Alcohol Feeding Protocol

Bing Gui 20120604

Jump below to

• Supplies
• Liquid diet preparation and feeding procedure
• Product #F1259SP; Liquid Rat Diet, LD '82; Shake & Pour; Control
• Product #F1258SP; Liquid Rat Diet, LD'82; Shake & Pour--Ethanol

Supplies

1. Determine the total number of mice maintained according to the experimental design.
2. Order Ethanol and Control food as well as Maltose Dextrin, tubes, tube holders and some spare caps from BioServ.
   • Liquid Diet Feeding Tubes and Holders
   • Rodent Liquid Diet Lieber-DeCarli '82, Shake and Pour

3. Use the feeding chart below to make ready-to-use liquid diet. The ready-to-use liquid diet has a shelf life of 3 days if kept in a refrigerator.

4. Make daily record of feeding amount to monitor the consumption. Ideally mice in the Control group and mice in the Ethanol group should consume similar amount of liquid food per day.

5. Use Fisher Colored Label Tapes to seal the feeding tubes before refill the tubes. Change the food tubes and wash the dirty tubes daily.
6. Feed the mice to 8 or 12 weeks according to the experimental design. Sacrifice the mice and collect the tissues at the end of the feeding course.

Liquid diet preparation and feeding procedure

1. Control diet:
   1. Dry mix: 225.5g
   2. Water: 860 ml
2. Alcohol diet:

   Pre-feeding	ETOH KCAL/L (G/L)	Dry mix (g)	Water (ml)	95% EtOH (ml/L)	Maltose dextrin (g)
   2 days	0	133	957	0
   2 days	220	133	916	41	34
   2 days	260	133	908	49	23
   2 days	300	133	901	56	14
   Pre-feeding	ETOH KCAL/L (G/L)	Dry mix (g)	Water (ml)	95% EtOH (ml/L)	Maltose dextrin (g)
   Week 1	338 (48)	133	894	63	4
   Week 2, 3	352 (50)	133	891	66	0
   Week 4, 5	366 (52)	133	889	68	0
   Week 6, 7	380 (54)	133	886	71	0
   Week 8	400 (57)	133	882	75	0

Product #F1259SP; Liquid Rat Diet, LD '82; Shake & Pour; Control

Directions for preparing 4 liters

1. Add 1/3 of the dry mix, Product F1259SP (902.2 gm/bag) into the Shake & Pour Bottle.
2. Measure 3440 ml. of warm tap water. Pour 113 of the warm tap water to dry mix into the Shake & Pour Bottle.
3. Shake approximately 30 seconds.
4. Add additional 113 of the diet and additional 1/3 of warm tap water into the Shake & Pour Bottle.
5. Shake approximately 30 seconds.
6. Add the remaining 113 of the diet and 113 warm tap water.
7. Shake approximately 30 seconds.
8. Refrigerate any unused portion.

For best results

1. As a matter of convenience, the mixing directions and package quantities may be proportionately reduced or increased. For added convenience, use a bottle with pre-measured water level markings.
2. If lumps occur, add the dry mix to the water in four increments of 25% each and shake the diet for one minute between each addition of dry mix.
3. Under refrigeration, the sealed packages of dry mix have a shelf life of up to six (6) months.
4. Shelf life of liquid diet: The prepared liquid diet should be maintained under refrigeration and should be dispensed within three (3) days. The prepared diet should not be kept longer than 48 hours at room temperature to avoid any spoilage. Invert bottle several times prior to dispensing to guard against possible settling in some complex formulations.

Product #F1258SP; Liquid Rat Diet, LD'82; Shake & Pour--Ethanol

Directions for preparing 4 liters

1. Add II3 of the dry mix, Product Fl258SP (533.6gm/bag) into the Shake & Pour Bottle.
2. Measure 3560 ml of warm tap water. Add II3 of warm water to dry mix in Shake & Pour Bottle. Shake approximately 30 seconds.
3. Add additional 1/3 of the diet and then 1/3 of warm tap water into the Shake & Pour Bottle.
4. Shake approximately 30 seconds.
5. Add the remaining 1/3 of the diet and the remaining water into the Shake & Pour Bottle and shake approximately 30 seconds.
6. Measure 268 m!. of 95% Ethanol (not included) and add to the Shake & Pour Bottle.
7. Shake approximately 30 seconds.
8. Refrigerate any unused portion.

NOTE: These directions are for preparing diets that yield 350 Kcal per liter of ETOH.

For best results

1. As a matter of convenience, the mixing directions and package quantities may be proportionately reduced or increased. For added convenience, use a bottle with pre-measured water level markings. Use a scoop to remove the product.
2. If lumps occur, add the dry mix to the water in four increments of 25% each and shake the diet for one minute between each addition of dry mix.
3. Under refrigeration, the sealed packages of dry mix have a shelf life of up to six (6) months.
4. Shelf life and feeding method:
   2. The prepared liquid diet should be maintained under refrigeration and should be dispensed within three (3) days.

   3. It is recommended that ETOH animals be gradually introduced to the ethanol diet as follows:

      1st--3rd Day 1/3 ETOH & 2/3 Control
      4th--7th Day 2/3 ETOH & l/3 Control
      8th Day on Pure ETOH diet.