This is a summary of my first week attachment in school. During the 1st week, there was no SIP involved due to delay in assigning TSO to us. So we basically focus on our MP. My MP is on the effect of functional food on the murine B & T cells.
One week before SIP commence, the 20 mice models arrived. They were to be isolated in the quarantine room, away from the other animals in the animal holding unit (AHU). This is to prevent the spread of potential infectious disease to the healthy animals. Everyday, we had to go back to school to check on the mice health and update the daily environmental record. We also had to ensure that there was sufficient food and water supplied to the mice. As we were dealing with live animals, it is necessary to check on them everyday i.e Mon to Sun. Sometimes on weekends, we had to wake up early just to go school and do a 15min check on them. It was kind of tiring at first but we have 5mths to let us get used to it.
First week of SIP - We prepared the solutions for the negative/postive control group and test group. Autoclaved water was used as a negative control with basal production of lymphocytes. Vitamin C was used as a positive control which will enhanced the production of lymphocytes. The method used to adminster the article/solution was by gavaging. Gavaging refers to the force feeding of article directly into the stomach with the insertion of gavage tubes from the mouth. This process was to ensure that each mouse recieved the specific amount of article administerd to prevent any potential pre-analytical variations. It had to be done carefully to avoid inappopriate insertion into the lungs instead. One of the identification method we used was cage cards for each group. It indicates the protocol number, strain, sex, age, supplier, investigator and contact person. We had done ear punching to ease the identification of every mouse. It involve the use of a special punch to produce a small notch near the edge or in the middle of the ear.
The above content was what I'd done for this week. If u have any questions, pls feel free to ask. Take care!
Taken from: http://www.jhu.edu/animalcare/images/rat_james_mouse_identification.gif
24 comments:
Hey there,
Just wondering, how do you perform gavaging - what equipment is involved? Also, is there a chance that the rat will throw up the solution since techinically, it's being force down it's throat? I mean, if i was force fed something, I'd probably throw it up by reflex due to the discomfort.
- Debra, TG02
Hi Joanne.. Can i ask what is functional food? Are there any specific examples? And also, you mentioned that you use gavaging to ensure that each mouse receive the specific amount of article administered right. But different mouse would have different physiology, what if different amount of article is being absorbed in the intestines, will it affect the results or something?
Ming Boon
Tg01
Although galvaging may ensure eual portion of food eaten by the mice,wouldn't force feeding violate animal rights? As the tubes are forced through the mouth, it would cause high amounts of discomfort. Won't this cause the mouse to be distressed, lowering the immunity of the mouse? And from DDCT, distressed animals make poor models of research.....
Thanks.
Have a nice day,
yeng ting
hihi.. juz to ask.. is there any other special reasons that Vitamin C is used instead of others which might also cause increase in lymphocytes production? why not use other known type of substance that might also raise the lymphocytes lvl. Another qn is.. are there some things that is being especially checked on during checks on weekends? And my last qn would be rather similar to wad debra mentioned earlier.. Are there procedures to ensure that this gavaging thingy is done appropriately and correctly.. that would especially make the mice felt less discomfort.. thanks..
Jia Hao
TG01
Hi joan, what instrument do you use to perform gavaging? If you accidentally insert the food into the lungs, will the mice die straightaway? Will it affect teh whole project?
Sally, TG02
hi there! wah very long never see u guys le, how's your SIP in TP?
ok some qns i wana ask, how do u actualy dispose the rats if they are found dead? Or what u will do if one of the rats is ill, will u give it treatment?
And like what most of us are asking, how do u ensure that all the rats consume the same amount of solution? Some might just spit it out.
Regards,
Chaur lee TG01
Hey Joanne
Just want to ask when you mention keep them in a quarantine area right? do you mean prevent the spread of infectious disease from those 20 new mice models or from the other animals in the facility?
Johanna
Hi joanne!!!
it seem like you are organising a 'food fest' for the mice. As you mention in the blog, the mice are number via ear punching. Can i ask how do you labelled the no. 26th( or any double digits) mouse?and how many time you have to gavage the mice( eg, daily, weekly or monthly)?
regards,
Avery TG02
HELLO! i see you in school every morning but i still have questions to ask. =D
Is there any other food or nutrients that can enhance the production of lymphocytes? Also, does Vitamin C increase the production of lymphocyte in human too after consumption?
