Page 1 Examining Behavioral Compatibility in Zebra Finches Joy Watkins University of Wyoming Mentor: Dr. Sarah Benson-Amram and Lisa Barrett Department of Zoology and Physiology Animal Behavior & Cognition Lab Mentor Approval:______ ________________________ Date:____8/30/18____________ Page 2 Introduction The zebra finch (Taeniopygia guttata) is a common Estrildid finch of Central Australia and Indonesia. These monogamous birds are easily kept in captivity[1.] and exhibit complex, adaptive social behaviors, making them ideal candidates for controlled studies of behavior. [2.] For example, zebra finches mate for life and coordinate their parenting abilities which increases their overall reproductive success.[3.] However, we lack an understanding about the factors affecting coordination in this species. Animals differ in their ability to problem solve and work together, and one way that they differ is in their individual personality. Animal personality is defined as individual differences in behavior that are consistent across time or context.[4.] One way we can assess coordination in animals is by presenting them with a cognitive task, such as a puzzle that they can solve by working together.[3.] A puzzle box is an apparatus with parts that must be moved or opened for the animal to obtain a food reward.[5.,6.] Personality traits can affect ability to access novel resources. For example, a bolder or more dominant individual may approach a novel object more quickly, and they might also solve a novel task more quickly than a shyer individual. However, there is a speed-accuracy trade-off where a quicker individual may not be as accurate at detecting seeds as a slower individual.[11.] Thus, it is not necessarily beneficial to be bold in all contexts. For monogamous species this could have importance in mate choice: It could be beneficial to form a pair bond with a mate who is dissimilar from your own personality.[7.8.] On the other hand, there are benefits to being similar—and perhaps more coordinated—with your partner.[9.10.] Therefore, we investigated which personality combinations would improve pair coordination by comparing pair bonds with similar personalities to those with dissimilar personalities on two novel coordination tasks: 1) a maze, and 2) a two-solution puzzle-box task. Another important attribute in pair coordination is communication. Recent studies have shown that zebra finches that vocalize more with their partners are more likely to successfully reproduce.[19.] For example, birds give an increased frequency and rapid series of tet calls to coordinate a flight take-off.[1.] Tet calls are some of the most common within-pair vocalizations in zebra finches. To identify how vocal communication is used during problem solving, and whether pair- bonded birds use vocal communication to facilitate coordination, we recorded tet vocalizations during coordination testing. Research Aims & Hypotheses We aim to: 1. Determine which personality combinations of zebra finch pair bonds perform best on novel coordination tasks. Null Hypothesis: There will be no difference in coordination task performance between personality combinations. Prediction: If there is no difference in coordination task performance between personality combinations, then the rate at which pair bond members solve a two-step puzzle task will be the same among pairs. Alternative Hypothesis: Similar or dissimilar personality pairs will solve coordination tasks faster than the other depending on the behavioral trait combination. Prediction: Pairs with compatible personalities will be more successful and solve the task faster than birds with incompatible personalities. 2. Identify how vocal communication is used during problem solving on novel tasks between members of a monogamous pair bond, and how vocalization might differ between pair bonds of differing personality types. Null Hypothesis: There will be no difference in coordination task performance between varying rate of vocal production of tet calls. Prediction: If there is no difference in coordination task performance between varying levels of vocal communication, then the rate at which pair bond members and familiar non-mate members vocalize during a trial will be the same, and there will be no difference in success between pairs that vary in their rate of vocalization. Alternative Hypothesis 1: Pairs use tet vocalizations to communicate their next and prior steps to their mate. Page 3 Prediction 1: If pairs use tet vocalizations to communicate their next and prior steps to their mate, then we would see a greater amount of tet vocalizing just prior to and following movement in the maze. Equipment and Methodology In a pilot study (Spring 2017), we allowed 10 birds to pair freely in a large flight cage. From this, four pair bonds were formed. Before pairing, birds were housed in same-sex flight cages, and after pairing they were housed with their pair bond member in a breeding cage, at the University of Wyoming (IACUC