Short communication: Molecular genetic characterization of ovine αS1-casein allele H caused by alternative splicing

Sequencing of ovine CSN1S1*H cDNA showed an absence of exon 8 in comparison with GenBank sequences; the absence was confirmed by protein sequencing. We demonstrated that this allelic aberration is the result of a deletion of 4 nucleotides, the last 3 of exon 8 and the first 1 of intron 8, which are replaced by an insertion of 13 nucleotides in the DNA sequence. The insertion is a precise duplication of a part of the adjacent intronic sequence of CSN1S1*C″. These sequence differences result in an inactivation of the splice donor sequence distal to exon 8, leading to upstream exon skipping during the serial splice reactions of the ovine CSN1S1*H pre-mRNA, and may affect the specific casein expression as well as protein characteristics.     

Biochemical and molecular characterization of polymorphisms of αs1-casein in Sudanese camel (Camelus dromedarius) milk

This work was designed to detect occurrence of biochemical polymorphism of α_s1-casein in two ecotypes of Sudanese camel (Camelus dromedarius) and to characterize these variants on molecular level. Milk samples were screened for α_s1-casein variability by isoelectric focusing, using skimmed milk, as well as isolated α_s1-casein. Two protein patterns, named α_s1-casein A and C, were identified, whereas the major allele A revealed frequencies of 0.8214 and 0.8615 in the two ecotypes. CSN1S1*A and CSN1S1*C are both characterized by missing of exon 16 on mRNA-level compared with the previously described CSN1S1*B. However, the sequence of exon 16 occurs on DNA-level in both. Therefore, this exon seems to be skipped out during mRNA-processing. Furthermore, CSN1S1*C shows a single G > T nucleotide substitution in exon 5, leading to a non-synonymous amino acid exchange (p.Glu30 > Asp30; GenBank ID: JF429138). A polymerase chain reaction-restriction fragment length polymorphism-method was established as a DNA-based test for this nucleotide substitution.     

Short communication: Carora cattle show high variability in alpha(s1)-casein

The objective of this study was to analyze the genetic variability of milk proteins of the Carora, a shorthorned Bos taurus cattle breed in Venezuela and in other Southern American countries that is primarily used for milk production. A total of 184 individual milk samples were collected from Carora cattle in 5 herds in Venezuela. The milk protein genes α_s1-casein (CN) (CSN1S1), β-CN (CSN2), κ-CN (CSN3), and β-lactoglobulin (LGB) were typed at the protein level by isoelectrofocusing. It was necessary to further analyze CSN1S1 at the DNA level by a PCR-based method to distinguish CSN1S1*G from B. Increased variation was found in particular at the CSN1S1 gene, where 4 variants were identified. The predominant variant was CSN1S1*B (frequency = 0.8). The second most common CSN1S1 variant was CSN1S1*G (0.101), followed by CSN1S1*C (0.082). Moreover, a new isoelectrofocusing pattern was identified, which may result from a novel CSN1S1 variant, named CSN1S1*I, migrating at an intermediate position between CSN1S1*B and CSN1S1*C. Six cows carried the variant at the heterozygous condition. For the other loci, predominance of CSN2*A² (0.764), CSN3*B (0.609), and LGB*B (0.592) was observed. Haplotype frequencies (AF) at the CSN1S1-CSN2-CSN3 complex were also estimated by taking association into account. Only 7 haplotypes showed AF values >0.05, accounting for a cumulative frequency of 0.944. The predominant haplotype was B-A²-B (frequency = 0.418), followed by B-A²-A (0.213). The occurrence of the G variant is at a rather high frequency, which is of interest for selection within the Carora breed because of the negative association of this variant with the synthesis of the specific protein. From a cheese-making point of view, this variant is associated with improved milk-clotting parameters but is negatively associated with cheese ripening. Thus, milk protein typing should be routinely carried out in the breed, with particular emphasis on using a DNA test to detect the CSN1S*G variant. The CSN1S*G allele is likely to have descended from the Brown Swiss, which contributed to the Carora breed and also carries this allele.     

