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The aim of this study was to estimate effects of CSN1S1-CSN3 (α(S1)-κ-casein) composite genotypes on milk production traits and milk coagulation properties (MCP) in Mediterranean water buffalo. Genotypes at CSN1S1 and CSN3 and coagulation properties [rennet clotting time (RCT), curd firming time (K₂₀), and curd firmness (A₃₀)] were assessed by reversed-phase HPLC and computerized renneting meter analysis, respectively, using single test-day milk samples of 536 animals. Alternative protein variants of α(S1)-CN and κ-CN were detected by HPLC, and identification of the corresponding genetic variants was carried out by DNA analysis. Two genetic variants were detected at CSN1S1 (A and B variants) and 2 at CSN3 (X1 and X2 variants). Statistical inference was based on a linear model including the CSN1S1-CSN3 composite genotype effect (7 genotypes), the effects of herd-test-day (8 levels), and a combined days in milk (DIM)-parity class. Composite genotype AB-X2X2 was associated with decreased test-day milk yield [-0.21 standard deviation (SD) units of the trait] relative to genotype BB-X2X2. Genotypes did not affect milk protein content, but genotype AB-X1X1 was associated with increased fat content compared with genotype BB-X2X2 (+0.28 SD units of the trait) and AB-X1X1 (+0.43 SD units of the trait). For RCT, the largest difference (+1.91 min; i.e., 0.61 SD units of the trait) was observed between genotype AA-X1X2 and AB-X1X1. Direction of genotype effects on K(20) was consistent with that for RCT. The maximum variation in K₂₀ due to genotype effects (between AA-X1X2 and AB-X1X1 genotypes) was almost 0.9 SD units of the trait. Magnitude of genotype effects was smaller for A₃₀ than for RCT and K₂₀, with a maximum difference of 0.5 SD units of the trait between genotype AA-X1X2 and AA-X1X1. The B allele at CSN1S1 was associated with increased RCT and K₂₀ and with weaker curds compared with allele A. Allele X2 at CSN3 exerted opposite effects on MCP relative to CSN1S1 B. Because of linkage disequilibrium, allele B at CSN1S1 and allele X2 at CSN3 tend to be associated and this likely makes their effects cancel each other. This study indicates a role for casein genes in variation of MCP of buffalo milk. Further studies are necessary to estimate the effects of casein genetic variants on variation of cheese yield.

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The aim of this study was to investigate the effects exerted by the content of casein and whey protein fractions on variation of pH, rennet-coagulation time (RCT), curd-firming time (K20), and curd firmness of Mediterranean buffalo individual milk. Measures of milk protein composition and assessment of genotypes at CSN1S1 and CSN3 were obtained by reversed-phase HPLC analysis of 621 individual milk samples. Increased content of αS1-casein (CN) was associated with delayed coagulation onset and increased K20, whereas average pH, RCT, and K20 decreased when β-CN content increased. Milk with low κ-CN content exhibited low pH and RCT relative to milk with high content of κ-CN. Increased content of glycosylated κ-CN was associated with unfavorable effects on RCT. Effects of milk protein composition on curd firmness were less important than those on pH, RCT, and K20. Likely, this occurred as a consequence of the very short RCT of buffalo milk, which guaranteed a complete strengthening of the curd even in the restricted 31 min time of analysis of coagulation properties and for samples initially showing soft curds. Effects of CSN1S1-CSN3 genotypes on coagulation properties were not to be entirely ascribed to existing variation in milk protein composition associated with polymorphisms at CSN1S1 and CSN3 genes. Although the role of detailed milk protein composition in variation of cheese yield needs to be further investigated, findings of this study suggest that modification of the relative content of specific CN fractions can relevantly influence the behavior of buffalo milk during processing.