This technique is non-invasive and minimizes disruption to the social group. Animals are easily habituated to the collection and exhibit no ill effects from this procedure. The easiest most efficient way to collect urine for a ctop, is to wake them up in the morning. All primates have a full bladder upon awakening and you can get a fresh sample very quickly. This technique works the best, if you have your animals on a controlled light cycle. Ideally, you want to turn the lights on when you enter the cage. Make sure you have your bucket ready, because they will urinate within a few minutes of waking up!! If you can’t control the light cycle in your exhibit, I would strongly urge you to consider collecting feces (see attached protocol).

Materials you will need:

• 1 clean urine bucket per animal. You should label each bucket with the animal's name and use only that bucket for that animal.

• A few treats to be given upon awakening and following urination (sunflower seeds)

• 1 female cotton-top (more if you have them)

• collection tubes for the freezer (Sarstedt tubes work the best) - contact Anne if you need tubes

Steps for Collecting Samples:

1. Turn on the lights and enter cage with the urine bucket and a few treats. Feed the female and her cagemates a few treats to wake them up. Make her walk to you to get the food. The activity will stimulate her to urinate.

2. The female will generally lift her tail slightly before she urinates. It has been our experience that females will urinate 5-15 minutes after the light has been turned on.

3. While the female is urinating, place her clean bucket under her. You do not need to place the bucket very close to her. In fact the closer you get the bucket to her genitals the more likely she is to move making urine collection difficult. If you position the bucket under the tip of her tail you should not have any difficulty obtaining a sample. It's best to avoid collecting urine that is running down branches since it may be contaminated by other steroids. However, if that is the only available means of collection do not pass up the opportunity.

4. Females will urinate approximately 4-6 mls in the morning! We only need approximately 1 ml for analysis so don't worry if you miss a few drops here and there. (Note: Even though we only need 1 ml for the analysis, we store the rest)

Urine Sample Preparation

• Aliquot approximately 4 mls ( but not more) of urine into the Sarstedt tube. Greater than 4 mls will expand when frozen and pop off the lid.

• Label the vials with the date, CTT (for cotton-top tamarin), name of animal, and your zoo name's initials, e.g.:

4 May 98                                                                                                                                       CTT Sue RWPZ

Please make sure you write out the month, do not record the date as 5/4/94 -- it's too confusing as to whether it is day or month first.

If you make a mistake when labeling the tube, it's not the end of the world, but please don't just cross out the mistake. Instead, THROW THE TUBE WITH THE MISTAKE AWAY, and make a new one. If you don't, several months from now when the urine is analyzed, people may not understand what is written on the tube and will have to throw it away, and not analyze the urine. Thus wasting the fact that you woke up at an ungodly hour this morning to collect urine from a bunch of monkeys.

• Record the information on a daily log sheet eg.

Date                     Number of Vials             Total Urine Volume

3 Sept 91                      1                                      4

4 Sept 91                      0**                                  0***

5 Sept 91                      1                                      4

*** Note: If urine is not collected on one day make sure it is documented in the log

5. Wash the urine buckets with soap (Nolvasan) and water. Make sure that it is rinsed clean and dry before you collect urine. Use the same urine bucket for each individual. If you mix buckets between animals you can potentially contaminate the sample.

Freezing Samples

The urine needs to be placed in the freezer. Please keep the urine frozen until it is shipped.

The more samples you can collect the easier it will be to interpret your results. We recommend a minimum of 3 samples per week for 3 weeks to determine if your female is pregnant. If you want to determine if you female has regular cycles, it is important that you collect 3-4 cycles worth of samples (note: ovarian cycle is 18 days, roughly 4-6 weeks). If you can collect more samples during the week, please do so!! For example, you may want to schedule 5 days per week, hoping that you will easily get 3 samples per week. If you schedule 3 collections and you miss one during the week, it makes it significantly harder for us to interpret what is happening. Please note that these sample numbers are our best estimates for "typical females". Somehow, you might be the lucky one that has an atypical female and we may need more samples to give you a definite answer!

Samples should be shipped on ice (dry ice is preferred but not essential) in an approved styrofoam shipping container overnight air (Fed Ex, Airborne). Please call Anne Savage the day before you ship the samples 407 938-2837. We are happy to receive packages Monday - Friday but not on the weekends.

Ship to:

Dr. Anne Savage

Disney’s Animal Kingdom

Animal Programs Administration

1200 North Savannah Circle East

Bay Lake, FL 32818

(407) 938-2837

PLEASE BE SURE TO INCLUDE A SPECIMEN REPORT FOR EACH INDIVIDUAL ANIMAL FOR WHOM YOU HAVE COLLECTED FECES AND THE CONTACT NAME AND PHONE NUMBER SO THAT WE CAN SEND YOU THE RESULTS!