Molecular characterization of bovine CSN1S2*B and extensive distribution of zebu-specific milk protein alleles in European cattle

The B allele of the bovine α_S2-casein gene (CSN1S2) was characterized at the molecular level and the distribution of zebu-specific milk protein alleles was determined in 26 cattle breeds originating from 3 continents. The CSN1S2 *B allele is characterized by a C → T transition affecting nucleotide 17 of exon 3, which leads to a change in the eighth amino acid of the mature protein, from Ser to Phe (i.e., TCC →TCC). DNA-based methods were developed to identify carriers of CSN1S2*B and the other alleles (CSN1S2 *A, C, and D) at the same locus. CSN1S2*B and other zebu-specific milk protein alleles and casein haplotypes are widely distributed in European cattle breeds, particularly those of southeastern origin. Alleles CSN1S2 *B and CSN3*H are important in searching for zebu imprints in European cattle breeds. Diversity estimates at the milk protein loci were highest in the zebus followed by southeastern European taurines. Anatolian Black had the highest number of zebu alleles among European taurines. Common, group, and intergroup haplotypes occurred in the breeds and demonstrated relationships that concurred with developmental histories, genetic makeup, and, in particular, exposed the extent of zebu influence on southeastern European cattle.     

The occurrence of noncoagulating milk and the association of bovine milk coagulation properties with genetic variants of the caseins in 3 Scandinavian dairy breeds

Substantial variation in milk coagulation properties has been observed among dairy cows. Consequently, raw milk from individual cows and breeds exhibits distinct coagulation capacities that potentially affect the technological properties and milk processing into cheese. This variation is largely influenced by protein composition, which is in turn affected by underlying genetic polymorphisms in the major milk proteins. In this study, we conducted a large screening on 3 major Scandinavian breeds to resolve the variation in milk coagulation traits and the frequency of milk with impaired coagulation properties (noncoagulation). In total, individual coagulation properties were measured on morning milk collected from 1,299 Danish Holstein (DH), Danish Jersey (DJ), and Swedish Red (SR) cows. The 3 breeds demonstrated notable interbreed differences in coagulation properties, with DJ cows exhibiting superior coagulation compared with the other 2 breeds. In addition, milk samples from 2% of DH and 16% of SR cows were classified as noncoagulating. Furthermore, the cows were genotyped for major genetic variants in the α_S1- (CSN1S1), β- (CSN2), and κ-casein (CSN3) genes, revealing distinct differences in variant frequencies among breeds. Allele I of CSN2, which had not formerly been screened in such a high number of cows in these Scandinavian breeds, showed a frequency around 7% in DH and DJ, but was not detected in SR. Genetic polymorphisms were significantly associated with curd firming rate and rennet coagulation time. Thus, CSN1S1 C, CSN2 B, and CSN3 B positively affected milk coagulation, whereas CSN2 A², in particular, had a negative effect. In addition to the influence of individual casein genes, the effects of CSN1S1-CSN2-CSN3 composite genotypes were also examined, and revealed strong associations in all breeds, which more or less reflected the single gene results. Overall, milk coagulation is under the influence of additive genetic variation. Optimal milk for future cheese production can be ensured by monitoring the frequency of unfavorable variants and thus preventing an increase in the number of cows producing milk with impaired coagulation. Selective breeding for variants associated with superior milk coagulation can potentially increase raw milk quality and cheese yield in all 3 Scandinavian breeds.     

Polymorphic AP-1 binding site in bovine CSN1S1 shows quantitative differences in protein binding associated with milk protein expression

Polymorphisms in 5′-flanking regions of milk protein encoding genes can influence the binding activity of the affected response elements and thus have an impact on the expression of the gene products. However, precise quantitative data concerning the binding properties of such variable response elements have so far not been described. In this study we present the results of a quantitative fluorescent electromobility shift assay comparing the allelic variants of a polymorphic activator protein-1 binding site in the promoter region of the bovine α_s1-casein encoding gene (CSN1S1), which is affected by an A→G exchange at −175 bp (CSN1S1^−175bp). A supershift assay using a commercial c-jun antibody was carried out to verify the specificity of protein binding. The gel shift analysis revealed specific and significantly reduced protein binding of oligonucleotides containing the G variant of the CSN1S1^−175bp binding site. Further investigations comprised genotyping of the variable CSN1S1^−175bp activator protein-1 element by an NmuCl restriction fragment length polymorphism in 62 cows of the breed Simmental and 80 cows of the breed German Holstein. Single milk proteins from at least 4 milk samples per cow were quantified by alkaline urea polyacrylamide gel electrophoresis. Homozygotes for CSN1S1^−175bp*G were not observed, and the allele frequencies were 0.19 in Simmental and 0.05 in German Holstein. Carriers of CSN1S1^−175bp*G showed higher content (%) as well as quantity (g/d) of α_s1-casein than CSN1S1^−175bp*A homozygotes, independent of breed. We assume that the positive association of the CSN1S1^−175bp*G variant with CSN1S1 expression is likely to be caused by a reduced affinity of the affected response element to a c-jun-containing CSN1S1 dimer with repressor properties.     

