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Additives to the Drinking Water for Rats and Mice Research Committees Boston University

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  Introduction It is often convenient as well as less interruptive to the animals to add medication as well asexperimental compounds to the drinking water, rather than administering them by gavage or parenterally. A critical factor to be determined when any compound is added to the drinking water isthe animals’ fluid intake. It must be assured that fluid intake will not be diminished and the animals willnot become dehydrated because of any new taste, smell, texture or other factor altering the drinkingwater. This policy outlines procedures to be implemented for any and all drinking water additives. Policies  A. Any compound added to the drinking water must be approved in the IACUC protocol. B.  Ad lib  water consumption must be determined before any compound is added to the drinkingwater.C. Compounds that are commonly used and with which the PI is familiar and can provide data or references that the water intake will not be reduced are exempted from pre-determination of ad lib water consumption. These include, but are not limited to, sulfatrim and trimethoprim-sulfamethoxazole. For compounds that have not been previously administered and for which data are not available, PImust provide data as described in II.A. LASC/LACF will attempt to maintain a database allowing theinformation of tested additives to drinking water to be shared between investigators.D. If animals will receive a choice of unadulterated drinking water in addition to a water bottle withadditive, as in a preference study, making a choice of drinking water possible, the study is exemptedfrom pre-determination of ad lib  water consumption.E. Monitoring for hydration by evaluating skin turgor, food consumption and body weight must be donefor novel compounds and documented daily for minimum of 3 days. F. The PI is responsible for labeling all drinking water with additives.G. A compound such as sucrose may be added to the drinking water in order to make theexperimental compound or medication more palatable (II. D. 2&3). Procedures  A. Evaluation of ad lib water consumption 1. Prior to introducing a new compound in the drinking water PI must determine the strain’stypical 24 hour water consumption.2. The mean daily (24 hour) water consumption varies between species, stocks and strains. It istherefore mandatory to determine the ad lib  water intake for the same strain, sex, age and weightrodents as will be used in the study. As an example, the mean daily water intake in mice hasbeen reported averaging 7.7 +/- 0.3 ml/g body weight and varying 2-fold across strains. 1 3. Once the ad lib  water consumption has been determined for one strain of mice or rats or other rodent species, ad lib  water intake need not be repeated.4. Published data, or data from the planned LASC/LACF database, if available, may be used. B. Method for determining 24 hour ad lib water consumption. Alternate Additives to the Drinking Water for Rats and Mice » Research Committe...http://www.bu.edu/orccommittees/iacuc/policies-and-guidelines/additi...1 of 329.09.2014 13:12  methods may be considered as reviewed and approved by the IACUC. 1. Rodents are housed in their usual cage. 2. A Special Care Instruction card stating  “water is being measured DO NOT CHANGEWATER BOTTLE ” is placed on the cage to prevent animal care staff from changing the water bottle or interfering with the study.3. The water bottle is filled (~ 200 ml-400 ml) and weighed using a digital scale to 0.1 gaccuracy. 4. The same sipper tube as the animals are used to is used for the study. 5. The bottle with water is weighed 3 days in a row at the same time of day to determine 24hour ad lib  water consumption.6. The 3 measurements are averaged and then divided by the # of rodents in the cage. C. Adding the novel compound to the drinking water anddocumentation of fluid intake 1.   Volume of the water with additive consumed must be measured each day, as describedabove, and documented over a period of at least three (3) days.2.   When a novel compound is added to the water, daily observations of the rodents to evaluatehydration and fluid and food consumption must be documented for the first three (3) days.3. If rodents drink less than 90% of their previous intake, become dehydrated or lose weight, PImust adjust the drinking water with additive so as to make it palatable for the animal. If problempersists, PI must consult the veterinary services. D. Antibiotics added to the drinking water   1. SULFAMETHOXAZOLE/TRIMETHOPRIM (SULFATRIM) Stock solution 200 mg Sulfamethoxazole +40 mg Trimethoprim /5 mlMOUSE Dose 95 mg/kg/24 hours 5.3 ml of stock in 400 ml drinking water. 2 RAT Dose 53 mg/kg/24 hours 4.4 ml of stock in 400 ml drinking water. 2  Suspension: Shake bottle dailyLight sensitive: Cover bottle with foil or dispense in colored bottle 2. DOXYCYCLINE (AS ANTIBIOTIC)* MOUSE Dose 5-10 mg/kg/24 hours 5.3 ml of stock in 400 ml drinking water RAT Dose 5-10 mg/kg/24 hours 4.4 ml of stock in 400 ml drinking water Suspension: YesLight sensitive: YesDoxycycline is bitter and consequently it is advisable to add 2.0 – 5.0 % sucrose to thewater.*Doxycycline is also used as genetic modifier.   Dose to be determined by PI. 3. TETRACYCLINE Additives to the Drinking Water for Rats and Mice » Research Committe...http://www.bu.edu/orccommittees/iacuc/policies-and-guidelines/additi...2 of 329.09.2014 13:12  MOUSE 50-60 mg in liter drinking water Tetracycline is bitter and consequently it is advisable to add 2.0 – 5.0 % sucrose to thewater. REFERENCES 1. Bachmanov, AA, Reed, DR, Beauchamp, GK and Tordoff, MG. Food intake, water intake, anddrinking spout preference of 28 mouse strains. Behav. Genet. 2002. 32:435-443. For detailedinformation please see http://phenome.jax.org/pub-cgi/phenome/mpdcgi?rtn=meas/catlister&req=Cmetabolismqqqwater%20intake .2. Hawk, C. Terrence et.al. Eds. Formulary for Laboratory Animals. Third Ed. 2005. BlackwellPublishing. Rat and mouse doses calculated from allometric correction from large animal dose (30mg/kg/day).  BU IACUC approved May 2011 Additives to the Drinking Water for Rats and Mice » Research Committe...http://www.bu.edu/orccommittees/iacuc/policies-and-guidelines/additi...3 of 329.09.2014 13:12
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