Protocol #20150507SB00161-02). Throughout the experiment the birds are fed a mixture of finch seed, cuttlebone, water, egg food supplement, fresh greens, and vitamin drops ad libitum. In order to create a control group of familiar, non-pair-bonded relationships between pairs of birds, two pair-bonded birds were housed in the same cage, separated by a mesh barrier. Pairs will be tested on two novel coordination tasks: a maze and a two-step puzzle box. Thus far, the pairs have already undergone pilot maze testing (Fall 2017) and are have finished with the training phase of the two-step puzzle box (Spring 2018). The pilot pairs were Figure 1: Birds that took longer to tested on the two-step puzzle box in Summer 2018. resume feeding after startle also tended to be more neophobic, on average. Personality Assessment All birds involved in this research were first assessed for personality differences. Personality traits of individual zebra finches were assessed (2015-2016) by evaluating dominance, exploration tendency, and neophobia (fear of novel objects). The three different personality tests were evaluated on 10 female and 10 male birds. Our preliminary data suggests that more neophobic individuals take longer to resume feeding after being startled than less neophobic birds (F1, 18=19.44, p=0.0003; Fig. 1). Additionally, more neophobic individuals were significantly more aggressive compared to less neophobic birds (p=0.026). These personality measures have had an impact on performance in cognitive tasks. Specifically, it was found that less neophobic individuals, as measured by the latency to perch near a feeder in the startle tests, tended to Figure 1: A schematic of the novel maze task, where solve a greater total number of individual problem-solving members must coordinate their behavior to achieve the tasks (string-pulling and wire-pulling tasks, p=0.08). food reward: The male zebra finch must travel through the blue square doorway and the female must travel through the red circle doorway. Maze Task All pairs of zebra finches performed two maze tasks, one with a pair-bonded mate, and one with a familiar non-mate, to assess whether pair bonding affects coordination (Fig. 1). In these tasks, each individual in the pair only has partial knowledge of the whole maze, such that members must coordinate their actions in order to solve the task efficiently. The two mazes differ in the symbols that are used to differentiate the compartments and in the location of the food. Individuals were food deprived for one hour prior to a trial to increase and standardize their motivation and were trained on their respective doorway two consecutive weeks prior to testing. Trials lasted one hour and were video-recorded for later data- extraction. Birds were considered to have succeeded at solving the task if they enter the compartment with food during the trial. Pilot testing of the maze task was carried out in Fall Figure 2: Two-step puzzle 2017 and Spring 2018. box: One member must remove a wire, and then Two-Step Puzzle Box the other member must To address the possibility that birds could follow one another throughout the maze without remove the bottom knob to necessarily combining their knowledge, we assessed pair bond coordination on a two-step obtain the food reward. Page 4 puzzle box. This assay required that each individual learns to solve one step of the two-step task such that in order to solve the complete task, both members of a pair must combine their knowledge (Fig. 2). Food is placed at the top of the apparatus where one partner must pull a wire to unhinge the platform. Once the wire has been extracted, the food reward falls to the top of a plug-like lid. The second member of the pair bond must then dislodge the lid’s knob to release the food from the apparatus. Each member of the pair was trained to solve one of the two solutions and was then tested together on the two- step apparatus as a pair and individually (to test for the possibility of social learning of the second step). Pilot testing of the two-step puzzle box was carried out in Summer 2018. Results and Discussion Our two-step puzzle-box task training protocol required that birds solve consecutively (i.e., Solve across three consecutive days) in Dominance and Latency to Learn order to proceed to testing with their mate. During Summer 2018, 164 learning ability of the pilot birds was assessed over the duration 162 of 130 days where each of the eight individuals were trained on their solution. Six of the individuals solved consistently for at 160 least three days and moved forward to being tested as a pair and 158 R² = 0.9144 individually to assess social learning. Interestingly, individuals 156 that were more neophobic in novel object tests experienced 154 decreased learning ability on the two-step task training when 152 compared to bolder individuals. Therefore, less neophobic 150 individuals experience greater solve frequency than more 148 neophilic individuals, perhaps because the