Short communication: the beta-casein (CSN2) silent allele C1 is highly spread in goat breeds

Several single nucleotide polymorphisms have been identified in the goat milk casein genes, most of them modifying the amino acid sequence of the coded protein. At least 9 variants have been found in goat β-CN (CSN2); 6 of them were characterized at the DNA level (A, A1, C, E, 0, and 0′), whereas the other 3 variants were described only at the protein level. The recently identified silent A1 allele is characterized by a C→T transition at the 180th nucleotide of the ninth exon. In the present work, typing results from different breeds (3 Italian, 3 German, and a composite of African breeds for a total of 335 samples) demonstrated that the same mutation is carried by the CSN2*C allele. In addition, the T nucleotide at the 180th nucleotide of the ninth exon was always associated with CSN2*C in all the breeds analyzed. Thus, another silent allele occurs at goat CSN2 and can be named CSN2*C1. The much wider distribution of C1 with respect to the A1 allele indicates that the single nucleotide polymorphisms characterizing the silent mutation originated from CSN2*C. A method for the identification of this allele simultaneously with 5 of the 6 DNA-characterized alleles is also proposed. The mutation involved codifies for the same protein of the C allele; nevertheless, its location in the 3′ untranslated region of the gene might affect the specific casein expression.     

Short communication: Milk protein genetic variation and casein haplotype structure in the Original Pinzgauer cattle

Milk protein genetic polymorphisms are often used for characterizing domesticated mammalian species and breeds, and for studying associations with economic traits. The aim of this work was to analyze milk protein genetic variation in the Original Pinzgauer, a dual-purpose (dairy and beef) cattle breed of European origin that was influenced in the past by human movements from different regions as well as by crossbreeding with Red Holstein. A total of 485 milk samples from Original Pinzgauer from Austria (n = 275) and Germany (n = 210) were typed at milk proteins α_S1-casein, β-casein, κ-casein, α-lactalbumin, and β-lactoglobulin by isoelectrofocusing to analyze the genetic variation affecting the protein amino acid charge. The Original Pinzgauer breed is characterized by a rather high genetic variation affecting the amino acid charge of milk proteins, with a total of 15 alleles, 12 of which were found at a frequency >0.05. The most polymorphic protein was β-casein with 4 alleles detected. The prevalent alleles were CSN1S1*B, CSN2*A², CSN1S2*A, CSN3*A, LGB*A, and LAA*B. A relatively high frequency of CSN1S2*B (0.202 in the whole data set) was found, mainly occurring within the C-A²-B-A haplotype (in the order CSN1S1-CSN2-CSN1S2-CSN3), which seems to be peculiar to the Original Pinzgauer, possibly because the survival of an ancestral haplotype or the introgression of Bos indicus.     

Technical note: simultaneous identification of CSN1S2 A, B, C, and E alleles in goats by polymerase chain reaction-single strand conformation polymorphism

Most variability in goat caseins originates from the high number of genetic polymorphisms often affecting the specific protein expression, with strong effects on milk composition traits and technological properties. At least 7 alleles have been found in the goat α_S2-CN gene (CSN1S2). Five of them (CSN1S2*A, CSN1S2*B, CSN1S2*C, CSN1S2*E, and CSN1S2*F) are widespread in most breeds, whereas the other 2 (CSN1S2*D and CSN1S2*0) are rarer alleles. Four different PCR-RFLP tests are needed to detect all of these variants at the DNA level. The objective of this study was to develop and validate a rapid method for typing 4 of the 5 most-common goat CSN1S2 alleles by means of PCR-single strand conformation polymorphism (SSCP). The method was validated by analyzing 37 goat samples at the protein and DNA level, respectively, by milk isoelectrofocusing and PCR-RFLP methods already described. The genotypes obtained using the PCR-SSCP approach were in full agreement with those obtained by the validation analyses. The newly developed PCR-SSCP approach provides an accurate and inexpensive assay highly suitable for genotyping goat CSN1S2.     

