBB pp 720-
Q23 Answers:Regulations
1. In the US, the Animal Welfare Act (AWA) was passed in 1966. What were the 3 purposes of the Act?
The Act (Public Law 89-544) was intended to prevent pets being stolen and sold for research. Specifically "To authorize the secretary of agriculture to regulate the 1)transportation, 2)sale and 3)handling of dogs, cats and certain other animals intended for the purposes of research or experimentation or other purposes. [History of the AWA here]
(bonus -what was the name of the English Pointer whose emaciated picture in Life Magazine Illustrated ignited a storm of public opinion?) Both Sports Illustrated and Life Magazine featured articles about pets being stolen and held in deplorable conditions at dealers before being sold for research. The pointer pictured in the Life magazine article was called Lucky and was later rescued. Pepper was the dalmation who was recognized by his owner in a newspaper picture after having been stolen from the yard of his home, and was featured in the Sports Illustrated article that started the public outcry.
2. The Public Health Service Act pre-dated the AWA. When was it first passed? 1944. What was its purpose? Preventing the introduction, transmission and spread of communicable diseases from foreign countries into the United States. nb.Why did I include this? - because there can be confusion between the names - it is The Health Research Extension act of 1985 (PL 99-158) that establishes guidelines for the proper care and treatment of research animals and is the legislative mandate for the Public Health Service Policy, and hence the Guide for the Care and Use of Laboratory Animals. PL 99-158 is administered through the Office of Laboratory Animal Welfare (OLAW)
3. Which government agency is responsible for enforcement of the AWA? Animal and Plant Health Inspection Service (Animal Care)
4. Which species are specifically excluded from the AWA? "farm animals used for food or fiber (fur, hide, etc.); coldblooded species (amphibians and reptiles); horses not used for research purposes; fish; invertebrates (crustaceans, insects, etc.); or birds, rats of the genus Rattus, and mice of the genus Mus that are bred for use in research. Birds (other than those bred for research) are covered under the AWA but the regulatory standards have not yet been established." From the APHIS website accessed 11/2017
5. True or false: The AWA regulates...
- Importation of dogs and cats for resale purposes
- Animals used for exhibition
- Animals sold for pets
- Animals transported commercially
- Animals used in research
- Cock- and bull-fighting
- Living conditions for regulated animals
- Swine and cattle used for biomedical research
- Birds bred for research
- Dealers
- Animal Care Committees
- Psychological well being
All true except birds bred for research. Note - cock fighting is specifically prohibited and although bull fighting is not specifically itemized, the AWA does prohibit individuals from sponsoring or exhibiting an animal in a fighting venture.
6. True or false: The AWA is enforcible through criminal penalties False, the penalties are civil penalties. What is the difference? Civil penalties are fines, whereas criminal penalties can be fines, imprisonment or even execution. However the civil fines are not insignificant: they can be $10,000 per violation per animal per day!
7. What are the 3 R's, when were they first described, by who, and in what document?
Replacement (interpreted to mean replacing with animals lower on the phylogenetic scale or non-animal methods), reduction (of numbers of animals), and refinement (so as to minimize suffering). They were first described in "The Principles of Humane Experimental Technique" published in 1959 by WMS Russell and RL Burch.
Replacement (interpreted to mean replacing with animals lower on the phylogenetic scale or non-animal methods), reduction (of numbers of animals), and refinement (so as to minimize suffering). They were first described in "The Principles of Humane Experimental Technique" published in 1959 by WMS Russell and RL Burch.
8. What is the purpose of Policy 12? It requires investigators to consider alternatives to procedures that cause more than momentary or slight pain or distress and requires a written narrative describing methods used to to determine the availability of alternatives (as defined by the 3 R's)
9. What is the Act that allows enforcement of the PHS Policy? Health Research Extension Act (PL 99-158) of 1985 - this should come immediately to mind as I already gave you this answer!
10. How do the requirements for IACUC composition differ between the AWA and the PHS policy:
PHS : 5 members including one veterinarian, one practicing scientist, one non-scientist, one not affiliated with the institution
AWA: 3 members including a chairman, a veterinarian and a non-affiliate, and if there are more than 3 members, no more than 3 can come from the same administrative unit.
11. Give examples of research animals that are NOT covered by either the AWA or the PHS policy. AWA regulates all warm blooded animals with specific exceptions (see above). PHS policy regulates all vertebrate animals that are supported by the PHS. Therefore animals excluded under the AWA such as rats mice or birds bred for research, or cold blooded animals, or farm animals used in food or fiber research) that are not supported on PHS or NIH grants are NOT covered.
Q22 Answers:New World Primate Classification
 |
| From Wikipedia |
- List the superfamilies included in the suborders prosimii, and anthropoidea.
- Prosimians: Lemurs, Lorises, Tarsiers
- Anthropoids: Monkeys, Apes, Humans
- Which suborder is classified either with the less- or more- advanced primates depending on which classification criteria are used?
