Colloid Mill Modified Zein Suspension Technology for Plant Protein Rich Bird Food: First Clutch Performance Compared with Egg Based Food in Fiorino Canaries

Background The present study evaluated a technology-driven approach for producing a plant-protein-rich bird food based on colloid-mill-modified zein suspension and compared its first-clutch reproductive performance with an egg-based food in Fiorino canaries. The formulation strategy was informed by literature showing that intensive mechanical processing can reduce aggregation and improve matrix homogeneity in zein-rich maize protein systems [16]. ZEIN16 was therefore evaluated primarily as a processed-zein technology platform and bird-food prototype. Methods Forty first-time Fiorino canary pairs were allocated to two isoprotein dietary groups, with 20 pairs per group: EGG16, an egg-based bird food, and ZEIN16, a modified zein suspension-based bird food. Both diets contained approximately 16% crude protein and were fed for 12 weeks before pairing. ZEIN16 was produced using colloid milling as the zein-modification step, guided by zein-processing literature. Each breeding pair was the independent experimental unit. First-clutch egg number, hatched chick number, unhatched eggs, and hatchability were analysed using pair-level data; pooled hatchability was treated as descriptive egg-level information. Results EGG16 produced significantly more eggs than ZEIN16 (3.65 ± 0.75 vs. 2.90 ± 0.55 eggs per pair; Mann-Whitney p = 0.0012; mean difference = 0.75 eggs per pair, 95% CI 0.33 to 1.17; Cliff's delta = 0.55). Hatched chick number was numerically higher in EGG16 (2.95 ± 1.15 vs. 2.50 ± 0.69 chicks per pair), but the complete-data comparison did not reach the 5% significance threshold (p = 0.0627; mean difference = 0.45 chicks per pair, 95% CI -0.16 to 1.06). Individual hatchability did not differ between groups (79.42 ± 25.42% vs. 86.25 ± 17.79%; p = 0.5759). Descriptive pooled hatchability was 80.82% for EGG16 and 86.21% for ZEIN16 (Fisher exact p = 0.4850). Supporting count and binomial sensitivity analyses did not change the qualitative interpretation. Conclusions EGG16 improved first-clutch egg production. ZEIN16 did not show a statistically detectable reduction in hatchability after eggs were laid, but equivalence or non-inferiority to EGG16 was not tested. ZEIN16 should therefore be positioned as a preliminary plant-protein-rich bird egg food analogue produced through colloid-mill-based zein suspension technology, rather than as a reproductive replacement for egg-based food.