Caprine CSN1S1 haplotype effect on gene expression and milk composition measured by Fourier transform infrared spectroscopy

The Norwegian dairy goat population has a high frequency of a CSN1S1 (α_S1-casein) haplotype with negative effects on protein and fat content. It is characterized by a single point deletion in exon 12 of CSN1S1, leading to a truncated protein and hence a low content of α_S1-casein in the milk. This haplotype together with another haplotype with a deletion in exon 9 are called “weak” haplotypes. “Strong” haplotypes, on the other hand, have positive effects on important milk production traits. We show that expression of CSN1S1 in the mammary gland of lactating goats is significantly lower in animals with 2 weak haplotypes. Moreover, the effects of defective alleles were not detected in animals having 1 strong and 1 weak haplotype. Expression levels of other genes in the casein cluster were not affected by the CSN1S1 haplotypes investigated. Milk samples from goats with 2 weak haplotypes could be distinguished from the other milk samples using Fourier transform infrared (FTIR) spectroscopy and partial least squares discriminant analysis (PLS-DA). The PLS-DA components were related to spectra of pure caseins and whey proteins, hence FTIR has a potential for identifying milk samples with low α_S1-casein content and different protein composition. The results indicate that FTIR-based measurements can be incorporated into breeding plans, or for selection of milk samples with high casein content, which in turn may improve cheese-making properties of the milk.     

Genetic variation in the αS1-casein of Chinese yak (Bos grunniens)

The aim of this study was to investigate diversity of the αS1-casein gene in Maiwa yak. A polymorphism of αS1-casein gene has been identified in Chinese Maiwa yak by PCR single-strand conformation polymorphism protocol. Comparing the X59856 sequence, some non-coding and coding DNA variants of yak CSN1S1 gene were detected in this present study, including KJ397913 (promoter region, 9969A/G, 9984 A/G, 10153 C/T, 10175 A/C, 10261 G/C), KJ397914 (promoter region 10261 G/C), KJ397896 (intron III, 14862 T/C), KJ397897 (intron III, 14862 T/C, and deleted 14944-14946 CTT), KJ397899 (intron IV, 15297 A/C), KJ397901 (intron VIII, 17503 A/C, 17666 A/C), KJ397902 (exon IX, 18901C/A, intron IX, 18970 T/G, 19088 add A, 19120 add A, 19202 add T, 19296 G/T), KJ397903 (intron IX, 18970 T/G, 19088 add A, 19120 add A, 19202 add T, 19296 G/T), KJ397904 (intron IX, 19537 T/G, 19539T/G, 19599G/A, 19638 C/T), KJ397905 (intron XIV, 23402 T/G, 23417 C/T, 23480 T/C, 23511 G/A), and KJ397907 (intron XVI, 25976 G/T; exon XVII, 26181A/G). The polymorphism of αS1-casein gene may be helpful for future studies on genetic variation within and between yak populations or on associated traits.     

Detection of β-casein variants in yak (Bos grunniens) by PCR-SSCP

The aim of this study was to investigate diversity of the β-casein gene in Maiwa yak. A polymorphism of β-casein gene exon VII has been identified in Chinese Maiwa yak by PCR-single-strand conformation polymorphism protocol. Two new variants have been identified in Maiwa yak by resequencing analysis and entered into GenBank with accession numbers JN655524 and JN655525. Comparing the X14711 sequence, the new variant JN655524 consists of 4 single nucleotide mutations at position 66, 74, 234 and 385 of the exon VII of CSN2 gene, resulting in three synonymous mutations and a non-synonymous mutation (codon 67 His/Pro). Compared with JN655524, the new variant JN655525 showed a nucleotide difference in position number 193 in exon VII of CSN2 gene. The new variants may be helpful for future studies on genetic variation within and between yak populations or on associated traits.     

