Estrus Record-Keeping System (San Diego Zoo)

Instructions for Giant Panda Estrus 1-2-3 Record-Keeping System

In addition to your detailed behavioral observations, you may be asked to assist in completing this simple daily checksheet designed to monitor the female’s estrus.  For an example of a completed 1-2-3 checksheet for estrus, click here.

• If you don't know for certain whether a particular behavior or condition was present, please do not guess. Record a "/" for that entry to indicate that you do not know. When analyzing the data, this will be treated as a "missing value" rather than a "0." For example, on rainy days it may be difficult to determine whether a female engaged in water play or to estimate the number of urinations. Also, before the onset of peak estrus, you will be less likely to observe the vulva to assess changes. Or you may simply not have observed that female on that day, or not spent as much time observing her as you normally do. If , for example, you observed a female for only a few minutes in the morning and she was eating bamboo, you really have no idea whether she bleated much on that day or not.

• If it was impossible for a particular behavior or condition to occur on that day, please record a "/" for that entry. For example, if a female was not housed next to a male, she would not be capable of showing any interest in a male, and males would not be capable of showing any interest in her. Also, copulation and mounting cannot occur unless the female is placed together with a male. Similarly, a female locked inside her bedroom for most of the day may not be able to express many behaviors, and, for example, the level of bleating, chirping, etc. may be artificially low due to lack of social stimulation.

• If a particular behavior or condition was not observed that day, despite a "normal effort" to observe that animal, record a "0."

• If a behavior/condition changes slightly from baseline for that animal, record a "1." For example, if the appetite is a little decreased or the vulva are a little more pink, record a "1." If the female shows a little more interest in neighboring males than is normal for her, record a 1. If the female lifts her tail only occasionally and does not hold it up long when touched, record a "1." If lordosis, backwards walking or rolling are seen, but only rarely, record a "1." If bleating and chirping increase above the "normal" amount for that panda, record a "1." For example, some pandas bleat frequently year-round, but it is associated with being fed. Only if bleating increases above this level, or the context of its occurrence changes (i.e., the female bleats socially with other pandas in addition to her usual food-related bleating), should you score a "1."

• Record a "2" if the behavioral changes are moderately elevated (or decreased in the case of appetite), and a "3" if they are dramatically elevated, that is, close to the maximum you have observed for this behavior/condition.

Trends associated with estrus:

1. Decreased appetite. Please monitor the female's food intake, both bamboo and other provisioned foods. If the female's consumption decreases as a result of mucus stool, stomach upset, or some other illness, please place an asterisk (*) by the entry on the record sheet and make a note of this in the comments.
2. Water play.  Definition: Splashing and rolling in water. You may also infer water play from wet pelage and evidence of water splashed out of the pool.
3. Urination. Females nearing peak estrus tend to urinate more frequently and in smaller amounts. This is often accompanied by scent marking. You may use a combination of direct behavioral observation as well as counting urine puddles in the enclosure.
4. Activity. Females in estrus tend to show a higher level of activity, though activity may fall again as peak estrus is reached. Definition: Any social or locomotor activity or movement of the body (e.g., scratching, rolling)
5. Scent marking. Scent marking, usually in the squatting or reverse position, increases markedly in estrus, though again it may decrease when peak estrus is reached. Definition: Rubs anogenital region (tail up) against object or substrate.
6. Vulva color. The vulva become progressively more red during estrus, though it may become a little white as peak estrus is reached. Please note this whitening in the "Notes" section at the bottom of the datasheet.
7. The vulva become progressively more swollen and more open as peak estrus approaches.
8. Secretions from the vulva may make the vulva appear wet.
9. Interest in male. An estrous female generally exhibits increased interest in neighboring males. Definition: Female interest should be determined first and foremost by a tendency for her to seek out the male, for example, spending more time adjacent to his pen and observing him. Their interactions need not be "friendly" for her to show such interest. Simply seeking the male out should usually receive a score of "1" or "2," but if she also shows heightened interest by greeting, bleating, chirping, raising her tail, presenting her anogenital region, etc., this should receive a score of "3." The occurrence of moans, barks, and paw swats appear to be a normal part of the courtship pattern, especially when the female is in early estrus. Therefore, the occurrence of these behaviors should not be used to indicate a "lack of interest."
10. Nearby male(s) interested in female. A capable male is probably the best estrus-detecting machine yet developed. Definition: As above, except the male exhibits typical male courtship behavior: elevated bleating, investigation of the anogenital region, and perhaps rapid pacing back and forth approaching the female and/or her pen.
11. Females approaching peak estrus often raise their tail when it is touched. If she raises it, but "reluctantly" or infrequently, record a "1," but if she raises it more readily and it remains up, score a "2" or "3."
12. Females approaching peak estrus will begin to exhibit lordosis, or the sexual posture, more frequently. This may occur in the absence of any apparent stimulus, or may occur when touched for the tail-up test or when presenting to a male.  Definition: Tail up, hind quarters raised, and kneeling on forelegs; head often on the ground. Back is often arched.
13. Bleating. The frequency of bleating increases during the estrous period, and as peak estrus is reached, the duration of individual bleats increases markedly.  Definition: A twittering, goat-like call of variable length (1-3s). Contact call, appeasing, non-aggressive, “friendly.”
14. Chirping. The frequency of chirping increases during estrus, especially peak estrus.  Definition: Short, tonal, high-pitched, descending in pitch toward end. Friendly, promotes social proximity. Do not confuse with a truncated bleat.
15. Backwards walking is seen more frequently during estrus, especially peak estrus.  Definition: Female walks backward for several steps. Often toward male and may be followed by anogenital present. Often accompanied by bleating and chirping.
16. Rolling is seen more frequently during estrus, especially peak estrus. Definition: Animal lies on back and rolls and writhes (as if back is itching). Looks different from normal dirt bathing, and is often accompanied by bleating and chirping.
17. Sexual mount by male. The male mounts the female and attempts to copulate. Obviously, this can occur only if the pair are put together for a breeding introduction.
18. Copulation/AI. Please note if the female copulates or if artificial insemination is performed on her.
19. % Cornefied cells (cells with broken nuclei) appears to increase to over 70% when peak estrus is reached. Usually more than 80% of the cells are cornified, but some females have been known to reproduce with less than 60%.
20. Please note if the female is housed next to an adult male and they are allowed to interact through some sort of barrier.