neophobic individuals 93 94 95 96 were inhibited by fear of a novel apparatus. In terms of Latency to Learn (days) dominance, there was a statistically significant positive correlation between dominance and learning ability on the task, with more dominant individuals tending to learn their step faster Figure 3: There was a significant positive relationship than less dominant individuals (R=0.95, p <0.05 (Fig. 3; between the number of dominant individuals to learn their Appendix, Table 3)). This supports our prediction that similar task and the decreased latency to learn their solution pair bonds would be better coordinated on novel tasks. (R(2)= .91, p-value < .05). Individuals took 93 to 127 days to learn their respective step of the two-step puzzle task. After testing with their mate, four individuals performed their mate’s solution in addition to their own when tested individually, suggesting that social learning of the second solution occurred. These four individuals differed in how long it took them to perform the newly acquired solution (range: 36-401 minutes). Anecdotally, he females were more latent than males to learn their mate’s solution. Vocal communication was recorded during maze testing, and all trials were video- recorded for later data extraction. RavenLite (Fig. 5) and Audacity acoustical software were utilized to analyze calling types so that we could determine which calls were used at different points in the maze. The ‘warbleR’ package in R was employed with Autodetect to analyze the frequency of tet Figure 5. Song harmonics of a male zebra finch (6 elements, fundamental frequency of .53 and and song calls made during duration of 4.66 seconds) produced with RavenLite acoustical software.. testing in the maze. Pair Dominance Score (minutes) Page 5 bonds that produced a greater number of tet calls experienced a higher success rate highlighting the importance of vocal communication in coordination contexts in this species (Appendix Figs. 4 & 5). The proposed project involved completing our pilot study to help prepare for expansion to a cohort of 40 zebra finches. In future work, these 40 birds will be allowed to freely choose their mates according to Silcox and Evans’ (1982) criteria.[22.] Half of these pairs will be experimentally manipulated so that they are dissimilar or similar, based on a predetermined absolute value difference in personality scores of all birds, and all of the birds will be evaluated on correlation of solve success to that of individual and paired personality traits. In summary, pair testing during Summer 2018 indicates that pair bonds coordinate their actions to solve novel tasks. There is some evidence that personality plays an important role in learning to solve a novel problem and that more similar pair bonds are more successful on the tasks than dissimilar pair bonds. Furthermore tet calling could serve to facilitate coordination on the maze task. The ability individuals to learn their mate’s solution on the two-step puzzle task indicates social learning could be important in this species. We plan to expand on this work by increasing to a cohort of 40 zebra finches as previously mentioned in the discussion. The birds paired experimentally and non-experimentally with similar and dissimilar personality traits will be evaluated on correlation of solve success to that of individual and paired personality traits within the maze task and the two-step puzzle task. All testing trials will be audio- and video- recorded for data extraction and will be analyzed with ‘warbleR’, Audacity, and Raven Pro. References 1. Zann, R. A. (1996). 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Dominance scores were assessed by the duration of time (in seconds) that an individual spent feeding at a single feeder within a group of 10 individuals. Neophobia Dominance ID Sex Pair (sec) (sec) Athena F A 601 0.9 Kennedy M A 11 4.78 Cleopatra F B 301.5 2.4 Truman M B 1 5.56 Theresa F C 35.5 2.5 Jackson M C 344 2.56 Jane F D 90 2.7 Grant M D 12.5 2.67 Table 3. There was a significant positive relationship between dominance score and latency to learn a single portion of the two-step puzzle (p < 0.05). Regression Statistics Multiple R 0.956226 R Square 0.914367 Adjusted R Square 0.871551 Standard Error 2.012461 Observations 4 ANOVA Significance df SS MS F F Regression 1 86.49 86.49 21.35556 0.043774 Residual 2 8.1 4.05 Page 8 Total 3 94.59 Standard Upper Lower Upper Coefficients Error t Stat P-value Lower 95% 95% 95.0% 95.0% Intercept 449.4 63.40051 7.088271 0.019328 176.6096 722.1904 176.6096 722.1904 Latency to Learn (days) -3.1 0.67082 -4.62121 0.043774 -5.98631 -0.21369 -5.98631 -0.21369 Figure 5. Pair-bonds that communicated less with tet vocalizations experienced a lower success rate on the maze task and increased latency to solve the maze task. Maze Task Solvers 400 350 300 250 200 150 100 50 0 -50 -100 -150 Tet Vocalization Frequency (n) Latency to Solve Maze (minutes) Pair C 257 32 Pair D 153 54 Pair C Pair D