Jiaxin
You mention about gavaging have to be done carefully to avoid inappopriate insertion into the lungs instead. If there was an accident, and was insert into the lungs. How to savage it? What would happen to the mice?
hi joan, i see you in school too, but i have a qns for you also.
erm..is there other substances that can be used to enhance lymphocytes production?is yes right, why is vitamin C choosen among the rest?and also, won't your project be affected greatly if something happens to those mice?
Charmaine Yeo, TG01
HeyHey! wad do u mean by functional food and how do u go about checking the mice health?
June, TG02
Hey, hope I ain't too late to ask a qn. How do you monitor the mices' health? As you know, the absorption of vitamins will deteriorate with gastrointestinal disorders. Not forgetting an increase of lymphocytes in a disease condition as part of the body's normal immune reaction. All these are other possible pre-analytical variations.(correct me if I'm wrong)
Kent
Hey ger,
can i know if the mice reject the solution what can be done?? is there any detector to indicate how much was injected annd how much was rejected so that the amount inside these mice are constant?? one last question is how are the labelling of the mice mentioned in the pic works? hope i did not confuse u ya..
Regards
Ai tee- 0503160D
TG01
Hi Joan,
Does your lab have other humane ways to label the rats? Instead of punching holes in their ears, I suggest using a string tag to tie it around their necks or feet with a labeled identification?
What kind of functional food is used to inco-operate the vitamin C into the mice?
Loh Sharon , Tg 01
Hi there...
Just out of curiosity, how do you actually puncture the ears of the rats? Isnt it painful and isnt it some kinda violation against animal rights?
Last quest, why do you use autoclaved water?? I mean distilled water is also clean and can be used as control. But why specifically choose autoclaved water??
Vinodhini
TGO1
-What is functional food?-
It refers to food or dietary components that may provide a health benefit beyond the basic nutrition. It has either health-promoting or disease-preventing properties.
-examples-
Lingzhi: have antiatherosclerotic, anti-inflammatory, anti-tumor properties etc.
Oats: Consumption can reduce total and LDL cholestrol.
Garlic: have cancer chemopreventive, anti-hypertensive, cholestrol-lowering properties
Vitamins: have immunomodulating effects
-Gavaging Procedures-
(materials required)
Sterile, Single-use syringes
Sterile gavage tubes
(methods)
1.Measure the distance from the tip of the mouse's nose to the last rib (stomach is located just below the last rib). This is the length of the gavage tube to be used.
2.The air in the syringe is purged to prevent air embolism. A required amount of solution was aspirated and gavage needle is attached.
3.The tip of the needle is placed in the mouse's mouth and slided down the back of the mouth.
4.The needle is moved forward in one fluid motion through the esophagus. (usually the mouse will help itself with the swallowing of needle, therefore there's no need to apply pressure)
5.The insertion of needle is to be done slowly to avoid entering the trachea which before that a resistance will be felt. if u're in the correct path, it'll be very smooth.
6.When u reached the location after the last rib, the solution will be administered. (DO NOT ADMINISTER IF IT STARTS STRUGGLING!)
7.Withdraw the needle carefully and observe the mouse for 15 min.
Note: if it shows any abnormal signs like coughing, gapping, it may indicate the solution has entered the lungs. If the lungs are punctured, immediate death may occur. Euthanasia (the act of inducing humane death in an animal) is to be carried out to reduce the pain to the minimum.
> Picture of gavage tubes
http://research.uiowa.edu/animal/mp19.jpg
> Picture on the gavaging process
http://research.uiowa.edu/animal/mp15.jpg
-Extra comments on gavaging-
When the needle slide smoothly down the esophagus and no resistance is felt, most of the time they will not throw up the solution. We did not experience these when performing this procedure. To achieve good gavaging techniques, we have to continuously practice on the spare mice (not those used for the exp). Only until we are confident enough, we will then start to gavage the mice. Although this procedure may traumatise them a little, but with good techniques, their pain will be reduced to the minimal. And it is best to do only once! nobody want to be tramautised for the 2nd time! (we are also traumatised!) so practice is very important!
-Reasons for Choice of Vit.C-
1. Vitamin C is required for leukocyte function.
2. It boosts the lymphocytes & macrophages' activity and improve the migration and mobility of leukocytes to the site of infection.
3. Vit.C also increases the synthesis of PGE1 which in turn modulates the lymphocyte formation.
As you can see, vitamin C contributes to the optimal function of the immune function, therefore we decided to choose vitC as our +ve control.
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