- Tarsiers
a. May have prehensile tail platyrrhines
b. cheek pouches catarrhines
c. ischial callosities catarrhines
d. "toilet claw" prosimians (bonus - do you know which digit?)
e. largely nocturnal prosimians (bonus - do you know the exceptions? - and which platyrrhine is nocturnal?)
f. Fissured nose prosimians
g. fused mandible platyrrhines and catarrhines
h. Hindlimbs longer than forelimbs prosimians
i. Can be bipedal catarrhines (specifically the great apes)
j. All arboreal platyrrhines (catarrhines are terrestrial or arboreal or both)
k. All carnivorous tarsiers
l. Stereoscopic color vision all anthropoids
m. Flat nose with flared nares platyrrhines
n. Require vitamin D3 platyrrines
3. Platyrrhines can be divided into families Callitrichidae and Cebidae.
 |
| Aotus |
b. What does CITES stand for? the Convention on International Trade in Endangered Species of Wild Fauna and Flora What specifically does it place controls on international trade in listed species
c. What is the unique feature of the placenta in Callitrichidae? - they usually have twins, the hemochorial placentas have vascular anastomoses and undergo continuous hematopoiesis so there is stem cell transfer between the 2 fetuses which are thus chimeras
d. Which genus is often used for malaria research? Aotus Why? susceptible to human and other NHP malaria spp. Saimiri are also used for e.g. vaccine development.
e. Which genus has naturally occurring atherosclerosis? Saimiri
f. Which genus was sent into space? Saimiri
f. Squirrel monkeys are often classified according to whether the hair pattern on their face resembles a gothic arch or a roman arch. Which genera fall into each, and which of these is used in malaria research? oops - meant to say which species - of those commonly used, S sciureus and S oerstedii and S ustus have gothic (or pointed) arches and S boliviensis has a round or roman arch. S boliviensis is used in malaria research.
g. which genus has a semi-prehensile tail? Cebus Which genera have true prehensile tails? Alouatta (howler monkeys), Ateles (spider monkey) Brachyteles (woolly spider monkey), Lagothrix (woolly monkey), What is the difference between prehensile and pseudo-prehensile tails anyway? True prehensile tails have a tactile pad on the underside for better grip
h. What is the regulatory impact of brachiation? Brachiating species require larger cage sizes
i. Which genus is used to study gout? Lagothrix
j. What is special about the hallux in callitrichidae? It is opposable whereas the thumb is not And what is a hallux? big toe - bonus - what is scientific name of the thumb?
k. Which genus gets Vitamin E responsive hemolytic anemia? Aotus
Q21: MOUSE BREEDING ANSWERS
1. Mouse pups: at what age does...
- The milk spot disappear d6
- Hair first appear d7
- Ears start to come away from head d3
- Teeth erupt d11
- Eyes are fully open d12
- Eat solid food d12
Check HERE if you need a reminder
2. Timed breeding. How would you time breeding without the use of drugs? ( there are good summaries HERE and HERE)
- Use group-housed females between 8 and 15 wks old (should be in diestrus - Lee Boot)
- Introduce male urine soaked bedding ( pheromonal stimulation - returns to cycling in sync - Whitten effect)
- Check for proestrus or estrus on 3rd night following exposure
- Introduce 1-2 females to single-housed male on 3rd night following )
- Check for plug early following morning =day 0.5
- Use group-housed females between 8 and 15 wks old (should be in diestrus - Lee Boot)
- Introduce male urine soaked bedding ( pheromonal stimulation - returns to cycling in sync - Whitten effect)
- Check for proestrus or estrus on 3rd night following exposure
- Introduce 1-2 females to single-housed male on 3rd night following )
- Check for plug early following morning =day 0.5
3. If you need to superovulate to produce many eggs (eg for embryo freezing) the protocol is...
- Take group housed female mice aged __21-35_____ days
- Inject 2-5 IU PMSG between _1pm - 4 pm__(time of day) on day 1.
- Inject 2-5 IU HCG __42-50 hours later on day _3____
- Mate with stud (8 weeks old individually housed)__immediately_(when?)
- Ovulation occurs _____12 hrs_____ hours later.
(And HERE is the protocol)
4. Estrus Cycle: visual external characteristics (ie vaginal opening appearance)
- What distinguishes proestrus from estrus?
- proestrus - swollen, pink, moist, +/- striations, open
- estrus - less swollen, less pink, drier, open
- Metestrus from Diestrus?
- metestrus - not swollen pink or moist. Maybe slightly open with cellular debris
- diestrus - not swollen pink or moist. Opening small and closed
- What distinguishes proestrus from estrus?
- proestrus - mostly nucleated epithelial cells (if early, with leukocytes, if late, with cornified epithelial cells
- estrus - mostly cornified epithelial cells (if late, with leukocytes)
- Metestrus from diestrus?
- mestestrus - leukocytes and cornified epithelial cells
- diestrus - mostly leukocytes (if late with some nucleated cells)
A really nice review for questions 4 and 5 can be found HERE
6. Early Pregnancy Diagnosis
And here are a few easy ones... -
7. Delayed implantation generally occurs when __the female becomes pregnant as a result of the postpartum estrus
8. For breeding, always add the female to the male's cage (otherwise he will waste his time marking territory rather than mating)
9. Gestation generally lasts between 19___ and _21___ days
10. B6 mouse pups should weigh approximately __10___grams on weaning at 3 weeks.
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