Focusing on the goat casein complex

The analysis of casein polymorphisms in goat species is rather difficult, because of a large number of mutations at each locus, and the tight linkage involving the 4 casein genes. Three goat breeds from Northern Italy, Orobica, Verzasca, and Frisa, were analyzed at the casein complex by milk isoelectrofocusing and analyses at the DNA level to identify the majority of all known polymorphisms. The casein gene structure of the 3 local breeds at α_S1-casein (CSN1S1), β-casein (CSN2), α_S2-casein (CSN1S2), and κ-casein (CSN3) was compared with that of Camosciata, a more widely distributed breed. A new allele was identified and characterized at CSN2 gene, which seemed to be specific to the Frisa breed. It was named CSN2*E, and was characterized by a transversion TCT → TAT responsible for the amino acid exchange Ser₁₆₆ → Tyr₁₆₆ in the mature protein. The casein haplotype structure is highly different among breeds. A total of 26 haplotypes showed a frequency higher than 0.01 in at least 1 of the 4 breeds considered, with 12, 3, 5, and 19 haplotypes in Frisa, Orobica, Verzasca, and Camosciata breeds, respectively. Only 13 haplotypes occurred at a frequency higher than 0.05 in at least 1 breed. With the molecular knowledge of each locus, the ancestral haplotype coding for CSN1S1*B, CSN2*A, CSN1S2*A, and CSN3*B protein variants can be postulated. A protein evolutionary model considering the whole casein haplotype is proposed.     

Genetic variation in the kappa-casein gene (CSN3) of Chinese yak (Bos grunniens) and phylogenetic analysis of CSN3 sequences in the genus Bos

Variants of κ-casein (CSN3) have been extensively studied in cattle and 13 alleles have been identified at the protein and DNA levels to date. Evolution of some of these alleles and a possible common ancestor remain unclear. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis of CSN3 exon IV in domesticated yak revealed a 2-allele polymorphism showing migration patterns different from known cattle variants. The PCR products of both yak CSN3 SSCP alleles were sequenced. All yak had nucleotide sequences coding for Thr in AA position 136 (identical to bovine CSN3*A) and Ala in position 148 (identical to bovine CSN3*B). The sequencing results were confirmed by PCR-RFLP analysis using HindIII and TaqI. A 12-bp insertion in the coding region, representing a repeated nucleotide and AA motif, was found in 1 yak allele. The duplication corresponds to the codons for AA 147 to 150 (Glu-Ala-Ser-Pro) or 148 to 151 (Ala-Ser-Pro-Glu), which are repeated identically. In 21 yak samples genotyped by PCR-SSCP analysis, frequencies for the insertion variant and the short variant were about 68 and 32%, respectively. The loss of the insertion may have led to the ancestral CSN3 allele from which all currently known variants of CSN3 in the genus Bos evolved. This is the first report of polymorphisms in the yak CSN3 gene and may be helpful for future studies on genetic variation within and between yak populations or on associated traits.     

Role of CSN2, CSN3, and BLG genes and the polygenic background in the cattle milk protein profile

To devise better selection strategies in dairy cattle breeding programs, a deeper knowledge of the role of the major genes encoding for milk protein fractions is required. The aim of the present study was to assess the effect of the CSN2, CSN3, and BLG genotypes on individual protein fractions (α_S1-CN, α_S2-CN, β-CN, κ-CN, β-LG, α-LA) expressed qualitatively as percentages of total nitrogen content (% N), quantitatively as contents in milk (g/L), and as daily production levels (g/d). Individual milk samples were collected from 1,264 Brown Swiss cows reared in 85 commercial herds in Trento Province (northeast Italy). A total of 989 cows were successfully genotyped using the Illumina Bovine SNP50 v.2 BeadChip (Illumina Inc.), and a genomic relationship matrix was constructed using the 37,519 SNP markers obtained. Milk protein fractions were quantified and the β-CN, κ-CN, and β-LG genetic variants were identified by reversed-phase HPLC (RP-HPLC). All protein fractions were analyzed through a Bayesian multitrait animal model implemented via Gibbs sampling. The effects of days in milk, parity order, and the CSN2, CSN3, and BLG genotypes were assigned flat priors in this model, whereas the effects of herd and animal additive genetic were assigned Gaussian prior distributions, and inverse Wishart distributions were assumed for the respective co-variance matrices. Marginal posterior distributions of the parameters of interest were compared before and after the inclusion of the effects of the 3 major genes in the model. The results showed that a high portion of the genetic variance was controlled by the major genes. This was particularly apparent in the qualitative protein profile, which was found to have a higher heritability than the protein fraction contents in milk and their daily yields. When the genes were included individually in the model, CSN2 was the major gene controlling all the casein fractions except for κ-CN, which was controlled directly by the CSN3 gene. The BLG gene had the most influence on the 2 whey proteins. The genetic correlations showed the major genes had only a small effect on the relationships between the protein fractions, but through comparison of the correlation coefficients of the proteins expressed in different ways they revealed potential mechanisms of regulation and competitive synthesis in the mammary gland. The estimates for the effects of the CSN2 and CSN3 genes on protein profiles showed overexpression of protein synthesis in the presence of the B allele in the genotype. Conversely, the β-LG B variant was associated with a lower concentration of β-LG compared with the β-LG A variant, independently of how the protein fractions were expressed, and it was followed by downregulation (or upregulation in the case of the β-LG B) of all other protein fractions. These results should be borne in mind when seeking to design more efficient selection programs aimed at improving milk quality for the efficiency of dairy industry and the effect of dairy products on human health.     

