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Simple Summary: The poultry industry recognizes that handling day-old chicks in commercial hatcheries can lead to long-lasting changes in their behavior. However, these hatchery-related stresses are more intense and complex than those measured by traditional behavioral tests. This study developed a new behavioral handling test for day-old chickens by incorporating concepts from behavioral tests used with both young and adult birds. In the new test, 96 two-day-old chicks from seven breeds of native Japanese and Western chickens were used to evaluate 10 behavioral traits, including vocalization frequency and responses to human interaction. The results classified chicken temperaments into three categories: bustle, aggression, and timidity. These categories were used to classify the seven breeds. This new handling test provides a valuable tool for understanding the behavior of various chicken breeds and offers insights into their complex behaviors. It is well known in the poultry industry that fear and stress experienced during the handling of day-old chicks in commercial hatcheries can have long-lasting effects on their behavior later in life. These hatchery-related stresses are more intense and complex than those encountered in traditional behavioral tests. Consequently, a single behavioral test may not be sufficient to measure hatchery stresses and chicken temperament. In this study, we developed a new behavioral handling test for day-old chickens, which incorporated concepts from established behavioral tests used with both young and adult birds. The new test assessed 10 behavioral traits, including vocalization frequency and responses to human interaction. It was conducted on 96 two-day-old chicks from seven breeds of native Japanese and Western chickens. The results of the principal component analysis classified chicken temperaments into three distinct categories: bustle, aggression, and timidity. Using these categories, the seven breeds were classified into five groups, each with distinct temperaments. This study highlights the reliability and value of the new handling test in characterizing the temperaments of various chicken breeds and provides insights into the complex behaviors of chickens. [ABSTRACT FROM AUTHOR]