Effects of β-κ-casein (CSN2-CSN3) haplotypes and β-lactoglobulin (BLG) genotypes on milk production traits and detailed protein composition of individual milk of Simmental cows

The aim of this study was to investigate the effects of CSN2-CSN3 (β-κ-casein) haplotypes and BLG (β-lactoglobulin) genotypes on milk production traits, content of protein fractions, and detailed protein composition of individual milk of Simmental cows. Content of the major protein fractions was measured by reversed-phase HPLC in individual milk samples of 2,167 cows. Protein composition was measured as percentage of each casein (CN) fraction to total CN and as percentage of β-lactoglobulin (β-LG) to total whey protein. Genotypes at CSN2, CSN3, and BLG were ascertained by reversed-phase HPLC, and CSN2-CSN3 haplotype probabilities were estimated for each cow. Traits were analyzed by using a linear model including the fixed effects of herd-test-day, parity, days in milk, and somatic cell score class, linear regressions on haplotype probabilities, class of BLG genotype, and the random effect of the sire of the cow. Effects of haplotypes and BLG genotypes on yields were weak or trivial. Genotype BB at BLG and haplotypes carrying CSN2 B and CSN3 B were associated with increased CN content and CN number. Haplotypes including CSN3 B were associated with increased κ-CN content and percentage of κ-CN to total CN and with decreased percentages of α_S1- and γ-CN to total CN. Allele CSN2 B had the effect of increasing β-CN content and decreasing content of α_S1-CN. Haplotypes including allele CSN2 A¹ exhibited decreased β-, α_S2-, and γ-CN concentrations and increased α_S1- and κ-CN contents, whereas CSN2 I had positive effects on β-CN concentration and trivial effects on content of other protein fractions. Effects of haplotypes on CN composition were similar to those exerted on content of CN fractions. Allele BLG A was associated with increased β-LG concentration and percentage of β-LG to total whey protein and with decreased content of other milk proteins, namely β-CN and α_S1-CN. Estimated additive genetic variance for investigated traits ranged from 14 to 39% of total variance. Increasing the frequency of specific genotypes or haplotypes by selective breeding might be an effective way to change milk protein composition.     

Effect of CSN1S1-CSN3 (α(S1)-κ-casein) composite genotype on milk production traits and milk coagulation properties in Mediterranean water buffalo

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 K₂₀ 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.     

Short communication: Effect of αS1-casein (CSN1S1) and κ-casein (CSN3) genotypes on milk composition in Murciano-Granadina goats

The effects of the caprine α_S1-casein (CSN1S1) polymorphisms on milk quality have been widely demonstrated. However, much less is known about the consequences of the κ-casein (CSN3) genotype on milk composition in goats. Moreover, the occurrence of interactions between CSN3 and CSN1S1 genotypes has not been investigated. In this study, an association analysis between CSN1S1 and CSN3 genotypes and milk quality traits was performed in 89 Murciano-Granadina goats. Total milk yield as well as total protein, fat, solids-not-fat, lactose, α_S1-casein (CSN1S1), and α_S2-casein (CSN1S2) contents were recorded every other month during a whole lactation (316 observations). Data analysis using a linear mixed model for repeated observations revealed no interaction between the CSN1S1 and CSN3 genotypes. With regard to the effect of the CSN3 locus, AB and BB genotypes were significantly associated with higher levels of total casein and protein content compared with the AA CSN3 genotype. In strong contrast with French breeds, the CSN1S1 genotype did not affect protein, casein, and fat concentrations in Murciano-Granadina goats. These results highlight the importance of taking into consideration the CSN3 genotype when performing selection for milk composition in dairy goats.